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Showing 1 - 200 of 1720 Journals sorted alphabetically
AAPS Journal     Hybrid Journal   (Followers: 22)
Achievements in the Life Sciences     Open Access   (Followers: 5)
ACS Synthetic Biology     Hybrid Journal   (Followers: 25)
Acta Biologica Colombiana     Open Access   (Followers: 7)
Acta Biologica Hungarica     Full-text available via subscription   (Followers: 4)
Acta Biologica Sibirica     Open Access   (Followers: 1)
Acta Biomaterialia     Hybrid Journal   (Followers: 27)
Acta Biotheoretica     Hybrid Journal   (Followers: 4)
Acta Chiropterologica     Full-text available via subscription   (Followers: 6)
acta ethologica     Hybrid Journal   (Followers: 4)
Acta Fytotechnica et Zootechnica     Open Access   (Followers: 1)
Acta Limnologica Brasiliensia     Open Access   (Followers: 3)
Acta Médica Costarricense     Open Access   (Followers: 2)
Acta Musei Silesiae, Scientiae Naturales     Open Access  
Acta Neurobiologiae Experimentalis     Open Access  
Acta Parasitologica     Hybrid Journal   (Followers: 10)
Acta Scientiarum. Biological Sciences     Open Access   (Followers: 2)
Acta Scientifica Naturalis     Open Access   (Followers: 3)
Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis     Open Access  
Actualidades Biológicas     Open Access   (Followers: 1)
Advanced Health Care Technologies     Open Access   (Followers: 4)
Advanced Journal of Graduate Research     Open Access  
Advanced Studies in Biology     Open Access  
Advances in Antiviral Drug Design     Full-text available via subscription   (Followers: 2)
Advances in Bioinformatics     Open Access   (Followers: 17)
Advances in Biological Regulation     Hybrid Journal   (Followers: 4)
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Advances in Biosensors and Bioelectronics     Open Access   (Followers: 7)
Advances in Cell Biology/ Medical Journal of Cell Biology     Open Access   (Followers: 25)
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Advances in Virus Research     Full-text available via subscription   (Followers: 5)
African Journal of Range & Forage Science     Hybrid Journal   (Followers: 6)
AFRREV STECH : An International Journal of Science and Technology     Open Access   (Followers: 1)
Ageing Research Reviews     Hybrid Journal   (Followers: 11)
Aging Cell     Open Access   (Followers: 16)
Agrokémia és Talajtan     Full-text available via subscription   (Followers: 2)
Agrokreatif Jurnal Ilmiah Pengabdian kepada Masyarakat     Open Access  
AJP Cell Physiology     Hybrid Journal   (Followers: 15)
AJP Endocrinology and Metabolism     Hybrid Journal   (Followers: 24)
AJP Lung Cellular and Molecular Physiology     Hybrid Journal   (Followers: 3)
Al-Kauniyah : Jurnal Biologi     Open Access  
Alasbimn Journal     Open Access   (Followers: 1)
Alces : A Journal Devoted to the Biology and Management of Moose     Open Access  
AMB Express     Open Access   (Followers: 1)
Ambix     Hybrid Journal   (Followers: 3)
American Biology Teacher     Full-text available via subscription   (Followers: 14)
American Fern Journal     Full-text available via subscription   (Followers: 1)
American Journal of Agricultural and Biological Sciences     Open Access   (Followers: 8)
American Journal of Bioethics     Hybrid Journal   (Followers: 12)
American Journal of Human Biology     Hybrid Journal   (Followers: 14)
American Journal of Medical and Biological Research     Open Access   (Followers: 8)
American Journal of Plant Sciences     Open Access   (Followers: 19)
American Journal of Primatology     Hybrid Journal   (Followers: 15)
American Malacological Bulletin     Full-text available via subscription   (Followers: 3)
American Naturalist     Full-text available via subscription   (Followers: 74)
Amphibia-Reptilia     Hybrid Journal   (Followers: 6)
Anadolu University Journal of Science and Technology : C Life Sciences and Biotechnology     Open Access  
Anaerobe     Hybrid Journal   (Followers: 4)
Analytical Methods     Full-text available via subscription   (Followers: 11)
Anatomical Science International     Hybrid Journal   (Followers: 3)
Animal Cells and Systems     Hybrid Journal   (Followers: 4)
Animal Models and Experimental Medicine     Open Access  
Annales de Limnologie - International Journal of Limnology     Hybrid Journal   (Followers: 1)
Annales françaises d'Oto-rhino-laryngologie et de Pathologie Cervico-faciale     Full-text available via subscription   (Followers: 3)
Annales Henri Poincaré     Hybrid Journal   (Followers: 3)
Annals of Applied Biology     Hybrid Journal   (Followers: 7)
Annals of Biomedical Engineering     Hybrid Journal   (Followers: 18)
Annals of Human Biology     Hybrid Journal   (Followers: 5)
Annals of Science and Technology     Open Access  
Annual Review of Biomedical Engineering     Full-text available via subscription   (Followers: 14)
Annual Review of Biophysics     Full-text available via subscription   (Followers: 23)
Annual Review of Cancer Biology     Full-text available via subscription   (Followers: 2)
Annual Review of Cell and Developmental Biology     Full-text available via subscription   (Followers: 37)
Annual Review of Food Science and Technology     Full-text available via subscription   (Followers: 13)
Annual Review of Genomics and Human Genetics     Full-text available via subscription   (Followers: 23)
Annual Review of Phytopathology     Full-text available via subscription   (Followers: 12)
Anthropological Review     Open Access   (Followers: 23)
Anti-Infective Agents     Hybrid Journal   (Followers: 3)
Antibiotics     Open Access   (Followers: 9)
Antioxidants     Open Access   (Followers: 4)
Antioxidants & Redox Signaling     Hybrid Journal   (Followers: 8)
Antonie van Leeuwenhoek     Hybrid Journal   (Followers: 5)
Anzeiger für Schädlingskunde     Hybrid Journal   (Followers: 1)
Apidologie     Hybrid Journal   (Followers: 4)
Apmis     Hybrid Journal   (Followers: 1)
APOPTOSIS     Hybrid Journal   (Followers: 8)
Applied Bionics and Biomechanics     Open Access   (Followers: 8)
Applied Vegetation Science     Full-text available via subscription   (Followers: 10)
Aquaculture Environment Interactions     Open Access   (Followers: 3)
Aquaculture International     Hybrid Journal   (Followers: 24)
Aquaculture Reports     Open Access   (Followers: 3)
Aquaculture, Aquarium, Conservation & Legislation - International Journal of the Bioflux Society     Open Access   (Followers: 7)
Aquatic Biology     Open Access   (Followers: 5)
Aquatic Ecology     Hybrid Journal   (Followers: 34)
Aquatic Ecosystem Health & Management     Hybrid Journal   (Followers: 15)
Aquatic Science and Technology     Open Access   (Followers: 3)
Aquatic Toxicology     Hybrid Journal   (Followers: 23)
Archaea     Open Access   (Followers: 3)
Archiv für Molluskenkunde: International Journal of Malacology     Full-text available via subscription   (Followers: 3)
Archives of Biological Sciences     Open Access  
Archives of Microbiology     Hybrid Journal   (Followers: 8)
Archives of Natural History     Hybrid Journal   (Followers: 6)
Archives of Oral Biology     Hybrid Journal   (Followers: 2)
Archives of Virology     Hybrid Journal   (Followers: 5)
Archivum Immunologiae et Therapiae Experimentalis     Hybrid Journal   (Followers: 2)
Arid Ecosystems     Hybrid Journal   (Followers: 2)
Arquivos do Instituto Biológico     Open Access   (Followers: 1)
Arquivos do Museu Dinâmico Interdisciplinar     Open Access  
Arthropod Structure & Development     Hybrid Journal   (Followers: 2)
Arthropods     Open Access   (Followers: 1)
Artificial DNA: PNA & XNA     Hybrid Journal   (Followers: 3)
Asian Bioethics Review     Full-text available via subscription   (Followers: 3)
Asian Journal of Biodiversity     Open Access   (Followers: 4)
Asian Journal of Biological Sciences     Open Access   (Followers: 3)
Asian Journal of Cell Biology     Open Access   (Followers: 5)
Asian Journal of Developmental Biology     Open Access   (Followers: 2)
Asian Journal of Medical and Biological Research     Open Access   (Followers: 4)
Asian Journal of Nematology     Open Access   (Followers: 4)
Asian Journal of Poultry Science     Open Access   (Followers: 4)
Atti della Accademia Peloritana dei Pericolanti - Classe di Scienze Medico-Biologiche     Open Access  
Australian Life Scientist     Full-text available via subscription   (Followers: 2)
Australian Mammalogy     Hybrid Journal   (Followers: 6)
Autophagy     Hybrid Journal   (Followers: 2)
Avian Biology Research     Full-text available via subscription   (Followers: 4)
Avian Conservation and Ecology     Open Access   (Followers: 11)
Bacteriology Journal     Open Access   (Followers: 1)
Bacteriophage     Full-text available via subscription   (Followers: 3)
Bangladesh Journal of Bioethics     Open Access  
Bangladesh Journal of Plant Taxonomy     Open Access  
Bangladesh Journal of Scientific Research     Open Access   (Followers: 1)
Batman Üniversitesi Yaşam Bilimleri Dergisi     Open Access  
Berita Biologi     Open Access   (Followers: 1)
Between the Species     Open Access   (Followers: 1)
Bio Tribune Magazine     Hybrid Journal  
BIO Web of Conferences     Open Access  
BIO-Complexity     Open Access  
Bio-Grafía. Escritos sobre la Biología y su enseñanza     Open Access  
Bioanalytical Reviews     Hybrid Journal   (Followers: 2)
Biocatalysis and Biotransformation     Hybrid Journal   (Followers: 6)
BioCentury Innovations     Full-text available via subscription   (Followers: 1)
Biochemistry and Cell Biology     Hybrid Journal   (Followers: 15)
Biochimie     Hybrid Journal   (Followers: 7)
BioControl     Hybrid Journal   (Followers: 5)
Biocontrol Science and Technology     Hybrid Journal   (Followers: 5)
Biodemography and Social Biology     Hybrid Journal  
BioDiscovery     Open Access   (Followers: 2)
Biodiversitas : Journal of Biological Diversity     Open Access  
Biodiversity Data Journal     Open Access   (Followers: 4)
Biodiversity Informatics     Open Access   (Followers: 1)
Biodiversity Information Science and Standards     Open Access  
Biodiversity: Research and Conservation     Open Access   (Followers: 26)
Bioedukasi : Jurnal Pendidikan Biologi FKIP UM Metro     Open Access  
Bioeksperimen : Jurnal Penelitian Biologi     Open Access  
Bioelectrochemistry     Hybrid Journal   (Followers: 2)
Bioelectromagnetics     Hybrid Journal   (Followers: 1)
Bioenergy Research     Hybrid Journal   (Followers: 2)
Bioengineering and Bioscience     Open Access   (Followers: 1)
BioEssays     Hybrid Journal   (Followers: 10)
Bioethics     Hybrid Journal   (Followers: 15)
BioéthiqueOnline     Open Access  
Biofabrication     Hybrid Journal   (Followers: 5)
Biofilms     Full-text available via subscription   (Followers: 1)
Biogeosciences (BG)     Open Access   (Followers: 10)
Biogeosciences Discussions (BGD)     Open Access   (Followers: 2)
Bioinformatics     Hybrid Journal   (Followers: 305)
Bioinformatics and Biology Insights     Open Access   (Followers: 11)
Bioinspiration & Biomimetics     Hybrid Journal   (Followers: 7)
Biointerphases     Open Access   (Followers: 1)
Biojournal of Science and Technology     Open Access  
BioLink : Jurnal Biologi Lingkungan, Industri, Kesehatan     Open Access  
Biologia     Hybrid Journal  
Biologia on-line : Revista de divulgació de la Facultat de Biologia     Open Access  
Biological Bulletin     Partially Free   (Followers: 6)
Biological Control     Hybrid Journal   (Followers: 4)
Biological Invasions     Hybrid Journal   (Followers: 20)
Biological Journal of the Linnean Society     Hybrid Journal   (Followers: 18)
Biological Letters     Open Access   (Followers: 5)
Biological Procedures Online     Open Access  
Biological Psychiatry     Hybrid Journal   (Followers: 48)
Biological Psychology     Hybrid Journal   (Followers: 6)
Biological Research     Open Access  
Biological Rhythm Research     Hybrid Journal   (Followers: 2)

        1 2 3 4 5 6 7 8 | Last

Journal Cover
Acta Biomaterialia
Journal Prestige (SJR): 1.967
Citation Impact (citeScore): 7
Number of Followers: 27  
  Hybrid Journal Hybrid journal (It can contain Open Access articles)
ISSN (Print) 1742-7061
Published by Elsevier Homepage  [3162 journals]
  • Radical polymerization-crosslinking method for improving extracellular
           matrix stability in bioprosthetic heart valves with reduced potential for
           calcification and inflammatory response
    • Abstract: Publication date: Available online 14 October 2018Source: Acta BiomaterialiaAuthor(s): Gaoyang Guo, Linhe Jin, Wanyu jin, Liang Chen, Yang Lei, Yunbing Wang In recent years, the number of heart valve replacements has multiplied with valve diseases because of aging populations and the surge in rheumatic heart disease in young people. Among them, bioprosthetic heart valves (BHVs) have become increasingly popular. Transcatheter aortic valve implantation (TAVI) valve as an emerging BHV has been increasingly applied to patients. However, the current commercially used BHVs treated with glutaraldehyde (Glut) still face the problem of durability. BHVs derived from Glut-treated xenogenetic tissues would undergo structural degeneration and calcification sometimes even as short as less than 10 years. This issue has already become a big challenge considering more and more young patients at the age of 50-60s are receiving the BHV replacement. In our study, an approach that is totally different from the previous techniques named by us as the radical polymerization-crosslinking (RPC) method was developed to improve extracellular matrix stability, prevent calcification, and reduce inflammatory response in BHVs. The porcine pericardium (PP) tissue was decellularized, functionalized with methacryloyl groups, and subsequently crosslinked by radical polymerization. We found that high-density RPC treatment remarkably improved the stability of collagen and elastin of PP, enhanced its endothelialization potential, and provided reliable biomechanical performance as compared to Glut treatment. The in vivo rat model also confirmed the increased componential stability and reduced the inflammatory response of RPC-treated PP. Moreover, the RPC-treated PP showed better in vivo anticalcification potential than Glut-treated PP.Graphical abstractGraphical abstract for this article
  • Bioactive effects of silica nanoparticles on bone cells are size, surface,
           and composition dependent
    • Abstract: Publication date: Available online 14 October 2018Source: Acta BiomaterialiaAuthor(s): Shin-Woo Ha, Manjula Viggeswarapu, Mark M Habib, George R. Beck Silica based nanoparticles have been demonstrated to have intrinsic biologic activity towards the skeleton and to function by promoting the differentiation of bone forming osteoblasts while inhibiting the differentiation of bone resorbing osteoclasts. The excitement surrounding nanomedicine in part revolves around the almost unlimited possibilities for varying the physicochemical properties including size, composition, and surface charge. To date few studies have attempted to manipulate these characteristics in concert to optimize a complex biologic outcome. Towards this end, spherical silica nanoparticles of various sizes (50-450 nm), of different surface properties (OH, CO2H, NR4+, mNH2), and of different composition (silica, gold, and polystyrene) were synthesized and evaluated for biological activity toward skeletal cells. Osteoblast activity was most influenced by composition and size variables, whereas osteoclasts were most affected by surface property variation. The study also establishes nanoparticle mediated suppression of Nfatc1, a key transcriptional regulator for osteoclast differentiation, identifying a novel mechanism of action. Collectively, the study highlights how during the design of bioactive nanoparticles, it is vital to consider not only the myriad of physical properties that can be manipulated, but also that the characteristics of the target cell plays an equally integral role in determining biological outcome.Graphical abstractGraphical abstract for this article
  • Pore Size Directs Bone Marrow Stromal Cell Fate and Tissue Regeneration in
           Nanofibrous Macroporous Scaffolds by Mediating Vascularization
    • Abstract: Publication date: Available online 13 October 2018Source: Acta BiomaterialiaAuthor(s): Melanie J. Gupte, W. Benton Swanson, Jiang Hu, Xiaobing Jin, Haiyun Ma, Zhanpeng Zhang, Zhongning Liu, Kai Feng, Ganjun Feng, Guiyong Xiao, Nan Hatch, Yuji Mishina, Peter X. Ma In the U.S., 30% of adults suffer joint pain, most commonly in the knee, which severely limits mobility and is often attributed to injury of cartilage and underlying bone in the joint. Current treatment methods such as microfracture result in less resilient fibrocartilage with eventual failure; autografting can cause donor site morbidity and poor integration. To overcome drawbacks in treatment, tissue engineers can design cell-instructive biomimetic scaffolds using biocompatible materials as alternate therapies for osteochondral defects. Nanofibrous poly (l-lactic acid) (PLLA) scaffolds of uniform, spherical, interconnected and well-defined pore sizes that are fabricated using a thermally-induced phase separation and sugar porogen template method create an extracellular matrix-like environment which facilitates cell adhesion and proliferation. Herein we report that chondrogenesis and endochondral ossification of rabbit and human bone marrow stromal cells (BMSCs) can be controlled by scaffold pore architecture, particularly pore size. Small-pore scaffolds support enhanced chondrogenic differentiation in vitro and cartilage formation in vivo compared to large-pore scaffolds. Endochondral ossification is prevented in scaffolds with very small pore sizes; pore interconnectivity is critical to promote capillary ingrowth for mature bone formation. These results provide a novel strategy to control tissue regenerative processes by tunable architecture of macroporous nanofibrous scaffolds.Statement of SignificanceProgress in understanding the relationship between cell fate and architectural features of tissue engineering scaffolds is critical for engineering physiologically functional tissues. Sugar porogen template scaffolds have uniform, spherical, highly interconnected macropores. Tunable pore-size guides the fate of bone marrow stromal cells (BMSCs) towards chondrogenesis and endochondral ossification, and is a critical design parameter to mediate neotissue vascularization. Preventing vascularization favors a chondrogenic cell fate while allowing vascularization results in endochondral ossification and mineralized bone formation. These results provide a novel strategy to control tissue regenerative processes by tunable architecture of macroporous nanofibrous scaffolds.Graphical abstractGraphical abstract for this article
  • Microstructure, mechanical properties, biocompatibility, and in vitro
           corrosion and degradation behavior of a new Zn–5Ge alloy for
           biodegradable implant materials
    • Abstract: Publication date: Available online 11 October 2018Source: Acta BiomaterialiaAuthor(s): Xian Tong, Dechuang Zhang, Xiaotuan Zhang, Yingchao Su, Zimu Shi, Kun Wang, Jianguo Lin, Yuncang Li, Jixing Lin, Cuie Wen Zinc (Zn)-based alloys are considered a new class of biodegradable implant materials due to their superior chemical stability and processability compared to biodegradable magnesium (Mg) alloys. In this study, we report a new biodegradable Zn–5Ge alloy with highly desirable mechanical, corrosion, and biological properties. Microstructural characterization revealed the effective grain-refining effect of germanium (Ge) on the Zn alloy. Tensile test results indicated that the hot-rolled Zn–5Ge alloy showed an ultimate tensile strength of 237.0 MPa, a yield strength of 175.1 MPa, and an elongation of 21.6%; while as-cast pure Zn showed an ultimate tensile strength of 33.6 MPa, a yield strength of 29.3 MPa, and an elongation of 1.2%. The corrosion rates measured by potentiodynamic polarization tests in Hank’s solution in ascending order are: as-cast Zn–5Ge (0.1272 mm/y) < as-cast pure Zn (0.1567 mm/y) < hot-rolled Zn–5Ge (0.2255 mm/y) < hot-rolled pure Zn (0.3057 mm/y). Immersion tests revealed that the degradation rate of as-cast Zn–5Ge is 0.042 mm/y, less than half of that of hot-rolled pure Zn and ∼62% of that of as-cast pure Zn. Moreover, the Zn–5Ge alloy showed excellent in vitro hemocompatibility and the addition of 5% Ge effectively enhanced the hemocompatibility of pure Zn. CCK-8 assay using murine preosteoblast MC3T3-E1 cells indicated that the diluted extracts at a concentration < 12.5% of both the as-cast Zn–5Ge alloy and pure Zn showed grade 0 cytotoxicity; the diluted extracts at the concentrations of 50% and 25% of Zn-5Ge alloy showed a significantly higher cell viability than those of pure Zn.Statement of SignificanceZinc (Zn)-based alloys are currently considered a new class of biodegradable implant materials due to their excellent processability. Here, we report a novel Zn–5Ge alloy with highly desirable mechanical, corrosion and biological properties. The tensile test results indicated that the hot-rolled Zn–5Ge alloy showed an ultimate tensile strength of 237.0 MPa, a yield strength of 175.1 MPa and an elongation of 21.6%; while as-cast pure Zn showed an ultimate tensile strength of 33.6 MPa, a yield strength of 29.3 MPa and an elongation of 1.2 %. The corrosion rate measured by potentiodynamic polarization tests in Hank’s solution in the ascending order is: as-cast Zn–5Ge (0.1272 mm/y) < as-cast pure Zn (0.1567 mm/y) < hot-rolled Zn–5Ge (0.2255 mm/y) < hot-rolled pure Zn (0.3057 mm/y). Immersion tests revealed that the degradation rate of the as-cast Zn–5Ge is 0.042 mm/y, less than half of that of the hot-rolled pure Zn, ∼62% of that of as-cast pure Zn. Moreover, the Zn-5Ge alloy showed excellent in vitro biocompatibility.Graphical abstractGraphical abstract for this article
  • Improving functional re-endothelialization of acellular liver scaffold
           using REDV cell-binding domain
    • Abstract: Publication date: 15 September 2018Source: Acta Biomaterialia, Volume 78Author(s): Julie Devalliere, Yibin Chen, Kevin Dooley, Martin L. Yarmush, Basak E. Uygun Engineering of functional vascularized liver tissues holds great promise in addressing donor organ shortage for transplantation. Whole organ decellularization is a cell removal method that retains the native vascular structures of the organ such that it can be anastomosed with the recipient circulation after recellularization with healthy cells. However, a main hurdle to successful implantation of bioengineered organ is the inability to efficiently re-endothelialize the vasculature with a functional endothelium, resulting in blood clotting which is the primary cause of failure in early transplant studies. Here, we present an efficient approach for enhancing re-endothelialization of decellularized rat liver scaffolds by conjugating the REDV cell-binding domain to improve attachment of endothelial cells (EC) on vascular wall surfaces. In order to facilitate expression and purification of the peptide, REDV was fused with elastin-like peptide (ELP) that confers thermally triggered aggregation behavior to the fusion protein. After validating the adhesive properties of the REDV-ELP peptide, we covalently coupled REDV-ELP to the blood vasculature of decellularized rat livers and seeded EC using perfusion of the portal vein. We showed that REDV-ELP increased cell attachment, spreading and proliferation of EC within the construct resulting in uniform endothelial lining of the scaffold vasculature. We further observed that REDV-ELP conjugation dramatically reduced platelet adhesion and activation. Altogether, our results demonstrate that this method allowed functional re-endothelialization of liver scaffold and show great potential toward the generation of functional bioengineered liver for long-term transplantation.Statement of SignificanceThere is a critical need for novel organ replacement therapies as the grafts for transplantation fall short of demand. Recent advances in tissue engineering, through the use of decellularized scaffolds, have opened the possibility that engineered grafts could be used as substitutes for donor livers. However, successful implantation has been challenged by the inability to create a functional vasculature. Our research study reports a new strategy to increase efficiency of endothelialization by increasing the affinity of the vascular matrix for endothelial cells. We functionalized decellularized liver scaffold using elastin-like peptides grafted with REDV cell binding domain. We showed that REDV-ELP conjugation improve endothelial cell attachment and proliferation within the scaffold, demonstrating the feasibility of re-endothelializing a whole liver vasculature using our technique.Graphical abstractGraphical abstract for this article
  • Inhibition of the fibrillation of highly amyloidogenic human calcitonin by
           cucurbit[7]uril with improved bioactivity
    • Abstract: Publication date: 15 September 2018Source: Acta Biomaterialia, Volume 78Author(s): Hui Shang, Anna Zhou, Jian Jiang, Yanpeng Liu, Jing Xie, Sheyu Li, Yantao Chen, Xiaofeng Zhu, Hong Tan, Jianshu Li Protein/peptide fibrillation is an important challenge for biotechnological drug development. Salmon calcitonin (sCT) is currently used in the clinical treatment of bone-related diseases such as osteoporosis and hypercalcemia, but it still has the risk of immune responses. Although human calcitonin (hCT) would be a better choice in terms of immunogenicity, it has a strong tendency to irreversibly aggregate in aqueous solutions and form long amyloid fibrils, which significantly reduces its bioavailability and therapeutic potency. Here, we demonstrate that cucurbit[7]uril (CB[7]) can inhibit hCT fibrillation by supramolecular interaction with its aromatic groups (affinity: Phe16 > Tyr12 > Phe19 > Phe22). The hCT-CB[7] complex exhibits low cytotoxicity, even promotes osteoblast proliferation and osteogenic capacity of MC3T3 cells. Meanwhile the hCT-CB[7] complexes shows higher bioactivity compared to hCT in reducing blood calcium levels in rats, and also decreases the immunogenicity of hCT. These results suggest that CB[7] has the potential to improve the therapeutic potency of amyloidogenic protein/peptide drugs such as hCT.Graphical abstractGraphical abstract for this article
  • Unveiling the fate of adhering bacteria to antimicrobial surfaces:
           expression of resistance-associated genes and macrophage-mediated
    • Abstract: Publication date: 15 September 2018Source: Acta Biomaterialia, Volume 78Author(s): Diana F. Alves, Andreia P. Magalhães, Damian Neubauer, Marta Bauer, Wojciech Kamysz, Maria O. Pereira Since most antibacterial coatings reported to fight biomaterial-associated infections (BAI) fail in completely preventing bacterial colonization, it is crucial to know the impact of that small fraction of adhered bacteria in BAI recrudescence. This study aims to understand the fate of Staphylococcus aureus able to adhere to an antimicrobial coating previously developed, in terms of potential development of bacterial resistance and their macrophage-mediated phagocytosis. Antimicrobial coating comprised the co-immobilization of Palm peptide and DNase I onto polydimethylsiloxane. Expression of genes associated to resistance and virulence mechanisms showed that cells in contact with antimicrobial surfaces for a long period of 30 days, exhibit genes equally or less expressed, as compared to cells recovered from control surfaces. Recovered cells also exhibit the same susceptibility patterns, which strengthens the evidence of no resistance development. Remarkably, cells adhered to modified surfaces shows a reduced metabolic activity upon vancomycin treatment unlike the cells found on control surfaces, which can be identified as a clinical opportunity for prophylactically administration after implant surgery. Furthermore, results highlight that functionalization of PDMS with Palm and DNase I should not compromise the action of host immune cells. The overall results reinforce the potential of this antimicrobial strategy to fight BAI.Graphical abstractGraphical abstract for this article
  • Synergistic effect of dual targeting vaccine adjuvant with aminated
           β-glucan and CpG-oligodeoxynucleotides for both humoral and cellular
           immune responses
    • Abstract: Publication date: 15 September 2018Source: Acta Biomaterialia, Volume 78Author(s): Jing Wei Jin, Shun Qing Tang, Min Zhi Rong, Ming Qiu Zhang Presently, clinically approved adjuvants (such as aluminum salts) fail to induce cellular immune responses, which is crucial to defend against intracellular pathogens (including HIV, malaria, tuberculosis and Ebola) and cancer. However, Freund’s complete adjuvant potently stimulates both humoral and cellular immune responses, accompanying by high toxicity and severe side reactions. Here in this work, a CpG-oligodeoxynucleotides (CpG-OND) crosslinked aminated β-glucan-Ovalbumin dual targeting nanoparticle (CpG-OND-AG-OVA) is prepared through a simple and mild ionic complexation method. The aminated β-glucan plays dual roles as antigen presenting cells (APCs) targeted carrier and immunopotentiator (targeting and activating dectin-1 on APCs). Meanwhile, CpG-OND also plays dual roles as ionic crosslinker and immunopotentiator (targeting and activating Toll-like receptor 9 in APCs). The adjuvant activity of the particles is evaluated through in vitro and in vivo experiments. The particles significantly enhance uptake and sustained proteolytic processing of antigens, and result in APCs maturation, inducing robust Th1 and Th2-type immune responses comparable to Freund’s adjuvant without obvious toxicity. The potent adjuvant activity of the nanoparticles may originate from dual targeting synergistic effects between aminated β-glucan and CpG-OND. Accordingly, the dual targeting nanoparticles may be a promising vaccine adjuvant for inducing robust humoral and cellular immune responses against infectious diseases and cancers.Statement of SignificanceAn ideal adjuvant for subunit vaccine should act as both a carrier to enhance the uptake, sustained processing and cytosolic delivery of antigens, and an immunopotentiator to stimulate antigen presenting cells (APCs) for activation of naive T cells. Additionally, it should be easy to obtain and safe with negligible toxicity. Unfortunately, both synthetic and natural polymers that have been developed into antigen delivery system cannot completely fulfill the requirements. In the present study, the authors design nanoparticles with aminated β-glucan and CpG-oligodeoxynucleotides (CpG-OND) through a simple and mild method. β-Glucan (a dectin-1 and TLR2 targeted PAMP) and CpG-OND (a TLR9 targeted PAMP) are readily accessible. Aminated β-glucan plays dual roles in the nanoparticle as APCs targeted carrier and immunopotentiator. Meanwhile, CpG-OND also plays dual roles as crosslinker and APCs targeted immunopotentiator. By making use of synergistic effect of the dual targeting vaccine adjuvant with aminated β-glucan and CpG-OND, the nanoparticles induce robust antigen specific immune responses comparable to Freund’s adjuvant without obvious toxicity.Graphical abstractGraphical abstract for this article
  • Development of an Innervated Tissue-engineered Skin with Human Sensory
           Neurons and Schwann Cells Differentiated from iPS Cells
    • Abstract: Publication date: Available online 11 October 2018Source: Acta BiomaterialiaAuthor(s): Quentin Muller, Marie-Josée Beaudet, Thiéry De Serres-Bérard, Sabrina Bellenfant, Vincent Flacher, François Berthod Cutaneous innervation is increasingly recognized as a major element of skin physiopathology through the neurogenic inflammation driven by neuropeptides that are sensed by endothelial cells and the immune system. To investigate this process in vitro, models of innervated tissue-engineered skin (TES) were developed, yet exclusively with murine sensory neurons extracted from dorsal root ganglions. In order to build a fully human model of innervated TES, we used induced pluripotent stem cells (iPSC) generated from human skin fibroblasts. Nearly 100% of the iPSC differentiated into sensory neurons were shown to express the neuronal markers BRN3A and β3-tubulin after 19 days of maturation. In addition, these cells were also positive to TRPV1 and neurofilament M, and some of them expressed Substance P, TrkA and TRPA1. When stimulated with molecules inducing neuropeptide release, iPSC-derived neurons released Substance P and CGRP, both in conventional monolayer culture and after seeding in a 3D fibroblast-populated collagen sponge model. Schwann cells, the essential partners of neurons for function and axonal migration, were also successfully differentiated from human iPSC as shown by their expression of the markers S100, GFAP, p75 and SOX10. When cultured for one additional month in the TES model, iPSC-derived neurons seeded at the bottom of the sponge formed a network of neurites spanning the whole TES up to the epidermis, but only when combined with mouse or iPSC-derived Schwann cells. This unique model of human innervated TES should be highly useful for the study of cutaneous neuroinflammation.Statement of SignificanceThe purpose of this work was to develop in vitro an innovative fully human tissue-engineered skin enabling the investigation of the influence of cutaneous innervation on skin pathophysiology. To reach that aim, neurons were differentiated from human induced pluripotent stem cells (iPSCs) generated from normal human skin fibroblasts. This innervated tissue-engineered skin model will be the first one to show iPSC-derived neurons can be successfully used to build a 3D nerve network in vitro. Since innervation has been recently recognized to play a central role in many human skin diseases, such as psoriasis and atopic dermatitis, this construct promises to be at the forefront to model these diseases while using patient-derived cells.Graphical abstractGraphical abstract for this article
  • Transferrin/aptamer conjugated mesoporous ruthenium nanosystem for
           redox-controlled and targeted chemo-photodynamic therapy of glioma
    • Abstract: Publication date: Available online 11 October 2018Source: Acta BiomaterialiaAuthor(s): Xufeng Zhu, Hui Zhou, Yanan Liu, Yayu Wen, Chunfang Wei, Qianqian Yu, Jie Liu The blood-brain barrier (BBB) and low targeting are major obstacles for the treatment of gliomas. Accordingly, overcoming the BBB and enhancing the targeting of drugs to the glioma area are key to achieving a good therapeutic effect. Here, we have developed the mesoporous ruthenium nanosystem RBT@MRN-SS-Tf/Apt with dual targeting function. Transferrin (Tf) and aptamer AS1411(Apt) are grafted on the surfaces of mesoporous ruthenium nanoparticles (MRN) with high loading capacity. This is achieved via redox-cleavable disulfide bonds, serving as both a capping agent and a targeting ligand, enabling the effective penetration of the blood-brain barrier and targeting the glioma. In addition, RBT@MRN-SS-Tf/Apt can specifically kill glioma cells in vitro and in vivo. Moreover, anti-tumor drugs [Ru(bpy)2(tip)]2+ (RBT) will produce reactive oxygen species and induce apoptosis of tumor cells under laser irradiation, providing photodynamic therapy(PDT) for the treatment of gliomas, and further prolonging the median survival period. The study shows that this chemical photodynamic therapy nanosystem can be used as an efficient and powerful synergistic system for the treatment of brain tumors and other brain diseases of the central nervous system.Statement of significanceIn order to overcome the blood-brain barrier and low targeting, and enhance the anti-glioma activities of nanodrugs. We have developed RBT@MRN-SS-Tf/Apt with dual targeting function.It is achieved release drug via redox-cleavable disulfide bonds, and enable the effective penetration of the blood-brain barrier and targeting the glioma. Moreover, anti-tumor drugs RBT will produce reactive oxygen species and induce apoptosis of tumor cells under laser irradiation, providing photodynamic therapy(PDT)for the treatment of gliomas, and further prolonging the median survival period. Therefore, this chemical photodynamic therapy nanosystem can be used as an efficient and powerful synergistic system for the treatment of brain tumors and other brain diseases of the central nervous system.Graphical abstractGraphical abstract for this article
  • 725Colistin Nanoparticle Assembly by Coacervate Complexation with
           Polyanionic Peptides for Treating Drug-Resistant Gram-Negative Bacteria
    • Abstract: Publication date: Available online 11 October 2018Source: Acta BiomaterialiaAuthor(s): Yu-Han Liu, Shu-Chen Kuo, Bing-Yu Yao, Zih-Syun Fang, Yi-Tzu Lee, Yuan-Chih Chang, Te-Li Chen, Che-Ming Jack Hu Amidst the ever-rising threat of antibiotics resistance, colistin, a decade-old antibiotic with lingering toxicity concern, is increasingly prescribed to treat many drug-resistant, gram-negative bacteria. With the aim of improving the safety profile while preserving the antimicrobial activity of colistin, a nanoformulation is herein developed through coacervate complexation with polyanionic peptides. Upon controlled mixing of cationic colistin with polyglutamic acids, formation of liquid coacervates was demonstrated. Subsequent stabilization by DSPE-PEG and homogenization through micro-fluidization of the liquid coacervates yielded nanoparticles 8 nm in diameter. In vitro assessment showed that the colistin antimicrobial activity against multiple drug-resistant bacterial strains was retained and, in some cases, enhanced following the nanoparticle assembly. In vivo administration in mice demonstrated improved safety of the colistin nanoparticle, which has a maximal tolerated dose of 12.5 mg/kg compared to 10 mg/kg of free colistin. Upon administration over a 7-day period, colistin nanoparticles also exhibited reduced hepatotoxicity as compared to free colistin. In mouse models of Klebsiella pneumoniae bacteremia and Acinetobacter baumannii pneumonia, treatment with colistin nanoparticles showed equivalent efficacy to free colistin. These results demonstrate coacervation-induced nanoparticle assembly as a promising approach toward improving colistin treatments against bacterial infections.Statement of significanceImproving the safety of colistin while retaining its antimicrobial activity has been a highly sought-after objective toward enhancing antibacterial treatments. Herein, we demonstrate formation of stabilized colistin nanocomplexes in the presence of anionic polypeptides and DSPE-PEG stabilizer. The nanocomplexes retain colistin’s antimicrobial activity while demonstrating improved safety upon in vivo administration. The supramolecular nanoparticle assembly of colistin presents a unique approach toward designing antimicrobial nanoparticles.Graphical abstractGraphical abstract for this article
  • Multi-Cellular Transitional Organotypic Models to Investigate Liver
    • Abstract: Publication date: Available online 11 October 2018Source: Acta BiomaterialiaAuthor(s): Sophia M. Orbach, Andrew J. Ford, Scott-Eugene Saverot, Padmavathy Rajagopalan Hepatic fibrosis is the result of wound healing and inflammation resulting in organ dysfunction. Hepatocytes, liver sinusoidal endothelial cells (LSECs), Kupffer cells (KCs), and hepatic stellate cells (HSCs) play critical roles in fibrogenesis. As the liver undergoes fibrosis, there are populations of cells that are healthy, fibrotic as well as those undergoing fibrosis. We investigated how a varying mechanical environment could induce changes in hepatic cells. In this study, a gradient in the mechanical properties of the microenvironment resulted in transitioning phenotypes in hepatic cells. We have designed detachable polyelectrolyte multilayers (PEMs) whose elastic moduli ranged from 21 – 43 kPa to serve as Space of Disse mimics. We assembled novel 3D organotypic liver models comprised of hepatocytes, LSECs, HSCs, KCs, and the Space of Disse mimic. We demonstrate how cells in contact with a mechanical gradient exhibit different properties compared to cells cultured using non-gradient PEMs. Significant differences were observed in HSC and KC proliferation between 3D cultures assembled with gradient and non-gradient PEMs. While HSCs on the stiffer regions of the gradient PEMs expressed both GFAP and α-SMA, cells in cultures assembled with homogeneous 43 kPa multilayers primarily expressed α-SMA. Over an 8-day culture, the elastic modulus in the 21 and 43 kPa regions of the gradient PEMs increased by 1.6 and 3.7- fold, respectively. This was accompanied by a 4-fold increase in hydroxyproline. Such in vitro tissues can be used to investigate the effects of liver fibrosis.Statement of SignificanceWe have assembled a liver model assembled with four major primary hepatic cell types to investigate how a varying mechanical environment induces changes in hepatic cells. In this study, a gradient in the mechanical properties of the microenvironment results in transitioning phenotypes in hepatic cells. Our goal was to investigate the interplay between mechanical properties and a multi-cellular engineered liver tissue. In these models, Kupffer cell proliferation and hepatic stellate cell activation occurred due to mechanical cues and inter-cellular signaling across a distance of 2000 μm. These models are unique, in that, fibrosis was initiated purely through changes to the microenvironment. These models were not exposed to fibrogenic factors nor were the models assembled with cells from fibrotic rats. To the best of our knowledge, these are the first liver models that capture how a gradient microenvironment can result in transitioning cellular phenotypes.Graphical abstractGraphical abstract for this article
  • Effects of Bisphosphonate Ligands and PEGylation on Targeted Delivery of
           Gold Nanoparticles for Contrast-Enhanced Radiographic Detection of Breast
    • Abstract: Publication date: Available online 11 October 2018Source: Acta BiomaterialiaAuthor(s): Lisa E. Cole, Tracie L. McGinnity, Lisa E. Irimata, Tracy Vargo-Gogola, Ryan K. Roeder A preclinical murine model of hydroxyapatite (HA) breast microcalcifications (µcals), which are an important clinical biomarker for breast cancer detection, was used to investigate the independent effects of high affinity bisphosphonate (BP) ligands and a polyethylene glycol (PEG) spacer on targeted delivery of gold nanoparticles (Au NPs) for contrast-enhanced radiographic detection. The addition of BP ligands to PEGylated Au NPs (BP-PEG-Au NPs) resulted in five-fold greater binding affinity for targeting HA µcals, as expected, due to the strong binding affinity of BP ligands for calcium. Therefore, BP-PEG-Au NPs were able to target HA µcals in vivo after intramammary delivery, which enabled contrast-enhanced radiographic detection of µcals in both normal and radiographically dense mammary tissues similar to previous results for BP-Au NPs, while PEG-Au NPs did not. The addition of a PEG spacer between the BP targeting ligand and Au NP surface enabled improved in vivo clearance. PEG-Au NPs and BP-PEG-Au NPs were cleared from all mammary glands (MGs) and control MGs, respectively, within 24-48 h after intramammary delivery, while BP-Au NPs were not. PEGylated Au NPs were slowly cleared from MGs by lymphatic drainage and accumulated in the spleen. Histopathology revealed uptake of PEG-Au NPs and BP-PEG-Au NPs by macrophages in the spleen, liver, and MGs; there was no evidence of toxicity due to the accumulation of NPs in organs and tissues compared with untreated controls for up to 28 days after delivery.Statement of SignificanceAu NP imaging probes and therapeutics are commonly surface functionalized with PEG and/or high affinity targeting ligands for delivery. However, direct comparisons of PEGylated Au NPs with and without a targeting ligand, or ligand-targeted Au NPs with and without a PEG spacer, on in vivo targeting efficiency, biodistribution, and clearance are limited. Therefore, the results of this study are important for the rationale design of targeted NP imaging probes and therapeutics, including the translation of BP-PEG-Au NPs which enable improved sensitivity and specificity for the radiographic detection of abnormalities (e.g., µcals) in women with dense breast tissue.Graphical abstractGraphical abstract for this article
  • Early angiogenesis detected by PET imaging with 64Cu-NODAGA-RGD is
           predictive of bone critical defect repair
    • Abstract: Publication date: Available online 10 October 2018Source: Acta BiomaterialiaAuthor(s): Anne-Margaux Collignon, Julie Lesieur, Nadège Anizan, Rana Ben Azzouna, Anne Poliard, Caroline Gorin, Didier Letourneur, Catherine Chaussain, Francois Rouzet, Gael Y. Rochefort Therapies using stem cells may be applicable to all fields of regenerative medicine, including craniomaxillofacial surgery. Dental pulp stem cells (DPSCs) have demonstrated in vitro and in vivo osteogenic and proangiogenic properties. The aim of the study was to evaluate whether early angiogenesis investigated by nuclear imaging can predict bone formation within a mouse critical bone defect. Two symmetrical calvarial critical-sized defects were created. Defects were left empty or filled with i) DPSC-containing dense collagen scaffold, ii) 5% hypoxia-primed DPSC-containing dense collagen scaffold, iii) acellular dense collagen scaffold, or iv) left empty. Early angiogenesis assessed by PET using 64Cu-NODAGA-RGD as a tracer was found to be correlated with bone formation determined by micro-CT within the defects from day 30, and to be correlated to the late calcium apposition observed at day 90 using 18F-Na PET. These results suggest that nuclear imaging of angiogenesis, a technique applicable in clinical practice, is a promising approach for early prediction of bone grafting outcome, thus potentially allowing to anticipate alternative regenerative strategies.Statement of SignificanceBone defects are a major concern in medicine. As life expectancy increases, the number of bone lesions grows, and occurring complications lead to a delay or even lack of consolidation. Therefore, to be able to predict healing or the absence of scarring at early times would be very interesting. This would not “waste time” for the patient.We report here that early nuclear imaging of angiogenesis, using 64Cu-NODAGA-RGD as a tracer, associated with nuclear imaging of mineralization, using 18F-Na as a tracer, is correlated to late bone healing objectivized by classical histology and microtomography.This nuclear imaging represents a promising approach for early prediction of bone grafting outcome in clinical practice, thus potentially allowing to anticipate alternative regenerative strategies.Graphical abstractGraphical abstract for this article
  • Bioengineering a novel 3D in vitro model of gastric mucosa for
           stomach permeability studies
    • Abstract: Publication date: Available online 9 October 2018Source: Acta BiomaterialiaAuthor(s): Bianca N. Lourenço, Tiago dos Santos, Carla Oliveira, Cristina C. Barrias, Pedro L. Granja The field of stomach-directed therapeutics and diagnosis is still hampered by the lack of reliable in vitro models that closely mimic the gastric mucosa where gastric cancer cells are generally confined. Here we propose a rapid, complex, and innovative 3D in vitro model of the gastric mucosa, by extending a conventional gastric monolayer model to an inner stratum of the mucosa – the lamina propria. The developed model comprises normal stomach fibroblasts embedded in a 3D alginate-RGD hydrogel prepared on the basolateral side of a Transwell® insert, mimicking the extracellular matrix and cellular component of the lamina propria, onto which a moderately differentiated adenocarcinoma stomach cell line (MKN74) was seeded, reproducing the physiological conditions of the gastric barrier. The integrity and functionality of the in vitro model was evaluated through permeability studies of FITC-dextran and 200 nm fluorescent polystyrene nanoparticles at gastric conditions. Nanoparticle transport was pH-dependent and strongly impacted by the biomimetic lamina propria, highlighting that a gastric ECM-like microenvironment should be integrated in an in vitro permeability model to be adopted as a reliable evaluation tool of innovative therapeutics and diagnosis of gastric diseases.Statement of SignificanceCurrent in vitro models of the gastric mucosa are limited to simplistic 2D cell culture systems, which ignore the dimensionality of the stomach wall and make it difficult to reliably test new therapeutic approaches to gastric pathologies. By combining stomach fibroblasts embedded within a 3D RGD-modified alginate hydrogel and epithelial gastric cancer cells in a transwell® system, we established a new biomimetic model of the stomach mucosa. Epithelial cells recreate the gastric epithelium, while the cell-laden 3D hydrogel recapitulates both the cellular composition and dimensionality of the extracellular matrix of gastric lamina propria. This cellularized 3D model stands as a promising evaluation platform to assist the development of new strategies for the treatment and diagnosis of gastric diseases.Graphical abstractGraphical abstract for this article
  • Design and development of pH-responsive polyurethane membranes for
           intravaginal release of nanomedicines
    • Abstract: Publication date: Available online 6 October 2018Source: Acta BiomaterialiaAuthor(s): Seungil Kim, Yannick Leandre Traore, Emmanuel A Ho, Muhammad Shafiq, Soo-Hyun Kim, Song Liu The objective of this study was to develop and characterize a novel intravaginal membrane platform for pH-triggered release of nanoparticles (NPs), which is essential for efficient intravaginal delivery of certain effective but acid-labile therapeutic agents for sexually transmitted infections, such as small interfering RNA (siRNA). pH-responsive polyurethane (PU) was electrospun into a porous nanofibrous membrane. The diameters of the fibers, as well as the thickness and pore sizes of the membrane under dry and wet conditions (pH 4.5 and 7.0), were determined from scanning electron microscopy (SEM) micrographs. pH-dependent zeta-potential (ζ) of the membrane was evaluated using a SurPASS electrokinetic analyzer. VisiblexTM color-dyed polystyrene NPs (PSNs, 200 nm, -COOH) and CCR5 siRNA-encapsulated solid lipid NPs (SLNs) were used for in vitro NP release studies in a vaginal fluid simulant (VFS) at pH 4.5 (normal physiological vaginal pH) and 7.0 (vaginal pH neutralization by semen). During 24 hours of incubation in VFS, close-to-zero PSNs (2 ± 1%) and 28 ± 4% SLNs were released through the PU membrane at pH 4.5, whereas the release of PSNs and SLNs significantly increased to 60 ± 6% and 59 ± 8% at pH 7.0, respectively. The pH-responsive release of NPs hinged on the electrostatic interaction between the pH-responsive membranes and the anionic NPs, and the change in pH-responsive morphology of the membrane. In vitro biocompatibility studies of the membranes showed no significant cytotoxicity to VK2/E6E7 human epithelial cells and Sup-T1 human T-cells and no significant changes in the expression of pro-inflammatory cytokines (IL-6, IL-8, and IL-1β). Overall, these porous pH-responsive PU membranes demonstrated their potential in serving as “window” membranes of reservoir-IVRs for pH-responsive intravaginal release of NPs.Statement of SignificanceStimuli-responsive intravaginal nanoparticle release is achieved for the first time through new electrospun pH-responsive polyurethane (PU) semi-permeable membranes, which serve as a “window” membrane of the reservoir-type intravaginal ring (IVR) for the prevention of human immunodeficiency virus (HIV) transmission. Almost no release of nanoparticles was observed at normal pH in the female genital tract (in vaginal fluid simulant [VFS], at pH 4.5); however, a continuous release of nanoparticles was observed at elevated pH in the female genital tract (in VFS, at pH 7.0). This pH-responsive intravaginal release can reduce side effect and drug resistance by avoiding unnecessary exposure. The PU semi-permeable membrane demonstrated potential use as biomaterials for “smart” intravaginal nanoparticle release and has great potential to protect women from HIV.Graphical abstractGraphical abstract for this article
  • Cartilaginous extracellular matrix derived from decellularized chondrocyte
           sheets for the reconstruction of osteochondral defects in rabbits
    • Abstract: Publication date: Available online 6 October 2018Source: Acta BiomaterialiaAuthor(s): Zhifa Wang, Zhiye Li, Zhijin Li, Buling Wu, Yanpu Liu, Wei Wu Cartilaginous extracellular matrix (ECM) materials derived from decellularized native articular cartilage are widely used in cartilage regeneration. However, it is difficult for endogenous cells to migrate into ECM derived from native cartilage owing to its nonporous structure and dense nature. Moreover, current decellularization approaches frequently lead to architectural breakdown and potential loss of surface composition of ECM. To solve this problem, we aimed to establish a novel biological ECM scaffold from chondrocyte sheets for cartilage regeneration. We cultured chondrocytes harvested from the auricular cartilage of 4-week-old New Zealand rabbits and enabled them to form cell sheets. These sheets were decellularized using sodium dodecyl sulfate (SDS) with three different concentrations, namely, 1%, 5%, and 10%, followed by 1% Triton X-100 and deoxyribonuclease enzyme solution. In vitro microstructural examination and mechanical tests demonstrated that 1% SDS not only removed chondrocytes completely but also maintained the native architecture and composition of ECM, thus avoiding the use of high-concentration SDS. Application of decellularized chondrocyte sheets for osteochondral defects in rabbits resulted in substantial host remodeling and variant regeneration of osteochondral tissues. One percent SDS-treated decellularized chondrocyte sheets contributed to the superior reconstruction of osteochondral defects as compared with 5% and 10% SDS groups, which includes vascularized subchondral bone, articular cartilage with adequate thickness, and integration with host tissues. Furthermore, ECM from 1% SDS significantly increased the migrating potential of bone marrow mesenchymal stem cells (BMSCs) in vitro. RT-PCR and western blot also revealed that ECM increased the expression of SOX-9 in BMSCs, whereas it decreased COL-X expression. In conclusion, our results suggested that the chondrocyte sheets decellularized with 1% SDS preserved the integrity and bioactivity, which favored cell recruitment and enabled osteochondral regeneration in the knee joints of rabbits, thus offering a promising approach for articular cartilage reconstruction without cell transplantation.Statement of SignificanceAlthough biological extracellular matrix (ECM) derived from decellularized native cartilage has been widely used in cartilage regeneration, it is difficult for endogenous cells to migrate into ECM owing to its dense nature. Moreover, current decellularization approaches lead to architectural breakdown of ECM. This study established a novel biological ECM from decellularized chondrocyte sheets for cartilage regeneration. Our results suggested that cartilaginous ECM favored cell recruitment and enabled osteochondral regeneration in rabbits, thus offering a promising approach for articular cartilage reconstruction without cell transplantation. SDS 1% adequately decellularized the chondrocytes in cell sheets, whereas it maintained the native architecture and composition of ECM, thereby avoiding the use of high-concentration SDS and providing a new way to acquire cartilaginous ECM.Graphical abstractGraphical abstract for this article
  • Vitronectin promotes the vascularization of porous polyethylene
    • Abstract: Publication date: Available online 6 October 2018Source: Acta BiomaterialiaAuthor(s): Maximilian E.T. Hessenauer, Kirsten Lauber, Gabriele Zuchtriegel, Bernd Uhl, Timon Hussain, Martin Canis, Sebastian Strieth, Alexander Berghaus, Christoph A. Reichel Rapid implant vascularization is a prerequisite for successful biomaterial engraftment. Vitronectin (VN) is a matricellular glycoprotein well known for its capability to interact with growth factors, proteases, and protease inhibitors/receptors. Since such proteins are highly relevant for angiogenic processes, we hypothesized that VN contributes to the tissue integration of biomaterials. Employing different in vivo and ex vivo microscopy techniques, engraftment of porous polyethylene (PPE) implants was analyzed in the dorsal skinfold chamber model in wild-type (WT) and VN-/- mice. Upon PPE implantation, vascularization of this biomaterial was severely compromised in animals lacking this matricellular protein. Proteome profiling revealed that VN deficiency does not cause major changes in angiogenic protein composition in the implants suggesting that VN promotes PPE vascularization via mechanisms modulating the activity of angiogenic factors rather than by directly enriching them in the implant. Consequently, surface coating with recombinant VN (embedded in Matrigel®) accelerated implant vascularization in WT mice by enhancing the maturation of a vascular network. Thus, VN contributes to the engraftment of PPE implants by promoting the vascularization of this biomaterial. Surface coating with VN might provide a promising strategy to improve the vascularization of PPE implants without affecting the host’s integrity.Statement of significancePorous polyethylene (PPE) is a biomaterial frequently used in reconstructive surgery. The proper vascularization of PPE implants is a fundamental prerequisite for its sucessful engraftment in host tissue. Although the overall biocompatibility of PPE is good, there are less favorable application sites for its use in tissue reconstruction mostly characterized by low blood supply. Employing advanced in vivo microscopy methods and proteomic analyses in genetically engineered mice, we here describe a previously unrecognized function of vitronectin (VN) that enables this abundantly present glycoprotein to particularly promote the vascularization of PPE biomaterial. These properties of VN specifically facilitate the formation of a dense vessel network within the implant which relies on modulating the activity of angiogenic mediators rather than on the enrichment of these factors in the implant. Consequently, surface coating with this matricellular protein effectively accelerated and intensified implant vascularization which might be beneficial for its implementation at unfavorable sites for implantation without affecting the host’s integrity.Graphical abstractEndogenous vitronectin (VN) promotes the vascularization of PPE biomaterial (left, mid). Additional surface coating with recombinant VN accelerates and intensifies this process (right).Graphical abstract for this article
  • Characterization of a Cell-Assembled extracellular Matrix and the effect
           of the devitalization process
    • Abstract: Publication date: Available online 6 October 2018Source: Acta BiomaterialiaAuthor(s): Laure Magnan, Gaëlle Labrunie, Sylvie Rey, Nathalie Dusserre, Nicolas L'Heureux, Sébastien Marais, Sabrina Lacomme, Etienne Gontier, Marc Bonneu We have previously shown that the Cell-Assembled extracellular Matrix (CAM) synthesized by normal, human, skin fibroblasts in vitro can be assembled in a completely biological vascular graft that was successfully tested in the clinic. The goal of this study was to perform a detailed analysis of the composition and the organization of this truly bio-material. In addition, we investigated whether the devitalization process (dehydration) used to store the CAM, and thus, make the material available “off-the-shelf,” could negatively affect its organization and mechanical properties. We demonstrated that neither the thickness nor the mechanical strength of CAM sheets were significantly changed by the dehydration/freezing/rehydration cycle. The identification of over 50 extracellular matrix proteins highlighted the complex composition of the CAM. Histology showed intense collagen and glycosaminoglycan staining throughout the CAM sheet. The distribution of collagen I, collagen VI, thrombospondin-1, fibronectin-1, fibrillins-1, biglycan, decorin, lumican and versican showed various patterns that were not affected by the devitalization process. Transmission electron microscopy analysis revealed that the remarkably dense collagen network was oriented in the plane of the sheet and that neither fibril density nor diameter was changed by devitalization. Second harmonic generation microscopy revealed an intricate, multi-scale, native-like collagen fiber orientation. In conclusion, this bio-material displayed many tissue-like properties that could support normal cell-ECM interactions and allow implantation without triggering degradative responses from the host's innate immune system. This is consistent with its success in vivo. In addition, the CAM can be devitalized without affecting its mechanical or unique biological architecture.Statement of significanceThe extracellular matrix (ECM) defines biological function and mechanical properties of tissues and organs. A number of promising tissue engineering approaches have used processed ECM from cadaver/animal tissues or cell-assembled ECM in vitro combined with scaffolds. We have shown the clinical potential of a scaffold-free approach based on an entirely biological material produced by human cells in culture without chemical processing. Here, we perform a comprehensive analysis of the properties of what can truly be called a bio-material. We also demonstrate that this material can be stored dried without losing its remarkable biological architecture.Graphical abstractGraphical abstract for this article
  • Micro Patterning of Hydroxyapatite by Soft Lithography on Hydrogels for
           Selective Osteoconduction
    • Abstract: Publication date: Available online 5 October 2018Source: Acta BiomaterialiaAuthor(s): Ryuji Kiyama, Takayuki Nonoyama, Susumu Wada, Shingo Semba, Nobuto Kitamura, Tasuku Nakajima, Takayuki Kurokawa, Kazunori Yasuda, Shinya Tanaka, Jian Ping Gong Mechanically robust hydrogels are promising biomaterials as artificial supportive tissue. These applications require selective and robust bonding of the hydrogels to living tissue. Recently, we achieved strong in vivo bone bonding of a tough double network (DN) hydrogel, a potential material for use as artificial cartilage and tendon, by hybridizing osteoconductive hydroxyapatite (HAp) in the gel surface layer. In this work, we report micro patterning of HAp at the surface of the DN hydrogel for selective osteoconduction. Utilizing the dissolution of HAp in an acidic environment, the soft lithography technique using an acid gel stamp was adopted to form a high-resolution HAp pattern on the gel. The HAp-patterned gel showed well-regulated migration and adhesion of cells in vitro. Moreover, the HAp-patterned gel showed selective and robust bonding to the rabbit bone tissue in vivo. This HAp soft lithography technique allows for simple and quick preparation of tailor-made osteoconductive hydrogels and can be applied to other hydrogels for selective bone bonding.Statement of SignificanceHydrogels, preserving large amount of water, have been studied for next-generation artificial soft tissues. However, fixation of hydrogels to living tissue was unsolved issue for clinical application. Recently, we achieved robust bonding of a tough Double Network gel to bone in vivo by coating of osteoconductive hydroxyapatite in the gel surface layer. For further progress for practical use, we report the micro patterning of HAp at the surface of the DN hydrogel by using soft lithography technique, to perform selective bonding to only objective area without unnecessary coalescence. The HAp lithography technique is simple, quick and non-toxic method to prepare tailor-made osteoconductive hydrogels and has universality of species of hydrogels.Graphical abstractGraphical abstract for this article
  • A ready-to-use acidic, brushite-forming calcium phosphate cement
    • Abstract: Publication date: Available online 4 October 2018Source: Acta BiomaterialiaAuthor(s): Jun Luo, Håkan Engqvist, Cecilia Persson Premixed calcium phosphate cements have been developed to simplify the usage of traditional calcium phosphate cements and reduce the influence of the setting reaction on the delivery process. However, difficulties in achieving a good cohesion, adequate shelf life and sufficient mechanical properties have so far impeded their use in clinical applications, especially for the more degradable acidic calcium phosphate cements.In this study, a brushite cement was developed from a series of ready-to-use calcium phosphate pastes. The brushite cement paste was formed via mixing of a monocalcium phosphate monohydrate (MCPM) paste and a β-tricalcium phosphate (β-TCP) paste with good injectability and adequate shelf life. The MCPM paste was based on a water-immiscible liquid with two surfactants and the β-TCP paste on a sodium hyaluronate aqueous solution. The effect of citric acid as a retardant was assessed. Formulations with suitable amounts of citric acid showed good cohesion and mechanical performance with potential for future clinical applications.Statement of SignificanceAcidic calcium phosphate cements have attracted extensive attention as bone substitute materials due to their ability to resorb faster than basic calcium phosphate cements in vivo. However, traditionally, short working times and low mechanical strength has limited their clinical application. Premixed cements could simplify the clinical use as well as improve property reproducibility, but short shelf lives, low cohesion and low mechanical properties have restricted the development.In this study, an injectable ready-to-use two-phase system consisting of an MCPM paste and a β-TCP paste was developed based on acidic cement. It shows good cohesion, compressive strength and adequate shelf life, which has the potential to be used in a dual chamber system for simplified and fast filling of bone defects in a minimally invasive manner. This will reduce surgery time, decrease the risk of contamination and ensure repeatable results.Graphical abstractGraphical abstract for this article
  • Development of photosynthetic sutures for the local delivery of oxygen and
           recombinant growth factors in wounds
    • Abstract: Publication date: Available online 1 October 2018Source: Acta BiomaterialiaAuthor(s): Carolina Centeno-Cerdas, Montserrat Jarquín-Cordero, Myra Noemi Chávez, Ursula Hopfner, Christopher Holmes, Daniel Schmauss, Hans-Günther Machens, Jörg Nickelsen, José Tomás Egaña Surgical sutures represent the gold standard for wound closure, however, their main purpose is still limited to a mechanical function rather than playing a bioactive role. Since oxygen and pro-regenerative growth factors have been broadly described as key players for the healing process, in this study we evaluated the feasibility of generating photosynthetic sutures that, in addition to mechanical fixation, could locally and stably release oxygen and recombinant human growth factors (VEGF, PDGF-BB, or SDF-1α) at the wound site. Here, photosynthetic genetically modified microalgae were seeded in commercially available sutures and their distribution and proliferation capacity was evaluated. Additionally, the mechanical properties of seeded sutures were compared to unseeded controls that showed no significant differences. Oxygen production, as well as recombinant growth factor release was quantified in vitro over time, and confirmed that photosynthetic sutures are indeed a feasible approach for the local delivery of bioactive molecules. Finally, photosynthetic sutures were tested in order to evaluate their resistance to mechanical stress and freezing. Significant stability was observed in both conditions, and the feasibility of their use in the clinical practice was therefore confirmed. Our results suggest that photosynthetic gene therapy could be used to produce a new generation of bioactive sutures with improved healing capacities.Statement of significanceDisruption of the vascular network is intrinsic to trauma and surgery, and consequently, wound healing is characterized by diminished levels of blood perfusion. Among all the blood components, oxygen and pro-regenerative growth factors have been broadly described as key players for the healing process. Therefore, in this study we evaluated the feasibility of generating photosynthetic sutures that, in addition to mechanical fixation, could locally and stably release oxygen and recombinant human growth factors at the wound site. This novel concept has never been explored before for this type of material and represents the first attempt to create a new generation of bioactive sutures with improved regenerative capabilities.Graphical abstractGraphical abstract for this article
  • Micro-patterned endogenous stroma equivalent induces polarized
           crypt-villus architecture of human small intestinal epithelium
    • Abstract: Publication date: Available online 30 September 2018Source: Acta BiomaterialiaAuthor(s): Vincenza De Gregorio, Giorgia Imparato, Francesco Urciuolo, Paolo A. Netti The small intestine is the major site for digestion, drug and nutrient absorption, as well as a primary site for many diseases. Current in vitro gut models fail in reproducing the complex intestinal extracellular matrix (ECM) network of the lamina propria and the peculiar architecture of the crypt-villus axis. Here we proposed a novel in vitro human intestine model that mimics the intestinal stromal topography and composition and strictly reproduces the tissue polarity with the crypt-villus architecture. First we developed a 3D human intestinal stromal equivalent (3D-ISE) composed of human intestinal subepithelial myofibroblasts (ISEMFs) embedded in their own extracellular matrix. Then, we seeded human colon carcinoma-derived cells (Caco-2) onto flat or patterned cell-synthetized stromal equivalent structure and cultured them until the formation of a well-oriented epithelium. We demonstrated that the patterned stroma increases the absorbing surface area, the epithelial proliferation rate, and the density of microvilli. In addition it induces changes in the biological functions of the epithelial cells such as enzymes and mucus production, polarization and tightness showing a physiological cell-lineage compartmentalization along the crypt/villi axes with the undifferentiated phenotypes at the base. At last, we reproduced an inflamed intestinal tissue model in which we identified the contribution of the stromal microenvironment by molecular (cytokines release and MMPs production) and immunofluorescence analyses and the effects of the epithelial-stromal cross-talk in the intestinal innate immunity by multiphoton investigation that revealed differences in the collagen network architecture.Statement of significanceThe intestinal stroma morphology and composition has a fundamental role in crypt-villus development and appropriate epithelial cell-lineage compartmentalization. On this base, here we develop an engineered organotypic model of human intestine equivalent in which a functional epithelial/ECM crosstalk is recapitulated. Due to its accessible luminal surface it provides a new platform for preclinical studies of mucosal immunology and bowel inflammation as well as the assessment of pharmaco-toxicity studies.Graphical abstractGraphical abstract for this article
  • The Inflammasome in Host Response to Biomaterials: Bridging Inflammation
           and Tissue Regeneration
    • Abstract: Publication date: Available online 29 September 2018Source: Acta BiomaterialiaAuthor(s): Daniela P. Vasconcelos, Artur P. Águas, Mário A. Barbosa, Pablo Pelegrín, Judite N. Barbosa The development of new biomaterials to be used in tissue engineering applications is creating new solutions for a range of healthcare problems. The trend in biomaterials research has shifted from biocompatible “immune-evasive” biomaterials to “immune-interactive” materials that modulate the inflammatory response supporting implant integration as well as improving healing and tissue regeneration.Inflammasomes are large intracellular multiprotein complexes that are key players in host defence during innate immune responses and assemble after recognition of pathogens or danger signals. The process of biomaterial implantation causes injury to tissues that will consequently release danger signals that could be sensed by the inflammasome. There are increasing evidences that the inflammasome has a role in several inflammatory processes, from pathogen clearance to chronic inflammation or tissue repair. Thus, modulation of the inflammasome activity appears as an important target in the development of effective approaches in regenerative medicine.In this review, we discuss the main points of the current understanding on the host response to implanted biomaterials and how the paradigm of “immune-evasive” biomaterials has shifted over the last years; the significance of the inflammasome in the inflammatory response to biomaterials; and the growing idea that the immune system is of key importance in an effective tissue repair and regeneration.Statement of significanceWe herein discuss the main points of the current understanding on the host response to implanted biomaterials and how the paradigm of “immune-evasive” biomaterials has shifted to “immune-interactive” over the last years; the significance of the inflammasome in the inflammatory response to biomaterials; and the growing idea that the immune system is of key importance in an effective tissue repair and regeneration, supporting the emerging concept of Regenerative Immunology. The inflammasome is a recent and central concept in immunology research. Since the beginning of this century the inflammasome is viewed as key platform of the innate immune response. We believe that, successful modulation of the inflammasome activity will become a milestone in the fields of tissue engineering and regenerative medicine.Graphical abstractGraphical abstract for this article
  • Graphene oxide induces p62/SQSTM-dependent apoptosis through the
           impairment of autophagic flux and lysosomal dysfunction in PC12 cells
    • Abstract: Publication date: Available online 29 September 2018Source: Acta BiomaterialiaAuthor(s): Xiaoli Feng, Lu Chen, Weihong Guo, Yaqing Zhang, Xuan Lai, Longquan Shao, Yiping Li Graphene oxide (GO), as a two-dimensional carbon nanosheet, has been extensively studied for potential biomedical applications due to its notable properties. Although a growing number of studies have investigated the adverse effects of GO nanosheets, the available toxicity data concerning GO’s effect on the neuronal cells remain highly limited. In this work, we systematically investigated the toxic responses of commercially available GO on a rat pheochromocytoma-derived PC12 cell line, which was an ideal in vitro model to study the neurotoxicity of GO. GO exerted a significant toxic effect on PC12 cells in a dose- and time-dependent manner. GO treatments under doses of 40, 50, and 60 μg/mL triggered an autophagic response and the blockade of autophagic flux via disrupting lysosome degradation capability. Caspase 9-mediated apoptosis was also observed in GO-treated cells. Moreover, GO-induced apoptosis was relevant to the aberrant accumulation of autophagy substrate p62/SQSTM. Inhibition of the accumulation of autophagic substrate alleviated GO-caused apoptotic cell death. Our findings raise a concern for the putative biomedical applications of GO in the form of diagnostic and therapeutic tools, where its systematic biocompatibility should be thoroughly explored.Statement of significanceGraphene oxide (GO) has attracted considerable interests in biomedical fields, which also resulted in numerous safety risks to human bodies. It is urgently required to establish a paradigm for accurately evaluating their adverse effects in biological systems. This study thoroughly explored the neurotoxicity of GO in PC12 cells. We found GO triggered an increased autophagic response and the impairment of autophagic flux, which was was functionally involved in cell apoptosis. Inhibition of excessive accumulation of autophagic cargo attenuated apoptotic cell death. Our findings highlight deep considerations on the regulation mechanism of autophagy-lysosomes-apotosis-axis, which will contribute to a better understanding of the neurotoxicity of graphene-family nanomaterials, and provide a new insight in the treatment of cancer cells at nanoscale levels.Graphical abstractGraphical abstract for this article
  • Bactericidal effects of nanopatterns: a systematic review
    • Abstract: Publication date: Available online 29 September 2018Source: Acta BiomaterialiaAuthor(s): Khashayar Modaresifar, Sara Azizian, Mahya Ganjian, Lidy E. Fratila-Apachitei, Amir A. Zadpoor We systematically reviewed the currently available evidence on how the design parameters of surface nanopatterns (e.g. height, diameter, and interspacing) relate to their bactericidal behavior. The systematic search of the literature resulted in 46 studies that satisfied the inclusion criteria of examining the bactericidal behavior of nanopatterns with known design parameters in absence of antibacterial agents. Twelve of the included studies also assessed the cytocompatibility of the nanopatterns. Natural and synthetic nanopatterns with a wide range of design parameters were reported in the included studies to exhibit bactericidal behavior. However, most design parameters were in the following ranges: heights of 100-1000 nm, diameters of 10-300 nm, and interspacings of < 500 nm. The most commonly used type of nanopatterns were nanopillars, which could kill bacteria in the following range of design parameters: heights of 100-900 nm, diameters of 20-207 nm, and interspacings of 9-380 nm. The vast majority of the cytocompatibility studies (11 out of 12) showed no adverse effects of bactericidal nanopatterns with the only exception being nanopatterns with extremely high aspect ratios. The paper concludes with a discussion on the evidence available in the literature regarding the killing mechanisms of nanopatterns and the effects of other parameters including surface affinity of bacteria, cell size, and extracellular polymeric substance (EPS) on the killing efficiency.Statement of significanceThe use of nanopatterns to kill bacteria without the need for antibiotics represents a rapidly growing area of research. However, the optimum design parameters to maximize the bactericidal behavior of such physical features need to be fully identified. The present manuscript provides a systematic review of the bactericidal nanopatterned surfaces. Identifying the effective range of dimensions in terms of height, diameter, and interspacings, as well as covering their impact on mammalian cells, has enabled a comprehensive discussion including the bactericidal mechanisms and the factors controlling the bactericidal efficiency. Overall, this review helps the readers have a better understanding of the state-of-the-art in the design of bactericidal nanopatterns, serving as a design guideline and contributing to the design of future experimental studies.Graphical abstractGraphical abstract for this article
  • Correlating diameter, mechanical and structural properties of poly
           (L-lactide) fibres from needleless electrospinning
    • Abstract: Publication date: Available online 29 September 2018Source: Acta BiomaterialiaAuthor(s): A. Morel, S. Domaschke, V. Urundolil Kumaran, D. Alexeev, A. Sadeghpour, S.N. Ramakrishna, S.J. Ferguson, R.M. Rossi, E. Mazza, A.E. Ehret, G. Fortunato The development and application of nanofibres requires a thorough understanding of the mechanical properties on a single fibre level including respective modelling tools for precise fibre analysis. This work presents a mechanical and morphological study of poly-L-lactide nanofibres developed by needleless electrospinning. Atomic force microscopy (AFM) and micromechanical testing (MMT) were used to characterise the mechanical response of the fibres within a diameter range of 200 to 1400 nm. Young’s moduli E determined by means of both methods are in sound agreement and show a strong increase for thinner fibres below a critical diameter of 800 nm. Similar increasing trends for yield stress and hardening modulus were measured by MMT. Finite element analyses show that the common practice of modelling three-point bending tests with either double supported or double clamped beams is prone to significant bias in the determined elastic properties, and that the latter is a good approximation only for small diameters. Therefore, an analytical formula based on intermediate boundary conditions is proposed that is valid for the whole tested range of fibre diameters, providing a consistently low error in axial Young’s modulus below 10%. The analysis of fibre morphology by differential scanning calorimetry and 2D wide-angle X-ray scattering revealed increasing polymer chains alignment in the amorphous phase and higher crystallinity of fibres for decreasing diameter. The combination of these observations with the mechanical characterisation suggests a linear relationship between Young’s modulus and both crystallinity and molecular orientation in the amorphous phase.Statement of significanceFibrous membranes have rapidly growing use in various applications, each of which comes with specific property requirements. However, the development and production of nanofibre membranes with dedicated mechanical properties is challenging, in particular with techniques suitable for industrial scales such as needleless electrospinning. It is therefore a key step to understand the mechanical and structural characteristics of single nanofibres developed in this process, and to this end, the present work presents changes of internal fibre structure and mechanical properties with diameter, based on dedicated models. Special attention was given to the commonly used models for analyzing Young’s modulus of single nanofibers in three-point bending tests, which are shown to be prone to large errors, and an improved robust approach is proposed.Graphical abstractGraphical abstract for this article
  • Mesoporous bioactive glasses (MBGs): promising platforms for antibacterial
    • Abstract: Publication date: Available online 28 September 2018Source: Acta BiomaterialiaAuthor(s): Saeid Kargozar, Maziar Montazerian, Sepideh Hamzehlou, Hae-Won Kim, Francesco Baino The control of bacterial infections is of particular importance in the field of tissue engineering. Recently, much attention has been addressed toward the use of mesoporous bioactive glasses (MBGs) for antibacterial strategies, primarily because of their capability of acting as carriers for the local release of antimicrobial agents. The incorporation of antibacterial metallic ions including silver (Ag+), zinc (Zn2+), copper (Cu+ and Cu2+), cerium (Ce3+ and Ce4+), and gallium (Ga3+) cations into the MBG structure and their controlled release is proposed as one of the most attractive strategies for inhibiting bacterial growth and reproduction. Moreover, the possibility of loading and delivering various antibacterial biomolecules (e.g., antibiotics) through the porous structure of MBGs makes them as ideal candidates for antibacterial applications. In this review, we aim to present a comprehensive evaluation of MBG potential regarding antibacterial activities. For this purpose, different types of antibacterial ion-doped and drug-loaded MBGs are introduced and discussed in the light of existing knowledge, along with the significant challenges ahead.Statement of significancePrevention and treatment of infections is one of the today’s greatest challenges in medical sciences, also considering the well-known issues related to increased bacterial resistance to antibiotics. The advent of mesoporous glasses led to the birth of a new class of multifunctional biomaterials acting as bioactive platforms for the local release of organic or inorganic agents eliciting an antimicrobial effect. This reviews summarizes the state of the art of MBGs in this field, highlighting the latest evolutions and the specific role played by metallic antimicrobial ions that can be incorporated in the glass composition and then properly released. Perspective for tissue engineering applications are also discussed to provide an up-to-date contribution that is useful to both experienced scientists and early-stage researchers.Graphical abstractGraphical abstract for this article
  • Antimicrobial activity of commercial calcium phosphate based materials
           functionalized with vanillin
    • Abstract: Publication date: Available online 28 September 2018Source: Acta BiomaterialiaAuthor(s): Lorena Polo, Borja Díaz de Greñu, Elena Della Bella, Stefania Pagani, Paola Torricelli, José Luis Vivancos, María Ruiz-Rico, José M. Barat, Elena Aznar, Ramón Martínez-Máñez, Milena Fini, Félix Sancenón Infections represent one of the most frequent causes of arthroplasty revision. Thus, design of new antimicrobial scaffolds to reduce implant rejections, bone infections and associated medical costs is highly desired. In recent years, essential oil components (EOCs) have merged as compounds with significant antimicrobial activity that can be attached to specific surfaces to enhance and prolong their antimicrobial effect. Herein calcium phosphate CaP regenerative materials have been coated with a vanillin derivative to combine its original bone regeneration properties with antimicrobial action of EOCs. Materials in form of microparticles and blocks were prepared and fully characterized. Clonogenic viability tests demonstrated that low concentrations of material (10 mg·mL-1) resulted effective to kill 100% of E.coli DH5α bacteria. Additionally, vanillin containing scaffolds did not display any toxic effect over cells, yet they preserve the ability to express alkaline phosphatase (ALPL), collagen type 1, chain α1 (COL1A1) and bone gamma-carboxyglutamic acid-containing protein or osteocalcin (BGLAP), which are genes typically expressed by osteoblasts. These results demonstrate that commercially available scaffolds can be functionalized with EOCs, achieving antimicrobial activity and open up a new approach for the treatment and prevention of infection.Statement of significanceDuring the last years, the interest in bone regenerative materials with antibiotic properties has increased, since prosthesis infection is one of the most usual complications in implant surgery. In this work, we report a hybrid system composed by a calcium phosphate material (powders and scaffolds) functionalized with the derivative of an essential oil component (EOC). Our purpose was to provide the calcium phosphate material with antimicrobial activity without harming its bone regenerative capability. The obtained results were encouraging, which opens up the possibility of developing new modified materials for the prevention and treatment of bone infection.Graphical abstractGraphical abstract for this article
  • Gene Expression and Protein Synthesis of Esterase from Streptococcus
           mutans are affected by Biodegradation By-product from Methacrylate Resin
           Composites and Adhesives
    • Abstract: Publication date: Available online 28 September 2018Source: Acta BiomaterialiaAuthor(s): Bo Huang, Lida Sadeghinejad, Olabisi I.A. Adebayo, Ma D, Yizhi Xiao, Walter L. Siqueira, Dennis G. Cvitkovitch, Yoav Finer An esterase from S. mutans UA159, SMU_118c, was shown to hydrolyze methacrylate resin-based dental monomers.ObjectiveTo investigate the association of SMU_118c to the whole cellular hydrolytic activity of S. mutans toward polymerized resin composites, and to examine how the bacterium adapts its hydrolytic activity in response to environmental stresses triggered by the presence of a resin composites and adhesives biodegradation by-product (BBP).Materials and MethodsBiofilms of S. mutans UA159 parent wild strain, SMU_118c knockout strain (ΔSMU_118c), and SMU_118c complemented strain were incubated with photo-polymerized resin composite. High performance liquid chromatography was used to quantify the amount of a universal 2,2-Bis[4-(2-hydroxy-3-methacryloxypropoxy)phenyl]propane (bisGMA)-derived BBP, bishydroxy-propoxy-phenyl-propane (bisHPPP) in the media. Fluorescence in situ hybridization (FISH) and quantitative proteomic analysis were used to measure SMU_118c gene expression and production of SMU_118c protein, respectively, from biofilms of S. mutans UA159 wild strain that were cultured with bisHPPP.ResultsThe levels of bisHPPP released from composite were similar for ΔSMU_118c and media control, and these were significantly lower compared to the parent wild-strain UA159 and complemented strain (ΔSMU_118cC) (p
  • Mechanically stimulated osteochondral organ culture for evaluation of
           biomaterials in cartilage repair studies
    • Abstract: Publication date: Available online 28 September 2018Source: Acta BiomaterialiaAuthor(s): ML Vainieri, D. Wahl, M. Alini, GJVM van Osch, S. Grad Surgical procedures such as microfracture or autologous chondrocyte implantation have been used to treat articular cartilage lesions; however, repair often fails in terms of matrix organization and mechanical behaviour. Advanced biomaterials and tissue engineered constructs have been developed to improve cartilage repair; nevertheless, their clinical translation has been hampered by the lack of reliable in vitro models suitable for pre-clinical screening of new implants and compounds.In this study, an osteochondral defect model in a bioreactor that mimics the multi-axial motion of an articulating joint, was developed. Osteochondral explants were obtained from bovine stifle joints, and cartilage defects of 4 mm diameter were created. The explants were used as an interface against a ceramic ball applying dynamic compressive and shear loading. Osteochondral defects were filled with chondrocytes-seeded fibrin-polyurethane constructs and subjected to mechanical stimulation. Cartilage viability, proteoglycan accumulation and gene expression of seeded chondrocytes were compared to free swelling controls. Cells within both cartilage and bone remained viable throughout the 10-day culture period. Loading did not wear the cartilage, as indicated by histological evaluation and glycosaminoglycan release. The gene expression of seeded chondrocytes indicated a chondrogenic response to the mechanical stimulation. Proteoglycan 4 and cartilage oligomeric matrix protein were markedly increased, while mRNA ratios of collagen type II to type I and aggrecan to versican were also enhanced. This mechanically stimulated osteochondral defect culture model provides a viable microenvironment and will be a useful pre-clinical tool to screen new biomaterials and biological regenerative therapies under relevant complex mechanical stimuli.Statement of SignificanceArticular cartilage lesions have a poor healing capacity and reflect one of the most challenging problems in orthopedic clinical practice. The aim of current research is to develop a testing system to assess biomaterials for implants, that can permanently replace damaged cartilage with the original hyaline structure and can withstand the mechanical forces long term.Here, we present an osteochondral ex vivo culture model within a cartilage bioreactor, which mimics the complex motion of an articulating joint in vivo. The implementation of mechanical forces is essential for pre-clinical testing of novel technologies in the field of cartilage repair, biomaterial engineering and regenerative medicine. Our model provides a unique opportunity to investigate healing of articular cartilage defects in a physiological joint-like environment.Graphical abstractGraphical abstract for this article
  • Peroxidase-Immobilized Porous Silica Particles for In Situ Formation of
           Peroxidase-Free Hydrogels with Attenuated Immune Responses
    • Abstract: Publication date: Available online 28 September 2018Source: Acta BiomaterialiaAuthor(s): Li Li, Ki Hyun Bae, Shengyong Ng, Atsushi Yamashita, Motoichi Kurisawa Enzymatic crosslinking chemistry using horseradish peroxidase (HRP) has been widely utilized as an effective approach to fabricating injectable hydrogels with high efficiency under mild reaction conditions. However, their clinical applications are limited by the immunogenicity of the plant-derived enzyme. Herein we report the design, synthesis and characterization of HRP-immobilized porous silica particles (HRP-particles) and their use for in situ formation of HRP-free hydrogels. HRP was immobilized on aminopropyl-modified porous silica particles of 70–140 µm in diameter via poly(ethylene glycol) spacers of different molecular weights by reductive amination reaction. Two different HRP-free hydrogels based on dextran-tyramine and gelatin-hydroxyphenylpropionic acid (GHPA) conjugates were produced by passing a solution containing the conjugates and H2O2 through a syringe packed with HRP-particles. The storage modulus and gelation rate of both hydrogels were tunable by varying the contact time between the polymer solution and HRP-particles. Our in vitro study revealed that HRP-free GHPA hydrogel was less stimulatory to activated mouse macrophages than HRP-containing GHPA hydrogel with the same stiffness. Furthermore, HRP-free GHPA hydrogel exhibited remarkably lower levels of local and systemic inflammation than HRP-containing one upon subcutaneous injection in immunocompetent C57BL/6J mice. The attenuated immunogenicity of HRP-free GHPA hydrogels makes them an attractive platform for tissue engineering applications.Statement of SignificanceThe immunogenicity of HRP is a significant issue for clinical application of HRP-catalyzed in situ forming hydrogels. HRP-particles are developed to overcome the safety concerns by fabricating HRP-free hydrogels. The porosity of silica particles and molecular weight of poly(ethylene glycol) spacers are discovered as important factors determining the catalytic ability of HRP-particles to induce the in situ crosslinking of polymer-phenol conjugates. Although several articles speculate the potential of HRP to trigger immune responses when administered as a part of hydrogel formulation, no literature has attempted to investigate the immunogenicity of HRP-containing hydrogels in comparison with HRP-free hydrogels. Our findings suggest that the immunogenicity issue should be carefully considered before clinical translation of HRP-containing hydrogels.Graphical abstractGraphical abstract for this article
  • Antibacterial and immunogenic behavior of silver coating on additively
           manufactured porous titanium
    • Abstract: Publication date: Available online 28 September 2018Source: Acta BiomaterialiaAuthor(s): M. Croes, S. Bakhshandeh, I.A.J. van Hengel, K. Lietaert, K.P. van Kessel, B. Pouran, B.C.H. van der Wal, H.C. Vogely, W. Van Hecke, A.C. Fluit, C.H.E. Boel, J. Alblas, A.A. Zadpoor, H. Weinans, S. Amin Yavari Implant-associated infections (IAI) are often recurrent, expensive to treat, and associated with high rates of morbidity, if not mortality. We biofunctionalized the surface of additively manufactured volume-porous titanium implants using electrophoretic deposition (EPD) as a way to eliminate the peri-operative bacterial load and prevent IAI. Chitosan-based (Ch) coatings were incorporated with different concentrations of silver (Ag) nanoparticles or vancomycin. A full-scale in vitro and in vivo study was then performed to evaluate the antibacterial, immunogenic, and osteogenic activity of the developed implants. In vitro, Ch+vancomycin or Ch+Ag coatings completely eliminated, or reduced the number of planktonic and adherent Staphylococcus aureus by up to 4 orders of magnitude, respectively. In an in vivo tibia osteomyelitis model, Ch+Ag coatings caused no adverse immune or bone response under aseptic conditions. Following Staphylococcus aureus inoculation, Ch+vancomycin coatings reduced the implant infection rate as compared to chitosan-only coatings. Ch+Ag implants did not demonstrate antibacterial effects in vivo and even aggravated infection-mediated bone remodeling including increased osteoclast formation and inflammation-induced new bone formation. As an explanation for the poor antibacterial activity of Ch+Ag implants, it was found that antibacterial Ag concentrations were cytotoxic for neutrophils, and that non-toxic Ag concentrations diminished their phagocytic activity. This study shows the potential of EPD coating to biofunctionalize porous titanium implants with different antibacterial agents. Using this method, Ag-based coatings seem inferior to antibiotic coatings, as their adverse effects on the normal immune response could cancel the direct antibacterial effects of Ag nanoparticles.Statement of significanceImplant-associated infections (IAI) are a clinical, societal, and economical burden. Surface biofunctionalization approaches can render complex metal implants with strong local antibacterial action. The antibacterial effects of inorganic materials such as silver nanoparticles (Ag NPs) are often highlighted under very confined conditions in vitro. As a novelty, this study also reports the antibacterial, immunogenic, and osteogenic activity of Ag NP-coated additively-manufactured titanium in vivo. Importantly, it was found that the developed coatings could impair the normal function of neutrophils, the most important phagocytic cells protecting us from acute bacterial infections. Not surprisingly, the Ag NP-based coatings were outperformed by an antibiotic-based coating. This emphasizes the importance of also targeting implant immune-modulatory functions in future coating strategies against IAI.Graphical abstractGraphical abstract for this article
  • Hydration-Induced Nano- to Micro-Scale Self-Recovery of the Tooth Enamel
           of the Giant Panda
    • Abstract: Publication date: Available online 28 September 2018Source: Acta BiomaterialiaAuthor(s): Zengqian Liu, Zhaoyong Weng, Zhao-Feng Zhai, Nan Huang, Zhen-Jun Zhang, Jun Tan, Chuanbin Jiang, Da Jiao, Guoqi Tan, Jian Zhang, Xin Jiang, Zhefeng Zhang, Robert O. Ritchie The tooth enamel of vertebrates comprises a hyper-mineralized bioceramic, but is distinguished by an exceptional durability to resist impact and wear throughout the lifetime of organisms; however, enamels exhibit a low resistance to the initiation of large-scale cracks comparable to that of geological minerals based on fracture mechanics. Here we reveal that the tooth enamel, specifically from the giant panda, is capable of developing durability through counteracting the early stage of damage by partially recovering its innate geometry and structure at nano- to micro- length-scales autonomously. Such an attribute results essentially from the unique architecture of tooth enamel, specifically the vertical alignment of nano-scale mineral fibers and micro-scale prisms within a water-responsive organic-rich matrix, and can lead to a decrease in the dimension of indent damage in enamel introduced by indentation. Hydration plays an effective role in promoting the recovery process and improving the indentation fracture toughness of enamel (by ∼73%), at a minor cost of micro-hardness (by ∼5%), as compared to the dehydrated state. The nano-scale mechanisms that are responsible for the recovery deformation, specifically the reorientation and rearrangement of mineral fragments and the inter- and intra-prismatic sliding between constituents that are closely related to the viscoelasticity of organic matrix, are examined and analyzed with respect to the structure of tooth enamel. Our study sheds new light on the strategies underlying Nature’s design of durable ceramics which could be translated into man-made systems in developing high-performance ceramic materials.Statement of significanceTooth enamel plays a critical role in the function of teeth by providing a hard surface layer to resist wear/impact throughout the lifetime of organisms; however, such enamel exhibits a remarkably low resistance to the initiation of large-scale cracks, of hundreds of micrometers or more, comparable to that of geological minerals. Here we reveal that tooth enamel, specifically that of the giant panda, is capable of partially recovering its geometry and structure to counteract the early stages of damage at nano- to micro-scale dimensions autonomously. Such an attribute results essentially from the architecture of enamel but is markedly enhanced by hydration. Our work discerns a series of mechanisms that lead to the deformation and recovery of enamel and identifies a unique source of durability in the enamel to accomplish this function. The ingenious design of tooth enamel may inspire the development of new durable ceramic materials in man-made systems.Graphical abstractGraphical abstract for this article
  • Nanoparticulate-based drug delivery systems for small molecule
           anti-diabetic drugs: an emerging paradigm for effective therapy
    • Abstract: Publication date: Available online 28 September 2018Source: Acta BiomaterialiaAuthor(s): Siddharth Uppal, Kishan S Italiya, Deepak Chitkara, Anupama Mittal Emergence of nanoparticulate drug delivery systems in diabetes has facilitated improved delivery of small molecule drugs which could dramatically improve the quality of life for diabetics. Conventional dosage forms of the anti-diabetic drugs exhibit variable/less bioavailability and short half-life, demanding frequent dosing and causing increased side-effects resulting in ineffectiveness of therapy and non-compliance with the patients. Considering the chronic nature of diabetes, nanotechnology-based approaches are more promising in terms of providing site-specific delivery of drugs with higher bioavailability and reduced dosage regimen. Nanomedicines act at the cellular and molecular levels to enhance the uptake of the drug into the cells or block the efflux mechanisms thus retaining the drug inside the cell for a longer duration of time. Many studies have hinted at the possibility of administering peptide drugs like glucagon like peptides orally by encapsulation into nanoparticles. Nanoparticles also allow further modifications including their encapsulation into microparticles, polyethylene glycol (PEG)- PEGylation- or functionalization with ligands for active targeting. Nevertheless, such remarkable benefits are fraught with their long-term safety concerns, regulatory hurdles, limitations of scale-up and ineffective patent protection which have hindered their commercialization. This review summarizes the latest advances in the area of nanoformulations as applied to the delivery of anti-diabetics.Statement of significanceThe present work describes the latest advancements in the area of nanoformulations for anti-diabetic therapy along with highlighting the advantages that these nanoformulations offer at molecular level for diabetes.Although several potent orally active anti-hyperglycemic agents are available, the current challenges in efficient management of diabetes include optimization of the present therapies to ensure an optimum and stable level of glucose, and also to reduce the occurrence of long term complications associated with diabetes. Nanoformulations because of their high surface area to volume ratio provide improved efficacy, targeting their delivery to the desired site of action tends to minimize adverse effects and administration of peptide drugs by oral route is also possible by encapsulating them in nanoparticles. As we reflect on the success and failures of latest research on nanoformulations for the treatment of diabetes, it is important not to dwell on lack of FDA approvals but rather define future directions that guarantee more effective anti-diabetic treatment. In proposed review we have explored the latest advancement in anti-diabetic nanotechnology based formulations.Graphical abstractGraphical abstract for this article
  • Optimal biomaterials for tracheal epithelial grafts: an in vitro
           systematic comparative analysis
    • Abstract: Publication date: Available online 27 September 2018Source: Acta BiomaterialiaAuthor(s): Ratna Varma, Fabio G. Aoki, Kayla Soon, Golnaz Karoubi, Thomas K. Waddell Tracheal injury, stenosis, and malignancy demand tracheal reconstruction, which often fails due to the lack of a functioning epithelium. We performed an extensive comparative analysis to determine optimal biomaterials for developing tracheal epithelial grafts with mucociliary function. We screened Hyaluronan-Poly(Ethylene Glycol), Chitosan-Collagen, Collagen Vitrigel Membrane, Fibrin Glue, Silk Fibroin, and Gelatin based on various parameters including mechanical strength, bulk degradation, cell attachment, spreading, metabolic activity, focal adhesion formation, and differentiation into ciliated and goblet cells. Silk Fibroin had significantly higher tensile strength (21.23 ± 4.42 MPa), retained 50% of its mass across 5 weeks, allowed 80-100% cell spreading and increasing metabolic activity across 10 days, focal adhesion formation within 2 hours, and differentiation into 5.9 ± 2.6% goblet cells. Silk Fibroin, however, led to poor ciliation, producing 5.5 ± 3.9% ciliated cells, whereas Collagen Vitrigel Membrane promoted excellent ciliation. To capitalize on the mechanical and differentiation benefits of its respective components, we developed a composite biomaterial of Silk Fibroin and Collagen Vitrigel Membrane (SF-CVM), which demonstrated enhanced maturation into 20.6 ± 1.7% ciliated and 5.6 ± 1.0% goblet cells. Development of biomaterials-based airway epithelial grafts that provide desirable mechanics and differentiation is a major step towards treatment of airway disease.Statement of SignificanceTracheal blockage, injury, and malignancy greater than 50% of the adult tracheal length cannot be safely resected. Tracheal replacement is one approach, but a major cause of transplant failure is the lack of a functioning epithelium. While tissue engineering for tracheal regeneration using biomaterials is promising, there is currently no gold standard. Therefore, we performed a systematic comparative study to characterize relevant materials for generating a biomaterials-based airway epithelial graft. We developed a composite biomaterial intended for surgical implantation providing tensile strength, slow biodegradation, and optimal support for differentiation of mature epithelia. This is a significant step augmenting current state-of-the-art methods for airway surgeries, laryngeal reconstruction, and tracheal tissue engineering.Graphical abstractGraphical abstract for this article
  • Metal-organic frameworks, NH2-MIL-88(Fe), as carriers for ophthalmic
           delivery of brimonidine
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Se-Na Kim, Chun Gwon Park, Beom Kang Huh, Seung Ho Lee, Chang Hee Min, Yun Young Lee, Young Kook Kim, Ki Ho Park, Young Bin Choy We have proposed a metal-organic framework (MOF), NH2-MIL-88(Fe), as a novel carrier for topical drug delivery to the eye. The NH2-MIL-88(Fe) particles were prepared via a solvothermal synthesis method and their structure was confirmed by powder X-ray diffraction, Fourier transform infrared analysis, thermogravimetric analysis, electron microscopy, and N2 adsorption-desorption measurements. When brimonidine, an anti-glaucoma medicine, was encapsulated into NH2-MIL(Fe)-88 (i.e., NH2-MIL-88(Fe)/Br), the drug was loaded at 121.3 µg/mg and released in a sustained manner for up to 12 h. The NH2-MIL-88(Fe)/Br exhibited mucoadhesive properties and remained on rabbit eyes for a period of up to 4 h. Consequently, a high concentration of brimonidine was found in tears for a prolonged period after the administration of NH2-MIL-88(Fe)/Br, which resulted in a greater than two-fold increase in drug bioavailability and activity period compared with those of Alphagan P, which are brimonidine eye drops already approved for clinical use. Hence, NH2-MIL-88(Fe) is suggested to be a promising carrier for topical delivery to the eye that provides enhanced bioavailability of ocular drugs.Statement of SignificanceWe suggest NH2-MIL(Fe)-88, a type of metal-organic frameworks (MOFs), as delivery carriers of an ophthalmic drug, brimonidine. The NH2-MIL(Fe)-88 particles possess a mucoadhesive property, hence prolonged retention in the preocular space when topically administered to the eye. The particles can also encapsulate the drug in their micro-pores, through which the drug can be released in a sustained manner. Therefore, when tested to rabbit eyes in vivo, the drug-loaded NH2-MIL(Fe)-88 particles were shown to enhance the ocular drug bioavailability, as compared with Alphagan P, the marketed eye drops of brimonidine.Graphical abstractGraphical abstract for this article
  • Nanocarriers with conjugated antimicrobials to eradicate pathogenic
           biofilms evaluated in murine in vivo and human ex vivo infection models
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Yong Liu, Yijin Ren, Yuanfeng Li, Linzhu Su, Yumin Zhang, Fan Huang, Jinjian Liu, Jianfeng Liu, Theo G. van Kooten, Yingli An, Linqi Shi, Henny C. van der Mei, Henk J. Busscher Conventional antimicrobials are becoming increasingly ineffective for treating bacterial infection due to the emergence of multi-drug resistant (MDR) pathogens. In addition, the biofilm-mode-of-growth of infecting bacteria impedes antimicrobial penetration in biofilms. Here, we report on poly(ethylene)glycol-poly(β-amino esters) (PEG-PAE) micelles with conjugated antimicrobials, that can uniquely penetrate biofilms, target themselves to bacterial cell surfaces once inside the low-pH environment of a biofilm and release conjugated antimicrobials through degradation of their ester-linkage with PAE by bacterial lipases. In vitro, PEG-PAE micelles with conjugated Triclosan (PEG-PAE-Triclosan) yielded no inadvertent leakage of their antimicrobial cargo and better killing of MDR Staphylococcus aureus, Escherichia coli and oral streptococcal biofilms than Triclosan in solution. In mice, PEG-PAE-Triclosan micelles with conjugated Triclosan yielded better eradication efficacy towards a MDR S. aureus-infection compared with Triclosan in solution and Triclosan-loaded micelles at equal Triclosan-equivalent concentrations. Ex vivo exposure of multi-species oral biofilms collected from orthodontic patients to PEG-PAE-Triclosan micelles, demonstrated effective bacterial killing at 30–40 fold lower Triclosan-equivalent concentrations than achieved by Triclosan in solution. Importantly, Streptococcus mutans, the main causative organism of dental caries, was preferentially killed by PEG-PAE-Triclosan micelles. Thus PEG-PAE-Triclosan micelles present a promising addendum to the decreasing armamentarium available to combat infection in diverse sites of the body.Statement of SignificancepH-adaptive polymeric micelles with conjugated antimicrobials can efficiently eradicate infectious biofilms from diverse body sites in mice and men. An antimicrobial was conjugated through an ester-linkage to a poly(ethylene glycol) (PEG)/poly(β-amino ester) block copolymer to create micellar nanocarriers. Stable micelle structures were formed by the hydrophobic poly(β-amino ester) inner core and a hydrophilic PEG outer shell. Thus formed PEG-PAE-Triclosan micelles do not lose their antimicrobial cargo underway to an infection site through the blood circulation, but penetrate and accumulate in biofilms to release antimicrobials once inside a biofilm through degradation of its ester-linkage by bacterial lipases, to kill biofilm-embedded bacteria at lower antimicrobial concentrations than when applied in solution. PEG-PAE-Triclosan micelles effectively eradicate biofilms of multi-drug-resistant pathogens and oral bacteria, most notably highly cariogenic Streptococcus mutans, in mice and men respectively, and possess excellent clinical translation possibilities.Graphical abstractGraphical abstract for this article
  • CX-5461-loaded nucleolus-targeting nanoplatform for cancer therapy through
           induction of pro-death autophagy
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Yanhong Duo, Min Yang, Zhenya Du, Chuhan Feng, Chen Xing, Yanping Wu, Zhenhua Xie, Fang Zhang, Laiqiang Huang, Xiaowei Zeng, Hongbo Chen Various drugs have been designed in the past to act on intracellular targets. For the desired effects to be exerted, these drugs should reach and accumulate in specific subcellular organelles. CX-5461 represents a potent small-molecule inhibitor of rRNA synthesis that specifically inhibits the transcription driven by RNA polymerase (Pol) I and induces tumor cell death through triggering a pro-death autophagy. In the current study an innovative kind of CX-5461-loaded mesoporous silica nano-particles enveloped by polyethylene glycol (PEG), polydopamine (PDA) and AS-1411 aptamer (MSNs-CX-5461@PDA-PEG-APt) with the aim of treating cancer cells was constructed, in which the high-surface-area MSNs allowed for high drug loading, PDA acted as gatekeeper to prevent the leakage of CX-5461 from MSNs, PEG grafts on PDA surfaces increased the stable and biocompatible property in physiological condition, and AS-1411 aptamer promoted the nucleolar accumulation of CX-5461. MSNs-CX-5461@PDA-PEG-APt was characterized regarding releasing characteristics, steadiness, encapsulation of drugs, phase boundary potential as well as sizes of particles. Expectedly, In vitro assays showed that aptamer AS-1411 significantly increased the nucleolar accumulation of CX-5461. The aptamer-tagged CX-5461-loaded MSNs demonstrated to be more cytotoxic to cervical cancer cells compared to the control MSNs, due to relatively strong inhibition of rRNA transcription and induction of pro-death autophagy. The in vivo treatment with AS-1411-tagged CX-5461-loaded MSNs showed a stronger distribution in tumor tissues by animal imaging assay and a significantly higher inhibition effect on the growth of HeLa xenografts compared to AS-1411-untagged CX-5461-loaded MSNs. In addition, histology analysis indicated that MSNs-CX-5461@PDA-PEG-APt did not exhibit any significant toxicity on main organs. These results collectively suggested that MSNs-CX-5461@PDA-PEG-APt represents both a safe and potentially nucleolus-targeting anti-cancer drug.Statement of SignificanceMany drugs function in specific subcellular organelles. CX-5461 is a specific inhibitor of nucleolar rRNA synthesis. Here, we reported a novel aptamer-tagged nucleolus-targeting CX-5461-loaded nanoparticle, which specifically accumulated in nucleoli and significantly inhibited the tumor growth in vitro and in vivo through inhibiting rRNA transcription and triggering a pro-death autophagy.Graphical abstractGraphical abstract for this article
  • A positron emission tomography image-guidable unimolecular micelle
           nanoplatform for cancer theranostic applications
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Jia Yang, Weifei Lu, Jinling Xiao, Qi Zong, Haixing Xu, Yihua Yin, Hao Hong, Wenjin Xu Unimolecular micelles based on hyperbranched polyamidoamine (PAMAM) dendrimer were synthesized as both a cargo delivery vector and an imaging agent for triple-negative breast tumors, and the chemical synthesis procedures are detailed in this study. With the chemical conjugation of a peptide (F3, against cellular nucleolin) to increase its cellular internalization, these micelles can accumulate potently and specifically in breast cancer cells (e.g., MDA-MB-231). The size and morphology of these PAMAM-based micelles have been measured by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The hydrazone bond (responsive to pH alteration) between the loaded doxorubicin (DOX, as a model drug here) and PAMAM micelles enables cargo release following pH changes. Flow cytometry and confocal fluorescence microscopy revealed that PAMAM micelles with F3 attachment (PAMAM-DOX-F3) had stronger internalization into MDA-MB-231 cells (nucleolin-positive) than PAMAM micelles without F3 conjugation (PAMAM-DOX), whereas both of them have minimal interactions with L929 fibroblasts (nucleolin-negative). The positron-emitting isotope 64Cu was added into PAMAM micelles by chelation to track their pharmacokinetic behavior (organ distribution profile) in vivo by positron emission tomography (PET) imaging. Serial PET imaging demonstrated that the accumulation of 64Cu-PAMAM-DOX-F3 in MDA-MB-231 tumors was fast, potent, and persistent (tumor uptake: 6.1 ± 1.2% injection dose per gram [%ID/g] at 24 h p.i.), significantly higher than that of 64Cu-PAMAM-DOX (2.5 ± 0.4%ID/g at the same time). Their distribution profiles in other organs/tissues were quite similar, with a relatively short circulation time. In addition, ex vivo fluorescence imaging confirmed that DOX can be delivered efficiently by these PAMAM micelles to MDA-MB-231 tumors. Deducing from these data, we believe that PAMAM-based micelles can be useful for selective combinational treatment of cancer.Statement of SignificanceMicelles are a very useful biomaterial for theranostic purposes, and one of the major hurdles for micelles (particularly those from self-assembling) is their relatively low stability, especially when administered in vivo. In this study, we have attempted to overcome this limitation by designing unimolecular micelles (based on the concept of “one micelle is composed of one macromolecule”) from polyamidoamine (PAMAM) dendrimers, in which the drug cargos (e.g., doxorubicin) are chemically attached to PAMAM through a hydrazone bond; hence, they can be used as a tumor-selective diagnostic/therapeutic platform. These unimolecular micelles possess superior stability compared to conventional micelles and can undertake stimulus (pH)-responsive cargo release for more “targeted” cancer therapy. With the incorporation of a tumor-targeting peptide sequence (F3) and a positron-emitting isotope (copper-64), the pharmacokinetic behavior of these micelles can be readily monitored by positron emission tomography imaging technique to confirm their specificity against cancer tissues. With further optimization, this micellar platform can have a broad clinical applicability owing to its biocompatibility, selectivity, and stability.Graphical abstractGraphical abstract for this article
  • Ultrasmall gold nanosatellite-bearing transformable hybrid nanoparticles
           for deep tumor penetration
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Soyoung Son, Veerasikku G. Deepagan, Sol Shin, Hyewon Ko, Jiwoong Min, Wooram Um, Jueun Jeon, Seunglee Kwon, Eun Sook Lee, Minah Suh, Doo Sung Lee, Jae Hyung Park Since delivering drugs to an entire tumoral region leads to high therapeutic efficacy and good prognosis, achieving deep tumoral penetration of drugs is a major issue in cancer treatment. In this regard, conventional nanomedicines (>50 nm) have shown limitations in cancer therapy, primarily attributed to the heterogeneous distribution of drugs because of the physiological barrier of the tumor interstitial space. To address this issue, we prepared transformable hybrid nanoparticles (TNPs) consisting of a pH-responsive nanocarrier (PEG-PBAE) and doxorubicin (DOX)-conjugated ultrasmall (
  • Generic and reversible opto-trapping of biomolecules
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Hannes M. Beyer, Oliver S. Thomas, Nils Riegel, Matias D. Zurbriggen, Wilfried Weber, Maximilian Hörner Molecular traps can control activity and abundance of many biological factors. Here, we report the development of a generic opto-trap to reversibly bind and release biomolecules with high spatiotemporal control by illumination with non-invasive and cell-compatible red and far-red light. We use the Arapidopsis thaliana photoreceptor phytochrome B to regulate the release of diverse proteins from a variety of material scaffolds. Fusion of a short 100 amino acids “PIF-tag”, derived from the phytochrome interacting factor 6, renders arbitrary molecules opto-trap-compatible. Reversible opto-trapping of target molecules enables novel possibilities for future developments in diagnostics, therapeutics, and basic research.Statement of SignificanceThe investigation of cellular signaling events or the development of complex therapeutics and integrative diagnostic devices requires the deliberate control of biomolecule abundance and activity. During recent years, the use of natural photoreceptors within cells leveraged the control of diverse cellular events, benefiting from the superior spatial and temporal control characteristics of light as compared to conventional chemical stimuli. Concurrently, biological switches entailing intrinsic compatibility toward biological environments increasingly found application in biohybrid materials. We employ the plant red/far-red photoreceptor phytochrome B, which reversibly interacts with its phytochrome interacting factors (PIFs), for developing a generic opto-trap. This platform allows the use of red and far-red light to spatiotemporally control binding and release of arbitrary PIF-fused biomolecules from various material scaffolds.Graphical abstractGraphical abstract for this article
  • Dual-functional 3D-printed composite scaffold for inhibiting bacterial
           infection and promoting bone regeneration in infected bone defect models
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Ying Yang, Linyang Chu, Shengbing Yang, Hongbo Zhang, Ling Qin, Olivier Guillaume, David Eglin, R. Geoff Richards, Tingting Tang Infection is one of the pivotal causes of nonunion in large bone defect after trauma or tumor resection. Three-dimensional (3D) composite scaffold with multifunctional-therapeutic properties offer many advantages over allogenic or xenogenic bone grafting for the restoration of challenging infected bone defects. In the previous study, we demonstrated that quaternized chitosan (HACC)-grafted polylactide-co-glycolide (PLGA)/hydroxyapatite (HA) scaffold (PLGA/HA/HACC) via 3D-printing technique exhibited significantly improved antimicrobial and osteoconductive property in vitro, together with good biocompatibility in vivo. Hence, the present study further investigated whether such an innovative bone substitute could effectively inhibit the bacterial biofilm formation and promote bone regeneration in vivo. To evaluate the bone repairing effects of the 3D-printed scaffolds on infected cortical and cancellous bone defects scenarios, eighty female Sprague Dawley rats and thirty-six female New Zealand white rabbits were used to establish infected femoral shaft defect and condyle defect model, respectively. X-ray, micro-CT, microbiological and histopathological analyses were used to assess the anti-infection and bone repairing potential of the dual-functional porous scaffolds. We observed that HACC-grafted PLGA/HA scaffolds exhibited significantly enhanced anti-infection and bone regeneration capability in different infected bone defect models. In addition, the degradation rate of the scaffolds appeared to be closely related to the progress of infection, influencing the bone repairing potential of the scaffolds in infected bone defects models. In general, this investigation is of great significance as it demonstrates promising applications of the 3D-printed dual-functional PLGA/HA/HACC scaffold for repairing different types of bone defect under infection.Statement of SignificanceCurrently, it is clinically urgent to exploit bone substitutes with potential of bacterial inhibition and bone regeneration. However, bone scaffolds with relatively low risks of bacterial resistance and tissue toxicity used for combating infected bone defects remain to be developed. We have reported that quaternized chitosan (HACC)-grafted 3D-printed PLGA/HA composite scaffold had enhanced in vitro antimicrobial and osteoconductive property, and well cytocompatibility in our published study. This continuing study further confirmed that HACC-grafted PLGA/HA scaffolds exhibited significantly enhanced anti-infection and bone regeneration efficacy in both cortical bone defect in rat and cancellous bone defect in rabbit under infection. Meanwhile, we also found that the degradation rate of the scaffolds seemed to be closely related to the progress of infection, influencing the bone repairing potential of the scaffolds in infected bone defects models. In conclusion, this study provides significant opportunities to develop a 3D-printed bone scaffold with dual functions used for infected bone defects in future plastic and orthopaedic surgery.Graphical abstractGraphical abstract for this article
  • Graft-specific immune tolerance is determined by residual antigenicity of
           xenogeneic extracellular matrix scaffolds
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Ailsa J. Dalgliesh, Mojtaba Parvizi, Manuela Lopera-Higuita, Jeny Shklover, Leigh G. Griffiths Antigenicity remains the primary barrier towards expanding the use of unfixed xenogeneic biomaterials in clinical applications. An unfixed xenogeneic biomaterial devoid of antigenicity, with maintained structural and mechanical integrity, has potential to overcome the limitations of current clinically utilized glutaraldehyde-fixed xenogeneic biomaterials, such as heart valve bioprostheses. Unfortunately, the threshold level of residual antigenicity necessary to overcome graft-specific immune responses in unfixed xenogeneic tissue has yet to be determined. Furthermore, little information is known regarding the extent to which in vitro disruption of native ECM properties, resulting from decellularization or antigen removal procedures, are tolerated following in vivo implantation. This manuscript demonstrates that humoral adaptive immune responses are more sensitive to residual xenogeneic biomaterial antigen content than are cell-mediated adaptive responses. Critically, the threshold for tolerable residual antigenicity is identified, with removal of ≥92% of lipophilic antigens required to reduce adaptive immune responses to levels equivalent to glutaraldehyde fixed tissue. Finally, the results demonstrated that the innate immune system tolerates minor changes in protein organization provided that molecular structure is maintained. Antigen removed xenogeneic biomaterials achieving these in vitro success criteria induce in vivo adaptive and innate tolerance, while modulating pro-regenerative constructive remodeling.Statement of SignificanceRemoval of antigenic components from candidate xenogeneic biomaterials is the primary success criteria for development of extracellular matrix (ECM) scaffolds in tissue engineering applications. Currently, the threshold level of residual biomaterial antigenicity required to overcome recipient graft-specific adaptive immune responses is unknown. Additionally, the extent to which the innate immune response tolerates changes to the native ECM, resulting from the ECM scaffold production process, has yet to be determined. This manuscript not only establishes the threshold for tolerable residual antigenicity, but also demonstrates that deviations in protein organization are tolerated by the innate immune system, provided macromolecular structure remains intact. In doing so, we provide the foundation of an immunologically-acceptable unfixed xenogeneic biomaterial for use in clinical applications.Graphical abstractGraphical abstract for this article
  • Fabrication of the FGF1-functionalized sericin hydrogels with cell
           proliferation activity for biomedical application using genetically
           engineered Bombyx mori (B. mori) silk
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Feng Wang, Yuancheng Wang, Chi Tian, Sheng Xu, Riyuan Wang, Kai Hou, Wenjing Chen, Ping Zhao, Ling Yu, Zhisong Lu, David L. Kaplan, Qingyou Xia Sericin, as the major component of Bombyx mori silk, is a useful biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Here, we report the fabrication of a human acidic fibroblast growth factor (FGF1)-functionalized sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its sericin layer. Sericin, together with FGF1, were simultaneously extracted from the silk fiber and then exposed to cold-induced hydrogel formation without additional crosslinking. The fabricated FGF1 sericin hydrogels demonstrated injectability, useful mechanical properties and a porous microstructure, which contributed to cell adhesion and survival. In addition, FGF1 achieved long-term storage in the sericin hydrogels over a wide range of temperatures. Further, the sericin-FGF1 demonstrated sustained release to promote cell proliferation and wound healing. Furthermore, cellular inflammatory responses showed that the FGF1 sericin hydrogels exhibited biocompatibility and no immunogenicity. This study revealed the successful exploration of FGF1-functionalized sericin hydrogels as a new protein-based biomaterial to expand applications of FGF1 and sericin in tissue and medical engineering. Further, we demonstrated a strategy for the predesign of exogenous protein-functionalized sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale.Statement of SignificanceSericin from the Bombyx mori silk, is regarded as a desirable biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Genetically engineering the sericin with functional exogenous proteins would enhance its biofunctions and further expand its application in tissue engineering. In this study, we demonstrated a method to fabricate a human acidic fibroblast growth factor (FGF1)-functionalized sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its sericin layer. The fabricated FGF1 sericin hydrogels demonstrated injectability, porous microstructure, biocompatibility and no immunogenicity which contributed to cell adhesion and survival. Remarkably, FGF1 could achieve a long-term stability in the sericin hydrogels over a wide range of temperatures and sustained release to promote cell proliferation and wound healing. This study revealed the successful exploration of FGF1-functionalized sericin hydrogels as a new protein-based biomaterial in tissue and medical engineering application, and provided a strategy for the predesign of exogenous protein-functionalized sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale.Graphical abstractGraphical abstract for this article
  • In vivo annular repair using high-density collagen gel seeded with annulus
           fibrosus cells
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Yu Moriguchi, Brandon Borde, Connor Berlin, Christoph Wipplinger, Stephen R. Sloan, Sertac Kirnaz, Brenton Pennicooke, Rodrigo Navarro-Ramirez, Thamina Khair, Peter Grunert, Eliana Kim, Lawrence Bonassar, Roger Härtl ObjectiveThe aim is assessing the in vivo efficacy of annulus fibrosus (AF) cells seeded into collagen by enhancing the reparative process around annular defects and preventing further degeneration in a rat-tail model.Summary of background dataTreating disc herniation with discectomy may relieve the related symptoms but does not address the underlying pathology. The persistent annular defect may lead to re-herniation and further degeneration. We recently demonstrated that riboflavin crosslinked high-density collagen gels (HDC) can facilitate annular repair in vivo.Methods42 rats, tail disc punctured with an 18-gauge needle, were divided into 3 groups: untreated (n = 6), injected with crosslinked HDC (n = 18), and injected with AF cell-laden crosslinked HDC (n = 18). Ovine AF cells were mixed with HDC gels prior to injection. X-rays and MRIs were conducted over 5 weeks, determining disc height index (DHI), nucleus pulposus (NP) size, and hydration. Histological assessments evaluated the viability of implanted cells and degree of annular repair.ResultsAlthough average DHIs of both HDC gel groups were higher than those of the puncture control group at 5 weeks, the retention of disc height, NP size and hydration at 1 and 5 weeks was significant for the cellular group compared to the punctured, and at 5 weeks to the acellular group. Histological assessment indicated that AF cell-laden HDC gels have accelerated reparative sealing compared to acellular HDC gels.ConclusionsAF cell-laden HDC gels have the ability of better repairing annular defects than acellular gels after needle puncture.Statement of SignificanceThis project addresses the compelling demand of a sufficient treatment strategy for degenerative disc disease (DDD) perpetuated by annulus fibrosus (AF) injury, a major cause of morbidity and burden to health care systems. Our study is designed to answer the question of whether injectable, photo-crosslinked, high density collagen gels can seal defects in the annulus fibrosus of rats and prevent disc degeneration. Furthermore, we investigated whether the healing of AF defects will be enhanced by the delivery of AF cells (fibrochondrocytes) to these defects.The use of cell-laden collagen gels in spine surgery holds promise for a wide array of applications, from current discectomy procedures to future nucleus pulposus reparative therapies, and our group is excited about this potential.Graphical abstractGraphical abstract for this article
  • Endowing polyetheretherketone with synergistic bactericidal effects and
           improved osteogenic ability
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Jianglong Yan, Wenhao Zhou, Zhaojun Jia, Pan Xiong, Yangyang Li, Pei Wang, Qiyao Li, Yan Cheng, Yufeng Zheng Biomedical associated infections (BAI) are difficult to treat and may even lead to amputation and death, especially after the emergence of drug-resistant bacteria. The aim of this study was to harness the potential synergistic effects of multiple bactericidal agents to endow polyetheretherketone (PEEK) with the ability of achieving full eradication of planktonic and adherent bacteria while maintaining acceptable biocompatibility. In this work, a mussel inspired, silver nanoparticles (AgNPs) incorporated silk fibroin (SF)/gentamicin sulfate (GS) coating was constructed upon porous PEEK surface. The obtained coating greatly enhanced the bactericidal efficiency to Gram-positive bacteria and Gram-negative bacteria. The number of bacteria survived in the culture medium after treated with this coating was 106 fold lower than that survived after treated with PEEK sample, while the number of viable bacteria adhered to this coating was 105 lower than that adhered to PEEK sample. Furthermore, release of Ag+ and GS increased with decreasing pH, indicating great potential of this coating to be a “smart” bacteria-triggered self-defensive coating. Meanwhile, this functional coating shows favorable cytocompatibility and osteogenic ability. The mechanism behind this dual function is also partially revealed. Expectedly, this “smart” dual function coating can give a promise for PEEK to become a solution to increasingly deteriorated BAI.Statement of SignificanceIn this study, a mussel inspired, silver nanoparticles (AgNPs) incorporated silk fibroin (SF)/gentamicin sulfate (GS) coating was constructed upon porous polyetheretherketone (PEEK) surface. This design was aimed to provide a solution to the increasingly deteriorated biomedical associated infections (BAI). Actually, this design endowed PEEK with dual function: bacteria-triggered synergistic bactericidal effect and improved osteogenic ability. The combination of silver and GS exhibited synergistic bacteria killing effect on both Gram-positive and Gram-negative bacteria, which showed 106 times higher in releasing-killing and 105 times higher in anti-adhesion than that of untreated PEEK. Furthermore, release of bactericidal agents increased with decreasing pH, indicating great potential of this coating to be a bacteria-triggered self-defensive coating. More interestingly, this study revealed the mechanism of synergistic effect between silver and GS.Graphical abstractGraphical abstract for this article
  • Cartilage regeneration using arthroscopic flushing fluid-derived
           mesenchymal stem cells encapsulated in a one-step rapid cross-linked
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Jun Li, Yongcan Huang, Jun Song, Xiaolin Li, Xintao Zhang, Zhiyu Zhou, Di Chen, Peter X. Ma, Wenjing Peng, Wenxin Wang, Guangqian Zhou Many attempts have been made to repair articular cartilage defects, including mesenchymal stem cell (MSC)-based tissue engineering strategies. Although this approach shows promise, optimizing MSC sources and their delivery is challenging. This study was designed to test the feasibility of using MSCs found in the human arthroscopic flushing fluid (AFF) for cartilage regeneration, by incorporating them into a newly developed one-step rapid cross-linking hyper-branched polyPEGDA/HA hydrogel. AFF-MSCs were isolated from the original intra-articular flushing fluid of 10 patients prior to arthroscopic procedures. The hydrogel was fabricated with hyper-branched polyPEGDA and thiolated hyaluronic acid (HA). In vitro assays demonstrated that AFF-MSCs possessed the typical MSC morphology and phenotype, and maintained chondrogenic differentiation properties when encapsulated within the hydrogel. The AFF-MSC/hydrogel composite could significantly repair full-thickness cartilage defects generated in a rat model after 8 weeks of implantation; smooth cartilage was formed with evidence of hyaline cartilage formation. These data suggest that human AFF-MSCs are a novel and abundant MSC source that have high therapeutic value for cartilage regeneration.Statement of SignificanceMany attempts have been made to repair the defects of articular cartilage, including mesenchymal stem cell (MSC)-based tissue engineering strategies. Optimizing MSC sources and their delivery approaches still remain clinically challenging. Recent studies determined that MSCs derived from synovium and synovial fluid exhibited superior chondrogenic potential. However, no feasible methods to harvest these human tissues and cells have been impeding them for clinical application. Hereby, we explored a simple and easy accessible approach to obtain a new stem cell source from arthroscopic flushing fluid (AFF-MSCs), which probably contains plenty of MSCs from synovium and synovial fluid. Further experiments demonstrated that encapsulation of these stem cells with one-step rapid cross-linked polyPEGDA/HA hydrogel held very encouraging potential for cartilage regeneration.Graphical abstractGraphical abstract for this article
  • Highly flexible and degradable dual setting systems based on PEG-hydrogels
           and brushite cement
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Michaela Rödel, Jörg Teßmar, Jürgen Groll, Uwe Gbureck With respect to the composition of natural bone, we established a degradable dual setting system of different poly(ethylene glycol) (PEG)-based hydrogels combined with a brushite cement. The idea was to reinforce the inorganic calcium phosphate mineral phase with an organic, polymeric phase to alter the cement’s properties towards ductility and elasticity. Extremely flexible samples were produced via this dual setting approach with a fully reversible elasticity of the samples containing high molecular weight PEG-based hydrogel precursors. Using the decalcifying agent EDTA, the whole inorganic phase was dissolved due to Ca2+-complexation and dimensionally stable hydrogels were obtained, indicating a homogenous polymeric phase within the composites. This was also confirmed by SEM-analysis, where no discontinuities or agglomerations of the phase were observed. Additional XRD-measurements proved a significant influence of the coherent polymeric matrix on the conversion from β-TCP/MCPA to brushite with a decrease in signal intensity. The results confirmed a parallelly running process of setting reaction and gelation without an inhibition of the conversion to brushite and the formation of interpenetrating networks of hydrogel and cement. The strengths of this newly developed dual setting system are based on the material degradability as well as flexibility, which can be a promising tool for bone regeneration applications in non-load bearing craniomaxillofacial defects.Statement of SignificanceBrushite based calcium phosphate cements (CPCs) are known as bone replacement materials, which degrade in vivo and are replaced by native bone. However, the pure inorganic material shows a brittle fracture behavior. Here, the addition of a polymeric phase can influence the mechanical properties to create more ductile and flexible materials. This polymeric phase should ideally form during cement setting by a polymerization reaction to achieve high polymer loads without altering cement viscosity and it should be degradable in vivo similar to the cement itself. Therefore, we developed a dual setting system based on simultaneous cement setting of brushite and lactide modified poly(ethylene glycol) dimethacrylate (PEG-PLLA-DMA)-based hydrogel. It was evident that the gels form a continuous phase within the cement after radical polymerization with a strong reduction of cement brittleness.Graphical abstractGraphical abstract for this article
  • Fabrication of heterogeneous porous bilayered nanofibrous vascular grafts
           by two-step phase separation technique
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Weizhong Wang, Wei Nie, Xiaojun Zhou, Wei Feng, Liang Chen, Qianqian Zhang, Zhengwei You, Qiusheng Shi, Chen Peng, Chuanglong He Innterconnected porous architecture is critical for tissue engineering scaffold as well as biomimetic nanofibrous structure. In addition, a paradigm shift is recently taking place in the scaffold design from homogeneous porous scaffold to heterogeneous porous scaffold for the complex tissues. In this study, a versatile and simple one-pot method, dual phase separation, is developed to fabricate macroporous nanofibrous scaffold by phase separating the mixture solutions of immiscible polymer blends without using porogens. The macropores in the scaffold are interconnected, and their size can be tuned by the polymer blend ratio. Moreover, benefiting from the easy operation of dual phase separation technique, an innovative, versatile and facile two-step phase separation method is developed to fabricate heterogeneous porous layered nanofibrous scaffolds with different shapes, such as bilayered tubular scaffold and tri-layered cylindrical scaffold. The bilayered tubular nanofibrous scaffold composed of poly(l-lactic acid) (PLLA)/poly(l-lactide-co-ε-caprolactone) (PLCL) microporous inner layer and PLLA/poly(ε-caprolactone) (PCL) macroporous outer layer matches simultaneously the functional growth of endothelial cells (ECs) and smooth muscle cells (SMCs), and shows the favorable performance for potential small diameter blood vessel application. Therefore, this study provides the novel and facile strategies to fabricate macroporous nanofibrous scaffold and heterogeneous porous layered nanofibrous scaffold for tissue engineering applications.Statement of SignificanceThe fabrication of porous tissue engineering scaffold made of non-water-soluble polymer commonly requires the use of porogen materials. This is complex and time-consuming, resulting in greater difficulty to prepare heterogeneous porous layered scaffold for multifunctional tissues repair, such as blood vessel and osteochondral tissue. Herein, a novel, versatile and simple one-pot dual phase separation technique is developed for the first time to fabricate porous scaffold without using porogens. Simultaneously, it also endows the resultant scaffold with the biomimetic nanofibrous architecture. Based on the easy operation of this dual phase separation technique, a facile two-step phase separation method is also put forward for the first time and applied in fabricating heterogeneous porous layered nanofibrous scaffold for tissue engineering applications.Graphical abstractGraphical abstract for this article
  • Design of a migration assay for human gingival fibroblasts on
           biodegradable magnesium surfaces
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): R. Amberg, A. Elad, D. Rothamel, T. Fienitz, G. Szakacs, S. Heilmann, F. Witte A novel regenerative approach to Guided Bone Regeneration (GBR) in dental surgery is based on the development of biodegradable and volume stable barrier membranes made of metallic magnesium. Currently used volume stable barrier membranes are made of titanium-reinforced PTFE or titanium-reinforced collagen membranes, both, however, are accompanied by a high incidence of wound dehiscence resulting in membrane exposure, which leads to an increased infection risk. An exposed membrane could also occur directly after insertion due to insufficient soft tissue coverage of the membrane. In both cases, fast wound margin regeneration is required. As a first step of soft-tissue regeneration, gingival fibroblasts need to migrate over the barrier membrane and close the dehiscent wound. Based on this aim, this study investigated the migration behaviour of human gingival fibroblasts on a magnesium surface. Major experimental challenges such as formation of hydrogen bubbles due to initial magnesium corrosion and non-transparent material surfaces have been addressed to allow cell adhesion and to follow cell migration. The designed scratch-based cell migration assay involved vital fluorescent cell staining on a pre-corroded magnesium membrane to simulate in vivo wound dehiscence. The assay has been used to compare cell migration on pre-corroded magnesium to titanium surfaces and tissue culture plastic as control substrates. First results of this assay showed that human gingival fibroblasts migrate slower on pre-corroded magnesium compared to plastic and titanium. However, the scratch was finally closed on all materials. Compared to titanium surfaces and tissue culture plastic, the surface roughness and the surface free energy (SFE) could not explain slower cell migration on magnesium surfaces. Immunohistological investigations of cellular structure revealed, that magnesium ions increased focal adhesion at concentration of additionally 75 mM MgCl2 in cell culture medium.The use of our designed cell migration assay has shown that ionic medium alterations due to magnesium corrosion has a higher impact on the cell migration rate than surface alterations.Statement of SignificanceThe design of a migration assay on non-transparent magnesium surfaces will add the option to study cell response to surface modifications, coatings and the corrosion process itself under life view conditions.Graphical abstractGraphical abstract for this article
  • Rapid and area-specific coating of fluoride-incorporated apatite layers by
           a laser-assisted biomimetic process for tooth surface functionalization
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): A. Joseph Nathanael, Ayako Oyane, Maki Nakamura, Moumita Mahanti, Kenji Koga, Kanako Shitomi, Hirofumi Miyaji Surface functionalization of teeth with fluoride-incorporated apatite layers displays great potential in treatments and prevention of dental disorders. In this study, we used a sintered hydroxyapatite (sHA) substrate as a model material of teeth, and established a rapid and area-specific coating technique of fluoride-incorporated apatite layers by using a laser-assisted biomimetic (LAB) process. In this technique, a sHA substrate was irradiated on the surface with a Nd:YAG pulsed UV laser for 30 min in supersaturated calcium phosphate (CaP) solutions with various fluoride concentrations. The fluoride concentration in the CaP solution was varied to control morphology, crystalline structure, and fluoride content of the resulting layers. Without fluoride in the CaP solution, an octacalcium phosphate (OCP) layer with a flake-like structure was formed on the laser-irradiated surface of the substrate. The addition of fluoride (1000 µM and 3000 µM) to the CaP solution led to the formation of fluoride-incorporated apatite layers with an enamel-like needle-like nanostructure. The fluoride-incorporated apatite layers adhered firmly to the sHA surface and reduced acid dissolution of the sHA substrate by acting as a protective covering. Additionally, the layers released fluoride ions for more than 24 h, and exhibited antibacterial activity relative to a caries-causing bacterium, namely Streptococcus mutans. Thus, our LAB process can potentially act as a new tool for functionalization of tooth surfaces.Statement of SignificanceWe used a sintered hydroxyapatite (sHA) substrate as a model material of teeth, and established a rapid and area-specific coating technique of fluoride-incorporated apatite layers on the sHA surface by using our laser-assisted biomimetic (LAB) process. In this process, pulsed laser was utilized to accelerate seeded crystal growth in supersaturated calcium phosphate solutions supplemented with NaF. The thus-fabricated fluoride-incorporated apatite layers consisted of enamel-like needle-like nanocrystals with c-axis orientation. These fluoride-incorporated apatite layers adhered firmly to the sHA surface, reduced acid dissolution of the sHA substrate by acting as a protective covering, and exhibited antibacterial activity against Streptococcus mutans through the fluoride release. Thus, our LAB process can potentially act as a new tool for functionalization of tooth surfaces.Graphical abstractGraphical abstract for this article
  • Concentration-independent mechanics and structure of hagfish slime
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Gaurav Chaudhary, Douglas S. Fudge, Braulio Macias-Rodriguez, Randy H. Ewoldt The defense mechanism of hagfish slime is remarkable considering that hagfish cannot control the concentration of the resulting gel directly; they simply exude a concentrated material into a comparably “infinite” sea of water to form a dilute, sticky, cohesive elastic gel. This raises questions about the robustness of gel formation and rheological properties across a range of concentrations, which we study here for the first time. Across a nearly 100-fold change in concentration, we discover that the gel has similar viscoelastic time-dependent properties with constant power-law exponent (α=0.18±0.01), constant relative damping tanδ=G′′/G′≈0.2–0.3, and varying overall stiffness that scales linearly with the concentration (∼c0.99±0.05). The power-law viscoelasticity (fit by a fractional Kelvin-Voigt model) is persistent at all concentrations with nearly constant fractal dimension. This is unlike other materials and suggests that the underlying material structure of slime remains self-similar irrespective of concentration. This interpretation is consistent with our microscopy studies of the fiber network. We derive a structure-rheology model to test the hypothesis that the origins of ultra-soft elasticity are based on bending of the fibers. The model predictions show an excellent agreement with the experiments. Our findings illustrate the unusual and robust properties of slime which may be vital in its physiological use and provide inspiration for the design of new engineered materials.Statement of SignificanceHagfish produce a unique gel-like material to defend themselves against predator attacks. The successful use of the defense gel is remarkable considering that hagfish cannot control the concentration of the resulting gel directly; they simply exude a small quantity of biomaterial which then expands by a factor of 10,000 (by volume) into an “infinite” sea of water. This raises questions about the robustness of gel formation and properties across a range of concentrations. This study provides the first ever understanding of the mechanics of hagfish slime over a very wide range of concentration. We discover that some viscoelastic properties of slime are remarkably constant regardless of its concentration. Such a characteristic is uncommon in most known materials.Graphical abstractGraphical abstract for this article
  • Peripapillary sclera architecture revisited: A tangential fiber model and
           its biomechanical implications
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Andrew P. Voorhees, Ning-Jiun Jan, Yi Hua, Bin Yang, Ian A. Sigal The collagen fiber architecture of the peripapillary sclera (PPS), which surrounds the scleral canal, is a critical factor in determining the mechanical response of the optic nerve head (ONH) to variations in intraocular pressure (IOP). Experimental and clinical evidence point to IOP-induced deformations within the scleral canal as important contributing factors of glaucomatous neural tissue damage and consequent vision loss. Hence, it is imperative to understand PPS architecture and biomechanics. Current consensus is that the fibers of the PPS form a closed ring around the canal to support the delicate neural tissues within. We propose an alternative fiber architecture for the PPS, in which the scleral canal is supported primarily by long-running fibers oriented tangentially to the canal. We present evidence that this tangential model is consistent with histological observations in multiple species, and with quantitative measurements of fiber orientation obtained from small angle light scattering and wide-angle X-ray scattering. Using finite element models, we investigated the biomechanical implications of a tangential fiber PPS architecture. We found that the tangential arrangement of fibers afforded better mechanical support to the tissues within the scleral canal as compared to a simple circumferential ring of fibers or a combination of fibers oriented radially and circumferentially. We also found that subtle variations from a tangential orientation could reproduce clinically observed ONH behavior which has yet to be explained using current theories of PPS architecture and simulation, namely, the contraction of the scleral canal under elevated IOP.Statement of SignificanceIt is hypothesized that vision loss in glaucoma is due to excessive mechanical deformation within the neural tissue inside the scleral canal. This study proposes a new model for how the collagen of the peripapillary sclera surrounding the canal is organized to support the delicate neural tissue inside. Previous low-resolution studies of the peripapillary sclera suggested that the collagen fibers are arranged in a ring around the canal. Instead, we provide microscopic evidence suggesting that the canal is also supported by long-running interwoven fibers oriented tangentially to the canal. We demonstrate that this arrangement has multiple biomechanical advantages over a circular collagen arrangement and can explain previously unexplained experimental findings including contraction of the scleral canal under elevated intraocular pressure.Graphical abstractGraphical abstract for this article
  • Cell regulation of collagen fibril macrostructure during corneal
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Elena Koudouna, Eric Mikula, Donald J. Brown, Robert D. Young, Andrew J. Quantock, James V. Jester While tissue form and function is highly dependent upon tissue-specific collagen composition and organization, little is known of the mechanisms controlling the bundling of collagen fibrils into fibers and larger structural designs that lead to the formation of bones, tendons and other tissues. Using the cornea as a model system, our previous 3 dimensional mapping of collagen fiber organization has demonstrated that macrostructural organization of collagen fibers involving interweaving, branching and anastomosing plays a critical role in controlling mechanical stiffness, corneal shape and refractive power. In this work, the cellular and mechanical mechanisms regulating critical events in the assembly of collagen macrostructure are analysed in the developing chicken cornea. We elucidated the temporal events leading to adult corneal structure and determined the effects of intraocular pressure (IOP) on the organization of the collagen macrostructure. Our findings indicate that the complex adult collagen organization begins to appear on embryonic day 10 (E10) after deposition of the primary stroma and full invasion of keratocytes. Importantly, organizational changes in keratocytes appearing at E9 preceded and predicted later changes in collagen organization. Corneal collagen organization remained unaffected when the development of IOP was blocked at E4. These findings support a primary role for keratocytes in controlling stromal organization, mechanical stiffness and corneal shape that are not regulated by the IOP. Our findings also suggest that the avian cornea represents an excellent experimental model for elucidating key regulatory steps and mechanisms controlling the collagen fiber organization that is critical to determining tissue form and function.Statement of significanceThis work by using an ex ovo model system, begins to investigate the potential mechanisms controlling collagen fibril macrostructure. In particular, this work highlights a convergent role for the corneal keratocytes in organizing the complex collagen macrostructure, necessary to support high visual acuity. Our data supports that the intraocular pressure does not influence collagen fibril macrostructure and suggest that the avian cornea represents an excellent experimental model for elucidating key regulatory steps and mechanisms controlling the collagen fiber organization that is critical to determining tissue form and function. Clearly understanding the cellular and molecular mechanisms that underlie collagen fibril macrostructure will be highly beneficial for future tissue engineering and regenerative medicine applications.Graphical abstractGraphical abstract for this article
  • Substrate elasticity regulates adipose-derived stromal cell
           differentiation towards osteogenesis and adipogenesis through β-catenin
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Jing Xie, Demao Zhang, Chenchen Zhou, Quan Yuan, Ling Ye, Xuedong Zhou It is generally recognised that mesenchymal stem cells (MSCs) can differentiate into multiple lineages through guidance from the biophysical properties of the substrates. However, the precise biophysical mechanism that enables MSCs to respond to substrate properties remains unclear. In the current study, polydimethylsiloxane (PDMS) substrates with different stiffnesses were fabricated and the way in which the elastic modulus of the substrate regulated differentiation towards osteogenesis and adipogenesis in adipose-derived stromal cells (ASCs) was explored. Initially, a cell morphology change by SEM was observed between the stiff and soft substrates. The cytoskeleton stains including microfilament by F-actin and microtubule by α- and β-tubulin further showed a larger cell spreading area on the stiff substrate. Then the expression of vinculin, in charge for the linkage of adhesion molecules to the actin cytoskeleton, was enhanced on the stiff substrate. This change in focal adhesion plaque further triggered intracellular β-catenin signaling and promoted its nuclear translocation especially on the stiff substrate. The influence of β-catenin signaling on direct differentiation to osteogenic lineages was through direct binding between its downstream protein, Lef-1, and the osteogenic transcriptional factors, Runx2 and Osx, while on differentiation to adipogenic lineages was through modulating the expression of PPARγ. The imbalance of stiffness-induced β-catenin signaling finally induced a stronger osteogenesis and a weaker adipogenesis on the stiff substrate relative to those on the soft substrate. This study indicates the importance of stiffness on ASC differentiation and could help to increase understanding of the mechanism underlying molecular signal transduction from mechanosensing, mechanotransducing to stem cell differentiation.Statement of SignificanceMesenchymal stem cells can differentiate into multiple lineages, such as adipogenesis, myogenesis, neurogenesis, angiogenesis and osteogenesis, through influence of biophysical properties of the extracellular matrix. However, the precise bio-mechanism that triggers stem cell differentiation in response to matrix biophysical properties remains unclear. In the current study, we provide a series of experiments involving the characterization of cell morphology, microfilament, microtubule and adhesion capacity of adipose-derived stromal cells (ASCs) in response to substrate stiffness, and further elucidation of cytoplasmic β-catenin-dependent signal transduction, nuclear translocation and resultant promoter activation of transcriptional factors for osteogenesis and adipogenesis. This study provides an explanation on deeper understanding of bio-mechanism underlying substrate stiffness-triggered β-catenin signal transduction from active mechanosensing, mechanotransducing to stem cell differentiation.Graphical abstractGraphical abstract for this article
  • Elastic materials for tissue engineering applications: Natural, synthetic,
           and hybrid polymers
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Anna M.J. Coenen, Katrien V. Bernaerts, Jules A.W. Harings, Stefan Jockenhoevel, Samaneh Ghazanfari Elastin and collagen are the two main components of elastic tissues and provide the tissue with elasticity and mechanical strength, respectively. Whereas collagen is adequately produced in vitro, production of elastin in tissue-engineered constructs is often inadequate when engineering elastic tissues. Therefore, elasticity has to be artificially introduced into tissue-engineered scaffolds. The elasticity of scaffold materials can be attributed to either natural sources, when native elastin or recombinant techniques are used to provide natural polymers, or synthetic sources, when polymers are synthesized. While synthetic elastomers often lack the biocompatibility needed for tissue engineering applications, the production of natural materials in adequate amounts or with proper mechanical strength remains a challenge. However, combining natural and synthetic materials to create hybrid components could overcome these issues. This review explains the synthesis, mechanical properties, and structure of native elastin as well as the theories on how this extracellular matrix component provides elasticity in vivo. Furthermore, current methods, ranging from proteins and synthetic polymers to hybrid structures that are being investigated for providing elasticity to tissue engineering constructs, are comprehensively discussed.Statement of SignificanceTissue engineered scaffolds are being developed as treatment options for malfunctioning tissues throughout the body. It is essential that the scaffold is a close mimic of the native tissue with regards to both mechanical and biological functionalities. Therefore, the production of elastic scaffolds is of key importance to fabricate tissue engineered scaffolds of the elastic tissues such as heart valves and blood vessels. Combining naturally derived and synthetic materials to reach this goal proves to be an interesting area where a highly tunable material that unites mechanical and biological functionalities can be obtained.Graphical abstractGraphical abstract for this article
  • 3D-printed bioceramic scaffolds: From bone tissue engineering to tumor
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Hongshi Ma, Chun Feng, Jiang Chang, Chengtie Wu Toward the aim of personalized treatment, three-dimensional (3D) printing technology has been widely used in bone tissue engineering owing to its advantage of a fast, precise, and controllable fabrication process. Conventional bioceramic scaffolds are mainly used for bone tissue engineering; however, there has been a significant change in the application of bioceramic scaffolds during the past several years. Therefore, this review focuses on 3D-printed bioceramic scaffolds with different compositions and hierarchical structures (macro, micro, and nano scales), and their effects on the mechanical, degradation, permeability, and biological properties. Further, this review highlights 3D-printed bioceramic scaffolds for applications extending from bone tissue regeneration to bone tumor therapy. This review emphasizes recent developments in functional 3D-printed bioceramic scaffolds with the ability to be used for both tumor therapy and bone tissue regeneration. Considering the challenges in bone tumor therapy, these functional bioceramic scaffolds have a great potential in repairing bone defects induced by surgery and kill the possibly residual tumor cells to achieve bone tumor therapy. Finally, a brief perspective regarding future directions in this field was also provided. The review not only gives a summary of the research developments in bioceramic science but also offers a new therapy strategy by extending multifunctions of traditional biomaterials toward a specific disease.Statement of significanceThis review outlines the development tendency of 3D-printed bioceramic scaffolds for applications ranging from bone tissue regeneration to bone tumor therapy. Conventional bioceramic scaffolds are mainly used for bone tissue engineering; however, there has been a significant change in the application of bioceramic scaffolds during the past several years. Therefore, this review focuses on 3D-printed bioceramic scaffolds with different compositions and hierarchical structures (macro, micro, and nano scales), and their effects on the mechanical, degradation, permeability, and biological properties. Further, this review highlights 3D-printed bioceramic scaffolds for applications extending from bone tissue regeneration to bone tumor therapy. This review emphasizes recent developments in the functional 3D-printed bioceramic scaffolds with the ability to be used for both bone tumor therapy and bone tissue regeneration.Graphical abstractGraphical abstract for this article
  • Advances in functionalized polymer coatings on biodegradable magnesium
           alloys – A review
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Ling-Yu Li, Lan-Yue Cui, Rong-Chang Zeng, Shuo-Qi Li, Xiao-Bo Chen, Yufeng Zheng, M. Bobby Kannan Magnesium (Mg) and its alloys have become a research frontier in biodegradable materials owing to their superior biocompatibility and excellent biomechanical compatibility. However, their high degradation rate in the physiological environment should be well tackled prior to clinical applications. This review summarizes the latest progress in the development of polymeric coatings on biodegradable Mg alloys over the last decade, regarding preparation strategies for polylactic acid (PLA), poly (latic-co-glycolic) acid (PLGA), polycaprolactone (PCL), polydopamine (PDA), chitosan (CS), collagen (Col) and their composite, and their performance in terms of corrosion resistance and biocompatibility. Feasible perspectives and developing directions of next generation of polymeric coatings with respect to biomedical Mg alloys are briefly discussed.Statement of SignificanceMagnesium (Mg) and its alloys have become a research frontier in biodegradable materials owing to their superior biocompatibility and suitable biomechanical compatibility. However, the principal drawback of Mg-based implants is their poor corrosion resistance in physiological environments. Hence, it is vital to mitigate the degradation/corrosion behavior of Mg alloys for safe biomedical deployments. This review summarizes the latest progress in development of polymeric coatings on biomedical Mg alloys regarding preparation strategy, corrosion resistance and biocompatibility, including polylactic acid (PLA), poly (latic-co-glycolic) acid (PLGA), polycaprolactone (PCL), chitosan (CS), polydopamine (PDA), collagen (Col) and their composite. In addition, functionalized polymer coatings with Mg alloys exhibits a promising prospect owing to their ability of degradation along with biocompatibility, self-healing, drug-delivery and osteoinduction.Graphical abstractGraphical abstract for this article
  • A critical review of multifunctional titanium surfaces: New frontiers for
           improving osseointegration and host response, avoiding bacteria
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Silvia Spriano, Seiji Yamaguchi, Francesco Baino, Sara Ferraris Evolution of metal implants progressively shifted the focus from adequate mechanical strength to improved biocompatibility and absence of toxicity and, finally, to fast osseointegration. Recently, new frontiers and challenges of Ti implants have been addressed to improvement of bioactivity, fighting of bacterial infection and biofilm formation, as well as modulation of inflammation. This is closely related to the clinical demand of multifunctional implants able to simultaneously have a number of specific responses with respect to body fluids, cells (osteoblasts, fibroblasts, macrophages) and pathogenic agents (bacteria, viruses). This complex system of multiple biological stimuli and surface responses is a major arena of the current research on biomaterials and biosurfaces. This review covers the strategies explored to this purpose since 2010 in the case of Ti and Ti alloys, considering that the number of related papers doubled about in the last seven years and no review has comprehensively covered this engaging research area yet. The different approaches followed for producing multifunctional Ti-based surfaces involve the use of thick and thin inorganic coatings, chemical surface treatments, and functionalization strategies coupled with organic coatings.Statement of SignificanceAccording to the clinical demand of multifunctional implants able to simultaneously have a number of specific responses with respect to body fluids, cells and pathogenic agents, new frontiers of Ti implants have been addressed to improvement of bioactivity, fighting of bacterial infection and biofilm formation, as well as modulation of inflammation. Literature since 2010 is here reviewed. Several strategies for getting bioactive and antibacterial actions on Ti surfaces have been suggested, but they still need to be optimized with respect to several concerns. A further step will be to combine on the same surface a proven ability of modulation of inflammatory response. The achievement of multifunctional surfaces able to modulate inflammation and to promote osteogenesis is a grand challenge.Graphical abstractGraphical abstract for this article
  • Autophagic response to cellular exposure to titanium dioxide nanoparticles
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Lauren Popp, Vinh Tran, Risha Patel, Laura Segatori Titanium dioxide is “generally regarded as safe” and titanium dioxide nanoparticles (TiO2 NPs) are used in a wide variety of consumer products. Cellular exposure to TiO2 NPs results in complex effects on cell physiology including induction of oxidative stress and impairment of lysosomal function, raising concerns about the impact of TiO2 NPs on biological systems. We investigated the effects of TiO2 NPs (15, 50, and 100 nm in diameter) on the lysosome-autophagy system, the main cellular catabolic pathway that mediates degradation of nanomaterials. Specifically, we monitored a comprehensive set of markers of the lysosome-autophagy system upon cell exposure to TiO2 NPs, ranging from transcriptional activation of genes required for the formation of autophagic vesicles to clearance of autophagic substrates. This study reveals that uptake of TiO2 NPs induces a response of the lysosome-autophagy system mediated by the transcription factor EB and consequent upregulation of the autophagic flux. Prolonged exposure to TiO2 NPs, however, was found to induce lysosomal dysfunction and membrane permeabilization, leading to a blockage in autophagic flux. Results from this study will inform the design of TiO2 NP based devices with specific autophagy-modulating properties.Graphical abstractGraphical abstract for this article
  • Transparent titanium dioxide nanotubes: Processing, characterization, and
           application in establishing cellular response mechanisms
    • Abstract: Publication date: 1 October 2018Source: Acta Biomaterialia, Volume 79Author(s): Jevin G. Meyerink, Divya Kota, Scott T. Wood, Grant A. Crawford The therapeutic applications of titanium dioxide nanotubes as osteogenic surface treatments for titanium-based implants are largely due to the finely tunable physical characteristics of these nanostructures. As these characteristics change, so does the cellular response, yet the exact mechanisms for this relationship remains largely undefined. We present a novel TiO2 NT imaging platform that is suitable for use with live-cell imaging techniques, thereby enabling, for the first time, dynamic investigation of those mechanisms. In this work, fabrication methods for producing transparent TiO2 NTs with diameters of 56 ± 6 nm, 75 ± 7 nm, 92 ± 9 nm, and 116 ± 10 nm are described. To demonstrate the diagnostic potential of these TiO2 NT imaging platforms, the focal adhesion protein vinculin and actin cytoskeletal filaments were fluorescently tagged in osteoblasts and real-time, high-resolution fluorescent microscopy of live-cell interactions with TiO2 NT substrates were observed. The scope of such a platform is expected to extend far beyond the current proof-of-concept, with great potential for addressing the dynamic response of cells interacting with nanostructured substrates.Statement of significanceTitanium dioxide (TiO2) nanotubes are known to strongly enhance bone/mesenchymal stem cell behavior and, consequently, have gained attention as potential osteogenic surface treatments for titanium-bone implants. The exact mechanism by which TiO2 nanotubes influence cellular function remains controversial, partly due to limitations in existing cellular imaging methods with opaque substrates. This work identifies fabrication conditions for the successful production of transparent TiO2 nanotube arrays with tailorable diameters, as well as their functionality with pre-osteoblast mouse cells (MC3T3-E1) transfected with fluorescent focal adhesion protein vinculin and cytoskeletal filament actin. We demonstrate a means of recording live-cell, cell–substrate interaction mechanisms via high-resolution fluorescent microscopy and customizable, transparent TiO2 nanotubes to begin defining the relationship between TiO2 nanotube features and cell function.Graphical abstractGraphical abstract for this article
  • Fab-conjugated PLGA nanoparticles effectively target cancer cells
           expressing human CD44v6
    • Abstract: Publication date: Available online 27 September 2018Source: Acta BiomaterialiaAuthor(s): Patrick J. Kennedy, Flavia Sousa, Daniel Ferreira, Carla Pereira, Marika Nestor, Carla Oliveira, Pedro L. Granja, Bruno Sarmento Targeting of CD44 isoforms containing exon v6 (CD44v6) represents a viable strategy for the therapy and/or early diagnosis of metastatic cancers of the epithelium (e.g. gastric and colorectal cancer). We developed and characterized for the first time poly(lactic-co-glycolic acid) (PLGA)-based nanoparticles (NPs) modified with polyethylene glycol (PEG) and engrafted, by site-directed conjugation, with an engineered human Fab that specifically target human CD44v6 (v6 Fab-PLGA NPs). The v6 Fab-PLGA NPs display spherical morphology around 300 nm and are negatively charged. They strongly bound to a CD44v6-derived peptide and, more importantly, to cells that endogenously and exogenously express CD44v6, but not to non-expressing cells and cells expressing the standard isoform of CD44. The v6 Fab-PLGA NPs also recognized CD44v6 in tumor sections from cells grown subcutaneously within mice. The NPs had nominal cytotoxicity at 50 µg/mL and withstood simulated intestinal fluid exposure. Interestingly, v6 Fab-PLGA NPs cryopreserved in 10% trehalose and long-term stored maintained specific cell binding. In conclusion, we envision NPs targeting CD44v6 as potential in vivo diagnostic agents and/or as anti-cancer agents in patients previously stratified with CD44v6+ carcinomas.Statement of SignificanceThe v6 Fab-PLGA NPs displayed many favorable qualities as a potential CD44v6-targeted drug and/or diagnostic delivery agent. The NPs were designed for optimal ligand orientation and for immediate administration into humans.NPs strongly bind to cells that endogenously and exogenously express CD44v6, but not to non-expressing cells and cells expressing the standard isoform of CD44. Binding ability was retained after freeze-drying and long-term storage, providing evidences, for the first time, on the stability of Fab-functionalized NPs.These NPs can potentially be used as an in vivo diagnostic from parenteral or oral/rectal administration.Graphical abstractGraphical abstract for this article
  • Sensitizing bacterial cells to antibiotics by shape recovery triggered
           biofilm dispersion
    • Abstract: Publication date: Available online 27 September 2018Source: Acta BiomaterialiaAuthor(s): Sang Won Lee, Huan Gu, James Bryan Kilberg, Dacheng Ren Microbial biofilms are a leading cause of chronic infections in humans and persistent biofouling in industries due to extremely high-level tolerance of biofilm cells to antimicrobial agents. Eradicating mature biofilms is especially challenging because of the protection of the extracellular matrix and slow growth of biofilm cells. Recently, we reported that established biofilms can be effectively removed (e.g. 99.9% dispersion of 48 h Pseudomonas aeruginosa PAO1 biofilms) by shape memory polymer-based dynamic changes in surface topography. Here, we demonstrate that such biofilm dispersion also sensitizes biofilm cells to conventional antibiotics. For example, shape recovery in the presence of 50 µg/mL tobramycin reduced biofilm cell counts by more than 3 logs (2,479-fold) compared to the static flat control. The observed effects were attributed to the disruption of biofilm structure and increase in cellular activities as evidenced by an 11.8-fold increase in intracellular level of adenosine triphosphate (ATP), and 4.1-fold increase in expression of the rrnB gene in detached cells. These results can help guide the design of new control methods to better combat biofilm associated antibiotic-resistant infections.Statement of SignificanceMicrobial infections are challenging due to high-level antibiotic resistance of biofilm cells. The protection of an extracellular matrix and slow growth of biofilm cells render conventional antibiotics ineffective. Thus, it is important to develop new technologies that can remove mature biofilms and sensitize biofilm cells to antibiotics. Recently, we demonstrated that dynamic change in surface topography can remove 48 h Pseudomonas aeruginosa PAO1 biofilms by 99.9%. In this study, we investigated how shape recovery triggered dispersion affect the physiology of biofilm cells and associated antibiotic susceptibility. These results are helpful for understanding biofilm dispersion and developing more effective control methods.Graphical abstractGraphical abstract for this article
  • Size-Controlled Lipid Nanoparticle Production Using Turbulent Mixing to
           Enhance Oral DNA Delivery
    • Abstract: Publication date: Available online 27 September 2018Source: Acta BiomaterialiaAuthor(s): Zhiyu He, Yizong Hu, Tianqi Nie, Haoyu Tang, Jinchang Zhu, Kuntao Chen, Lixin Liu, Kam W. Leong, Yongming Chen, Hai-Quan Mao Lipid-based nanoparticles (LNPs) have been developed to address the transport and uptake barriers to enhance the delivery efficiency of plasmid DNA therapeutics. In these systems, plasmid DNA can be encapsulated through condensation by a cationic lipid to form lipo-complexes, or polycation following complexation into cationic liposomes to form lipo-polyplexes. Conventional methods for achieving these two DNA-delivering LNP vehicles suffer from significant batch-to-batch variation, poor scalability and complicated multi-step preparation procedures. Resultant nanoparticles often have uncontrollable size and surface charge with wide distribution, and poor stability when exposed to physiological media. Here we report a single-step flash nanocomplexation (FNC) process using turbulent mixing to prepare uniform lipo-complex or lipo-polyplex LNPs in a scalable manner, demonstrating excellent control over the nanoparticle size (from 40 to several hundred nm) and surface charge, with narrow size distribution. The FNC-produced LNPs could be purified and concentrated using a tangential flow filtration (TFF) process in a scalable manner. An optimized formulation of purified lipo-complex LNPs (DOTAP/Chol/DNA, 45 nm) showed significantly higher (5-fold in the lungs and 4-fold in the liver) transgene expression activity upon oral dosage than lipo-polyplex LNPs (DPPC/Chol/lPEI/DNA, 75 nm) or lPEI/DNA nanoparticles (43 nm). Repeated dosing (4 days, 150 μg/day) of the lipo-complex LNPs sustained the transgene activity over a period of one week without detectable toxicity in major organs, suggesting its potential for clinical translation.Statement of SignificanceWe report a new method to prepare uniform size-controlled lipid-based DNA-loaded nanoparticles by turbulent mixing delivered by a multi-inlet vortex mixer. Two distinct compositions were successfully prepared: (1) lipo-complexes, through condensation of the plasmid DNA by cationic lipids; (2) lipo-polyplexes, by encapsulation of DNA/PEI together with neutral lipids. Comparing with conventional methods, which use multi-step processes with high batch-to-batch variations and poor control over nanoparticle characteristics, this method offers a single-step, continuous and reproducible assembly methodology that would promote the translation of such gene medicine products. Effective purification and concentration of nanoparticles were achieved by adopted tangential flow filtration method. Following oral gavage in mice, the lipo-complex nanoparticles showed the highest level of transgene expression in the lung and liver.Graphical abstractGraphical abstract for this article
  • Directing fibroblast self-assembly to fabricate highly-aligned,
           collagen-rich matrices
    • Abstract: Publication date: Available online 27 September 2018Source: Acta BiomaterialiaAuthor(s): Benjamin T. Wilks, Elisabeth B. Evans, Morcos N. Nakhla, Jeffrey R. Morgan Extracellular matrix composition and organization play a crucial role in numerous biological processes ranging from cell migration, differentiation, survival and metastasis. Consequently, there have been significant efforts towards the development of biomaterials and in vitro models that recapitulate the complexity of native tissue architecture. Here, we demonstrate an approach to fabricating highly aligned cell-derived tissue constructs via the self-assembly of human dermal fibroblasts. By optimizing mold geometry, cell seeding density, and media composition we can direct human dermal fibroblasts to adhere to one another around a non-adhesive agarose peg to facilitate the development of cell-mediated circumferential tension. By removing serum and adding ascorbic acid and l-proline, we tempered fibroblast contractility to enable the formation of stable tissue constructs. Similarly, we show that the alignment of cells and the ECM they synthesize can be modulated by changes to seeding density and that constructs seeded with the lowest number of cells have the highest degree of fibrillar collagen alignment. Finally, we show that this highly aligned, tissue engineered construct can be decellularized and that when re-seeded with fibroblasts, it provides instructive cues which enable cells to adhere to and align in the direction of the remaining collagen fiber network.Statement of SignificanceCell and extracellular matrix organization is directly related to biological function including cell signaling and tissue mechanics. Changes to this organization are often associated with injury or disease. The majority of in vitro tissue engineering models investigating cell and matrix organization rely on the addition of stress-shielding exogenous proteins and polymers and, or the application of external forces to promote alignment. Here we present a completely cell-based approach that relies on the development of cell-mediated tension to direct anisotropic cellular alignment and matrix synthesis using human dermal fibroblasts. A major challenge with this approach is excessive cellular contractility that results in necking and failure of the tissue construct. While other groups have tried to overcome this challenge by simply adding more cells, here we show that matrix alignment is inversely related to cell seeding density. To engineer tissue constructs with the highest degree of alignment, we optimized media components to reduce cellular contractility and promote collagen synthesis such that fibroblast toroids remained stable for at least 28 days in culture. We subsequently showed that these collagen-rich tissue constructs could be decellularized while maintaining their collagen microstructure and that cells adhered to and responded to the decellularized cell-derived matrix by aligning and elongating along the collagen fibers. The complexity of cell-derived matrices has been shown to better recapitulate in vivo tissue architecture and composition. This study provides a straight-forward approach to fabricating instructive cell-derived matrices with a high degree of uniaxial alignment generated purely by cell-mediated tension.Graphical abstractGraphical abstract for this article
  • Injectable decellularized nucleus pulposus-based cell delivery system for
           differentiation of adipose-derived stem cells and nucleus pulposus
    • Abstract: Publication date: Available online 27 September 2018Source: Acta BiomaterialiaAuthor(s): Xiaopeng Zhou, Jingkai Wang, Xianpeng Huang, Weijing Fang, Yiqing Tao, Tengfei Zhao, Chengzhen Liang, Jianming Hua, Qixin Chen, Fangcai Li Stem cell-based tissue engineering is a promising treatment for intervertebral disc (IVD) degeneration. A bio-scaffold that can maintain the function of transplanted cells and possesses favorable mechanical properties is needed in tissue engineering. Decellularized nucleus pulposus (dNP) has the potential to be a suitable bio-scaffold because it mimics the native nucleus pulposus (NP) composition. However, matrix loss during decellularization and difficulty in transplantation limit the clinical application of dNP scaffolds. In this study, we fabricated an injectable dNP-based cell delivery system (NPCS) and evaluated its properties by assessing the microstructure, biochemical composition, water content, biosafety, biostability, and mechanical properties. We also investigated the stimulatory effects of the bio-scaffold on the NP-like differentiation of adipose-derived stem cells (ADSCs) in vitro and the regenerative effects of the NPCS on degenerated NP in an in vivo animal model. The results showed that approximately 68% and 43% of the collagen and sGAG, respectively, remained in the NPCS after 30 days. The NPCS also showed mechanical properties similar to those of fresh NP. In addition, the NPCS was biocompatible and able to induce NP-like differentiation and extracellular matrix (ECM) synthesis in ADSCs. The disc height index (almost 81%) and the MRI index (349.05 ± 38.48) of the NPCS-treated NP were significantly higher than those of the degenerated NP after 16 weeks. The NPCS also partly restored the ECM content and the structure of degenerated NP in vivo. Our NPCS has good biological and mechanical properties and has the ability to promote the regeneration of degenerated NP.Statement of significanceNucleus pulposus (NP) degeneration is usually the origin of intervertebral disc degeneration. Stem cell-based tissue engineering is a promising treatment for NP regeneration. Bio-scaffolds which have favorable biological and mechanical properties are needed in tissue engineering. Decellularized NP (dNP) scaffold is a potential choice for tissue engineering, but the difficulty in balancing complete decellularization and retaining ECM limits its usage. Instead of choosing different decellularization protocols, we complementing the sGAG lost during decellularization by cross-linking via genipin and fabricating an injectable dNP-based cell delivery system (NPCS) which has similar components as the native NP. We also investigated the biological and mechanical properties of the NPCS in vitro and verified its regenerative effects on degenerated IVDs in an animal model.Graphical abstractGraphical abstract for this article
  • Calcium-carbonate packaging magnetic polydopamine nanoparticles loaded
           with indocyanine green for near-infrared induced photothermal/photodynamic
    • Abstract: Publication date: Available online 27 September 2018Source: Acta BiomaterialiaAuthor(s): Peng Xue, Mengmeng Hou, Lihong Sun, Qian Li, Lei Zhang, Zhigang Xu, Yuejun Kang Indocyanine green (ICG) is an efficient photosensitizer that can facilitate producing cytotoxic reactive oxygen species (ROS). At the same time, ICG also has characteristic absorption of near infrared light and thus can induce a strong photothermal effect. Both of these important features of ICG may be applied for noninvasive light-induced tumor ablation. On the other hand, ICG is lack of stability in blood circulation and susceptible to aggregation or premature clearance from the body. These issues need to be effectively addressed before antitumor application of ICG becomes possible. Herein, a nanocomposite consisting of calcium carbonate modified magnetic polydopamine (PDA) nanoparticles and loaded with ICG, namely Fe3O4@PDA@CaCO3/ICG (FPCI) NPs, was developed to integrate the photothermal capability of PDA with the photodynamic capability of ICG. Particularly, calcium carbonate not only entrapped ICG in the form of stable aggregate to evade blood clearance, but also facilitated controlled release of ICG in response to acidic tumor microenvironment via self-decomposition. With the aid of magnetic guidance, this multifunctional therapeutic agent makes it possible to achieve the combination of photothermal (PTT) and photodynamic therapies (PDT) against tumors, which was demonstrated by this proof-of-concept study based on in vitro and in vivo tumor models.Statement of SignificanceCurrently, there is an ongoing trend of realizing precise and targeted tumor therapy using functional nanocomplexes. Magnetic particles, which can be manipulated by magnetic field, have attracted increasing attention for tumor therapy. This submitted work demonstrated that calcium carbonate nanoshell was precipitated onto magnetic nanocores mediated by polydopamine. Moreover, indocyanine green (ICG), as a potent photosensitizer, was embedded in this nanocomplex and protected by the calcium carbonate nanoshell, resulting in high drug loading efficiency and enhanced drug stability on the carrier. This new nanocomposite was demonstrated to achieve controlled and pH-responsive release of ICG in tumor environment. This work explored the relation between the physiochemical properties of the nanocomplex and their potential biomedical applications, aiming to inspire the development of analogous nanoplatforms featured with calcium carbonate blocks.Graphical abstractGraphical abstract for this article
  • Therapeutic enhancement of a cytotoxic agent using Photochemical
           internalisation in 3D compressed collagen constructs of ovarian cancer
    • Abstract: Publication date: Available online 27 September 2018Source: Acta BiomaterialiaAuthor(s): Layla Mohammad Hadi, Elnaz Yaghini, Katerina Stamati, Marilena Loizidou, Alexander J. MacRobert Photochemical internalisation (PCI) is a method for enhancing delivery of drugs to their intracellular target sites of action. In this study we investigated the efficacy of PCI using a porphyrin photosensitiser and a cytotoxic agent on spheroid and non-spheroid compressed collagen 3D constructs of ovarian cancer versus conventional 2D culture. The therapeutic responses of two human carcinoma cell lines (SKOV3 and HEY) were compared using a range of assays including optical imaging. The treatment was shown to be effective in non-spheroid constructs of both cell lines causing a significant and synergistic reduction in cell viability measured at 48 or 96 hours post-illumination. In the larger spheroid constructs, PCI was still effective but required higher saporin and photosensitiser doses. Moreover, in contrast to the 2D and non-spheroid experiments, where comparable efficacy was found for the two cell lines, HEY spheroid constructs were found to be more susceptible to PCI and a lower dose of saporin could be used. PCI treatment was observed to induce death principally by apoptosis in the 3D constructs compared to the mostly necrotic cell death caused by PDT. At low oxygen levels (1%) both PDT and PCI were significantly less effective in the constructs.Statement of significanceAssessment of new drugs or delivery systems for cancer therapy prior to conducting in vivo studies often relies on the use of conventional 2D cell culture, however 3D cancer constructs can provide more physiologically relevant information owing to their 3D architecture and the presence of an extracellular matrix. This study investigates the efficacy of Photochemical Internalisation mediated drug delivery in 3D constructs. In 3D cultures, both oxygen and drug delivery to the cells are limited by diffusion through the extracellular matrix unlike 2D models, and in our model we have used compressed collagen constructs where the density of collagen mimics physiological values. These 3D constructs are therefore well suited to studying drug delivery using PCI. Our study highlights the potential of these constructs for identifying differences in therapeutic response to PCI of two ovarian carcinoma lines.Graphical abstractGraphical abstract for this article
  • Corrigendum to “Synergistic interplay between the two major bone
           minerals, hydroxyapatite and whitlockite nanoparticles, for osteogenic
           differentiation of mesenchymal stem cells” [Acta Biomater. 69 (2018)
    • Abstract: Publication date: Available online 26 September 2018Source: Acta BiomaterialiaAuthor(s): Hao Cheng, Rosa Chabok, Xiaofei Guan, Aditya Chawla, Yuxiao Li, Ali Khademhosseini, Hae Lin Jang
  • Deciphering the combined effect of bone morphogenetic protein 6 (BMP6) and
           calcium phosphate on bone formation capacity of periosteum derived
           cells-based tissue engineering constructs
    • Abstract: Publication date: Available online 26 September 2018Source: Acta BiomaterialiaAuthor(s): Wei Ji, Greet Kerckhofs, Carla Geeroms, Marina Marechal, Liesbet Geris, Frank P. Luyten Cell based combination products with growth factors on optimal carriers represent a promising tissue engineering strategy to treat large bone defects. In this concept, bone morphogenetic protein (BMP) and calcium phosphate (CaP)-based scaffolds can act as potent components of the constructs to steer stem cell specification, differentiation and initiate subsequent in vivo bone formation. However, limited insight into BMP dosage and the cross-talk between BMP and CaP materials, hampers the optimization of in vivo bone formation and subsequent clinical translation. Herein, we combined human periosteum derived progenitor cells with different doses of BMP6 and with three types of clinical grade CaP-scaffolds with (ChronOs®, ReproBoneTM, & CopiOs®). Comprehensive cellular and molecular analysis was performed based on in vitro cell metabolic activity and signaling pathway activation, as well as in vivo ectopic bone forming capacity after 2 weeks and 5 weeks in nude mice. Our data showed that cells seeded on CaP scaffolds with an intermediate Ca2+ release rate combined with low or medium dosage of BMP6 demonstrated a robust new bone formation after 5 weeks, which was contributed by both donor and host cells. This phenomenon might be due to the delicate balance between Ca2+ and BMP pathways, allowing an appropriate activation of the canonical BMP signaling pathway that is required for in vivo bone formation. For high BMP6 dosage, we found that the BMP6 dosage overrides the effect of the Ca2+ release rate and this appeared to be a dominant factor for ectopic bone formation. Taken together, this study illustrates the importance of matching BMP dosage and CaP properties to allow an appropriate activation of canonical BMP signaling that is crucial for in vivo bone formation. It also provides insightful knowledge with regard to clinical translation of cell-based constructs for bone regeneration.Statement of significanceThe combination of bone morphogenetic proteins (BMP) and calcium phosphate (CaP)-based biomaterials with mesenchymal stromal cells represents a promising therapeutic strategy to treat large bone defects, an unmet medical need. However, there is limited insight into the optimization of these combination products, which hampers subsequent successful clinical translation. Our data reveal a delicate balance between Ca2+ and BMP pathways, allowing an appropriate activation of canonical BMP signaling required for in vivo bone formation. Our findings illustrate the importance of matching BMP dosage and CaP properties in the development of cell-based constructs for bone regeneration.Graphical abstractGraphical abstract for this article
  • Chondrogenic Differentiation of Synovial Fluid Mesenchymal Stem Cells on
           Human Meniscus-Derived Decellularized Matrix Requires Exogenous Growth
    • Abstract: Publication date: Available online 26 September 2018Source: Acta BiomaterialiaAuthor(s): Yan Liang, Enaam Idrees, Alexander Szojka, Stephen Andrews, Melanie Kunze, Aillette Mulet-Sierra, Nadr Jomha, Adetola B. Adesida The objective of this study was to investigate whether meniscus-derived decellularized matrix (DCM) has the capacity to induce differentiation of synovial fluid-derived mesenchymal stem cells (SF-MSCs) towards a meniscus fibrochondrocyte (MFC) phenotype. The potential roles of transforming growth factor beta-3 (TGF-β3) and insulin-like growth factor 1 (IGF-1) in the differentiation of SF-MSCs towards an MFC phenotype were also investigated.SF-MSCs were isolated via plastic adherence cell culture from the synovial fluid of five donors (5 male, average age 34.4 years). Porous DCM was generated by homogenizing and freeze-drying fresh normal human cadaveric meniscus tissue. SF-MSCs were seeded and cultured on the DCM scaffold in a defined serum-free media (SFM) supplemented with or without the combination of TGF-β3 and IGF-1. Cell pellets of SF-MSCs were cultured in SFM with either TGF-β3 or IGF-1 or their combination as controls. The duration of culture was 3 weeks for both experimental configurations. We assessed newly-formed tissues by biochemical assays, scanning electron microscopy (SEM), immunofluorescence and quantitative real-time PCR (qPCR).The combination of TGF-β3 and IGF1 induced production of the cartilaginous matrix in DCM and upregulated the expression of aggrecan, collagens I and II. Moreover, the SF-MSCs exhibited a round morphology in the DCM scaffolds in the presence of the growth factors. In pellets, combined IGF-1 and TGF-β3 synergistically enhanced cartilaginous matrix production. In contrast to bone marrow mesenchymal stem cells (BM-MSCs), the differentiated SF-MSCs showed little evidence of the expression of the hypertrophic differentiation marker, collagen X.In conclusion, meniscus-derived DCM appears to require exogenous growth factor supplementation to direct differentiation of SF-MSCs.Statement of SignificanceMeniscus tears are the most common injury of the knee joint. These tears pose a major risk factor for the early development of knee osteoarthritis. Unfortunately, the majority of these tears occur in the largest region of the meniscus and lacks blood supply with no reparative or regenerative capacity.The goal of this study was to determine if the native extracellular matrix (ECM) of human meniscus have the capacity to differentiate human knee synovial fluid resident mesenchymal stem cells (SF-MSCs) towards a meniscus phenotype as a potential strategy to repair avascular meniscal tears.Our findings show that human meniscus-derived ECM without supplementation with growth factors (IGF-1 and TGF-β3) cannot differentiate SF-MSCs towards a meniscus phenotype.The use of meniscus-derived scaffolds as a material to stimulate endogenous repair of meniscus tears via differentiation of SF-MSCs may require supplementation with IGF-1 and TGF-β3.Graphical abstractGraphical abstract for this article
  • Intracorneal Injection of a Detachable Hybrid Microneedle for Sustained
           Drug Delivery
    • Abstract: Publication date: Available online 26 September 2018Source: Acta BiomaterialiaAuthor(s): KangJu Lee, Hyun Beom Song, Wonwoo Cho, Jin Hyoung Kim, Jeong Hun Kim, WonHyoung Ryu There are increasing demands for long-term and controlled corneal drug delivery to treat various ocular diseases. Although biodegradable ocular inserts or contact lenses have been developed, the invasiveness and inefficiency of the approaches still need to be improved. Microneedle (MN) technology can deliver therapeutic molecules to the eye in a minimally invasive manner. However, the current ocular MN technology is limited to either short-term corneal drug delivery or retinal drug delivery by suprachoroidal injection. For long-term and minimally invasive corneal drug delivery, we have developed a detachable biodegradable MN that can be delivered to the inside of the cornea for sustained drug release. The detachable and biodegradable MN is a hybrid MN consisting of a drug-loaded biodegradable tip and a supporting base. The hybrid MN can be applied to the cornea by impact insertion, and it leaves only the drug-loaded biodegradable tip within the corneal tissue so that it can release the drug for a certain period. By concentration-controlled molding, the dimension of drug-loaded MN tips was precisely controlled and their detachability was optimized. The detachable tip and a supporting base were assembled to form a hybrid MN by pressure-assisted transfer molding. We carefully optimized the dimension of the drug-tip, injection dwell time, and insertion depth to achieve effective intracorneal injection of the drug-tip. The detachable hybrid MN was applied to an Acanthamoeba keratitis model wherein a biodegradable drug-tip was successfully delivered to the inside of the mouse cornea in vivo. Follow-up of the MN-treated cases for 7 days confirmed the therapeutic efficacy of the detachable biodegradable MN tips.Statement of SignificanceFor the treatment of infectious diseases in the cornea, such as keratitis, eye drops need to be applied topically every hour for a couple of days. This is extremely uncomfortable, and poor compliance to such tightly scheduled drug administration can result in permanent scar formation in the cornea. In this work, we demonstrate a simple and rapid injection of biodegradable microneedle tips in the corneal tissue wherein the tips can deliver antibacterial drugs for 4 days to treat keratitis. Unlike other patch-style microneedle technologies, this approach allows for insertion depth-controlled and highly localized injection of detachable individual microneedle tips to the diseased tissue for sustained drug delivery. This overcomes the limitations of patch-style microneedles such as short-term drug delivery and unnecessary blockage of tissue.Graphical abstractGraphical abstract for this article
  • Programmed pH/reduction-responsive nanoparticles for efficient delivery of
           antitumor agents in vivo
    • Abstract: Publication date: Available online 26 September 2018Source: Acta BiomaterialiaAuthor(s): Wei-liang Chen, Shu-di Yang, Fang Li, Chen-xi Qu, Yang Liu, Yu Wang, Dan-dan Wang, Xue-nong Zhang To bypass the biological barriers during the drug delivery process, it is desirable to develop smart nanoparticles (NPs) with flexible physical and chemical properties. In this study, a programmed NP delivery system with a pH-triggered detachable PEG layer and a lactobionic acid (Lac)-modified reduction-responsive core was developed to address the “PEG dilemma” and provide an on-demand intracellular release of doxorubicin (DOX). The positively charged DOX-loaded lactobionic acid-chitosan-lipoic acid (DOX/LCL) NPs were prepared and coated with a negatively charged dimethylmaleic acid-PEG-chitosan (PEG-CS-DA) layer to obtain a prolonged circulation time and improve the tumor-targeting effect. After reaching the tumor tissues through a targeted delivery effect, the surface charge of the PEG-CS-DA layer was reversed from negative to positive because of the trigger by the acidic microenvironment (pH 6.8), thus leading to the detachment of the PEG layer. The exposure of positive charges and the active targeting ligand enhanced cellular uptake and facilitated penetration into tumor tissues. Subsequently, the rapid release and diffusion of DOX into the nuclei was triggered by the intracellular high concentration of glutathione, thus leading to cell apoptosis. In conclusion, these programmed pH/reduction-responsive NPs provide a promising strategy for the delivery of antitumor agents in vivo.Statement of SignificanceIn this study, novel programmed pH/reduction-responsive NPs were developed for the delivery of DOX in vivo. These NPs were coated with a negatively charged PEG layer to improve the serum stability and tumor target effect. The PEG layer detached because of the trigger by tumor acidic microenvironment (pH 6.8), thus leading to the exposure of positive charges and the active targeting ligand, which enhanced cellular uptake and facilitated penetration into tumor tissues. Subsequently, the rapid release of DOX was triggered by the intracellular high concentration of glutathione, thereby resulting in enhanced cytotoxicity. These programmed pH/reduction-responsive NPs provide a promising strategy for the delivery of antitumor agents in vivo.Graphical abstractGraphical abstract for this article
  • Versatile biomimetic conductive polypyrrole films doped with hyaluronic
           acid of different molecular weights
    • Abstract: Publication date: Available online 25 September 2018Source: Acta BiomaterialiaAuthor(s): Semin Kim, Yohan Jang, Minsu Jang, Ahyoun Lim, John G. Hardy, Hyun S. Park, Jae Young Lee Electrically conductive polypyrrole (PPy) is an intriguing biomaterial capable of efficient electrical interactions with biological systems. Especially, biomimetic PPy-based biomaterials incorporating biomolecules, such as hyaluronic acid (HA), can impart the characteristic biological interactions with living cells/tissues to the conductive biomaterials. Here we report the effects of the molecular weight (MW) of HA on PPy-based biomaterials. We utilized HA of a wide range of MW (35 × 103 Da – 3 × 106 Da) as dopants during the electrochemical production of PPy/HA films and their characterization of materials and cellular interactions. With increases in the MWs of HA dopants, PPy/HA exhibited more hydrophilic, higher electrochemical activity and lower impedance. In vitro studies revealed that PPy films doped with low MW HA were supportive to cell adhesion and growth, while PPy films doped with high MW HA were resistant to cell attachment. Subcutaneous implantation of the PPy/HA films for 4 weeks revealed that all the PPy/HA films were tissue compatible. We successfully demonstrate the importance of HA dopant MWs in modulating the chemical and electrical properties of the materials and cellular responses to the materials. Such materials have potential for various biomedical applications, including as tissue engineering scaffolds and as electrodes for neural recording and neuromodulation.Statement of SignificanceHyaluronic acid (HA)-doped polypyrrole (PPy) films were electrochemically synthesized as novel biomimetic conductive materials capable of efficient electrical signaling and preferential biological interactions. Molecular weights (MWs) of HA varied in a wide range (35 x 103 – 2 x 106 Da) and critically determine chemical, electrochemical, and biological properties of PPy/HA. Especially, PPy films with low MW HA markedly support cell adhesion and growth, while PPy films with high MW HA are resistant to cell attachment. Furthermore, PPy/HA exhibits greatly improved tissue compatibility and in vivo EMG signal recording ability. We for the first time demonstrate that biomimetic PPy/HA-based biomaterials can serve as versatile and effective platforms for various biomedical applications, such as tissue engineering scaffolds and bioelectrodes.Graphical abstractGraphical abstract for this article
  • Cellulose-based scaffolds for fluorescence lifetime imaging-assisted
           tissue engineering
    • Abstract: Publication date: Available online 25 September 2018Source: Acta BiomaterialiaAuthor(s): Neil O'Donnell, Irina A. Okkelman, Peter Timashev, Tatyana I. Gromovykh, Dmitri B. Papkovsky, Ruslan I. Dmitriev Quantitative measurement of pH and metabolite gradients by microscopy is one of the challenges in the production of scaffold-grown organoids and multicellular aggregates. Herein, we used the cellulose-binding domain (CBD) of the Cellulomonas fimi CenA protein for designing biosensor scaffolds that allow measurement of pH and Ca2+ gradients by fluorescence intensity and lifetime imaging (FLIM) detection modes. By fusing CBD with pH-sensitive enhanced cyan fluorescent protein (CBD-ECFP), we achieved efficient labeling of cellulose-based scaffolds based on nanofibrillar, bacterial cellulose, and decellularized plant materials. CBD-ECFP bound to the cellulose matrices demonstrated pH sensitivity comparable to untagged ECFP (1.9-2.3 ns for pH 6-8), thus making it compatible with FLIM-based analysis of extracellular pH. By using 3D culture of human colon cancer cells (HCT116) and adult stem cell-derived mouse intestinal organoids, we evaluated the utility of the produced biosensor scaffold. CBD-ECFP was sensitive to increases in extracellular acidification, the results showed a decline in 0.2-0.4 pH units in response to membrane depolarization by the protonophore FCCP. With the intestinal organoid model, we demonstrated multiparametric imaging by combining extracellular acidification (FLIM) with phosphorescent probe-based monitoring of cell oxygenation. The described labeling strategy allows for the design of extracellular pH-sensitive scaffolds for multiparametric FLIM assays and their use in engineered live cancer and stem cell-derived tissues. Collectively, this research can help in achieving the controlled biofabrication of 3D tissue models with known metabolic characteristics.Statement of SignificanceWe designed biosensors consisting of a cellulose-binding domain (CBD) and pH- and Ca2+-sensitive fluorescent proteinsCBD-tagged biosensors efficiently label various types of cellulose matrices including nanofibrillar cellulose and decellularized plant materialsHybrid biosensing cellulose scaffolds designed in this study were successfully tested by multiparameter FLIM microscopy in 3D cultures of cancer cells and mouse intestinal organoidsGraphical abstractGraphical abstract for this article
  • Polyglutamic acid-trimethyl chitosan-based intranasal peptide nano-vaccine
           induces potent immune responses against group A streptococcus
    • Abstract: Publication date: Available online 25 September 2018Source: Acta BiomaterialiaAuthor(s): Reshma J. Nevagi, Zeinab G. Khalil, Waleed M. Hussein, Jessica Powell, Michael R. Batzloff, Robert J. Capon, Michael F. Good, Mariusz Skwarczynski, Istvan Toth Peptide-based vaccines have the potential to overcome the limitations of classical vaccines; however, their use is hampered by a lack of carriers and adjuvants suitable for human use. In this study, an efficient self-adjuvanting peptide vaccine delivery system was developed based on the ionic interactions between cationic trimethyl chitosan (TMC) and a peptide antigen coupled with synthetically defined anionic α-poly-(l-glutamic acid) (PGA). The antigen, possessing a conserved B-cell epitope derived from the group A streptococcus (GAS) pathogen and a universal T-helper epitope, was conjugated to PGA using cycloaddition reaction. The produced anionic conjugate formed nanoparticles (NP-1) through interaction with cationic TMC. These NP-1 induced higher systemic and mucosal antibody titers compared to antigen adjuvanted with standard mucosal adjuvant cholera toxin B subunit or antigen mixed with TMC. The produced serum antibodies were also opsonic against clinically isolated GAS strains. Further, a reduction in bacterial burden was observed in nasal secretions, pharyngeal surface and nasopharyngeal-associated lymphoid tissue of mice immunized with NP-1 in GAS challenge studies. Thus, conjugation of defined-length anionic polymer to peptide antigen as a means of formulating ionic interaction-based nanoparticles with cationic polymer is a promising strategy for peptide antigen delivery.Statement of significanceA self-adjuvanting delivery system is required for peptide vaccines to enhance antigen delivery to immune cells and generate systemic and mucosal immunity. Herein, we developed a novel self-adjuvanting nanoparticulate delivery system for peptide antigens by combining polymer-conjugation and complexation strategies. We conjugated peptide antigen with anionic α-poly-(L-glutamic acid) that in turn, formed nanoparticles with cationic trimethyl chitosan by ionic interactions, without using external crosslinker. On intranasal administration to mice, these nanoparticles induced systemic and mucosal immunity, at low dose. Additionally, nanoparticles provided protection to vaccinated mice against group A streptococcus infection. Thus, this concept should be particularly useful in developing nanoparticles for the delivery of peptide antigens.Graphical abstractGraphical abstract for this article
  • Peptide LL-37 coating on micro-structured titanium implants to facilitate
           bone formation in vivo via mesenchymal stem cells recruitment
    • Abstract: Publication date: Available online 25 September 2018Source: Acta BiomaterialiaAuthor(s): Ye He, Caiyun Mu, Xinkun Shen, Zhang Yuan, Ju Liu, Weizhen Chen, Chuanchuan Lin, Bailong Tao, Bin Liu, Kaiyong Cai Titanium (Ti) and Ti- alloys were widely used in clinic orthopedics, however, the insufficient bone formation surrounding Ti-based implants still limited their biological performances. Surface modification of Ti substrates is essential to improve their interactions with bone-forming cells and bone tissue. In this study, we modified Ti substrates by coating peptide LL-37 onto micro-structured Ti substrates and aimed to (i) induce mesenchymal stem cells (MSCs) migration both in vitro and in vivo, (ii) facilitate osteogenic differentiation of MSCs and new bone formation. The surface micro-structured Ti substrates with hydroxyapatite deposition were fabricated by a two-step method including micro-arc oxidation (MAO) and hydrothermal treatment. LL-37 was loaded on micro-structured Ti substrates with the assistance of polydopamine coating. We confirmed that surface-modified Ti substrates benefited viability, adhesion, migration and osteogenic differentiation of MSCs in vitro. In a femur-defect rat model, the surface-modified Ti implants effectively induced CD29+/CD90+ positive cells migration in one week after implantation. According to the results of H&E, Masson’s trichrome staining and immunohistochemical staining of OCN, OPN and collagen I, the targeted Ti implants exhibited significant new bone formation after implantation for 4 weeks. These results indicate that the surface modification of Ti samples facilitated bone formation through MSCs recruitment.Statement of SignificanceThe inherent surface bioinertness of titanium (Ti) and Ti-alloys still limits their biological performances in clinical applications. Recently, the strategy of mesenchymal stem cells (MSCs) recruitment has been proposed to improve the osteointegration of bone implants. Herein, we reports the surface modification of Ti implants from the point of MSCs recruitment. Peptide LL-37 was coated on micro-structured Ti substrates to (i) recruit MSCs, (ii) regulate bio-physiological performance of MSCs, and (iii) facilitate bone formation in vivo. Our results improve the understanding of the interaction between Ti implants and MSCs, and provide a promising strategy of MSCs recruitment in the design of bone repair related biomaterials.Graphical abstractGraphical abstract for this article
  • Current State of Fabrication Technologies and Materials for Bone Tissue
    • Abstract: Publication date: Available online 22 September 2018Source: Acta BiomaterialiaAuthor(s): Abiy Wubneh, Eleni Tsekoura, Cagri Ayranci, Hasan Uludağ A range of traditional and free-form fabrication technologies have been investigated and, in numerous occasions, commercialized for use in the field of regenerative tissue engineering (TE). The demand for technologies capable of treating bone defects inherently difficult to repair has been on the rise. This quest, accompanied by the advent of functionally tailored, biocompatible, and biodegradable materials, has garnered an enormous research interest in bone TE. As a result, different materials and fabrication methods have been investigated towards this end, leading to a deeper understanding of the geometrical, mechanical and biological requirements associated with bone scaffolds. As our understanding of the scaffold requirements expands, so do the capability requirements of the fabrication processes. The goal of this review is to provide a broad examination of existing scaffold fabrication processes and highlight future trends in their development. To appreciate the clinical requirements of bone scaffolds, a brief review of the biological process by which bone regenerates itself is presented first. This is followed by a summary and comparisons of commonly used implant techniques to highlight the advantages of TE-based approaches over traditional grafting methods. A detailed discussion on the clinical and mechanical requirements of bone scaffolds then follows. The remainder of the manuscript is dedicated to current scaffold fabrication methods, their unique capabilities and perceived shortcomings. The range of biomaterials employed in each fabrication method is summarized. Selected traditional and non-traditional fabrication methods are discussed with a highlight on their future potential from the authors’ perspective. This study is motivated by the rapidly growing demand for effective scaffold fabrication processes capable of economically producing constructs with intricate and precisely controlled internal and external architectures.Statement of significanceThe manuscript summarizes the current state of fabrication technologies and materials used for creating scaffolds in bone tissue engineering applications. A comprehensive analysis of different fabrication methods (traditional and free-form) were summarized in this review paper, with emphasis on recent developments in the field. The fabrication techniques suitable for creating scaffolds for tissue engineering was particularly targeted and their use in bone tissue engineering were articulated. Along with the fabrication techniques, we emphasized the choice of materials in these processes. Considering the limitations of each process, we highlighted the materials and the material properties critical in that particular process and provided a brief rational for the choice of the materials. The functional performance for bone tissue engineering are summarized for different fabrication processes and the choice of biomaterials. Finally, we provide a perspective on the future of the field, highlighting the knowledge gaps and promising avenues in pursuit of effective scaffolds for bone tissue engineering. This extensive review of the field will provide research community with a reference source for current approaches to scaffold preparation. We hope to encourage the researchers to generate next generation biomaterials to be used in these fabrication processes. By providing both advantages and disadvantage of each fabrication method in detail, new fabrication techniques might be devised that will overcome the limitations of the current approaches. These studies should facilitate the efforts of researchers interested in generating ideal scaffolds, and should have applications beyond the repair of bone tissue.Graphical abstractGraphical abstract for this article
  • Bio-inspired and optimized interlocking features for strengthening
           metal/polymer interfaces in additively manufactured prostheses
    • Abstract: Publication date: Available online 21 September 2018Source: Acta BiomaterialiaAuthor(s): Ammar A. Alsheghri, Omar Alageel, Mohamed Amine Mezour, Binhan Sun, Stephen Yue, Faleh Tamimi, Jun Song Biomedical and dental prostheses combining polymers with metals often suffer failure at the interface. The weak chemical bond between these two dissimilar materials can cause debonding and mechanical failure. This manuscript introduces a new mechanical interlocking technique to strengthen metal/polymer interfaces through optimized additively manufactured features on the metal surface. To reach an optimized design of interlocking features, we started with the bio-mimetic stress-induced material transformation (SMT) optimization method. The considered polymer and metal materials were cold-cured Poly(methyl methacrylate) (PMMA) and laser-sintered Cobalt-Chromium (Co-Cr), respectively. Optimal dimensions of the bio-inspired interlocking features were then determined by mesh adaptive direct search (MADS) algorithm combined with finite element analysis (FEA) and tensile experiments such that they provide the maximum interfacial tensile strength and stiffness while minimizing the stress in PMMA and the displacement of PMMA at the Co-Cr/PMMA interface. The SMT optimization process suggested a Y-shape as a more favorable design, which was similar to mangrove tree roots. Experiments confirmed that our optimized interlocking features increased the strength of the Co-Cr/PMMA interface from 2.3 MPa (flat interface) to 34.4±1 MPa, which constitutes 85% of the tensile failure strength of PMMA (40.2±1 MPa).Statement of SignificanceThe objective of this study was to improve metal/polymer interfacial strength in dental and orthopedic prostheses. This was achieved by additive manufacturing of optimized interlocking features on metallic surfaces using laser-sintering. The interlocking design of the features, which was a Y-shape similar to the roots of mangrove trees, was inspired by a bio-memetic optimization algorithm. This interlocking design lowered the PMMA displacement at the Co-Cr/PMMA interface by 70%, enhanced the interfacial strength by more than 12%, and increased the stiffness by 18% compared with a conventional bead design, meanwhile no significant difference was found in the toughness of both designs.Graphical abstractGraphical abstract for this article
  • Targeted cellular delivery of robust enzyme nanoparticles for the
           treatment of drug-induced hepatotoxicity and liver injury
    • Abstract: Publication date: Available online 19 September 2018Source: Acta BiomaterialiaAuthor(s): Min Sang Lee, Nak Won Kim, Jung Eun Lee, Myung Goo Kim, Yue Yin, Sun Young Kim, Bo Sung Ko, Aeseon Kim, Jong Han Lee, Su Yeon Lim, Dong Woo Lim, Sun Hwa Kim, Ji Won Park, Yong Taik Lim, Ji Hoon Jeong Direct delivery of proteins into cells has been considered an effective approach for treating the protein-related diseases. However, clinical use of proteins has still been limited due to their instability in the blood and poor membrane permeability. To achieve an efficient cellular delivery of the protein to target cells via a systemic administration, a multifunctional carrier system having desirable stability both in the blood stream and the cells, specific cell-targeting property and endosomal escape functions may be required. In this study, we prepared a catalytic nanoparticle containing an active enzyme by cross-tethering multiple superoxide dismutase (SOD) molecules with catechol-derivatized hyaluronic acid (HA). The permeable shell of hydrophilic HA chains effectively protects the enzyme from degradation in the blood after intravenous administration and provides an additional function for targeting hepatocytes expressing HA receptor (CD44). The structure and catalytic activity of the enzyme molecules in the nanoparticle were not significantly compromised in the nanoparticle. In addition, ultra-small calcium phosphate nanoparticles (USCaP, 2-5 nm) were crystalized and decorated on the surface of the nanoparticle for the efficient endosomal escape after cellular uptake. The SOD-containing nanoparticle fortified with USCaP was used for the treatment of acetaminophen (APAP)-induced fulminant hepatotoxicity and liver injury. The nanoparticle achieved the efficient hepatic cellular delivery of SOD via a systemic administration and resulted in efficient removal of reactive oxygen species (ROS) in the liver and remarkable improvement of APAP-induced hepatotoxicity and liver injury in animals.Statement of significanceDespite the enormous therapeutic potential, the intracellular delivery of proteins has been limited due to their poor membrane permeability and stability. In this study, we demonstrated an active enzyme-containing nanoparticle functionalized by hyaluronic acid and ultra-small size calcium phosphate nanoparticles (2∼5 nm) for targeted cellular delivery of superoxide dismutase (SOD). The nanoparticle was designed to integrate all the essential functions, including serum stability, target specificity, and endosomal escape capability, for a systemic delivery of a therapeutic protein to the cells of the liver tissue. The intravenous administration of the nanoparticle efficiently removes reactive oxygen species (ROS) in the liver and remarkably improves the drug-induced hepatotoxicity and the progress of fulminant liver injury in an acetaminophen-overdose animal model.Graphical abstractGraphical abstract for this article
  • Hyaluronic acid shell and disulfide-crosslinked core micelles for in vivo
           targeted delivery of bortezomib for the treatment of multiple myeloma
    • Abstract: Publication date: Available online 19 September 2018Source: Acta BiomaterialiaAuthor(s): Zhaoxin Gu, Xiuxiu Wang, Ru Cheng, Liang Cheng, Zhiyuan Zhong Bortezomib (BTZ) provides one of the best treatments for multiple myeloma (MM). The efficacy of BTZ is, nevertheless, restricted by its fast clearance, low selectivity, and dose limiting toxicities. Here, we report on targeted BTZ therapy of MM in vivo by hyaluronic acid-shelled and core-disulfide-crosslinked biodegradable micelles (HA-CCMs) encapsulating lipophilized BTZ, bortezomib-pinanediol (BP). HA-CCMs loaded with 7.3 BTZ equiv. wt.% exhibited a small size of 78 nm, good stability in 10% FBS, and glutathione-triggered drug release. MTT assays in CD44 positive LP-1 multiple myeloma cells revealed that BP encapsulated in HA-CCMs caused enhanced antiproliferative effect compared with free BP. Flow cytometry, confocal microscopy and MTT assays indicated BP-loaded HA-CCMs (HA-CCMs-BP) could actively target to LP-1 cells and induce high antitumor effect. Proteasome activity assays in vitro showed HA-CCMs-BP had a similar proteasome activity inhibition as compared to free BTZ at 18 h. The fluorescence imaging using Cy5-labeled HA-CCMs showed that HA-CCMs had a long elimination half-life and enhanced tumor accumulation via HA-mediated uptake mechanism. The therapeutic studies in LP-1 MM-bearing mice revealed better treatment efficacy of HA-CCMs-BP compared with free BTZ, in which HA-CCMs-BP at 3 mg BTZ equiv./kg brought about significant tumor growth inhibition and survival benefits. Loading of lipophilized BTZ into HA-shelled multifunctional micelles has emerged as an exciting approach for bortezomib therapy of MM.Statement of SignificanceMultiple myeloma (MM) is the second most common hematological malignancy. Bortezomib (BTZ), a potent proteasome inhibitor, provides one of the best treatments for MM. The clinical efficacy of BTZ is, however, limited by its quick clearance, poor selectivity, and significant side effects including myelosuppression and peripheral neuropathy. Here, we report on targeted BTZ therapy of MM in vivo by hyaluronic acid-shelled and core-disulfide-crosslinked biodegradable micelles (HA-CCMs) encapsulating lipophilized BTZ, bortezomib-pinanediol (BP). Our results showed that BP-loaded HA-CCMs exhibit markedly enhanced toleration, broadened therapeutic window, and significantly more effective growth suppression of CD44-overexpressed multiple myeloma in nude mice than free bortezomib. Lipophilized BTZ-loaded HA-CCMs has opened a new avenue for targeted bortezomib therapy of multiple myeloma.Graphical abstractGraphical abstract for this article
  • Energy dissipation in mammalian collagen fibrils: Cyclic strain-induced
           damping, toughening, and strengthening
    • Abstract: Publication date: Available online 19 September 2018Source: Acta BiomaterialiaAuthor(s): Julia Liu, Debashish Das, Fan Yang, Andrea G. Schwartz, Guy M. Genin, Stavros Thomopoulos, Ioannis Chasiotis As the fundamental structural protein in mammals, collagen transmits cyclic forces that are necessary for the mechanical function of tissues, such as bone and tendon. Although the tissue-level mechanical behavior of collagenous tissues is well understood, the response of collagen at the nanometer length scales to cyclical loading remains elusive. To address this major gap, we cyclically stretched individual reconstituted collagen fibrils, with average diameter of 141±53 nm, to small and large strains in the partially hydrated conditions of 60% RH. It is shown that cyclical loading results in large steady-state hysteresis that is reached immediately after the first loading cycle, followed thereafter by limited accumulation of inelastic strain and constant initial elastic modulus. Cyclic loading above 20% strain resulted in 70% increase in tensile strength, from 638±98 MPa to 1086±86 MPa, and 60% increase in toughness, while maintaining the ultimate tensile strain of collagen fibrils not subjected to cyclic loading. Throughout cyclic stretching, the fibrils maintained a steady-state hysteresis, yielding loss coefficients that are 5-10 times larger than those of known homogeneous materials in their modulus range, thus establishing damping of nanoscale collagen fibrils as a major component of damping in tissues.Statement of Significance:It is shown that steady-state energy dissipation occurs in individual collagen fibrils that are the building blocks of hard and soft tissues. To date, it has been assumed that energy dissipation in tissues takes place mainly at the higher length scales of the tissue hierarchy due to interactions between collagen fibrils and fibers, and in limited extent inside collagen fibrils. It is shown that individual collagen fibrils need only a single loading cycle to assume a highly dissipative, steady-state, cyclic mechanical response. Mechanical cycling at large strains leads to 70% increase in mechanical strength and values exceeding those of engineering steels. The same cyclic loading conditions lead to 60% increase in toughness and loss properties that are five times higher than those of engineering materials with comparable stiffness.Graphical abstractGraphical abstract for this article
  • Recipients of the 2017 Acta Student Awards
    • Abstract: Publication date: Available online 19 September 2018Source: Acta BiomaterialiaAuthor(s):
  • Lipopeptisomes: Anticancer Peptide-Assembled Particles for Fusolytic
    • Abstract: Publication date: Available online 19 September 2018Source: Acta BiomaterialiaAuthor(s): Matthew R. Aronson, Andrew W. Simonson, Lindsey M. Orchard, Manuel Llinás, Scott H. Medina Anticancer peptides (ACPs) are cationic amphiphiles that preferentially kill cancer cells through folding-dependent membrane disruption. Although ACPs represent attractive therapeutic candidates, particularly against drug-resistant cancers, their successful translation into clinical practice has gone unrealized due to their poor bioavailability, serum instability and, most importantly, severe hemolytic toxicity. Here, we exploit the membrane-specific interactions of ACPs to prepare a new class of peptide-lipid particle, we term a lipopeptisome (LP). This design sequesters loaded ACPs within a lipid lamellar corona to avoid contact with red blood cells and healthy tissues, while affording potent lytic destruction of cancer cells following LP-membrane fusion. Biophysical studies show ACPs rapidly fold at, and integrate into, liposomal membranes to form stable LPs with high loading efficiencies (>80%). Rational design of the particles to possess lipid combinations mimicking that of the aberrant cancer cell outer leaflet allows LPs to rapidly fuse with tumor cell membranes and afford localized assembly of loaded ACPs within the bilayer. This leads to preferential fusolytic killing of cancer cells with minimal collateral toxicity towards non-cancerous cells and erythrocytes, thereby imparting clinically relevant therapeutic indices to otherwise toxic ACPs. Thus, integration of ACPs into self-assembled LPs represents a new delivery strategy to improve the therapeutic utility of oncolytic agents, and suggests this technology may be added to targeted combinatorial approaches in precision medicine.Statement of SignificanceDespite their significant clinical potential, the therapeutic utility of many ACPs has been limited by their collateral hemolysis during administration. Leveraging the membrane-specific interactions of ACPs, here we prepare self-assembled peptide-lipid nanoparticles, or ‘lipopeptisomes’ (LPs), capable of preferentially fusing with and lysing cancer cell membranes. Key to this fusolytic action is the construction of LPs from lipids simulating the cancer cell outer leaflet. This design recruits the oncolytic peptide payload into the carrier lamella and allows for selective destruction of cancer cells without disrupting healthy cells. Consequently, LPs impart clinically relevant therapeutic indexes to previously toxic ACPs, and thus open new opportunities to improve the clinical translation of oncolytics challenged by narrow therapeutic windows.Graphical abstractGraphical abstract for this article
  • A light-controllable specific drug delivery nanoplatform for targeted
           bimodal imaging-guided photothermal/chemo synergistic cancer therapy
    • Abstract: Publication date: Available online 19 September 2018Source: Acta BiomaterialiaAuthor(s): Yuan Guo, Xing-Yue Wang, Yu-Li Chen, Feng-Qiu Liu, Mi-Xiao Tan, Meng Ao, Jin-Hong Yu, Hai-tao Ran, Zhao-Xia Wang Breast cancer is a severe threat to the health and lives of women due to its difficult early diagnosis and the unsatisfactory therapeutic efficacy of breast cancer treatments. The development of theranostic strategies to combat breast cancer with high accuracy and effectiveness is therefore urgently needed. In this study, we describe a near-infrared (NIR) light-controllable, targeted and biocompatible drug delivery nanoplatform (PFH-PTX@PLGA/SPIO-Her) for photoacoustic (PA)/ultrasound (US) bimodal imaging-guided photothermal (PTT) /chemo synergistic cancer therapy of breast cancer. Carboxyl-modified PEGylated poly (lactic-co-glycolic acid) (PLGA-PEG-COOH) constituted the skeleton of the nanoplatform. Especially, the antibody Herceptin was modified onto the surface of nanoplatform for active HER2-targing to facilitate the tumor accumulation of the nanoplatform. The encapsulated superparamagnetic iron oxide (SPIO) nanoparticles could be employed as an excellent PA imaging agent to guide tumor therapy. When exposed to NIR light, the SPIO also could transform NIR light into thermal energy for photothermal ablation of tumor. The NIR-induced thermal effect subsequently triggered the optical droplet vaporization (ODV) of perfluorohexane (PFH) to generate PFH gas bubbles, which not only achieved the US imaging enhancement, but also contributed to the release of loaded paclitaxel (PTX) from the nanoplatform for significantly improving PTT therapeutic efficacy. Our results demonstrated that the targeted tumor accumulation, accurate real-time bimodal imaging, and the abundant drug release at the tumor site were all closely associated with the PTT therapeutic efficacy. Therefore, the theranostic nanoplatform is a very promising strategy for targeted imaging-guided photothermal/chemo synergistic tumor therapy with high therapeutic efficacy and minimized side effects.Statement of SignificanceBreast cancer is the most frequent cancer in women. Herein, we successfully developed a light-controllable and HER2 targeted theranostic nanoparticels (PFH-PTX@PLGA/SPIO-Her) as a specific drug delivery nanoplatform to overcome the low accuracy of tumor detection and low specificity of traditional chemo-therapeutic protocols. The study demonstrated that PFH-PTX@PLGA/SPIO-Her could actively target to breast cancer cells with positive HER2 expression. The biocompatible PFH-PTX@PLGA/SPIO-Her nanoplatform as both photoacoustic/ultrasound bimodal imaging agents, photothermal-conversion nanomaterials (photothermal hyperthermia) and controllable drug delivery nanoagents (optical droplet vaporization) have completely eradicated the tumor without severe side effects. The theranostic strategy not only integrates strengthens of traditional imaging or therapeutic modalities, but also paves a new way for the efficient cancer treatment by taking the advantage of quickly-developing nanomedicine.Graphical abstractGraphical abstract for this article
  • Alternating block copolymer-based nanoparticles as tools to modulate the
           loading of multiple chemotherapeutics and imaging probes
    • Abstract: Publication date: Available online 18 September 2018Source: Acta BiomaterialiaAuthor(s): C. Mattu, G.Brachi, L. Menichetti, A. Flori, P. Armanetti, E. Ranzato, S. Martinotti, S. Nizzero, M. Ferrari, G. Ciardelli Cancer therapy often relies on the combined action of different molecules to overcome drug resistance and enhance patient outcome. Combined strategies relying on molecules with different pharmacokinetics often fail due to the lack of concomitant tumor accumulation and, thus, in the loss of synergistic effect. Due to their ability to enhance treatment efficiency, improve drug pharmacokinetics, and reduce adverse effects, polymer nanoparticles (PNPs) have been widely investigated as co-delivery vehicles for cancer therapies. However, co-encapsulation of different drugs and probes in PNPs requires a flexible polymer platform and a tailored particle design, in which both the bulk and surface properties of the carriers are carefully controlled.In this work, we propose a core-shell PNP design based on a polyurethane (PUR) core and a phospholipid external surface. The modulation of the hydrophilic/hydrophobic balance of the PUR core enhanced the encapsulation of two chemotherapeutics with dramatically different water solubility (Doxorubicin hydrochloride, DOXO and Docetaxel, DCTXL) and of Iron Oxide Nanoparticles for MRI imaging. The outer shell remained unchanged among the platforms, resulting in un-modified cellular uptake and in vivo biodistribution. We demonstrate that the choice of PUR core allowed a very high entrapment efficiency of all drugs, superior or comparable to previously reported results and that higher core hydrophilicity enhances the loading efficiency of the hydrophilic Doxorubicin and the MRI contrast. Moreover, we show that changing the PUR core did not alter the surface properties of the carriers, since all particles showed a similar behavior in terms of cell internalization and in vivo biodistribution. We also show that PUR PNPs have very high passive tumor accumulation and that they can efficient co-deliver the 2 drugs to the tumor, reaching an 11-fold higher DOXO/DCTXL ratio in tumor as compared to free drugs.Statement of significanceExploiting the synergistic action of multiple chemotherapeutics is a promising strategy to improve the outcome of cancer patients, as different agents can simultaneously engage different features of tumor cells and/or their microenvironment. Unfortunately, the choice is limited to drugs with similar pharmacokinetics that can concomitantly accumulate in tumors. To expand the spectrum of agents that can be delivered in combination, we propose a multi-compartmental core-shell nanoparticle approach, in which the core is made of biomaterials with high affinity for drugs of different physical properties. We successfully co-encapsulated Doxorubicin Hydrochloride, Docetaxel, and contrast agents and achieved a significantly higher concomitant accumulation in tumor versus free drugs, demonstrating that nanoparticles can improve synergistic cancer chemotherapy.Graphical abstractGraphical abstract for this article
  • A systematic electron microscopic study on the uptake of barium sulphate
           nano-, submicro-, microparticles by bone marrow-derived phagocytosing
    • Abstract: Publication date: Available online 18 September 2018Source: Acta BiomaterialiaAuthor(s): V. Sokolova, K. Loza, T. Knuschke, J. Heinen-Weiler, H. Jastrow, M. Hasenberg, J. Buer, A.M. Westendorf, M. Gunzer, M. Epple Nanoparticles can act as transporters for synthetic molecules and biomolecules into cells, also in immunology. Antigen-presenting cells like dendritic cells are important targets for immunotherapy in nanomedicine. Therefore, we have used primary murine bone marrow-derived phagocytosing cells (bmPCs), i.e. dendritic cells and macrophages, to study their interaction with spherical barium sulphate particles of different size (40 nm, 420 nm, and 1 µm) and to follow their uptake pathway. Barium sulphate is chemically and biologically inert (no dissolution, no catalytic effects), i.e. we can separate the particle uptake effect from potential biological reactions. The colloidal stabilization of the nanoparticles was achieved by a layer of carboxymethylcellulose (CMC) which is biologically inert and gives the particles a negative zeta potential (i.e. charge). The particles were made fluorescent by conjugating 6-aminofluoresceine to CMC. Their uptake was visualized by flow cytometry, confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and correlative light and electron microscopy (CLEM). Barium sulphate particles of all sizes were readily taken up by dendritic cells and even more by macrophages, with the uptake increasing with time and particle concentration. They were mainly localized inside phagosomes, heterophagosomes, and in the case of nanoparticles also in the nearby cytosol. No particles were found in the nucleus. In nanomedicine, inorganic nanoparticles from the nanometer to the micrometer size are therefore well suited as transporters of biomolecules, including antigens, into dendritic cells and macrophages. The presented model system may also serve to describe the aseptic loosening of endoprostheses caused by abrasive wear of inert particles and the subsequent cell reaction, a question which relates to the field of nanotoxicology.Statement of significanceThe interaction of particles and cells is at the heart of nanomedicine and nanotoxicology, including abrasive wear from endoprostheses. It also comprises the immunological reaction to different kinds of nanomaterials, triggered by an immune response, e.g. by antigen-presenting cells. However, it is often difficult to separate the particle effect from a chemical or biochemical reaction to particles or their cargo. We show how chemically inert barium sulphate particles with three different sizes (nano, sub-micro, and micro) interact with relevant immune cells (primary dendritic cells and macrophages). Particles of all three sizes are readily taken up into both cell types by phagocytosis, but the uptake by macrophages is significantly more prominent than that by dendritic cells. The cells take up particles until they are virtually stuffed, but without direct adverse effect. The uptake increases with time and particle concentration. Thus, we have an ideal model system to follow particles into and inside cells without the side effect of a chemical particle effect, e.g. by degradation or ion release.Graphical abstractGraphical abstract for this article
  • Biodegradation of ECM hydrogel promotes endogenous brain tissue
           restoration in a rat model of stroke
    • Abstract: Publication date: Available online 16 September 2018Source: Acta BiomaterialiaAuthor(s): Harmanvir Ghuman, Carrinton Mauney, Julia Donnelly, Andre R. Massensini, Stephen F. Badylak, Michel Modo The brain is considered to have a limited capacity to repair damaged tissue and no regenerative capacity following injury. Tissue lost after a stroke is therefore not spontaneously replaced. Extracellular matrix (ECM)-based hydrogels implanted into the stroke cavity can attract endogenous cells. These hydrogels can be formulated at different protein concentrations that govern their rheological and inductive properties. We evaluated histologically 0, 3, 4 and 8 mg/mL of porcine-derived urinary bladder matrix (UBM)-ECM hydrogel concentrations implanted in a 14-day old stroke cavity. Less concentrated hydrogels (3 and 4 mg/mL) were efficiently degraded with a 95% decrease in volume by 90 days, whereas only 32% of the more concentrated and stiffer hydrogel (8 mg/mL) was resorbed. Macrophage infiltration and density within the bioscaffold progressively increased in the less concentrated hydrogels and decreased in the 8 mg/mL hydrogels. The less concentrated hydrogels showed a robust invasion of endothelial cells with neovascularization. No neovascularization occurred with the stiffer hydrogel. Invasion of neural cells increased with time in all hydrogel concentrations. Differentiation of neural progenitors into mature neurons with axonal projections was evident, as well as a robust invasion of oligodendrocytes. However, relatively few astrocytes were present in the ECM hydrogel, although some were present in the newly forming tissue between degrading scaffold patches. Implantation of an ECM hydrogel partially induced neural tissue restoration, but a more complete understanding is required to evaluate its potential therapeutic application.Statement of SignificanceExtracellular matrix hydrogel promotes tissue regeneration in many peripheral soft tissues. However, the brain has generally been considered to lack the potential for tissue regeneration. We here demonstrate that tissue regeneration in the brain can be achieved using implantation of ECM hydrogel into a tissue cavity. We here demonstrate that a structure-function relationship is key to promote tissue regeneration in the brain. Specifically, weaker hydrogels that were retained in the cavity underwent an efficient biodegradation within 14 days post-implantation to promote a tissue restoration within the lesion cavity. In contrast, stiffer ECM hydrogel only underwent minor biodegradation and did not lead to a tissue restoration. Inductive hydrogels weaker than brain tissue provide the appropriate condition to promote an endogenous regenerative response that restores tissue in a cavity. This approach offers new avenues for the future treatment of chronic tissue damage caused by stroke and other acute brain injuries.Graphical abstractGraphical abstract for this article
  • Bacterial microbots for acid-labile release of hybrid micelles to promote
           the synergistic antitumor efficacy
    • Abstract: Publication date: 15 September 2018Source: Acta Biomaterialia, Volume 78Author(s): Songzhi Xie, Maohua Chen, Xiaojie Song, Zhao Zhang, Zhanlin Zhang, Zhoujiang Chen, Xiaohong Li Bacteria have inherent properties of self-propelled navigation and specific infiltration into solid tumors. In the current study, we investigate a novel type of bacterial microbots for delivery of hybrid micelles to promote the synergistic antitumor efficacy. Escherichia coli Nissle 1917 (EcN) is used as a bacterial carrier to immobilize amphiphilic copolymers through acid-labile 2-propionic-3-methylmaleic anhydride (CDM) linkers. Doxorubicin (DOX) and α-tocopheryl succinate (TOS) are conjugated with poly(ethylene glycol) through disulfide linkers to obtain amphiphilic promicelle polymers (PMTOS and PMDOX). Tetrazine and norbornene terminals are grafted on EcN and PMTOS/PMDOX copolymers, respectively, and the mild and site-specific bioorthogonal reaction between them maintains the viability, motion ability, and tumor accumulation capability of the conjugated EcN. The PMTOS/PMDOX copolymers are released from bacterial microbots in response to the slightly acidic tumor microenvironment, followed by in situ formation of these copolymers as hybrid micelles (MD/T). The self-assembled micelles from PMTOS/PMDOX with a ratio of 1:2 demonstrate the most significant synergistic efficacy, and the released MD/T hybrid micelles exhibit cellular uptake efficiency, glutathione (GSH)-sensitive drug release, and cytotoxicities similar to those exhibited by micelles prepared by solvent evaporation. Because of the consecutive process of the self-propelling nature of bacteria and preferential accumulation of EcN in tumors, in situ formation of MD/T hybrid micelles, and intracellular drug release, bacterial microbots have shown remarkable antitumor efficacy with regard to animal survival, tumor growth, and apoptosis induction in tumor cells. Therefore, we demonstrate a feasible strategy for the construction of bacterial microbots to achieve tumor accumulation and on-demand release of multiple therapeutic agents for synergistic antitumor efficacy.Statement of significanceChallenges remain in the targeted delivery of nanoparticles to solid tumors and the realization of synergistic efficacy in cancer chemotherapy. In the current study, we explore a novel class of bacterial microbots to load, deliver, and release hybrid micelles. Escherichia coli Nissle 1917 (EcN) is used as a bacterial carrier to immobilize amphiphilic copolymers through acid-labile linkers, and the released copolymers are self-assembled into micelles. The resulting bacterial microbots integrate self-propelling bacteria and self-assembling amphiphilic polymers into micelles and realize pH-responsive release of promicelle polymers from bacterial microbots and glutathione-responsive intracellular release of drugs. A synergistic antitumor efficacy is achieved using hybrid micelles, which release both doxorubicin and α-tocopheryl succinate to display toxicities in the nucleus and mitochondria, respectively.Graphical abstractGraphical abstract for this article
  • Additive manufactured biodegradable poly(glycerol sebacate methacrylate)
           nerve guidance conduits
    • Abstract: Publication date: 15 September 2018Source: Acta Biomaterialia, Volume 78Author(s): Dharaminder Singh, Adam J. Harding, Emad Albadawi, Fiona M. Boissonade, John W. Haycock, Frederik Claeyssens Entubulating devices to repair peripheral nerve injuries are limited in their effectiveness particularly for critical gap injuries. Current clinically used nerve guidance conduits are often simple tubes, far stiffer than that of the native tissue. This study assesses the use of poly(glycerol sebacate methacrylate) (PGSm), a photocurable formulation of the soft biodegradable material, PGS, for peripheral nerve repair. The material was synthesized, the degradation rate and mechanical properties of material were assessed and nerve guidance conduits were structured via stereolithography. In vitro cell studies confirmed PGSm as a supporting substrate for both neuronal and glial cell growth. Ex vivo studies highlight the ability of the cells from a dissociated dorsal root ganglion to grow out and align along the internal topographical grooves of printed nerve guide conduits. In vivo results in a mouse common fibular nerve injury model show regeneration of axons through the PGSm conduit into the distal stump after 21 days. After conduit repair levels of spinal cord glial activation (an indicator for neuropathic pain development) were equivalent to those seen following graft repair. In conclusion, results indicate that PGSm can be structured via additive manufacturing into functional NGCs. This study opens the route of personalized conduit manufacture for nerve injury repair.Statement of SignificanceThis study describes the use of photocurable of Poly(Glycerol Sebacate) (PGS) for light-based additive manufacturing of Nerve Guidance Conduits (NGCs). PGS is a promising flexible biomaterial for soft tissue engineering, and in particular for nerve repair. Its mechanical properties and degradation rate are within the desirable range for use in neuronal applications. The nerve regeneration supported by the PGS NGCs is similar to an autologous nerve transplant, the current gold standard. A second assessment of regeneration is the activation of glial cells within the spinal cord of the tested animals which reveals no significant increase in neuropathic pain by using the NGCs. This study highlights the successful use of a biodegradable additive manufactured NGC for peripheral nerve repair.Graphical abstractGraphical abstract for this article
  • Dual release of growth factor from nanocomposite fibrous scaffold promotes
           vascularisation and bone regeneration in rat critical sized calvarial
    • Abstract: Publication date: 15 September 2018Source: Acta Biomaterialia, Volume 78Author(s): Shruthy Kuttappan, Dennis Mathew, Jun-ichiro Jo, Ryusuke Tanaka, Deepthy Menon, Takuya Ishimoto, Takayoshi Nakano, Shantikumar V. Nair, Manitha B. Nair, Yasuhiko Tabata A promising strategy for augmenting bone formation involves the local delivery of multiple osteoinductive and vasculogenic growth factors. However, success depends on sustained growth factor release and its appropriate combination to induce stem cells and osteogenic cells at the bony site. Herein, we have developed a nanocomposite fibrous scaffold loaded with fibroblast growth factor 2 (FGF2), vascular endothelial growth factor (VEGF) and bone morphogenetic protein 2 (BMP2) and its ability to promote vascularisation and bone regeneration in critical sized calvarial defect was compared to the scaffold with VEGF + BMP2 and FGF2 + BMP2. Simple loading of growth factors on the scaffold could provide a differential release pattern, both in vitro and in vivo (VEGF release for 1 week where as BMP2 and FGF2 release for 3 weeks). Among all the groups, dual growth factor loaded scaffold (VEGF + BMP2 & FGF2 + BMP2) enhanced vascularisation and new bone formation, but there was no difference between FGF2 and VEGF loaded scaffolds although its release pattern was different. FGF2 mainly promoted stem cell migration, whereas VEGF augmented new blood vessel formation at the defect site. This study suggests that biomimetic nanocomposite scaffold is a promising growth factor delivery vehicle to improve bone regeneration in critical sized bone defects.Statement of SignificanceMany studies have shown the effect of growth factors like VEGF-BMP2 or FGF2-BMP2 in enhancing bone formation in critical sized defects, but there are no reports that demonstrate the direct comparison of VEGF-BMP2 and FGF2-BMP2. In this study, we have developed a nanocomposite fibrous scaffold that could differentially release growth factors like VEGF, BMP2 and FGF2 (VEGF release for 1 week where as BMP2 and FGF2 release for 3 weeks), which in turn promoted neovascularisation and new bone formation in critical sized defects. There was no difference in vascularisation and bone formation induced by VEGF + BMP2 or FGF2 + BMP2. The growth factor was loaded in a simple manner, which would ensure ease of use for the end-user, especially for the surgeons treating a patient in an operating room.Graphical abstractGraphical abstract for this article
  • Surface fluorination of polylactide as a path to improve platelet
           associated hemocompatibility
    • Abstract: Publication date: 15 September 2018Source: Acta Biomaterialia, Volume 78Author(s): Razieh Khalifehzadeh, Winston Ciridon, Buddy D. Ratner Surface-induced thrombosis is still a significant clinical concern for many types of blood-contacting medical devices. In particular, protein adsorption and platelet adhesion are important events due to their ability to trigger the coagulation cascade and initiate thrombosis. Poly(lactic acid) (PLA) has been the predominant polymer used for making bioresorbable stents. Despite long-term advantages, these stents are associated with higher rates of early thrombosis compared with permanent metallic stents. To address this issue, we modified the surface of PLA with a perfluoro compound facilitated by surface activation using radio frequency (RF) plasma. Fluoropolymers have been extensively used in blood contacting materials, such as blood vessel replacements due to their reduced thrombogenicity and reduced platelet reactivity. The compositions of plasma-treated surfaces were determined by electron spectroscopy for chemical analysis (ESCA). Also, contact angle measurements, cell cytotoxicity and the degradation profile of the treated polymers are presented. Finally, relevant blood compatibility parameters, including plasma protein adsorption, platelet adhesion and morphology, were evaluated. We hypothesized that tight binding of adsorbed albumin by fluoropolymers enhances its potential for blood-contacting applications.Statement of SignificanceAlthough bioresorbable stents made from poly(lactic acid) (PLA) may have long-term clinical advantages, they have shown higher rates of early thrombosis as compared with permanent metallic stents. To improve the thromboresistance of PLA, we developed a novel method for surface fluorination of this polymer with a perfluoro compound. Fluoropolymers (e.g., expanded polytetrafluoroethylene) have long been used in blood-contacting applications due to their satisfactory clinical performance. This is the first report of PLA surface fluorination which might be applied to the fabrication of a new generation of fluorinated PLA stents with improved platelet interaction, tunable degradability and drug release capabilities. Also, we describe a general strategy for improving the platelet interactions with biomaterials based on albumin retention.Graphical abstractGraphical abstract for this article
  • Osteogenesis by foamed and 3D-printed nanostructured calcium phosphate
           scaffolds: effect of pore architecture
    • Abstract: Publication date: Available online 6 September 2018Source: Acta BiomaterialiaAuthor(s): Albert Barba, Yassine Maazouz, Anna Diez-Escudero, Katrin Rappe, Montserrat Espanol, Edgar B. Montufar, Caroline Öhman-Mägi, Cecilia Persson, Pedro Fontecha, Maria-Cristina Manzanares, Jordi Franch, Maria-Pau Ginebra There is an urgent need of synthetic bone grafts with enhanced osteogenic capacity. This can be achieved by combining biomaterials with exogenous growth factors, which however can have numerous undesired side effects, but also by tuning the intrinsic biomaterial properties. In a previous study, we showed the synergistic effect of nanostructure and pore architecture of biomimetic calcium deficient hydroxyapatite (CDHA) scaffolds in enhancing osteoinduction, i.e. fostering the differentiation of mesenchymal stem cells to bone forming cells. This was demonstrated by assessing bone formation after implanting the scaffolds intramuscularly. The present study goes one step forward, since it analyzes the effect of the geometrical features of the same CDHA scaffolds, obtained either by 3D-printing or by foaming, on the osteogenic potential and resorption behaviour in a bony environment. After 6 and 12 weeks of intraosseous implantation, both bone formation and material degradation had been drastically affected by the macropore architecture of the scaffolds. Whereas nanostructured CDHA was shown to be highly osteoconductive both in the robocast and foamed scaffolds, a superior osteogenic capacity was observed in the foamed scaffolds, which was associated with their higher intrinsic osteoinductive potential. Moreover, they showed a significantly higher cell-mediated degradation than the robocast constructs, with a simultaneous and progressive replacement of the scaffold by new bone. In conclusion, these results demonstrate that the control of macropore architecture is a crucial parameter in the design of synthetic bone grafts, which allows fostering both material degradation and new bone formation.Statement of Significance3D-printing technologies open new perspectives for the design of patient-specific bone grafts, since they allow customizing the external shape together with the internal architecture of implants. In this respect, it is important to design the appropriate pore geometry to maximize the bone healing capacity of these implants. The present study analyses the effect of pore architecture of nanostructured hydroxyapatite scaffolds, obtained either by 3D-printing or foaming, on the osteogenic potential and scaffold resorption in an in vivo model. While nanostructured hydroxyapatite showed excellent osteoconductive properties irrespective of pore geometry, we demonstrated that the spherical, concave macropores of foamed scaffolds significantly promoted both material resorption and bone regeneration compared to the 3D-printed scaffolds with orthogonal-patterned struts and therefore prismatic, convex macropores.Graphical abstractGraphical abstract for this article
  • PEGylation of model drug carriers enhances phagocytosis by primary human
    • Abstract: Publication date: Available online 6 September 2018Source: Acta BiomaterialiaAuthor(s): William J. Kelley, Catherine A. Fromen, Genesis Lopez-Cazares, Omolola Eniola-Adefeso Targeted drug carriers are attractive for the delivery of therapeutics directly to the site of a disease, reducing systemic side effects and enhancing the efficacy of therapeutic molecules. However, the use of particulate carriers for drug delivery comes with its own set of challenges and barriers. Among these, a great deal of research effort has focused on protecting carriers from clearance by phagocytes via altering carrier surface chemistry, mostly with the use of polyethylene glycol (PEG) chain coatings. However, few papers have explored the effects of PEGylation on uptake by freshly obtained primary human phagocytes in physiological conditions. In this work, we investigate the effect of PEGylation on particle uptake by primary human neutrophils in vitro and compare these effects to several cell lines and other model phagocytic cells systems. We find that human neutrophils in whole blood preferentially phagocytose PEGylated particles (e.g., ∼40% particle positive neutrophils for PEGylated versus ∼20% for carboxylated polystyrene microspheres) and that this effect is linked to factors present in human plasma. Model phagocytes internalized PEGylated particles less efficiently or equivalently to carboxylated particles in culture medium but preferentially phagocytosed PEGylated particles in the human plasma (e.g., ∼86% versus ∼63% PEGylated versus carboxylated particle positive cells, respectively). These findings have significant implications for the efficacy of PEGylation in designing long-circulating drug carriers, as well as the need for thorough characterization of drug carrier platforms in a wide array of in vitro and in vivo assays.Statement of SignificanceThe work in this manuscript is highly significant to the field of drug delivery, as it explores in-depth the effects of polyethylene glycol (PEG) coatings, which are frequently used to prevent phagocytic clearance of particulate drug carriers, on the phagocytosis of such carriers by neutrophils, the most abundant leukocyte in blood circulation. Surprisingly, we find that PEGylation enhances uptake by primary human neutrophils, specifically in the presence of human plasma. This result suggests that PEGylation may not confer the benefits in humans once thought, and may help to explain why PEG has not become the “magic bullet” it was once thought to be in the field of particulate drug delivery.Graphical abstractGraphical abstract for this article
  • Toxicological assessment of additively manufactured methacrylates for
           medical devices in dentistry
    • Abstract: Publication date: Available online 10 August 2018Source: Acta BiomaterialiaAuthor(s): Frank Alifui-Segbaya, Jasper Bowman, Alan R. White, Sony Varma, Graham J. Lieschke, Roy George The paucity of information on the biological risks of photopolymers in additive manufacturing or three-dimensional printing (3DP) is a major challenge for the uptake of the technology in the construction of medical devices in dentistry. In this paper, the biocompatibility of additively manufactured methacrylates are evaluated using the innovative zebrafish (Danio rerio) embryo model. Data obtained confirmed gradations of toxicity primarily influenced by ethanol treatment, exposure scenarios, and extraction vehicles. In direct exposure tests, zebrafish (AB/Tü) embryos cultured on materials with ultrapure water showed accelerated toxicological effects compared to counterparts in transparent E3 medium. Similarly, toxic extracts induced mostly acute responses rather than cumulative chronic as observed in direct exposure. FTIR spectra was used to quantify the double bond conversion of the materials and those with>60 % BisEMA displayed relatively low degree of conversion; however, they were safe in zebrafish bioassays after ethanol treatment in contrast to untreated methacrylates. The study affirms that biocompatibility was influenced primarily by the physico-chemical characteristics, which subsequently influenced their residual monomer content before and after ethanol treatment. Given the precautionary implications of the study, we propose a 3-tiered approach i.e. using approved materials, apposite manufacturing parameters and post-processing techniques that together guarantee optimal results for all medical devices.Statement of significanceThis study is timely and relevant since there is limited published literature that precisely describes the toxicological properties of additively manufactured methacrylates despite their increased popularity for medical devices. While it is generally accepted that the zebrafish excels as a model system for developmental toxicity, a further examination of its utility in this study using different protocols provides basis for its consideration and adoption at a crucial time when there is a lack of consensus regarding the most suited biological assessment methods for medical devices.Graphical abstractGraphical abstract for this article
  • Lessons to be learned and future directions for intervertebral disc
    • Abstract: Publication date: Available online 6 August 2018Source: Acta BiomaterialiaAuthor(s): Matteo D'Este, David Eglin, Mauro Alini Biomaterials science has achieved significant advancements for the replacement, repair and regeneration of intervertebral disc tissues. However, the translation of this research to the clinic presents hurdles. The goal of this paper is to identify strategies to recapitulate the intrinsic complexities of the intervertebral disc, to highlight the unresolved issues in basic knowledge hindering the clinical translation, and finally to report on the emerging technologies in the biomaterials field. On this basis, we identify promising research directions, with the hope of stimulating further debate and advances for resolving clinical problems such as cervical and low back pain using biomaterial-based approaches.
  • Young’s modulus of trabecular bone at the tissue level: A review
    • Abstract: Publication date: Available online 4 August 2018Source: Acta BiomaterialiaAuthor(s): Dan Wu, Per Isaksson, Stephen J. Ferguson, Cecilia Persson The tissue-level Young’s modulus of trabecular bone is important for detailed mechanical analysis of bone and bone-implant mechanical interactions. However, the heterogeneity and small size of the trabecular struts complicate an accurate determination. Methods such as micro-mechanical testing of single trabeculae, ultrasonic testing, and nanoindentation have been used to estimate the trabecular Young’s modulus. This review summarizes and classifies the trabecular Young’s moduli reported in the literature. Information on species, anatomic site, and test condition of the samples has also been gathered. Advantages and disadvantages of the different methods together with recent developments are discussed, followed by some suggestions for potential improvement, for future work. In summary, this review provides a thorough introduction to the approaches used for determining trabecular Young’s modulus, highlights important considerations when applying these methods and summarizes the reported Young’s modulus for follow-up studies on trabecular properties.Statement of SignificanceThe spongy trabecular bone provides mechanical support while maintaining a low weight. A correct measure of its mechanical properties at the tissue level, i.e. at a single-trabecula level, is crucial for analysis of interactions between bone and implants, necessary for understanding e.g. bone healing mechanisms. In this study, we comprehensively summarize the Young’s moduli of trabecular bone estimated by currently available methods, and report their dependency on different factors. The critical review of different methods with recent updates is intended to inspire improvements in estimating trabecular Young’s modulus. It is strongly suggested to report detailed information on the tested bone to enable statistical analysis in the future.Graphical abstractGraphical abstract for this article
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