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BIOLOGY (1424 journals)                  1 2 3 4 5 6 7 8 | Last

Showing 1 - 200 of 1720 Journals sorted alphabetically
AAPS Journal     Hybrid Journal   (Followers: 20)
Achievements in the Life Sciences     Open Access   (Followers: 4)
ACS Synthetic Biology     Full-text available via subscription   (Followers: 21)
Acta Biologica Colombiana     Open Access   (Followers: 7)
Acta Biologica Hungarica     Full-text available via subscription   (Followers: 4)
Acta Biologica Sibirica     Open Access  
Acta Biomaterialia     Hybrid Journal   (Followers: 25)
Acta Biotheoretica     Hybrid Journal   (Followers: 5)
Acta Chiropterologica     Full-text available via subscription   (Followers: 6)
acta ethologica     Hybrid Journal   (Followers: 4)
Acta Limnologica Brasiliensia     Open Access   (Followers: 3)
Acta Médica Costarricense     Open Access   (Followers: 2)
Acta Musei Silesiae, Scientiae Naturales : The Journal of Silesian Museum in Opava     Open Access  
Acta Neurobiologiae Experimentalis     Open Access  
Acta Parasitologica     Hybrid Journal   (Followers: 9)
Acta Scientiarum. Biological Sciences     Open Access   (Followers: 2)
Acta Scientifica Naturalis     Open Access   (Followers: 2)
Actualidades Biológicas     Open Access   (Followers: 1)
Advanced Health Care Technologies     Open Access   (Followers: 4)
Advances in Antiviral Drug Design     Full-text available via subscription   (Followers: 3)
Advances in Bioinformatics     Open Access   (Followers: 18)
Advances in Biological Regulation     Hybrid Journal   (Followers: 4)
Advances in Biosensors and Bioelectronics     Open Access   (Followers: 6)
Advances in Cell Biology     Open Access   (Followers: 24)
Advances in Cellular and Molecular Biology of Membranes and Organelles     Full-text available via subscription   (Followers: 12)
Advances in Developmental Biology     Full-text available via subscription   (Followers: 11)
Advances in DNA Sequence-Specific Agents     Full-text available via subscription   (Followers: 5)
Advances in Ecological Research     Full-text available via subscription   (Followers: 42)
Advances in Environmental Sciences - International Journal of the Bioflux Society     Open Access   (Followers: 20)
Advances in Enzyme Research     Open Access   (Followers: 9)
Advances in Experimental Biology     Full-text available via subscription   (Followers: 7)
Advances in Genome Biology     Full-text available via subscription   (Followers: 11)
Advances in High Energy Physics     Open Access   (Followers: 19)
Advances in Human Biology     Open Access   (Followers: 1)
Advances in Life Science and Technology     Open Access   (Followers: 14)
Advances in Life Sciences     Open Access   (Followers: 4)
Advances in Marine Biology     Full-text available via subscription   (Followers: 16)
Advances in Molecular and Cell Biology     Full-text available via subscription   (Followers: 22)
Advances in Planar Lipid Bilayers and Liposomes     Full-text available via subscription   (Followers: 3)
Advances in Regenerative Biology     Open Access   (Followers: 1)
Advances in Structural Biology     Full-text available via subscription   (Followers: 8)
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: 8)
Aging Cell     Open Access   (Followers: 10)
Agrokémia és Talajtan     Full-text available via subscription   (Followers: 2)
Agrokreatif Jurnal Ilmiah Pengabdian kepada Masyarakat     Open Access  
AJP Cell Physiology     Full-text available via subscription   (Followers: 13)
AJP Endocrinology and Metabolism     Full-text available via subscription   (Followers: 22)
AJP Lung Cellular and Molecular Physiology     Full-text available via subscription   (Followers: 3)
Al-Kauniyah : Jurnal Biologi     Open Access  
Alasbimn Journal     Open Access   (Followers: 1)
AMB Express     Open Access   (Followers: 1)
Ambix     Hybrid Journal   (Followers: 3)
American Biology Teacher     Full-text available via subscription   (Followers: 12)
American Fern Journal     Full-text available via subscription   (Followers: 1)
American Journal of Agricultural and Biological Sciences     Open Access   (Followers: 10)
American Journal of Bioethics     Hybrid Journal   (Followers: 10)
American Journal of Biostatistics     Open Access   (Followers: 9)
American Journal of Human Biology     Hybrid Journal   (Followers: 12)
American Journal of Medical and Biological Research     Open Access   (Followers: 5)
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: 66)
Amphibia-Reptilia     Hybrid Journal   (Followers: 6)
Anaerobe     Hybrid Journal   (Followers: 4)
Analytical Methods     Full-text available via subscription   (Followers: 7)
Anatomical Science International     Hybrid Journal   (Followers: 2)
Animal Cells and Systems     Hybrid Journal   (Followers: 4)
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)
Annales UMCS, Biologia     Open Access   (Followers: 1)
Annals of Applied Biology     Hybrid Journal   (Followers: 8)
Annals of Biomedical Engineering     Hybrid Journal   (Followers: 18)
Annals of Human Biology     Hybrid Journal   (Followers: 4)
Annual Review of Biomedical Engineering     Full-text available via subscription   (Followers: 17)
Annual Review of Biophysics     Full-text available via subscription   (Followers: 25)
Annual Review of Cancer Biology     Full-text available via subscription   (Followers: 1)
Annual Review of Cell and Developmental Biology     Full-text available via subscription   (Followers: 39)
Annual Review of Food Science and Technology     Full-text available via subscription   (Followers: 14)
Annual Review of Genomics and Human Genetics     Full-text available via subscription   (Followers: 19)
Annual Review of Phytopathology     Full-text available via subscription   (Followers: 10)
Anthropological Review     Open Access   (Followers: 25)
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: 9)
Aquaculture Environment Interactions     Open Access   (Followers: 2)
Aquaculture International     Hybrid Journal   (Followers: 22)
Aquaculture Reports     Open Access   (Followers: 3)
Aquaculture, Aquarium, Conservation & Legislation - International Journal of the Bioflux Society     Open Access   (Followers: 6)
Aquatic Biology     Open Access   (Followers: 4)
Aquatic Ecology     Hybrid Journal   (Followers: 30)
Aquatic Ecosystem Health & Management     Hybrid Journal   (Followers: 13)
Aquatic Science and Technology     Open Access   (Followers: 3)
Aquatic Toxicology     Hybrid Journal   (Followers: 19)
Archaea     Open Access   (Followers: 3)
Archiv für Molluskenkunde: International Journal of Malacology     Full-text available via subscription   (Followers: 3)
Archives of Biomedical Sciences     Open Access   (Followers: 7)
Archives of Microbiology     Hybrid Journal   (Followers: 8)
Archives of Natural History     Hybrid Journal   (Followers: 7)
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: 3)
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: 2)
Artificial Photosynthesis     Open Access   (Followers: 1)
Asian Bioethics Review     Full-text available via subscription   (Followers: 1)
Asian Journal of Biodiversity     Open Access   (Followers: 5)
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: 2)
Asian Journal of Nematology     Open Access   (Followers: 3)
Asian Journal of Poultry Science     Open Access   (Followers: 4)
Australian Life Scientist     Full-text available via subscription   (Followers: 2)
Australian Mammalogy     Hybrid Journal   (Followers: 5)
Autophagy     Hybrid Journal   (Followers: 2)
Avian Biology Research     Full-text available via subscription   (Followers: 4)
Avian Conservation and Ecology     Open Access   (Followers: 12)
Bacteriology Journal     Open Access   (Followers: 2)
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)
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)
Biochemistry and Cell Biology     Hybrid Journal   (Followers: 14)
Biochimie     Hybrid Journal   (Followers: 7)
BioControl     Hybrid Journal   (Followers: 5)
Biocontrol Science and Technology     Hybrid Journal   (Followers: 5)
Biodemography and Social Biology     Hybrid Journal   (Followers: 1)
Biodiversidad Colombia     Open Access  
Biodiversity : Research and Conservation     Open Access   (Followers: 26)
Biodiversity and Natural History     Open Access   (Followers: 5)
Biodiversity Data Journal     Open Access   (Followers: 3)
Biodiversity Informatics     Open Access  
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: 14)
BioéthiqueOnline     Open Access  
Biofabrication     Hybrid Journal   (Followers: 3)
Biogeosciences (BG)     Open Access   (Followers: 10)
Biogeosciences Discussions (BGD)     Open Access   (Followers: 1)
Bioinformatics     Hybrid Journal   (Followers: 292)
Bioinformatics and Biology Insights     Open Access   (Followers: 15)
Bioinspiration & Biomimetics     Hybrid Journal   (Followers: 6)
Biointerphases     Open Access   (Followers: 1)
Biojournal of Science and Technology     Open Access  
Biologia     Hybrid Journal  
Biologia on-line : Revista de divulgació de la Facultat de Biologia     Open Access  
Biological Bulletin     Partially Free   (Followers: 4)
Biological Control     Hybrid Journal   (Followers: 5)
Biological Invasions     Hybrid Journal   (Followers: 16)
Biological Journal of the Linnean Society     Hybrid Journal   (Followers: 15)
Biological Letters     Open Access   (Followers: 4)
Biological Procedures Online     Open Access  
Biological Psychiatry     Hybrid Journal   (Followers: 42)
Biological Psychology     Hybrid Journal   (Followers: 6)
Biological Research     Open Access  
Biological Rhythm Research     Hybrid Journal   (Followers: 2)
Biological Theory     Hybrid Journal   (Followers: 1)
Biological Trace Element Research     Hybrid Journal  
Biologicals     Full-text available via subscription   (Followers: 9)
Biologics: Targets & Therapy     Open Access   (Followers: 1)
Biologie Aujourd'hui     Full-text available via subscription  
Biologie in Unserer Zeit (Biuz)     Hybrid Journal   (Followers: 42)
Biologija     Open Access  
Biology     Open Access   (Followers: 5)
Biology and Philosophy     Hybrid Journal   (Followers: 16)
Biology Bulletin     Hybrid Journal   (Followers: 1)
Biology Bulletin Reviews     Hybrid Journal  
Biology Direct     Open Access   (Followers: 7)
Biology Letters     Full-text available via subscription   (Followers: 36)
Biology Methods and Protocols     Hybrid Journal  

        1 2 3 4 5 6 7 8 | Last

Journal Cover Acta Biomaterialia
  [SJR: 2.02]   [H-I: 104]   [25 followers]  Follow
   Hybrid Journal Hybrid journal (It can contain Open Access articles)
   ISSN (Print) 1742-7061
   Published by Elsevier Homepage  [3043 journals]
  • Cell-laden hydrogels for osteochondral and cartilage tissue engineering
    • Authors: Jingzhou Yang; Yu Shrike Zhang; Kan Yue; Ali Khademhosseini
      Pages: 1 - 25
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Jingzhou Yang, Yu Shrike Zhang, Kan Yue, Ali Khademhosseini
      Despite tremendous advances in the field of regenerative medicine, it still remains challenging to repair the osteochondral interface and full-thickness articular cartilage defects. This inefficiency largely originates from the lack of appropriate tissue-engineered artificial matrices that can replace the damaged regions and promote tissue regeneration. Hydrogels are emerging as a promising class of biomaterials for both soft and hard tissue regeneration. Many critical properties of hydrogels, such as mechanical stiffness, elasticity, water content, bioactivity, and degradation, can be rationally designed and conveniently tuned by proper selection of the material and chemistry. Particularly, advances in the development of cell-laden hydrogels have opened up new possibilities for cell therapy. In this article, we describe the problems encountered in this field and review recent progress in designing cell-hydrogel hybrid constructs for promoting the reestablishment of osteochondral/cartilage tissues. Our focus centers on the effects of hydrogel type, cell type, and growth factor delivery on achieving efficient chondrogenesis and osteogenesis. We give our perspective on developing next-generation matrices with improved physical and biological properties for osteochondral/cartilage tissue engineering. We also highlight recent advances in biomanufacturing technologies (e.g. molding, bioprinting, and assembly) for fabrication of hydrogel-based osteochondral and cartilage constructs with complex compositions and microarchitectures to mimic their native counterparts. Statement of significance Despite tremendous advances in the field of regenerative medicine, it still remains challenging to repair the osteochondral interface and full-thickness articular cartilage defects. This inefficiency largely originates from the lack of appropriate tissue-engineered biomaterials that replace the damaged regions and promote tissue regeneration. Cell-laden hydrogel systems have emerged as a promising tissue-engineering platform to address this issue. In this article, we describe the fundamental problems encountered in this field and review recent progress in designing cell-hydrogel constructs for promoting the reestablishment of osteochondral/cartilage tissues. Our focus centers on the effects of hydrogel composition, cell type, and growth factor delivery on achieving efficient chondrogenesis and osteogenesis. We give our perspective on developing next-generation hydrogel/inorganic particle/stem cell hybrid composites with improved physical and biological properties for osteochondral/cartilage tissue engineering. We also highlight recent advances in biomanufacturing and bioengineering technologies (e.g. 3D bioprinting) for fabrication of hydrogel-based osteochondral and cartilage constructs.
      Graphical abstract image

      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.01.036
      Issue No: Vol. 57 (2017)
  • 3D bioprinting for drug discovery and development in pharmaceutics
    • Authors: Weijie Peng; Pallab Datta; Bugra Ayan; Veli Ozbolat; Donna Sosnoski; Ibrahim T. Ozbolat
      Pages: 26 - 46
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Weijie Peng, Pallab Datta, Bugra Ayan, Veli Ozbolat, Donna Sosnoski, Ibrahim T. Ozbolat
      Successful launch of a commercial drug requires significant investment of time and financial resources wherein late-stage failures become a reason for catastrophic failures in drug discovery. This calls for infusing constant innovations in technologies, which can give reliable prediction of efficacy, and more importantly, toxicology of the compound early in the drug discovery process before clinical trials. Though computational advances have resulted in more rationale in silico designing, in vitro experimental studies still require gaining industry confidence and improving in vitro-in vivo correlations. In this quest, due to their ability to mimic the spatial and chemical attributes of native tissues, three-dimensional (3D) tissue models have now proven to provide better results for drug screening compared to traditional two-dimensional (2D) models. However, in vitro fabrication of living tissues has remained a bottleneck in realizing the full potential of 3D models. Recent advances in bioprinting provide a valuable tool to fabricate biomimetic constructs, which can be applied in different stages of drug discovery research. This paper presents the first comprehensive review of bioprinting techniques applied for fabrication of 3D tissue models for pharmaceutical studies. A comparative evaluation of different bioprinting modalities is performed to assess the performance and ability of fabricating 3D tissue models for pharmaceutical use as the critical selection of bioprinting modalities indeed plays a crucial role in efficacy and toxicology testing of drugs and accelerates the drug development cycle. In addition, limitations with current tissue models are discussed thoroughly and future prospects of the role of bioprinting in pharmaceutics are provided to the reader. Statement of Significance Present advances in tissue biofabrication have crucial role to play in aiding the pharmaceutical development process achieve its objectives. Advent of three-dimensional (3D) models, in particular, is viewed with immense interest by the community due to their ability to mimic in vivo hierarchical tissue architecture and heterogeneous composition. Successful realization of 3D models will not only provide greater in vitro-in vivo correlation compared to the two-dimensional (2D) models, but also eventually replace pre-clinical animal testing, which has their own shortcomings. Amongst all fabrication techniques, bioprinting- comprising all the different modalities (extrusion-, droplet- and laser-based bioprinting), is emerging as the most viable fabrication technique to create the biomimetic tissue constructs. Notwithstanding the interest in bioprinting by the pharmaceutical development researchers, it can be seen that there is a limited availability of comparative literature which can guide the proper selection of bioprinting processes and associated considerations, such as the bioink selection for a particular pharmaceutical study. Thus, this work emphasizes these aspects of bioprinting and presents them in perspective of differential requirements of different pharmaceutical studies like in vitro predictive toxicology, high-throughput screening, drug delivery and tissue-specific efficacies. Moreover, since bioprinting techniques are mostly applied in regenerative medicine and tissue engineering, a comparative analysis of similarities and differences are also expounded to help researchers make informed decisions based on contemporary literature.
      Graphical abstract image

      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.025
      Issue No: Vol. 57 (2017)
  • 3D tumor microtissues as an in vitro testing platform for
           microenvironmentally-triggered drug delivery systems
    • Authors: Virginia Brancato; Filomena Gioiella; Martina Profeta; Giorgia Imparato; Daniela Guarnieri; Francesco Urciuolo; Pietro Melone; Paolo A. Netti
      Pages: 47 - 58
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Virginia Brancato, Filomena Gioiella, Martina Profeta, Giorgia Imparato, Daniela Guarnieri, Francesco Urciuolo, Pietro Melone, Paolo A. Netti
      Therapeutic approaches based on nanomedicine have garnered great attention in cancer research. In vitro biological models that better mimic in vivo conditions are crucial tools to more accurately predict their therapeutic efficacy in vivo. In this work, a new 3D breast cancer microtissue has been developed to recapitulate the complexity of the tumor microenvironment and to test its efficacy as screening platform for drug delivery systems. The proposed 3D cancer model presents human breast adenocarcinoma cells and cancer-associated fibroblasts embedded in their own ECM, thus showing several features of an in vivo tumor, such as overexpression of metallo-proteinases (MMPs). After demonstrating at molecular and protein level the MMP2 overexpression in such tumor microtissues, we used them to test a recently validated formulation of endogenous MMP2-responsive nanoparticles (NP). The presence of the MMP2-sensitive linker allows doxorubicin release from NP only upon specific enzymatic cleavage of the peptide. The same NP without the MMP-sensitive linker and healthy breast microtissues were also produced to demonstrate NP specificity and selectivity. Cell viability after NP treatment confirmed that controlled drug delivery is achieved only in 3D tumor microtissues suggesting that the validation of therapeutic strategies in such 3D tumor model could predict human response. Statement of Significance A major issue of modern cancer research is the development of accurate and predictive experimental models of human tumors consistent with tumor microenvironment and applicable as screening platforms for novel therapeutic strategies. In this work, we developed and validated a new 3D microtissue model of human breast tumor as a testing platform of anti-cancer drug delivery systems. To this aim, biodegradable nanoparticles responsive to physiological changes specifically occurring in tumor microenvironment were used. Our findings clearly demonstrate that the breast tumor microtissue well recapitulates in vivo physiological features of tumor tissue and elicits a specific response to microenvironmentally-responsive nanoparticles compared to healthy tissue. We believe this study is of particular interest for cancer research and paves the way to exploit tumor microtissues for several testing purposes.
      Graphical abstract image

      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.004
      Issue No: Vol. 57 (2017)
  • Heparin-based hydrogels induce human renal tubulogenesis in vitro
    • Authors: Heather M. Weber; Mikhail V. Tsurkan; Valentina Magno; Uwe Freudenberg; Carsten Werner
      Pages: 59 - 69
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Heather M. Weber, Mikhail V. Tsurkan, Valentina Magno, Uwe Freudenberg, Carsten Werner
      Dialysis or kidney transplantation is the only therapeutic option for end stage renal disease. Accordingly, there is a large unmet clinical need for new causative therapeutic treatments. Obtaining robust models that mimic the complex nature of the human kidney is a critical step in the development of new therapeutic strategies. Here we establish a synthetic in vitro human renal tubulogenesis model based on a tunable glycosaminoglycan-hydrogel platform. In this system, renal tubulogenesis can be modulated by the adjustment of hydrogel mechanics and degradability, growth factor signaling, and the presence of insoluble adhesion cues, potentially providing new insights for regenerative therapy. Different hydrogel properties were systematically investigated for their ability to regulate renal tubulogenesis. Hydrogels based on heparin and matrix metalloproteinase cleavable peptide linker units were found to induce the morphogenesis of single human proximal tubule epithelial cells into physiologically sized tubule structures. The generated tubules display polarization markers, extracellular matrix components, and organic anion transport functions of the in vivo renal proximal tubule and respond to nephrotoxins comparable to the human clinical response. The established hydrogel-based human renal tubulogenesis model is thus considered highly valuable for renal regenerative medicine and personalized nephrotoxicity studies. Statement of Significance The only cure for end stage kidney disease is kidney transplantation. Hence, there is a huge need for reliable human kidney models to study renal regeneration and establish alternative treatments. Here we show the development and application of an in vitro human renal tubulogenesis model using heparin-based hydrogels. To the best of our knowledge, this is the first system where human renal tubulogenesis can be monitored from single cells to physiologically sized tubule structures in a tunable hydrogel system. To validate the efficacy of our model as a drug toxicity platform, a chemotherapy drug was incubated with the model, resulting in a drug response similar to human clinical pathology. The established model could have wide applications in the field of nephrotoxicity and renal regenerative medicine and offer a reliable alternative to animal models.
      Graphical abstract image

      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.035
      Issue No: Vol. 57 (2017)
  • Photothermal and photodynamic activity of polymeric nanoparticles based on
           α-tocopheryl succinate-RAFT block copolymers conjugated to IR-780
    • Authors: Raquel Palao-Suay; Francisco M. Martín-Saavedra; María Rosa Aguilar; Clara Escudero-Duch; Sergio Martín-Saldaña; Francisco J. Parra-Ruiz; Nathan A. Rohner; Susan N. Thomas; Nuria Vilaboa; Julio San Román
      Pages: 70 - 84
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Raquel Palao-Suay, Francisco M. Martín-Saavedra, María Rosa Aguilar, Clara Escudero-Duch, Sergio Martín-Saldaña, Francisco J. Parra-Ruiz, Nathan A. Rohner, Susan N. Thomas, Nuria Vilaboa, Julio San Román
      The aim of this work was the generation of a multifunctional nanopolymeric system that incorporates IR-780 dye, a near-infrared (NIR) imaging probe that exhibits photothermal and photodynamic properties; and a derivate of α-tocopheryl succinate (α-TOS), a mitochondria-targeted anticancer compound. IR-780 was conjugated to the hydrophilic segment of copolymer PEG-b-polyMTOS, based on poly(ethylene glycol) (PEG) and a methacrylic derivative of α-tocopheryl succinate (MTOS), to generate IR-NP, self-assembled nanoparticles (NPs) in aqueous media which exhibit a hydrophilic shell and a hydrophobic core. During assembly, the hydrophobic core of IR-NP could encapsulate additional IR-780 to generate derived subspecies carrying different amount of probe (IR-NP-eIR). Evaluation of photo-inducible properties of IR-NP and IR-NP-eIR were thoroughly assessed in vitro. Developed nanotheranostic particles showed distinct fluorescence and photothermal behavior after excitation by a laser light emitting at 808nm. Treatment of MDA-MB-453 cells with IR-NP or IR-NP-eIR resulted in an efficient internalization of the IR-780 dye, while subsequent NIR-laser irradiation led to a severe decrease in cell viability. Photocytoxicity conducted by IR-NP, which could not be attributed to the generation of lethal hyperthermia, responded to an increase in the levels of intracellular reactive oxygen species (ROS). Therefore, the fluorescence imaging and inducible phototoxicity capabilities of NPs derived from IR-780-PEG-b-polyMTOS copolymer confer high value to these nanotheranostics tools in clinical cancer research. Statement of Significance Multifunctional polymeric nanoparticles (NPs) that combine imaging and therapeutic properties are highly valuable in cancer treatment. In this paper we describe the development of NPs that are fluorescent in the near-infrared (NIR). This is important for their visualization in living tissues that present low absorption and low autofluorescence in this wavelength region (between 700 and 1000nm). Moreover, NPs present photothermal and photodynamic properties when NIR irradiated: the NPs produce an efficient increment of temperature and increase the intracellular reactive oxygen species (ROS) when laser irradiated at 808nm. These tuneable photoinduced properties make the NPs highly cytotoxic after NIR irradiation and provide a new tool for highly precise cancer treatment.
      Graphical abstract image

      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.028
      Issue No: Vol. 57 (2017)
  • Athero-inflammatory nanotherapeutics: Ferulic acid-based
           poly(anhydride-ester) nanoparticles attenuate foam cell formation by
           regulating macrophage lipogenesis and reactive oxygen species generation
    • Authors: Rebecca A. Chmielowski; Dalia S. Abdelhamid; Jonathan J. Faig; Latrisha K. Petersen; Carol R. Gardner; Kathryn E. Uhrich; Laurie B. Joseph; Prabhas V. Moghe
      Pages: 85 - 94
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Rebecca A. Chmielowski, Dalia S. Abdelhamid, Jonathan J. Faig, Latrisha K. Petersen, Carol R. Gardner, Kathryn E. Uhrich, Laurie B. Joseph, Prabhas V. Moghe
      Enhanced bioactive anti-oxidant formulations are critical for treatment of inflammatory diseases, such as atherosclerosis. A hallmark of early atherosclerosis is the uptake of oxidized low density lipoprotein (oxLDL) by macrophages, which results in foam cell and plaque formation in the arterial wall. The hypolipidemic, anti-inflammatory, and antioxidative properties of polyphenol compounds make them attractive targets for treatment of atherosclerosis. However, high concentrations of antioxidants can reverse their anti-atheroprotective properties and cause oxidative stress within the artery. Here, we designed a new class of nanoparticles with anti-oxidant polymer cores and shells comprised of scavenger receptor targeting amphiphilic macromolecules (AMs). Specifically, we designed ferulic acid-based poly(anhydride-ester) nanoparticles to counteract the uptake of high levels of oxLDL and regulate reactive oxygen species generation (ROS) in human monocyte derived macrophages (HMDMs). Compared to all compositions examined, nanoparticles with core ferulic acid-based polymers linked by diglycolic acid (PFAG) showed the greatest inhibition of oxLDL uptake. At high oxLDL concentrations, the ferulic acid diacids and polymer nanoparticles displayed similar oxLDL uptake. Treatment with the PFAG nanoparticles downregulated the expression of macrophage scavenger receptors, CD-36, MSR-1, and LOX-1 by about 20–50%, one of the causal factors for the decrease in oxLDL uptake. The PFAG nanoparticle lowered ROS production by HMDMs, which is important for maintaining macrophage growth and prevention of apoptosis. Based on these results, we propose that ferulic acid-based poly(anhydride ester) nanoparticles may offer an integrative strategy for the localized passivation of the early stages of the atheroinflammatory cascade in cardiovascular disease. Statement of Significance Future development of anti-oxidant formulations for atherosclerosis applications is essential to deliver an efficacious dose while limiting localized concentrations of pro-oxidants. In this study, we illustrate the potential of degradable ferulic acid-based polymer nanoparticles to control macrophage foam cell formation by significantly reducing oxLDL uptake through downregulation of scavenger receptors, CD-36, MSR-1, and LOX-1. Another critical finding is the ability of the degradable ferulate-based polymer nanoparticles to lower macrophage reactive oxygen species (ROS) levels, a precursor to apoptosis and plaque escalation. The degradable ferulic acid-based polymer nanoparticles hold significant promise as a means to alter the treatment and progression of atherosclerosis.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.029
      Issue No: Vol. 57 (2017)
  • Affinity interactions drive post-implantation drug filling, even in the
           presence of bacterial biofilm
    • Authors: Erika L. Cyphert; Sean T. Zuckerman; Julius N. Korley; Horst A. von Recum
      Pages: 95 - 102
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Erika L. Cyphert, Sean T. Zuckerman, Julius N. Korley, Horst A. von Recum
      Current post-operative standard of care for surgical procedures, including device implantations, dictates prophylactic antimicrobial therapy, but a percentage of patients still develop infections. Systemic antimicrobial therapy needed to treat such infections can lead to downstream tissue toxicities and generate drug-resistant bacteria. To overcome issues associated with systemic drug administration, a polymer incorporating specific drug affinity has been developed with the potential to be filled or refilled with antimicrobials, post-implantation, even in the presence of bacterial biofilm. This polymer can be used as an implant coating or stand-alone drug delivery device, and can be translated to a variety of applications, such as implanted or indwelling medical devices, and/or surgical site infections. The filling of empty affinity-based drug delivery polymer was analyzed in an in vitro filling/refilling model mimicking post-implantation tissue conditions. Filling in the absence of bacteria was compared to filling in the presence of bacterial biofilms of varying maturity to demonstrate proof-of-concept necessary prior to in vivo experiments. Antibiotic filling into biofilm-coated affinity polymers was comparable to drug filling seen in same affinity polymers without biofilm demonstrating that affinity polymers retain ability to fill with antibiotic even in the presence of biofilm. Additionally, post-implantation filled antibiotics showed sustained bactericidal activity in a zone of inhibition assay demonstrating post-implantation capacity to deliver filled antibiotics in a timeframe necessary to eradicate bacteria in biofilms. This work shows affinity polymers can fill high levels of antibiotics post-implantation independent of biofilm presence potentially enabling device rescue, rather than removal, in case of infection. Statement of Significance Post-operative prophylactic antimicrobial therapy greatly reduces risk of infection, such as on biomedical implants, but does not totally eliminate infections, and the healthcare cost of these remaining infections remains a major concern. Systemic antimicrobial therapy to treat these infections can lead to tissue toxicity and drug-resistant bacteria. In order to treat only those patients who have developed infections, a customizable antimicrobial delivery system made of cyclodextrin-based affinity polymer has been developed that is capable of filling post-implantation and delivering the filled antibiotic in a sustained manner even when the delivery device covered in bacterial biofilm. These observations have the potential to be translated to a wide variety of applications, such as implanted or indwelling medical devices, and/or surgical site infections.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.015
      Issue No: Vol. 57 (2017)
  • Disruption of drug-resistant biofilms using de novo designed short
           α-helical antimicrobial peptides with idealized facial amphiphilicity
    • Authors: Jasmeet Singh Khara; Sybil Obuobi; Ying Wang; Melissa Shea Hamilton; Brian D. Robertson; Sandra M. Newton; Yi Yan Yang; Paul R. Langford; Pui Lai Rachel Ee
      Pages: 103 - 114
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Jasmeet Singh Khara, Sybil Obuobi, Ying Wang, Melissa Shea Hamilton, Brian D. Robertson, Sandra M. Newton, Yi Yan Yang, Paul R. Langford, Pui Lai Rachel Ee
      The escalating threat of antimicrobial resistance has increased pressure to develop novel therapeutic strategies to tackle drug-resistant infections. Antimicrobial peptides have emerged as a promising class of therapeutics for various systemic and topical clinical applications. In this study, the de novo design of α-helical peptides with idealized facial amphiphilicities, based on an understanding of the pertinent features of protein secondary structures, is presented. Synthetic amphiphiles composed of the backbone sequence (X1Y1Y2X2)n, where X1 and X2 are hydrophobic residues (Leu or Ile or Trp), Y1 and Y2 are cationic residues (Lys), and n is the number repeat units (2 or 2.5 or 3), demonstrated potent broad-spectrum antimicrobial activities against clinical isolates of drug-susceptible and multi-drug resistant bacteria. Live-cell imaging revealed that the most selective peptide, (LKKL)3, promoted rapid permeabilization of bacterial membranes. Importantly, (LKKL)3 not only suppressed biofilm growth, but effectively disrupted mature biofilms after only 2h of treatment. The peptides (LKKL)3 and (WKKW)3 suppressed the production of LPS-induced pro-inflammatory mediators to levels of unstimulated controls at low micromolar concentrations. Thus, the rational design strategies proposed herein can be implemented to develop potent, selective and multifunctional α-helical peptides to eradicate drug-resistant biofilm-associated infections. Statement of Significance Antimicrobial peptides (AMPs) are increasingly explored as therapeutics for drug-resistant and biofilm-related infections to help expand the size and quality of the current antibiotic pipeline in the face of mounting antimicrobial resistance. Here, synthetic peptides rationally designed based upon principles governing the folding of natural α-helical AMPs, comprising the backbone sequence (X1Y1Y2X2)n, and which assemble into α-helical structures with idealized facial amphiphilicity, is presented. These multifunctional peptide amphiphiles demonstrate high bacterial selectivity, promote the disruption of pre-formed drug-resistant biofilms, and effectively neutralize endotoxins at low micromolar concentrations. Overall, the design strategies presented here could provide a useful tool for developing therapeutic peptides with broad-ranging clinical applications from the treatment and prevention of drug-resistant biofilms to the neutralization of bacterial endotoxins.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.032
      Issue No: Vol. 57 (2017)
  • Polyurethane acrylates as effective substrates for sustained in vitro
           culture of human myotubes
    • Authors: Yosephine Andriani; Jason Min-Wen Chua; Benjamin Yan-Jiang Chua; In Yee Phang; Ng Shyh-Chang; Wui Siew Tan
      Pages: 115 - 126
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Yosephine Andriani, Jason Min-Wen Chua, Benjamin Yan-Jiang Chua, In Yee Phang, Ng Shyh-Chang, Wui Siew Tan
      Muscular disease has debilitating effects with severe damage leading to death. Our knowledge of muscle biology, disease and treatment is largely derived from non-human cell models, even though non-human cells are known to differ from human cells in their biochemical responses. Attempts to develop highly sought after in vitro human cell models have been plagued by early cell delamination and difficulties in achieving human myotube culture in vitro. In this work, we developed polyurethane acrylate (PUA) materials to support long-term in vitro culture of human skeletal muscle tissue. Using a constant base with modulated crosslink density we were able to vary the material modulus while keeping surface chemistry and roughness constant. While previous studies have focused on materials that mimic soft muscle tissue with stiffness ca. 12kPa, we investigated materials with tendon-like surface moduli in the higher 150MPa to 2.4GPa range, which has remained unexplored. We found that PUA of an optimal modulus within this range can support human myoblast proliferation, terminal differentiation and sustenance beyond 35days, without use of any extracellular protein coating. Results show that PUA materials can serve as effective substrates for successful development of human skeletal muscle cell models and are suitable for long-term in vitro studies. Statement of Significance We developed polyurethane acrylates (PUA) to modulate the human skeletal muscle cell growth and maturation in vitro by controlling surface chemistry, morphology and tuning material’s stiffness. PUA was able to maintain muscle cell viability for over a month without any detectable signs of material degradation. The best performing PUA prevented premature cell detachment from the substrate which often hampered long-term muscle cell studies. It also supported muscle cell maturation up to the late stages of differentiation. The significance of these findings lies in the possibility to advance studies on muscle cell biology, disease and therapy by using human muscle cells instead of relying on the widely used animal-based in vitro models.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.022
      Issue No: Vol. 57 (2017)
  • Development of a local anesthetic lidocaine-loaded redox-active injectable
           gel for postoperative pain management
    • Authors: Yukio Nagasaki; Yutaro Mizukoshi; Zhenyu Gao; Chitho P. Feliciano; Kyungho Chang; Hiroshi Sekiyama; Hiroyuki Kimura
      Pages: 127 - 135
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Yukio Nagasaki, Yutaro Mizukoshi, Zhenyu Gao, Chitho P. Feliciano, Kyungho Chang, Hiroshi Sekiyama, Hiroyuki Kimura
      Although local anesthesia is commonly applied for pain relief, there are several issues such as its short duration of action and low effectiveness at the areas of inflammation due to the acidic pH. The presence of excessive amount of reactive oxygen species (ROS) is known to induce inflammation and aggravate pain. To resolve these issues, we developed a redox-active injectable gel (RIG) with ROS-scavenging activity. RIG was prepared by mixing polyamine-b-poly(ethylene glycol)-b-polyamine with nitroxide radical moieties as side chains on the polyamine segments (PMNT-b-PEG-b-PMNT) with a polyanion, which formed a flower-type micelle via electrostatic complexation. Lidocaine could be stably incorporated in its core. When the temperature of the solution was increased to 37°C, the PIC-type flower micelle transformed to gel. The continuous release of lidocaine from the gel was observed for more than three days, without remarkable initial burst, which is probably owing to the stable entrapment of lidocaine in the PIC core of the gel. We evaluated the analgesic effect of RIG in carrageenan-induced arthritis mouse model. Results showed that lidocaine-loaded RIG has stronger and longer analgesic effect when administered in inflamed areas. In contrast, while the use of non-complexed lidocaine did not show analgesic effect one day after its administration. Note that no effect was observed when PIC-type flower micelle without ROS-scavenging ability was used. These findings suggest that local anesthetic-loaded RIG can effectively reduce the number of injection times and limit the side effects associated with the use of anti-inflammatory drugs for postoperative pain management. Statement of Significance 1. We have been working on nanomaterials, which effectively eliminate ROS, avoiding dysfunction of mitochondria in healthy cells. 2. We designed redox injectable gel using polyion complexed flower type micelle, which can eliminates ROS locally. 3. We could prepare local anesthesia-loaded redox injectable gel (lido@RIG). 4. Drug release could be extended by local administration of lido@RIG. 5. Deprotonation of lidocaine improved anesthetic effect because ROS were eliminated locally by RIG. 6. Local inflammation could be also suppressed by lido@RIG.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.031
      Issue No: Vol. 57 (2017)
  • Precise manipulation of biophysical particle parameters enables control of
           proinflammatory cytokine production in presence of TLR 3 and 4 ligands
    • Authors: Yoshinori Kakizawa; Jung Seok Lee; Brendan Bell; Tarek M. Fahmy
      Pages: 136 - 145
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Yoshinori Kakizawa, Jung Seok Lee, Brendan Bell, Tarek M. Fahmy
      The biophysical parameters governing nanoparticle (NP)-cell interactions significantly affect biological responses, particularly in the application of NP-based immunotherapeutics. Modulation of the surface biophysical character of NPs can be achieved via introduction of amino acids, which offer the ability to fine tune a range of biophysical parameters of interest. We employed this approach using monodisperse silica NPs coated with numerous poly(amino acid)s (PAAs). The NPs were incubated with dendritic cells (DCs) in conjunction with TLR ligands and production of IL-1β from DCs and IFNγ from T cells primed by these DCs were measured. These key cytokines can prognosticate the efficacy of the NP platform as a potential vaccine or active cellular immunotherapy carrier. IL-1β production showed a correlation with both NP size and degree of hydrophobicity. High IFNγ secretion from T cells was shown to be correlated with both the hydrophobicity and charge of the NPs used to activate the DCs. Other cytokines were also screened in order to compare the immune responses. The results of this study highlight the importance of nanoparticle biophysical parameters and the selection of TLR ligands to the rational design of nanoparticle-based vaccines and immunotherapies. Statement of Significance The manuscript describes a systematic investigation into the effects of biophysical parameters of nanoparticles (NPs) on immune cells. Modulation of the biophysical character of the NP surface can be achieved by introduction of amino acids on monodisperse silica NPs, introducing a range of tunable biophysical parameters of interest, i.e. distinct sizes, different surface charges and varying degrees of surface hydrophobicity. We examine internalization of the NP in dendritic cells (DCs) and measure a myriad of cytokines, including IL-1β and IFNγ, which prognosticate the efficacy of the NPs as a potential vaccine (IL-1β metric) or active cellular immunotherapy carrier (IFNγ metric). Two different TLR ligands (a viral TLR3 ligand and a bacterial TLR4 ligand) were used along with the PAA NPs to compare their costimulatory immunogenicity. We strongly believe that this study will provide crucial information to many readers of Acta Biomaterialia and further drive the use of nanoparticle platforms in modulating immune responses.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.01.025
      Issue No: Vol. 57 (2017)
  • Inhibition of allogeneic cytotoxic T cell (CD8+) proliferation via
           polymer-induced Treg (CD4+) cells
    • Authors: Ning Kang; Wendy M. Toyofuku; Xining Yang; Mark D. Scott
      Pages: 146 - 155
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Ning Kang, Wendy M. Toyofuku, Xining Yang, Mark D. Scott
      T cell-mediated immune rejection remains a barrier to successful transplantation. Polymer-based bioengineering of cells may provide an effective means of preventing allorecognition and the proliferation of cytotoxic (CD8+) T lymphocytes (CTL). Using MHC-disparate murine splenocytes modified with succinimidyl valerate activated methoxypoly(ethylene glycol) [SVA-mPEG] polymers, the effects of leukocyte immunocamouflage on CD8+ and CD4+ alloproliferation and T regulatory (Treg) cell induction were assessed in a mixed lymphocyte reaction (MLR) model. Polymer-grafting effectively camouflaged multiple leukocyte markers (MHC class I and II, TCR and CD3) essential for effective allorecognition. Consequent to the polymer-induced immunocamouflage of the cell membrane, both CD8+ and CD4+ T cell alloproliferation were significantly inhibited in a polymer dose-dependent manner. The loss of alloproliferation correlated with the induction of Treg cells (CD4+CD25+Foxp3+). The Tregs, surprisingly, arose primarily via differentiation of naive, non-proliferating, CD4+ cells. Of biologic importance, the polymer-induced Treg were functional and exhibited potent immunosuppressive activity on allogeneic CTL proliferation. These results suggest that immunocamouflage-mediated attenuation of alloantigen-TCR recognition can prevent the tissue destructive allogeneic CD8+ T cell response, both directly and indirectly, through the generation/differentiation of functional Tregs. Immunocamouflage induced tolerance could be clinically valuable in attenuating T cell-mediated transplant rejection and in the treatment of autoimmune diseases. Statement of Significance While our previous studies have demonstrated that polymer-grafting to MHC disparate leukocytes inhibits CD4+ cell proliferation, the effects of PEGylation on the alloproliferation of CD8+ cytotoxic T cells (CTL) was not examined. As shown here, PEGylation of allogeneic leukocytes prevents the generation of the CTL response responsible for acute rejection. The loss of CTL proliferation is consequent to the polymer-based attenuation of allorecognition and the induction of T regulatory cells (Tregs). Interestingly, the Tregs are primarily generated via the differentiation of non-proliferating naive T cells. Importantly, the Tregs are functional and effectively induce a tolerogenic environment when transferred to an alloresponsive environment. The use of polymer-modified leukocytes provides a unique approach to effectively maximize the biologic production of functional Tregs both in vitro and in vivo. By using this approach it may be possible to attenuate unwanted alloresponses (e.g., graft rejection) or to treat autoimmune diseases.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.025
      Issue No: Vol. 57 (2017)
  • Retro-inverso d-peptide-modified hyaluronic acid/bioreducible
           hyperbranched poly(amido amine)/pDNA core-shell ternary nanoparticles for
           the dual-targeted delivery of short hairpin RNA-encoding plasmids
    • Authors: Jijin Gu; Xinyi Chen; Xiaoling Fang; Xianyi Sha
      Pages: 156 - 169
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Jijin Gu, Xinyi Chen, Xiaoling Fang, Xianyi Sha
      The active targeting of gene carriers is a powerful strategy for improving tumour-specific delivery and therapy. Although numerous l-peptide ligands play significant roles in the active targeting of nanomedicine, retro-inverso d-peptides have been explored as targeting ligands due to their superior stability and bioactivity in vivo. In this study, retro-inverso d-peptide (RIF7)-modified hyaluronic acid (HA)/bioreducible hyperbranched poly(amido amine) (RHB)/plasmid DNA (pDNA) ternary nanoparticles were successfully developed using the layer-by-layer method for the CD44-positive tumour-specific delivery of short hairpin RNA (shRNA)-encoding pDNA through the combination of the Anxa1 (tumour vasculature) and CD44 (tumour cell-surface) receptors, which mediated the dual targeting. The potential of these newly designed nanoparticles was evaluated by examining the efficacy of their cellular uptake and transfection in cell monolayers, tumour spheroids, and malignant xenograft animal models. With negligible cytotoxicity, the spherical-shaped RIF7-HA/RHB/pDNA nanoparticles were the direct result of an electrostatic complex that had efficiently targeted CD44-positive tumour delivery, penetration, and cellular uptake in vitro. The nanoparticles showed excellent target-specific gene transfection even in the presence of serum. The in vivo therapeutic effect of RIF7-HA/RHB/pDNA-shRNA nanoparticle-mediated shRNA targeting of the Cyclin gene (shCyclin) was evaluated in tumour-bearing mice. The RIF7-HA/RHB/pDNA-shCyclin nanoparticles significantly increased the survival time of tumour-bearing mice and substantially reduced tumour growth due to their extremely specific tumour-targeting activity. These results suggested that the combination of HA and retro-inverso peptide RIF7 significantly increased the therapeutic effect of pDNA-shCyclin-loaded nanoparticles for CD44-positive tumours. Thus, RIF7-HA-mediated multi-target ternary gene vectors are an efficient and promising strategy for the delivery of pDNA-shRNA in the targeted treatment of malignant and metastatic cancers. Statement of Significance Although l-peptide ligands play significant roles in the active targeting of nanomedicine, retro-inverso d-peptides have been explored as targeting ligands due to their superior stability and bioactivity in vivo. Retro-inverso peptide RIF7 was designed as a ligand of Anxa1 receptor. The resultant peptide, RIF7, displayed high binding efficiency within Anxa1 receptor, which is highly expressed tumour vasculature cells and some tumour cells such as B16F10 and U87MG cells. The most important feature of RIF7 is its high stability in the blood, which is suitable and promising for application in vivo. Multifunctional RIF7-HA was then synthesized by conjugating the RIF7 peptide to HA, which was used to modify the surface of RHB/pDNA nanoparticles to prepare RIF7-HA/RHB/pDNA core-shell ternary nanoparticles for the dual-targeted delivery of shRNA-encoding plasmids in vitro and in vivo.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.024
      Issue No: Vol. 57 (2017)
  • Selective phenylalanine to proline substitution for improved antimicrobial
           and anticancer activities of peptides designed on phenylalanine heptad
    • Authors: Amit Kumar Tripathi; Tripti Kumari; Anshika Tandon; Mohd. Sayeed; Tayyaba Afshan; Manoj Kathuria; P.K. Shukla; Kalyan Mitra; Jimut Kanti Ghosh
      Pages: 170 - 186
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Amit Kumar Tripathi, Tripti Kumari, Anshika Tandon, Mohd. Sayeed, Tayyaba Afshan, Manoj Kathuria, P.K. Shukla, Kalyan Mitra, Jimut Kanti Ghosh
      Introducing cell-selectivity in antimicrobial peptides (AMPs) without compromising the antimicrobial and anti-endotoxin properties is a crucial step towards the development of new antimicrobial agents. A peptide designed on phenylalanine heptad repeat possesses significant cytotoxicity along with desired antimicrobial and anti-endotoxin properties. Amino acid substitutions at ‘a’ and/or ‘d’ positions of heptad repeats of AMPs could alter their helical structure in mammalian membrane-mimetic environments and cytotoxicity towards mammalian cells. Since proline is a helix breaker, effects of selective proline substitution(s) at ‘a’ and/or ‘d’ positions of a 15-residue peptide designed on phenylalanine heptad repeat (FR-15) were investigated. Proline-substituted FR-15 variants were highly selective toward bacteria and fungi over hRBCs and murine 3T3 cells and also retained their antibacterial activities at high salt, serum and elevated temperatures. These non-cytotoxic variants also inhibited LPS-induced production of pro-inflammatory cytokines/chemokines in human monocytes, THP-1, RAW 264.7 and in BALB/c mice. The two non-cytotoxic variants (FR8P and FR11P) showed potent anti-cancer activity against highly metastatic human breast cancer cell line MDA-MB-231 with IC50 values less than 10μM. At sub-IC50 concentrations, FR8P and FR11P also showed anti-migratory and anti-invasive effects against MDA-MB-231 cells. FR8P and FR11P induced cellular apoptosis by triggering intrinsic apoptotic pathway through depolarization of mitochondrial membrane potential and activation of caspases. Overall the results demonstrated the utilization of selective phenylalanine to proline substitution in a heptad repeat of phenylalanine residues for the design of cell-selective, broad-spectrum AMPs with significant anti-cancer properties. Statement of Significance We have demonstrated a methodology to design cell-selective potent antimicrobial and anti-endotoxin peptides by utilizing phenylalanine zipper as a template and replacement of phenylalanine residue(s) from “a” and/or “d” position(s) with proline residue(s) produced non-cytotoxic AMPs with improved antibacterial properties against the drug-resistant strains of bacteria. The work showed that the ‘a’ and ‘d’ positions of the phenylalanine heptad repeat could be replaced by an appropriate amino acid to control cytotoxicity of the peptide without compromising its potency in antimicrobial and anti-endotoxin properties. The direct bacterial membrane targeting mechanism of proline substituted analogs of parent peptide makes difficult for bacteria to grow resistance against them. The peptides designed could be lead molecules in the area of sepsis as they possess significant anti-LPS activities for in vitro and in vivo. Interestingly since cancer cells and bacterial cell membranes possess the structural resemblances, the cancer cells are also targets for these peptides making them lead molecules in this field. However, unlike in bacteria where the peptides showed membrane permeabilization property to lyse them, the peptides induced apoptosis in MDA-MB-231 breast cancer cells to inhibit their proliferation and growth. The results are significant because it reveals that “a” and “d” positions of a phenylalanine zipper can be utilized as switches to design cell-selective, antimicrobial, anti-endotoxin and anticancer peptides.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.007
      Issue No: Vol. 57 (2017)
  • pH-controllable cell-penetrating polypeptide that exhibits cancer
    • Authors: DaeYong Lee; Ilkoo Noh; Jisang Yoo; N. Sanoj Rejinold; Yeu-Chun Kim
      Pages: 187 - 196
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): DaeYong Lee, Ilkoo Noh, Jisang Yoo, N. Sanoj Rejinold, Yeu-Chun Kim
      Helical peptides were naturally-occurring ordered conformations that mediated various biological functions essential for biotechnology. However, it was difficult for natural helical polypeptides to be applied in biomedical fields due to low bioavailability. To avoid these problems, synthetic alpha-helical polypeptides have recently been introduced by further modifying pendants in the side chain. In spite of an attractive biomimetic helical motif, these systems could not be tailored for targeted delivery mainly due to nonspecific binding events. To address these issues, we created a conformation-transformable polypeptide capable of eliciting a pH-activated cell-penetrating property solely at the cancer region. The developed novel polypeptide showed that the bare helical conformation had a function at physiological conditions while the pH-induced helical motif provided an active cell-penetrating characteristic at a tumor extracellular matrix pH. The unusual conformation-transformable system can elicit bioactive properties exclusively at mild acidic pH. Statement of Significance We developed pH-controllable cell-penetrating polypeptides (PCCPs) undergoing pH-induced conformational transitions. Unlike natural cell-penetrating peptides, PCCPs was capable of penetrating the plasma membranes dominantly at tumor pH, driven by pH-controlled helicity. The conformation of PCCPs at neutral pH showed low helical propensity because of dominant electrostatic attractions within the side chains. However, the helicity of PCCPs was considerably augmented by the balance of electrostatic interactions, thereby inducing selective cellular penetration. Three polypeptides undergoing different conformational transitions were prepared to verify the selective cellular uptake influenced by their structures. The PCCP undergoing low-to-high helical conformation provided the tumor specificity and enhanced uptake efficiency. pH-induced conformation-transformable polypeptide might provide a novel platform for stimuli-triggered targeting systems.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.040
      Issue No: Vol. 57 (2017)
  • Preparation and in vivo evaluation of cationic elastic liposomes
           comprising highly skin-permeable growth factors combined with hyaluronic
           acid for enhanced diabetic wound-healing therapy
    • Authors: Jeong Uk Choi; Seong Wook Lee; Rudra Pangeni; Youngro Byun; In-Soo Yoon; Jin Woo Park
      Pages: 197 - 215
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Jeong Uk Choi, Seong Wook Lee, Rudra Pangeni, Youngro Byun, In-Soo Yoon, Jin Woo Park
      To enhance the therapeutic effects of exogenous administration of growth factors (GFs) in the treatment of chronic wounds, we constructed GF combinations of highly skin-permeable epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I), and platelet-derived growth factor-A (PDGF-A). We genetically conjugated a low-molecular-weight protamine (LMWP) to the N-termini of these GFs to form LMWP-EGF, LMWP-IGF-I, and LMWP-PDGF-A. Subsequently, these molecules were complexed with hyaluronic acid (HA). Combinations of native or LMWP-fused GFs significantly promoted fibroblast proliferation and the synthesis of procollagen, with a magnification of these results observed after the GFs were complexed with HA. The optimal proportions of LMWP-EGF, LMWP-IGF-I, LMWP-PDGF-A, and HA were 1, 1, 0.02, and 200, respectively. After confirming the presence of a synergistic effect, we incorporated the LMWP-fused GFs-HA complex into cationic elastic liposomes (ELs) of 107±0.757nm in diameter and a zeta potential of 56.5±1.13mV. The LMWP-fused GFs had significantly improved skin permeation compared with native GFs. The in vitro wound recovery rate of the LMWP-fused GFs-HA complex was 23% higher than that of cationic ELs composed of LMWP-fused GFs alone. Moreover, the cationic ELs containing the LMWP-fused GFs-HA complex significantly accelerated the wound closure rate in a diabetic mouse model and the wound size was maximally decreased by 65% and 58% compared to cationic ELs loaded with vehicle or native GFs-HA complex, respectively. Thus, topical treatment with cationic ELs loaded with the LMWP-fused GFs-HA complex synergistically enhanced the healing of chronic wounds, exerting both rapid and prolonged effects. Statement of Significance We believe that our study makes a significant contribution to the literature, because it demonstrated the potential application of cationic elastic liposomes as topical delivery systems for growth factors (GFs) that have certain limitations in their therapeutic effects (e.g., low percutaneous absorption of GFs at the lesion site and the requirement for various GFs at different healing stages). Topical treatment with cationic elastic liposomes loaded with highly skin-permeable low-molecular-weight protamine (LMWP)-fused GFs-hyaluronic acid (HA) complex synergistically enhanced the healing of diabetic wounds, exerting both rapid and prolonged effects.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.034
      Issue No: Vol. 57 (2017)
  • Halloysite and chitosan oligosaccharide nanocomposite for wound healing
    • Authors: Giuseppina Sandri; Carola Aguzzi; Silvia Rossi; Maria Cristina Bonferoni; Giovanna Bruni; Cinzia Boselli; Antonia Icaro Cornaglia; Federica Riva; Cesar Viseras; Carla Caramella; Franca Ferrari
      Pages: 216 - 224
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Giuseppina Sandri, Carola Aguzzi, Silvia Rossi, Maria Cristina Bonferoni, Giovanna Bruni, Cinzia Boselli, Antonia Icaro Cornaglia, Federica Riva, Cesar Viseras, Carla Caramella, Franca Ferrari
      Halloysite is a natural nanotubular clay mineral (HNTs, Halloysite Nano Tubes) chemically identical to kaolinite and, due to its good biocompatibility, is an attractive nanomaterial for a vast range of biological applications. Chitosan oligosaccharides are homo- or heterooligomers of N-acetylglucosamine and D-glucosamine, that accelerate wound healing by enhancing the functions of inflammatory and repairing cells. The aim of the work was the development of a nanocomposite based on HNTs and chitosan oligosaccharides, to be used as pour powder to enhance healing in the treatment of chronic wounds. A 1:0.05 wt ratio HTNs/chitosan oligosaccharide nanocomposite was obtained by simply stirring the HTNs powder in a 1% w/w aqueous chitosan oligosaccharide solution and was formed by spontaneous ionic interaction resulting in 98.6% w/w HTNs and 1.4% w/w chitosan oligosaccharide composition. Advanced electron microscopy techniques were considered to confirm the structure of the hybrid nanotubes. Both HTNs and HTNs/chitosan oligosaccharide nanocomposite showed good in vitro biocompatibility with normal human dermal fibroblasts up to 300μg/ml concentration and enhanced in vitro fibroblast motility, promoting both proliferation and migration. The HTNs/chitosan oligosaccharide nanocomposite and the two components separately were tested for healing capacity in a murine (rat) model. HTNs/chitosan oligosaccharide allowed better skin reepithelization and reorganization than HNTs or chitosan oligosaccharide separately. The results suggest to develop the nanocomposite as a medical device for wound healing. Statement of Significance The present work is focused on the development of halloysite and chitosan oligosaccharide nanocomposite for wound healing. It considers a therapeutic option for difficult to heal skin lesions and burns. The significance of the research considers two fundamental aspects: the first one is related to the development of a self-assembled nanocomposite, formed by spontaneous ionic interaction, while the second one is related to the possibility to find an effective treatment for cutaneous non healing lesions. The characterization of this hybrid system involves a multidisciplinary approach considering integrated techniques of solid state investigation and advanced electron microscopies, and in vitro/in vivo models to understand biocompatibility and proliferation properties (enhancement of in vitro fibroblast motility, proliferation and migration, and of in vivo burn healing), to understand safety and effectiveness of the developed nanocomposite.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.032
      Issue No: Vol. 57 (2017)
  • Controlled release of GAG-binding enhanced transduction (GET) peptides for
           sustained and highly efficient intracellular delivery
    • Authors: Hosam Al-Deen M. Abu-Awwad; Lalitha Thiagarajan; James E. Dixon
      Pages: 225 - 237
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Hosam Al-Deen M. Abu-Awwad, Lalitha Thiagarajan, James E. Dixon
      Controlled release systems for therapeutic molecules are vital to allow the sustained local delivery of their activities which direct cell behaviour and enable novel regenerative strategies. Direct programming of cells using exogenously delivered transcription factors can by-pass growth factor signalling but there is still a requirement to deliver such activity spatio-temporally. We previously developed a technology termed GAG-binding enhanced transduction (GET) to efficiently deliver a variety of cargoes intracellularly, using GAG-binding domains which promote cell targeting, and cell penetrating peptides (CPPs) which allow cell entry. Herein we demonstrate that GET system can be used in controlled release systems to mediate sustained intracellular transduction over one week. We assessed the stability and activity of GET peptides in poly(dl-lactic acid-co-glycolic acid) (PLGA) microparticles (MPs) prepared using a S/O/W double emulsion method. Efficient encapsulation (∼65%) and tailored protein release profiles could be achieved, however intracellular transduction was significantly inhibited post-release. To retain GET peptide activity we optimized a strategy of co-encapsulation of l-Histidine, which may form a complex with the PLGA degradation products under acidic conditions. Simulations of the polymer microclimate showed that hydrolytic acidic PLGA degradation products directly inhibited GET peptide transduction activity, and use of l-Histidine significantly enhanced released protein delivery. The ability to control the intracellular transduction of functional proteins into cells will facilitate new localized delivery methods and allow approaches to direct cellular behaviour for many regenerative medicine applications. Statement of Significance The goal for regenerative medicine is to restore functional biological tissue by controlling and augmenting cellular behaviour. Either Transcription (TFs) or growth factors (GFs) can be presented to cells in spatio-temporal gradients for programming cell fate and gene expression. Here, we have created a sustained and controlled release system for GET (Glycosaminoglycan-enhanced transducing)-tagged proteins using S/O/W PLGA microparticle fabrication. We demonstrated that PLGA and its acidic degradants inhibit GET-mediated transduction, which can be overcome by using pH-activated l-Histidine. l-Histidine inhibits the electrostatic interaction of GET/PLGA and allows enhanced intracellular transduction. GET could provide a powerful tool to program cell behaviour either in gradients or with sustained delivery. We believe that our controlled release systems will allow application of GET for tissue regeneration directly by TF cellular programming.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.028
      Issue No: Vol. 57 (2017)
  • Chondroitin sulfate-functionalized polyamidoamine as a tumor-targeted
           carrier for miR-34a delivery
    • Authors: Wenqi Chen; Yong Liu; Xiao Liang; Yu Huang; Quanshun Li
      Pages: 238 - 250
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Wenqi Chen, Yong Liu, Xiao Liang, Yu Huang, Quanshun Li
      Chondroitin sulfate (CS) was modified on a polyamidoamine dendrimer (PAMAM) through Michael addition to construct a tumor-targeted carrier CS-PAMAM for miR-34a delivery. The derivative CS-PAMAM was demonstrated to achieve an efficient cellular uptake of miR-34a in a CD44-dependent endocytosis way and further facilitate the endosomal escape of miR-34a after 4h. Through the miR-34a delivery, obvious inhibition of cell proliferation could be detected which was attributed to the enhancement of cell apoptosis and cell cycle arrest, and meanwhile the cell migration and invasion has been observed to be inhibited. Finally, the intravenous injection of CS-PAMAM/miR-34a formulation into mice bearing human lung adenocarcinoma cell A549 xenografts could efficiently inhibit the tumor growth and induce the tumor apoptosis owing to the enhanced accumulation of miR-34a in tumor tissue. Overall, CS-PAMAM is potential to be used as a tumor-targeted oligonucleotide carrier for achieving tumor gene therapy. Statement of Significance The cationic dendrimer PAMAM was modified by chondroitin sulfate (CS) through Michael addition to construct a tumor-targeted carrier CS-PAMAM for miR-34a delivery. The introduction of CS could achieve an efficient cellular uptake and intracellular transfection of miR-34a in a CD44-dependent endocytosis manner. The miR-34a delivery could execute the anti-proliferation activity by simultaneously inducing cell apoptosis and cell cycle arrest, and also the anti-migration activity. The CS-PAMAM-mediated systemic delivery of miR-34a showed significant inhibition of tumor growth and induction of tumor apoptosis using a mice model of subcutaneously implanted tumors.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.030
      Issue No: Vol. 57 (2017)
  • Folic acid-decorated polyamidoamine dendrimer exhibits high tumor uptake
           and sustained highly localized retention in solid tumors: Its utility for
           local siRNA delivery
    • Authors: Leyuan Xu; W. Andrew Yeudall; Hu Yang
      Pages: 251 - 261
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Leyuan Xu, W. Andrew Yeudall, Hu Yang
      The utility of folic acid (FA)-decorated polyamidoamine dendrimer G4 (G4-FA) as a vector was investigated for local delivery of siRNA. In a xenograft HN12 (or HN12-YFP) tumor mouse model of head and neck squamous cell carcinomas (HNSCC), intratumorally (i.t.) injected G4-FA exhibited high tumor uptake and sustained highly localized retention in the tumors according to near infrared (NIR) imaging assessment. siRNA against vascular endothelial growth factor A (siVEGFA) was chosen as a therapeutic modality. Compared to the nontherapeutic treatment groups (PBS solution or dendrimer complexed with nontherapeutic siRNA against green fluorescent protein (siGFP)), G4-FA/siVEGFA showed tumor inhibition effects in single-dose and two-dose regimen studies. In particular, two doses of G4-FA/siVEGFA i.t. administered eight days apart resulted in a more profound inhibition of tumor growth, accompanied with significant reduction in angiogenesis, as judged by CD31 staining and microvessel counts. Tumor size reduction in the two-dose regimen study was ascertained semi-quantitatively by live fluorescence imaging of YFP tumors and independently supported antitumor effects of G4-FA/siVEGFA. Taken together, G4-FA shows high tumor uptake and sustained retention properties, making it a suitable platform for local delivery of siRNAs to treat cancers that are readily accessible such as HNSCC. Statement of Significance Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide and is difficult to transfect for gene therapy. We developed folate receptor (FR)-targeted polyamidoamine (PAMAM) dendrimer for enhanced delivery of genes to HNSCC and gained in-depth understanding of how gene delivery and transfection in head and neck squamous cancer cells can be enhanced via FR-targeted PAMAM dendrimers. The results we report here are encouraging and present latest advances in using dendrimers for cancer therapies, in particular for HNSCC. Our work has demonstrated that localized delivery of FR-targeted PAMAM dendrimer G4 complexed with siVEGFA resulted in pronounced tumor suppression in an HN12 xenograft tumor model. Tumor suppression was attributed to enhanced tumor uptake of siRNA and prolonged nanoparticle retention in the tumor. Taken together, G4-FA shows high tumor uptake and sustained highly localized retention properties, making it a suitable platform for local delivery of siRNAs to treat cancers that are readily accessible such as HNSCC.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.023
      Issue No: Vol. 57 (2017)
  • Chemosensitizing indomethacin-conjugated chitosan oligosaccharide
           nanoparticles for tumor-targeted drug delivery
    • Authors: Jae-Young Lee; Ubonvan Termsarasab; Mee Yeon Lee; Dong-Hwan Kim; Song Yi Lee; Jung Sun Kim; Hyun-Jong Cho; Dae-Duk Kim
      Pages: 262 - 273
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Jae-Young Lee, Ubonvan Termsarasab, Mee Yeon Lee, Dong-Hwan Kim, Song Yi Lee, Jung Sun Kim, Hyun-Jong Cho, Dae-Duk Kim
      A chitosan oligosaccharide (CSO)-indomethacin (IDM) conjugate (CI) was synthesized to fabricate chemosensitizing nanoparticles (NPs) for tumor-targeted drug delivery. IDM was conjugated to a CSO backbone via amide bond formation, of which successful synthesis was confirmed by proton-nuclear magnetic resonance analyses. Doxorubicin (DOX)-loaded CI (CI10/DOX; CI:DOX=10:1 [w/w]) NPs with <75nm of mean diameter, polydispersity index of ∼0.2, and positive zeta potential were prepared. The release of DOX from the NPs was enhanced at acidic pH (pH 5.5 and 6.8) compared to physiological pH (pH 7.4). The release of IDM increased in the presence of A549 cell lysates. In A549 cells (human lung carcinoma cells), more efficient cellular uptake of CI10/DOX NPs than that of free DOX was observed by using confocal laser scanning microscopy and flow cytometry. The in vitro cytotoxicity of CI10/DOX NPs in A549 cells was higher than those of free DOX and CI NPs with free DOX groups. In vivo pharmacokinetic studies after intravenous administration in rats showed significantly lower clearance of DOX from NPs compared with the free DOX group. Tumor targetability of the developed CI NPs was also verified by a real-time optical imaging study. In summary, the chemosensitizing CI/DOX NP with enhanced anticancer activity, prolonged blood circulation, and passive tumor targeting can be a promising anticancer drug delivery system for tumor-targeted therapy. Statement of Significance Chemosensitizing nanoparticles (NPs) based on amphiphilic chitosan oligosaccharide-indomethacin (CSO-IDM; CI) conjugate were developed for tumor-targeted delivery of doxorubicin (DOX). IDM was introduced to the CSO backbone as a hydrophobic residue to synthesize an amphiphilic conjugate and a chemosenstizer of DOX for improving antitumor efficacies. IDM, conjugated to CSO, may inhibit the efflux of cellular uptaken DOX via multidrug resistance-associated protein (MRP) and subsequently augment the anti-proliferation potentials of DOX in A549 cells (MRP-expressed human lung cancer cells). Chemosensitizing properties of developed CI NPs were assessed in cell culture models and the tumor targetability of CI/DOX NPs was demonstrated in A549 tumor-xenografted mouse model by a real-time optical imaging. Developed CI NPs can be used as a multifunctional nanosystem for the therapy of MRP-expressed cancers.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.012
      Issue No: Vol. 57 (2017)
  • Augmented liver targeting of exosomes by surface modification with
           cationized pullulan
    • Authors: Ryo Tamura; Shinji Uemoto; Yasuhiko Tabata
      Pages: 274 - 284
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Ryo Tamura, Shinji Uemoto, Yasuhiko Tabata
      Exosomes are membrane nanoparticles containing biological substances that are employed as therapeutics in experimental inflammatory models. Surface modification of exosomes for better tissue targetability and enhancement of their therapeutic ability was recently attempted mainly using gene transfection techniques. Here, we show for the first time that the surface modification of exosomes with cationized pullulan, which has the ability to target hepatocyte asialoglycoprotein receptors, can target injured liver and enhance the therapeutic effect of exosomes. Surface modification can be achieved by a simple mixing of original exosomes and cationized pullulan and through an electrostatic interaction of both substances. The exosomes modified with cationized pullulan were internalized into HepG2 cells in vitro to a significantly greater extent than unmodified ones and this internalization was induced through the asialoglycoprotein receptor that was specifically expressed on HepG2 cells and hepatocytes. When injected intravenously into mice with concanavalin A-induced liver injury, the modified exosomes accumulated in the liver tissue, resulting in an enhanced anti-inflammatory effect in vivo. It is concluded that the surface modification with cationized pullulan promoted accumulation of the exosomes in the liver and the subsequent biological function, resulting in a greater therapeutic effect on liver injury. Statement of significance Exosomes have shown potentials as therapeutics for various inflammatory disease models. This study is the first to show the specific accumulation of exosomes in the liver and enhanced anti-inflammatory effect via the surface modification of exosomes using pullulan, which is specifically recognized by the asialoglycoprotein receptor (AGPR) on HepG2 cells and hepatocytes. The pullulan was expressed on the surface of PKH-labeled exosomes, and it led increased accumulation of PKH into HepG2 cells, whereas the accumulation was canceled by AGPR inhibitor. In the mouse liver injury model, the modification of PKH-labeled exosomes with pullulan enabled increased accumulation of PKH specifically in the injured liver. Furthermore the greater therapeutic effects against the liver injury compared with unmodified original exosomes was observed.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.013
      Issue No: Vol. 57 (2017)
  • Direct quantification of dual protein adsorption dynamics in three
           dimensional systems in presence of cells
    • Authors: Melika Sarem; Daniel Vonwil; Steffen Lüdeke; V. Prasad Shastri
      Pages: 285 - 292
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Melika Sarem, Daniel Vonwil, Steffen Lüdeke, V. Prasad Shastri
      Understanding the composition of the adsorbed protein layer on a biomaterial surface is of an extreme importance as it directs the primary biological response. Direct detection using labeled proteins and indirect detection based on enzymatic assays or changes to mass, refractive index or density of a surface have been so far established. Nevertheless, using current methodologies, detection of multiple proteins simultaneously and particularly in a three-dimensional (3D) substrates is challenging, with the exception of radiolabeling. Here using fluorescence molecular tomography (FMT), we present a non-destructive and versatile approach to quantify adsorption of multiple proteins within 3D environments and reveal the dynamics of adsorption of human serum albumin (HSA) and fibrinogen (Fib) on 3D polymeric scaffold. Furthermore, we show that serum starved human articular chondrocytes in 3D environment preferentially uptake HSA over Fib and to our knowledge this represents the first example of direct visualization and quantification of protein adsorption in a 3D cell culture system. Statement of Significance The biomaterial surface upon exposure to biological fluids is covered by a layer of proteins, which is modified over a period of time and dictates the fate of the biomaterial. In this study, we present and validate a new methodology for quantification of protein adsorption on to a three-dimensional polymer scaffold from unitary and binary systems, using fluorescence molecular tomography, an optical trans-illumination technique with picomolar sensitivity. In additional to being able to follow behavior of two proteins simultaneously, this methodology is also suitable for studying protein uptake in cells situated in a polymer environment. The ability to follow protein adsorption/uptake in a continuous manner opens up new possibilities to study the role of serum proteins in biomaterial compatibility.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.021
      Issue No: Vol. 57 (2017)
  • Preparation and evaluation of human choroid extracellular matrix scaffolds
           for the study of cell replacement strategies
    • Authors: Kathleen R. Chirco; Kristan S. Worthington; Miles J. Flamme-Wiese; Megan J. Riker; Joshua D. Andrade; Beatrix M. Ueberheide; Edwin M. Stone; Budd A. Tucker; Robert F. Mullins
      Pages: 293 - 303
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Kathleen R. Chirco, Kristan S. Worthington, Miles J. Flamme-Wiese, Megan J. Riker, Joshua D. Andrade, Beatrix M. Ueberheide, Edwin M. Stone, Budd A. Tucker, Robert F. Mullins
      Endothelial cells (ECs) of the choriocapillaris are one of the first cell types lost during age-related macular degeneration (AMD), and cell replacement therapy is currently a very promising option for patients with advanced AMD. We sought to develop a reliable method for the production of human choroidal extracellular matrix (ECM) scaffolds, which will allow for the study of choroidal EC (CEC) replacement strategies in an environment that closely resembles the native tissue. Human RPE/choroid tissue was treated sequentially with Triton X-100, SDS, and DNase to remove all native cells. While all cells were successfully removed from the tissue, collagen IV, elastin, and laminin remained, with preserved architecture of the acellular vascular tubes. The ECM scaffolds were then co-cultured with exogenous ECs to determine if the tissue can support cell growth and allow EC reintegration into the decellularized choroidal vasculature. Both monkey and human ECs took up residence in the choriocapillary tubes of the decellularized tissue. Together, these data suggest that our decellularization methods are sufficient to remove all cellular material yet gentle enough to preserve tissue structure and allow for the optimization of cell replacement strategies. Statement of Significance Age-related macular degeneration (AMD) is a devastating disease affecting more than 600 million people worldwide. Endothelial cells of the choriocapillaris (CECs) are among the first cell types lost in early AMD, and cell replacement therapy is currently the most promising option for restoring vision in patients with advanced AMD. In order to study CEC replacement strategies we have generated a 3D choroid scaffold using a novel decellularization method in human RPE/choroid tissue. To our knowledge, this is the first report describing decellularization of human RPE/choroid, as well as recellularization of a choroid scaffold with CECs. This work will aid in our development and optimization of cell replacement strategies using a tissue scaffold that is similar to the in vivo environment.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.011
      Issue No: Vol. 57 (2017)
  • Spatial distributions of pericellular stiffness in natural extracellular
           matrices are dependent on cell-mediated proteolysis and contractility
    • Authors: M. Keating; A. Kurup; M. Alvarez-Elizondo; A.J. Levine; E. Botvinick
      Pages: 304 - 312
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): M. Keating, A. Kurup, M. Alvarez-Elizondo, A.J. Levine, E. Botvinick
      Bulk tissue stiffness has been correlated with regulation of cellular processes and conversely cells have been shown to remodel their pericellular tissue according to a complex feedback mechanism critical to development, homeostasis, and disease. However, bulk rheological methods mask the dynamics within a heterogeneous fibrous extracellular matrix (ECM) in the region proximal to a cell (pericellular region). Here, we use optical tweezers active microrheology (AMR) to probe the distribution of the complex material response function (α = α′+ α″, in units of µm/nN) within a type I collagen ECM, a biomaterial commonly used in tissue engineering. We discovered cells both elastically and plastically deformed the pericellular material. α′ is wildly heterogeneous, with 1/α′ values spanning three orders of magnitude around a single cell. This was observed in gels having a cell-free 1/α′ of approximately 0.5nN/µm. We also found that inhibition of cell contractility instantaneously softens the pericellular space and reduces stiffness heterogeneity, suggesting the system was strain hardened and not only plastically remodeled. The remaining regions of high stiffness suggest cellular remodeling of the surrounding matrix. To test this hypothesis, cells were incubated within the type I collagen gel for 24-h in a media containing a broad-spectrum matrix metalloproteinase (MMP) inhibitor. While pericellular material maintained stiffness asymmetry, stiffness magnitudes were reduced. Dual inhibition demonstrates that the combination of MMP activity and contractility is necessary to establish the pericellular stiffness landscape. This heterogeneity in stiffness suggests the distribution of pericellular stiffness, and not bulk stiffness alone, must be considered in the study of cell-ECM interactions and design of complex biomaterial scaffolds. Statement of Significance Collagen is a fibrous extracellular matrix (ECM) protein widely used to study cell-ECM interactions. Stiffness of ECM has been shown to instruct cells, which can in turn modify their ECM, as has been shown in the study of cancer and regenerative medicine. Here we measure the stiffness of the collagen microenvironment surrounding cells and quantitatively measure the dependence of pericellular stiffness on MMP activity and cytoskeletal contractility. Competent cell-mediated stiffening results in a wildly heterogeneous micromechanical topography, with values spanning orders of magnitude around a single cell. We speculate studies must consider this notable heterogeneity generated by cells when testing theories regarding the role of ECM mechanics in health and disease.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.008
      Issue No: Vol. 57 (2017)
  • Multilayered membranes with tuned well arrays to be used as regenerative
    • Authors: Nádia I. Martins; Maria P. Sousa; Catarina A. Custódio; Vânia C. Pinto; Paulo J. Sousa; Graça Minas; Franck Cleymand; João F. Mano
      Pages: 313 - 323
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Nádia I. Martins, Maria P. Sousa, Catarina A. Custódio, Vânia C. Pinto, Paulo J. Sousa, Graça Minas, Franck Cleymand, João F. Mano
      Membranes have been explored as patches in tissue repair and regeneration, most of them presenting a flat geometry or a patterned texture at the nano/micrometer scale. Herein, a new concept of a flexible membrane featuring well arrays forming pore-like environments to accommodate cell culture is proposed. The processing of such membranes using polysaccharides is based on the production of multilayers using the layer-by-layer methodology over a patterned PDMS substrate. The detached multilayered membrane exhibits a layer of open pores at one side and a total thickness of 38±2.2µm. The photolithography technology used to produce the molds allows obtaining wells on the final membranes with a tuned shape and micro-scale precision. The influence of post-processing procedures over chitosan/alginate films with 100 double layers, including crosslinking with genipin or fibronectin immobilization, on the adhesion and proliferation of human osteoblast-like cells is also investigated. The results suggest that the presence of patterned wells affects positively cell adhesion, morphology and proliferation. In particular, it is seen that cells colonized preferentially the well regions. The geometrical features with micro to sub-millimeter patterned wells, together with the nano-scale organization of the polymeric components along the thickness of the film will allow to engineer highly versatile multilayered membranes exhibiting a pore-like microstructure in just one of the sides, that could be adaptable in the regeneration of multiple tissues. Statement of Significance Flexible multilayered membranes containing multiple micro-reservoirs are found as potential regenerative patches. Layer-by-layer (LbL) methodology over a featured PDMS substrate is used to produce patterned membranes, composed only by natural-based polymers, that can be easily detached from the PDMS substrate. The combination of nano-scale control of the polymeric organization along the thickness of the chitosan/alginate (CHT/ALG) membranes, provided by LbL, together with the geometrical micro-scale features of the patterned membranes offers a uniqueness system that allows cells to colonize 3-dimensionally. This study provides a promising strategy to control cellular spatial organization that can face the region of the tissue to regenerate.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.021
      Issue No: Vol. 57 (2017)
  • Towards rebuilding vaginal support utilizing an extracellular matrix
    • Authors: Rui Liang; Katrina Knight; Deanna Easley; Stacy Palcsey; Steven Abramowitch; Pamela A. Moalli
      Pages: 324 - 333
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Rui Liang, Katrina Knight, Deanna Easley, Stacy Palcsey, Steven Abramowitch, Pamela A. Moalli
      As an alternative to polypropylene mesh, we explored an extracellular matrix (ECM) bioscaffold derived from urinary bladder matrix (MatriStem™) in the repair of vaginal prolapse. We aimed to restore disrupted vaginal support simulating application via transvaginal and transabdominal approaches in a macaque model focusing on the impact on vaginal structure, function, and the host immune response. In 16 macaques, after laparotomy, the uterosacral ligaments and paravaginal attachments to pelvic side wall were completely transected (IACUC# 13081928). 6-ply MatriStem was cut into posterior and anterior templates with a portion covering the vagina and arms simulating uterosacral ligaments and paravaginal attachments, respectively. After surgically exposing the correct anatomical sites, in 8 animals, a vaginal incision was made on the anterior and posterior vagina and the respective scaffolds were passed into the vagina via these incisions (transvaginal insertion) prior to placement. The remaining 8 animals underwent the same surgery without vaginal incisions (transabdominal insertion). Three months post implantation, firm tissue bands extending from vagina to pelvic side wall appeared in both MatriStem groups. Experimental endpoints examining impact of MatriStem on the vagina demonstrated that vaginal biochemical and biomechanical parameters, smooth muscle thickness and contractility, and immune responses were similar in the MatriStem no incision group and sham-operated controls. In the MatriStem incision group, a 41% decrease in vaginal stiffness (P=0.042), a 22% decrease in collagen content (P=0.008) and a 25% increase in collagen subtypes III/I was observed vs. Sham. Active MMP2 was increased in both Matristem groups vs. Sham (both P=0.002). This study presents a novel application of ECM bioscaffolds as a first step towards the rebuilding of vaginal support. Statement of Significance Pelvic organ prolapse is a common condition related to failure of the supportive soft tissues of the vagina; particularly at the apex and mid-vagina. Few studies have investigated methods to regenerate these failed structures. The overall goal of the study was to determine the feasibility of utilizing a regenerative bioscaffold in prolapse applications to restore apical (level I) and lateral (level II) support to the vagina without negatively impacting vaginal structure and function. The significance of our findings is two fold: 1. Implantation of properly constructed extracellular matrix grafts promoted rebuilding of level I and level II support to the vagina and did not negatively impact the overall functional, morphological and biochemical properties of the vagina. 2. The presence of vaginal incisions in the transvaginal insertion of bioscaffolds may compromise vaginal structural integrity in the short term.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.015
      Issue No: Vol. 57 (2017)
  • Mechanical phenotyping of cells and extracellular matrix as grade and
           stage markers of lung tumor tissues
    • Authors: Valeria Panzetta; Ida Musella; Ida Rapa; Marco Volante; Paolo A. Netti; Sabato Fusco
      Pages: 334 - 341
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Valeria Panzetta, Ida Musella, Ida Rapa, Marco Volante, Paolo A. Netti, Sabato Fusco
      The mechanical cross-talk between cells and the extra-cellular matrix (ECM) regulates the properties, functions and healthiness of the tissues. When this is disturbed it changes the mechanical state of the tissue components, singularly or together, and cancer, along with other diseases, may start and progress. However, the bi-univocal mechanical interplay between cells and the ECM is still not properly understood. In this study we show how a microrheology technique gives us the opportunity to evaluate the mechanics of cells and the ECM at the same time. The mechanical phenotyping was performed on the surgically removed tissues of 10 patients affected by adenocarcinoma of the lung. A correlation between the mechanics and the grade and stage of the tumor was reported and compared to the mechanical characteristics of the healthy tissue. Our findings suggest a sort of asymmetric modification of the mechanical properties of the cells and the extra-cellular matrix in the tumor, being the more compliant cell even though it resides in a stiffer matrix. Overall, the simultaneous mechanical characterization of the tissues constituents (cells and ECM) provided new support for diagnosis and offered alternative points of analysis for cancer mechanobiology. Statement of significance When the integrity of the mechanical cross-talk between cells and the extra-cellular matrix is disturbed cancer, along with other diseases, may initiate and progress. Here, we show how a new technique gives the opportunity to evaluate the mechanics of cells and the ECM at the same time. It was applied on surgically removed tissues of 10 patients affected by adenocarcinoma of the lung and a correlation between the mechanics and the grade and stage of the tumor was reported and compared to the mechanical characteristics of the healthy tissue.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.002
      Issue No: Vol. 57 (2017)
  • A comprehensive study of layer-specific morphological changes in the
           microstructure of carotid arteries under uniaxial load
    • Authors: Witold Krasny; Claire Morin; Hélène Magoariec; Stéphane Avril
      Pages: 342 - 351
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Witold Krasny, Claire Morin, Hélène Magoariec, Stéphane Avril
      The load bearing properties of large blood vessels are principally conferred by collagen and elastin networks and their microstructural organization plays an important role in the outcomes of various arterial pathologies. In particular, these fibrous networks are able to rearrange and reorient spatially during mechanical deformations. In this study, we investigate for the first time whether these well-known morphological rearrangements are the same across the whole thickness of blood vessels, and subsequently if the underlying mechanisms that govern these rearrangements can be predicted using affine kinematics. To this aim, we submitted rabbit carotid samples to uniaxial load in three distinct deformation directions, while recording live images of the 3D microstructure using multiphoton microscopy. Our results show that the observed realignment of collagen and elastin in the media layer, along with elastin of the adventitia layer, remained limited to small angles that can be predicted by affine kinematics. We show also that collagen bundles of fibers in the adventitia layer behaved in significantly different fashion. They showed a remarkable capacity to realign in the direction of the load, whatever the loading direction. Measured reorientation angles of the fibers were significantly higher than affine predictions. This remarkable property of collagen bundles in the adventitia was never observed before, it shows that the medium surrounding collagen in the adventitia undergoes complex deformations challenging traditional hyperelastic models based on mixture theories. Statement of significance The biomechanical properties of arteries are conferred by the rearrangement under load of the collagen and elastin fibers making up the arterial microstructure. Their kinematics under deformation is not yet characterized for all fiber networks. In this respect we have submitted samples of arterial tissue to uniaxial tension, simultaneously to confocal imaging of their microstructure. Our method allowed identifying for the first time the remarkable ability of adventitial collagen fibers to reorient in the direction of the load, achieving reorientation rotations that exceeded those predicted by affine kinematics, while all other networks followed the affine kinematics. Our results highlight new properties of the microstructure, which might play a role in the outcomes of vascular pathologies like aneurysms.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.033
      Issue No: Vol. 57 (2017)
  • Dynamic fatigue measurement of human erythrocytes using dielectrophoresis
    • Authors: Yuhao Qiang; Jia Liu; E Du
      Pages: 352 - 362
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Yuhao Qiang, Jia Liu, E Du
      Erythrocytes must undergo severe deformation to pass through narrow capillaries and submicronic splenic slits for several hundred thousand times in their normal lifespan. Studies of erythrocyte biomechanics have been mainly focused on cell deformability and rheology measured from a single application of stress and mostly under a static or quasi-static state using classical biomechanical techniques, such as optical tweezers and micropipette aspiration. Dynamic behavior of erythrocytes in response to cyclic stresses that contributes to the membrane failure in blood circulation is not fully understood. This paper presents a new experimental method for dynamic fatigue analysis of erythrocytes, using amplitude modulated electrokinetic force field in a microfluidic platform. We demonstrate the capability of this new technique using a low cycle fatigue analysis of normal human erythrocytes and ATP-depleted erythrocytes. Cyclic tensile stresses are generated to induce repeated uniaxial stretching and extensional recovery of single erythrocytes. Results of morphological and biomechanical parameters of individually tracked erythrocytes show strong correlations with the number of the loading cycles. Under a same strength of electric field, after 180 stress cycles, for normal erythrocytes, maximum stretch ratio decreases from 3.80 to 2.86, characteristic time of cellular extensional recovery increases from 0.16s to 0.37s, membrane shear viscosity increases from 1.0(µN/m)s to 1.6(µN/m)s. Membrane deformation in a small number of erythrocytes becomes irreversible after large deformation for about 200 cyclic loads. ATP-depleted cells show similar trends in decreased deformation and increased characteristic time with the loading cycles. These results show proof of concept of the new microfluidics technique for dynamic fatigue analysis of human erythrocytes. Statement of significance Red blood cells (RBCs) experience a tremendous number of deformation in blood circulation before losing their mechanical deformability and eventually being degraded in the reticuloendothelial system. Prior efforts in RBC biomechanics have mostly focused on a single-application of stress, or quasi-static loading through physical contact to deform cell membranes, thus with limited capabilities in probing cellular dynamic responses to cyclic stresses. We present a unique electrokinetic microfluidic system for the study of dynamic fatigue behavior of RBCs subjected to cyclic loads. Our work shows quantitatively how the cyclic stretching loads cause membrane mechanical degradation and irreversibly deformed cells. This new technique can be useful to identify biomechanical markers for prediction of the mechanical stability and residual lifespan of circulating RBCs.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.037
      Issue No: Vol. 57 (2017)
  • Investigating mechanisms of tendon damage by measuring multi-scale
           recovery following tensile loading
    • Authors: Andrea H. Lee; Spencer E. Szczesny; Michael H. Santare; Dawn M. Elliott
      Pages: 363 - 372
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Andrea H. Lee, Spencer E. Szczesny, Michael H. Santare, Dawn M. Elliott
      Tendon pathology is associated with damage. While tendon damage is likely initiated by mechanical loading, little is known about the specific etiology. Damage is defined as an irreversible change in the microstructure that alters the macroscopic mechanical parameters. In tendon, the link between mechanical loading and microstructural damage, resulting in macroscopic changes, is not fully elucidated. In addition, tendon damage at the macroscale has been proposed to initiate when tendon is loaded beyond a strain threshold, yet the metrics to define the damage threshold are not determined. We conducted multi-scale mechanical testing to investigate the mechanism of tendon damage by simultaneously quantifying macroscale mechanical and microstructural changes. At the microscale, we observe full recovery of the fibril strain and only partial recovery of the interfibrillar sliding, indicating that the damage initiates at the interfibrillar structures. We show that non-recoverable sliding is a mechanism for tendon damage and is responsible for the macroscale decreased linear modulus and elongated toe-region observed at the fascicle-level, and these macroscale properties are appropriate metrics that reflect tendon damage. We concluded that the inflection point of the stress-strain curve represents the damage threshold and, therefore, may be a useful parameter for future studies. Establishing the mechanism of damage at multiple length scales can improve prevention and rehabilitation strategies for tendon pathology. Statement of Significance Tendon pathology is associated with mechanically induced damage. Damage, as defined in engineering, is an irreversible change in microstructure that alters the macroscopic mechanical properties. Although microstructural damage and changes to macroscale mechanics are likely, this link to microstructural change was not yet established. We conducted multiscale mechanical testing to investigate the mechanism of tendon damage by simultaneously quantifying macroscale mechanical and microstructural changes. We showed that non-recoverable sliding between collagen fibrils is a mechanism for tendon damage. Establishing the mechanism of damage at multiple length scales can improve prevention and rehabilitation strategies for tendon pathology.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.011
      Issue No: Vol. 57 (2017)
  • Protection of cortex by overlying meninges tissue during dynamic
           indentation of the adolescent brain
    • Authors: David B. MacManus; Baptiste Pierrat; Jeremiah G. Murphy; Michael D. Gilchrist
      Pages: 384 - 394
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): David B. MacManus, Baptiste Pierrat, Jeremiah G. Murphy, Michael D. Gilchrist
      Traumatic brain injury (TBI) has become a recent focus of biomedical research with a growing international effort targeting material characterization of brain tissue and simulations of trauma using computer models of the head and brain to try to elucidate the mechanisms and pathogenesis of TBI. The meninges, a collagenous protective tri-layer, which encloses the entire brain and spinal cord has been largely overlooked in these material characterization studies. This has resulted in a lack of accurate constitutive data for the cranial meninges, particularly under dynamic conditions such as those experienced during head impacts. The work presented here addresses this lack of data by providing for the first time, in situ large deformation material properties of the porcine dura-arachnoid mater composite under dynamic indentation. It is demonstrated that this tissue is substantially stiffer (shear modulus, μ=19.10±8.55kPa) and relaxes at a slower rate (τ1 =0.034±0.008s, τ2 =0.336±0.077s) than the underlying brain tissue (μ=6.97±2.26kPa, τ1 =0.021±0.007s, τ2 =0.199±0.036s), reducing the magnitudes of stress by 250% and 65% for strains that arise during indentation-type deformations in adolescent brains. Statement of Significance We present the first mechanical analysis of the protective capacity of the cranial meninges using in situ micro-indentation techniques. Force-relaxation tests are performed on in situ meninges and cortex tissue, under large strain dynamic micro-indentation. A quasi-linear viscoelastic model is used subsequently, providing time-dependent mechanical properties of these neural tissues under loading conditions comparable to what is experienced in TBI. The reported data highlights the large differences in mechanical properties between these two tissues. Finite element simulations of the indentation experiments are also performed to investigate the protective capacity of the meninges. These simulations show that the meninges protect the underlying brain tissue by reducing the overall magnitude of stress by 250% and up to 65% for strains.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.022
      Issue No: Vol. 57 (2017)
  • Site-specific characterization of beetle horn shell with micromechanical
           bending test in focused ion beam system
    • Authors: Hyun-Taek Lee; Ho-Jin Kim; Chung-Soo Kim; Kenji Gomi; Minoru Taya; Shûhei Nomura; Sung-Hoon Ahn
      Pages: 395 - 403
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Hyun-Taek Lee, Ho-Jin Kim, Chung-Soo Kim, Kenji Gomi, Minoru Taya, Shûhei Nomura, Sung-Hoon Ahn
      Biological materials are the result of years of evolution and possess a number of efficient features and structures. Researchers have investigated the possibility of designing biomedical structures that take advantage of these structural features. Insect shells, such as beetle shells, are among the most promising types of biological material for biomimetic development. However, due to their intricate geometries and small sizes, it is challenging to measure the mechanical properties of these microscale structures. In this study, we developed an in-situ testing platform for site-specific experiments in a focused ion beam (FIB) system. Multi-axis nano-manipulators and a micro-force sensor were utilized in the testing platform to allow better results in the sample preparation and data acquisition. The entire test protocol, consisting of locating sample, ion beam milling and micro-mechanical bending tests, can be carried out without sample transfer or reattachment. We used our newly devised test platform to evaluate the micromechanical properties and structural features of each separated layer of the beetle horn shell. The Young’s modulus of both the exocuticle and endocuticle layers was measured. We carried out a bending test to characterize the layers mechanically. The exocuticle layer bent in a brick-like manner, while the endocuticle layer exhibited a crack blunting effect. Statement of Significance This paper proposed an in-situ manipulation/test method in focused ion beam for characterizing micromechanical properties of beetle horn shell. The challenge in precise and accurate fabrication for the samples with complex geometry was overcome by using nano-manipulators having multi-degree of freedom and a micro-gripper. With the aid of this specially designed test platform, bending tests were carried out on cantilever-shaped samples prepared by focused ion beam milling. Structural differences between exocuticle and endocuticle layers of beetle horn shell were explored and the results provided insight into the structural advantages of each biocomposite structure.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.04.026
      Issue No: Vol. 57 (2017)
  • Biocompatible nanostructured solid adhesives for biological soft tissues
    • Authors: Masahiro Okada; Akira Nakai; Emilio Satoshi Hara; Tetsushi Taguchi; Takayoshi Nakano; Takuya Matsumoto
      Pages: 404 - 413
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Masahiro Okada, Akira Nakai, Emilio Satoshi Hara, Tetsushi Taguchi, Takayoshi Nakano, Takuya Matsumoto
      Over the past few years, the development of novel adhesives for biological soft tissue adhesion has gained significant interest. Such adhesives should be non-toxic and biocompatible. In this study, we synthesized a novel solid adhesive using nanostructured hydroxyapatite (HAp) and evaluated its physical adhesion properties through in vitro testing with synthetic hydrogels and mouse soft tissues. The results revealed that HAp-nanoparticle dispersions and HAp-nanoparticle-assembled nanoporous plates showed efficient adhesion to hydrogels. Interestingly, the HAp plates showed different adhesive properties depending upon the shape of their nanoparticles. The HAp plate made up of 17nm-sized nanoparticles showed an adhesive strength 2.2times higher than that of the conventional fibrin glue for mouse skin tissues. Statement of Significance The present study indicates a new application of inorganic biomaterials (bioceramics) as a soft tissue adhesive. Organic adhesives such as fibrin glues or cyanoacrylate derivatives have been commonly used clinically. However, their limited biocompatibility and/or low adhesion strength are some drawbacks that impair their clinical application. In this study, we synthesized a novel solid adhesive with biocompatible and biodegradable HAp nanoparticles without the aid of organic molecules, and showed a rapid and strong adhesion of mouse soft tissues compared to conventional fibrin glues. Given the importance of wet adhesion in biomedicine and biotechnology applications, our results will help not only in developing an efficient approach to close incised soft tissues, but also in finding novel ways to integrate soft tissues with synthetic hydrogels (such as drug reservoirs).
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.014
      Issue No: Vol. 57 (2017)
  • Avidin-conjugated calcium phosphate nanoparticles as a modular targeting
           system for the attachment of biotinylated molecules in vitro and in vivo
    • Authors: Selina Beatrice van der Meer; Torben Knuschke; Annika Frede; Nina Schulze; Astrid M. Westendorf; Matthias Epple
      Pages: 414 - 425
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): Selina Beatrice van der Meer, Torben Knuschke, Annika Frede, Nina Schulze, Astrid M. Westendorf, Matthias Epple
      Avidin was covalently conjugated to the surface of calcium phosphate nanoparticles, coated with a thin silica shell and terminated by sulfhydryl groups (diameter of the solid core about 50nm), with a bifunctional crosslinker connecting the amino groups of avidin to the sulfhydryl group on the nanoparticle surface. This led to a versatile nanoparticle system where all kinds of biotinylated (bio-)molecules can be easily attached to the surface by the non-covalent avidin-biotin-complex formation. It also permits the attachment of different biomolecules on the same nanoparticle (heteroavidity), creating a modular system for specific applications in medicine and biology. The variability of the binding to the nanoparticle surface of the was demonstrated with various biotinylated molecules, i.e. fluorescent dyes and antibodies. The accessibility of the conjugated avidin was demonstrated by a fluorescence-quenching assay. About 2.6 binding sites for biotin were accessible on each avidin tetramer. Together with a number of about 240 avidin tetramer units per nanoparticle, this offers about 600 binding sites for biotin on each nanoparticle. The uptake of fluorescently labelled avidin-conjugated calcium phosphate nanoparticles by HeLa cells showed the co-localization of fluorescent avidin and fluorescent biotin, indicating the stability of the complex under cell culture conditions. CD11c-antibody functionalized nanoparticles specifically targeted antigen-presenting immune cells (dendritic cells; DCs) in vitro and in vivo (mice) with high efficiency. Statement of significance Calcium phosphate nanoparticles have turned out to be very useful transporters for biomolecules into cells, both in vitro and in vivo. However, their covalent surface functionalization with antibodies, fluorescent dyes, or proteins requires a separate chemical synthesis for each kind of surface molecule. We have therefore developed avidin-terminated calcium phosphate nanoparticles to which all kinds of biotinylated molecules can be easily attached, also as a mixture of two or more molecules. This non-covalent bond is stable both in cell culture and after injection into mice in vivo. Thus, we have created a highly versatile system for many applications, from the delivery of biomolecules over the targeting of cells and tissue to in vivo imaging.
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      PubDate: 2017-06-24T05:51:15Z
      DOI: 10.1016/j.actbio.2017.05.049
      Issue No: Vol. 57 (2017)
  • Corrigendum to “Non-invasive tracking of hydrogel degradation using
           upconversion nanoparticles” [Acta Biomater. 55 (2017) 410–419]
    • Authors: Yuqing Dong; Guorui Jin; Changchun Ji; Rongyan He; Min Lin; Xin Zhao; Ang Li; Tian Jian Lu; Feng Xu
      Abstract: Publication date: Available online 22 July 2017
      Source:Acta Biomaterialia
      Author(s): Yuqing Dong, Guorui Jin, Changchun Ji, Rongyan He, Min Lin, Xin Zhao, Ang Li, Tian Jian Lu, Feng Xu

      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.015
  • Dragonfly wing nodus: a one-way hinge contributing to the asymmetric wing
    • Authors: H. Rajabi; N. Ghoroubi; K. Stamm; E. Appel; S.N. Gorb
      Abstract: Publication date: Available online 22 July 2017
      Source:Acta Biomaterialia
      Author(s): H. Rajabi, N. Ghoroubi, K. Stamm, E. Appel, S.N. Gorb
      Dragonfly wings are highly specialized locomotor systems, which are formed by a combination of several structural components. The wing components, also known as structural elements, are responsible for the various aspects of the wing functionality. Considering the complex interactions between the wing components, modelling of the wings as a whole is only possible with inevitable huge oversimplifications. In order to overcome this difficulty, we have recently proposed a new approach to model individual components of complex wings comparatively. Here, we use this approach to study nodus, a structural element of dragonfly wings which has been less studied to date. Using a combination of several imaging techniques including scanning electron microscopy (SEM), wide-field fluorescence microscopy (WFM), confocal laser scanning microscopy (CLSM) and micro-computed tomography (micro-CT) scanning, we aim to characterize the spatial morphology and material composition of fore- and hindwing nodi of the dragonfly Brachythemis contaminata. The microscopy results show the presence of resilin in the nodi, which is expected to help the deformability of the wings. The computational results based on three-dimensional (3D) structural data suggest that the specific geometry of the nodus restrains its displacements when subjected to pressure on the ventral side. This effect, resulting from an interlocking mechanism, is expected to contribute to the dorso-ventral asymmetry of wing deformation and to provide a higher resistance to aerodynamic forces during the downstroke. Our results provide an important step towards better understanding of the structure-property-function relationship in dragonfly wings. Statement of significance In this study, we investigate the wing nodus, a specialized wing component in dragonflies. Using a combination of modern imaging techniques, we demonstrate the presence of resilin in the nodus, which is expected to facilitate the wing deformability in flight. The specific geometry of the nodus, however, seems to restrain its displacements when subjected to pressure on the ventral side. This effect, resulting from an interlocking mechanism, is suggested to contribute to dorso-ventral asymmetry of wing deformations and to provide a higher resistance to aerodynamic forces during the downstroke. Our results provide an important step towards better understanding of the structure-property-function relationship in dragonfly wings and might help to design more efficient wings for biomimetic micro-air vehicles.
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      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.034
  • An optimized non-destructive protocol for testing mechanical properties in
           decellularized rabbit trachea
    • Authors: M Den Hondt; BM Vanaudenaerde; EF Maughan; CR Butler; C Crowley; EK Verbeken; SE Verleden; JJ Vranckx
      Abstract: Publication date: Available online 21 July 2017
      Source:Acta Biomaterialia
      Author(s): M Den Hondt, BM Vanaudenaerde, EF Maughan, CR Butler, C Crowley, EK Verbeken, SE Verleden, JJ Vranckx
      Successful tissue-engineered tracheal transplantation relies on the use of non-immunogenic constructs, which can vascularize rapidly, support epithelial growth, and retain mechanical properties to that of native trachea. Current strategies to assess mechanical properties fail to evaluate the trachea to its physiological limits, and lead to irreversible destruction of the construct. Our aim was to develop and evaluate a novel non-destructive method for biomechanical testing of tracheae in a rabbit decellularization model. To validate the performance of this method, we simultaneously analyzed quantitative and qualitative graft changes in response to decellularization, as well as in-vivo biocompatibility of implanted scaffolds. Rabbit tracheae underwent two, four and eight cycles of detergent-enzymatic decellularization. Biomechanical properties were analyzed by calculating luminal volume of progressively inflated and deflated tracheae with microCT. DNA, glycosaminoglycan and collagen contents were compared to native trachea. Scaffolds were prelaminated in vivo. Native, two- and four-cycle tracheae showed equal mechanical properties. Collapsibility of eight-cycle tracheae was significantly increased from -40 cmH2O (-3.9 kPa). Implantation of two- and four-cycle decellularized scaffolds resulted in favorable flap-ingrowth; eight-cycle tracheae showed inadequate integration. We showed a more limited detergent-enzymatic decellularization successfully removing non-cartilaginous immunogenic matter without compromising extracellular matrix content or mechanical stability. With progressive cycles of decellularization, important loss of functional integrity was detected upon mechanical testing and in-vivo implantation. This instability was not revealed by conventional quantitative nor qualitative architectural analyses. These experiments suggest that non-destructive, functional evaluation, e.g. by microCT, may serve as an important tool for mechanical screening of scaffolds before clinical implementation.
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      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.035
  • In-situ tissue regeneration through SDF-1α driven cell recruitment and
           stiffness-mediated bone regeneration in a critical-sized segmental femoral
    • Authors: Amaia Cipitria; Kathrin Boettcher; Sophia Schoenhals; Daniela S. Garske; Katharina Schmidt-Bleek; Agnes Ellinghaus; Anke Dienelt; Anja Peters; Manav Mehta; Christopher M. Madl; Nathaniel Huebsch; David J. Mooney; Georg N. Duda
      Abstract: Publication date: Available online 21 July 2017
      Source:Acta Biomaterialia
      Author(s): Amaia Cipitria, Kathrin Boettcher, Sophia Schoenhals, Daniela S. Garske, Katharina Schmidt-Bleek, Agnes Ellinghaus, Anke Dienelt, Anja Peters, Manav Mehta, Christopher M. Madl, Nathaniel Huebsch, David J. Mooney, Georg N. Duda
      In-situ tissue regeneration aims to utilize the body’s endogenous healing capacity through the recruitment of host stem or progenitor cells to an injury site. Stromal cell-derived factor-1 α (SDF-1 α) is widely discussed as a potent chemoattractant. Here we use a cell-free biomaterial-based approach to (i) deliver SDF-1 α for the recruitment of endogenous bone marrow-derived stromal cells (BMSC) into a critical-sized segmental femoral defect in rats and to (ii) induce hydrogel stiffness-mediated osteogenic differentiation in-vivo. Ionically crosslinked alginate hydrogels with a stiffness optimized for osteogenic differentiation were used. Fast-degrading porogens were incorporated to impart a macroporous architecture that facilitates host cell invasion. Endogenous cell recruitment to the defect site was successfully triggered through the controlled release of SDF-1 α. A trend for increased bone volume fraction (BV/TV) and a significantly higher bone mineral density (BMD) were observed for gels loaded with SDF-1 α, compared to empty gels at two weeks. A trend was also observed, albeit not statistically significant, towards matrix stiffness influencing BV/TV and BMD at two weeks. However, over a six week time-frame, these effects were insufficient for bone bridging of a segmental femoral defect. While mechanical cues combined with ex-vivo cell encapsulation have been shown to have an effect in the regeneration of less demanding in-vivo models, such as cranial defects of nude rats, they are not sufficient for a SDF-1 α mediated in-situ regeneration approach in segmental femoral defects of immunocompetent rats, suggesting that additional osteogenic cues may also be required. Statement of Significance Stromal cell-derived factor-1 α (SDF-1 α) is a chemoattractant used to recruit host cells for tissue regeneration. The concept that matrix stiffness can direct mesenchymal stromal cell (MSC) differentiation into various lineages was described a decade ago using in-vitro experiments. Recently, alginate hydrogels with an optimized stiffness and ex-vivo encapsulated MSCs were shown to have an effect in the regeneration of skull defects of nude rats. Here, we apply this material system, loaded with SDF-1 α and without encapsulated MSCs, to (i) recruit endogenous cells and (ii) induce stiffness-mediated osteogenic differentiation in-vivo, using as model system a load-bearing femoral defect in immunocompetent rats. While a cell-free approach is of great interest from a translational perspective, the current limitations are described.
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      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.032
  • In vitro degradation of calcium phosphates: effect of multiscale porosity,
           textural properties and composition
    • Authors: A. Díez-Escudero; M. Espanol; S. Beats; M.P. Ginebra
      Abstract: Publication date: Available online 21 July 2017
      Source:Acta Biomaterialia
      Author(s): A. Díez-Escudero, M. Espanol, S. Beats, M.P. Ginebra
      The capacity of calcium phosphates to be replaced by bone is tightly linked to their resorbability. However, the relative importance of some textural parameters on their degradation behaviour is still unclear. The present study aims to quantify the effect of composition, specific surface area (SSA), and porosity at various length scales (nano-, micro- and macroporosity) on the in vitro degradation of different calcium phosphates. Degradation studies were performed in an acidic medium to mimic the osteoclastic environment. Small degradations were found in samples with interconnected nano- and micropores with sizes below 3 µm although they were highly porous (35-65%), with maximum weight loss of 8 wt%. Biomimetic calcium deficient hydroxyapatite, with high SSA and low crystallinity, presented the highest degradation rates exceeding even the more soluble β-TCP. A dependence of degradation on SSA was indisputable when porosity and pore sizes were increased. The introduction of additional macroporosity with pore interconnections above 20 µm significantly impacted degradation, more markedly in the substrates with high SSA (>15 m2/g), whereas in sintered substrates with low SSA (<1 m2/g) it resulted just in a linear increase of degradation. Up to 30% of degradation was registered in biomimetic substrates, compared to 15% in β-TCP or 8% in sintered hydroxyapatite. The incorporation of carbonate in calcium deficient hydroxyapatite did not increase its degradation rate. Overall, the study highlights the importance of textural properties, which can modulate or even outweigh the effect of other features such as the solubility of the compounds. Statement of Significance The physicochemical features of calcium phosphates are crucial to tune biological events like resorption during bone remodeling. Understanding in vitro resorption can help to predict the in vivo behavior. Besides chemical composition, other parameters such as porosity and specific surface area have a strong influence on resorption. The complexity of isolating the contribution of each parameter lies in the close interrelation between them. In this work, a multiscale study was proposed to discern the extent to which each parameter influences degradation in a variety of calcium phosphates, using an acidic medium to resemble the osteoclastic environment. The results emphasize the importance of textural properties, which can modulate or even outweigh the effect of the intrinsic solubility of the compounds.
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      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.033
  • Scaffold curvature-mediated novel biomineralization process originates a
           continuous soft tissue-to-bone interface
    • Authors: Michael Paris; Andreas Götz; Inga Hettrich; Cécile M. Bidan; John W.C. Dunlop; Hajar Razi; Ivo Zizak; Dietmar W. Hutmacher; Peter Fratzl; Georg N. Duda; Wolfgang Wagermaier; Amaia Cipitria
      Abstract: Publication date: Available online 20 July 2017
      Source:Acta Biomaterialia
      Author(s): Michael Paris, Andreas Götz, Inga Hettrich, Cécile M. Bidan, John W.C. Dunlop, Hajar Razi, Ivo Zizak, Dietmar W. Hutmacher, Peter Fratzl, Georg N. Duda, Wolfgang Wagermaier, Amaia Cipitria
      A myriad of shapes are found in biological tissues, often naturally evolved to fulfill a particular function. In the field of tissue engineering, substrate geometry influences cell behavior and tissue formation in-vitro, yet little is known how this translates to an in-vivo scenario. Here we investigate scaffold curvature-induced tissue growth, without additional growth factors or cells, in an ovine animal model. We show that soft tissue formation follows a curvature-driven tissue growth model. The highly organized endogenous soft matrix, potentially under mechanical strain, leads to a non-standard form of biomineralization, whereby the pre-existing organic matrix is mineralized without collagen remodeling and without an intermediate cartilage ossification phase. Micro- and nanoscale characterization of the tissue microstructure using histology, backscattered electron (BSE) and second-harmonic generation (SHG) imaging and synchrotron small angle X-ray scattering (SAXS) revealed (i) continuous collagen fibers across the soft-hard tissue interface on the tip of mineralized cones, and (ii) bone remodeling by basic multicellular units (BMUs) in regions adjacent to the native cortical bone. Thus, features of soft tissue-to-bone interface resembling the insertion sites of ligaments and tendons into bone were created, using a scaffold that did not mimic the structural or biological gradients across such a complex interface at its mature state. This study provides fundamental knowledge for biomimetic scaffold design in the fields of bone regeneration and soft tissue-to-bone interface tissue engineering. Statement of significance Geometry influences cell behavior and tissue formation in-vitro. However, little is known how this translates to an in-vivo scenario. Here we investigate the influence of scaffold mean surface curvature on in-vivo tissue growth using an ovine animal model. Based on a multiscale tissue microstructure characterization, we show a seamless integration of soft tissue into newly formed bone, resembling the insertion sites of ligaments and tendons into bone. This interface was created using a scaffold without additional growth factors or cells that did not recapitulate the structural or biological gradients across such a complex tissue interface at its mature state. These findings have important implications for biomimetic scaffold design for bone regeneration and soft tissue-to-bone interface tissue engineering.
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      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.029
  • Manufacturing of hydrogel biomaterials with controlled mechanical
           properties for tissue engineering applications
    • Authors: Armin Vedadghavami; Farnaz Minooei; Mohammad Hossein Mohammadi; Sultan Khetani; Ahmad Rezaei; Shohreh Mashayekhan; Amir Sanati-Nezhad
      Abstract: Publication date: Available online 20 July 2017
      Source:Acta Biomaterialia
      Author(s): Armin Vedadghavami, Farnaz Minooei, Mohammad Hossein Mohammadi, Sultan Khetani, Ahmad Rezaei, Shohreh Mashayekhan, Amir Sanati-Nezhad
      Hydrogels have been recognized as crucial biomaterials in the field of tissue engineering, regenerative medicine, and drug delivery applications due to their specific characteristics. These biomaterials benefit from retaining a large amount of water, effective mass transfer, similarity to natural tissues and the ability to form different shapes. However, having relatively poor mechanical properties is a limiting factor associated with hydrogel biomaterials. Controlling the biomechanical properties of hydrogels is of paramount importance. In this work, firstly, mechanical characteristics of hydrogels and methods employed for characterizing these properties are explored. Subsequently, the most common approaches used for tuning mechanical properties of hydrogels including but are not limited to, interpenetrating polymer networks, nanocomposites, self-assembly techniques, and co-polymerization are discussed. The performance of different techniques used for tuning biomechanical properties of hydrogels is further compared. Such techniques involve lithography techniques for replication of tissues with complex mechanical profiles; microfluidic techniques applicable for generating gradients of mechanical properties in hydrogel biomaterials for engineering complex human tissues like intervertebral discs, osteochondral tissues, blood vessels and skin layers; and electrospinning techniques for synthesis of hybrid hydrogels and highly ordered fibers with tunable mechanical and biological properties. We finally discuss future perspectives and challenges for controlling biomimetic hydrogel materials possessing proper biomechanical properties. Statement of Significance Hydrogels biomaterials are essential constituting components of engineered tissues with the applications in regenerative medicine and drug delivery. The mechanical properties of hydrogels play crucial roles in regulating the interactions between cells and extracellular matrix and directing the cells phenotype and genotype. Despite significant advances in developing methods and techniques with the ability of tuning the biomechanical properties of hydrogels, there are still challenges regarding the synthesis of hydrogels with complex mechanical profiles as well as limitations in vascularization and patterning of complex structures of natural tissues which barricade the production of sophisticated organs. Therefore, in addition to a review on advanced methods and techniques for measuring a variety of different biomechanical characteristics of hydrogels, the new techniques for enhancing the biomechanics of hydrogels are presented. It is expected that this review will profit future works for regulating the biomechanical properties of hydrogel biomaterials to satisfy the demands of a variety of different human tissues.
      Graphical abstract image

      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.028
  • Protein-only, antimicrobial peptide-containing recombinant nanoparticles
           with inherent built-in antibacterial activity
    • Authors: Naroa Serna; Laura Sánchez-García; Alejandro Sánchez-Chardi; Ugutz Unzueta; Mónica Roldán; Ramón Mangues; Esther Vázquez; Antonio Villaverde
      Abstract: Publication date: Available online 19 July 2017
      Source:Acta Biomaterialia
      Author(s): Naroa Serna, Laura Sánchez-García, Alejandro Sánchez-Chardi, Ugutz Unzueta, Mónica Roldán, Ramón Mangues, Esther Vázquez, Antonio Villaverde
      The emergence of bacterial antibiotic resistances is a serious concern in human and animal health. In this context, naturally occurring cationic antimicrobial peptides (AMPs) might play a main role in a next generation of drugs against bacterial infections. Taking an innovative approach to design self-organizing functional proteins, we have generated here protein-only nanoparticles with intrinsic AMP microbicide activity. Using a recombinant version of the GWH1 antimicrobial peptide as building block, these materials show a wide antibacterial activity spectrum in absence of detectable toxicity on mammalian cells. The GWH1-based nanoparticles combine clinically appealing properties of nanoscale materials with full biocompatibility, structural and functional plasticity and biological efficacy exhibited by proteins. Because of the largely implemented biological fabrication of recombinant protein drugs, the protein-based platform presented here represents a novel and scalable strategy in antimicrobial drug design, that by solving some of the limitations of AMPs offers a promising alternative to conventional antibiotics. Statment of significance The low molecular weight antimicrobial peptide GWH1 has been engineered to oligomerize as self-assembling protein-only nanoparticles of around 50 nm. In this form, the peptide exhibits potent and broad antibacterial activities against both Gram-positive and Gram-negative bacteria, without any harmful effect over mammalian cells. As a solid proof-of-concept, this finding strongly supports the design and biofabrication of nanoscale antimicrobial materials with in-built functionalities. The protein-based homogeneous composition offer advantages over alternative materials explored as antimicrobial agents, regarding biocompatibility, biodegradability and environmental suitability. Beyond the described prototype, this transversal engineering concept has wide applicability in the design of novel nanomedicines for advanced treatments of bacterial infections.
      Graphical abstract image

      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.027
  • Translation of an Injectable Triple-Interpenetrating-Network Hydrogel for
           Intervertebral Disc Regeneration in a Goat Model
    • Authors: Sarah E. Gullbrand; Thomas P. Schaer; Prateek Agarwal; Justin R. Bendigo; George R. Dodge; Weiliam Chen; Dawn M. Elliott; Robert L. Mauck; Neil R. Malhotra; Lachlan J. Smith
      Abstract: Publication date: Available online 19 July 2017
      Source:Acta Biomaterialia
      Author(s): Sarah E. Gullbrand, Thomas P. Schaer, Prateek Agarwal, Justin R. Bendigo, George R. Dodge, Weiliam Chen, Dawn M. Elliott, Robert L. Mauck, Neil R. Malhotra, Lachlan J. Smith
      Degeneration of the intervertebral discs is a progressive cascade of cellular, compositional and structural changes that is frequently associated with low back pain. As the first signs of disc degeneration typically arise in the disc’s central nucleus pulposus (NP), augmentation of the NP via hydrogel injection represents a promising strategy to treat early to mid-stage degeneration. The purpose of this study was to establish the translational feasibility of a triple interpenetrating network hydrogel composed of dextran, chitosan, and teleostean (DCT) for augmentation of the degenerative NP in a preclinical goat model. Ex vivo injection of the DCT hydrogel into degenerated goat lumbar motion segments restored range of motion and neutral zone modulus towards physiologic values. To facilitate non-invasive assessment of hydrogel delivery and distribution, zirconia nanoparticles were added to the hydrogel to make the hydrogel radiopaque. Importantly, the addition of zirconia did not negatively impact viability or matrix producing capacity of goat mesenchymal stem cells or NP cells seeded within the hydrogel in vitro. In vivo studies demonstrated that the radiopaque DCT hydrogel was successfully delivered to degenerated goat lumbar intervertebral discs, where it distributed throughout both the NP and annulus fibrosus, and that the hydrogel remained contained within the disc space for two weeks without evidence of extrusion. These results demonstrate the translational potential of this hydrogel for functional regeneration of degenerate intervertebral discs. Statement of Significance The results of this work illustrate that a radiopaque hydrogel is capable of normalizing the mechanical function of the degenerative disc, is supportive of disc cell and mesenchymal stem cell viability and matrix production, and can be maintained in the disc space without extrusion following intradiscal delivery in a preclinical large animal model. These results support evaluation of this hydrogel as a minimally invasive disc therapeutic in long-term preclinical studies as a precursor to future clinical application in patients with disc degeneration and low back pain.
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      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.025
  • Peptide Modification of Polyimide-Insulated Microwires: Towards Improved
           Biocompatibility through Reduced Glial Scarring
    • Authors: Sangita Sridar; Matthew A. Churchward; Vivian K. Mushahwar; Kathryn G. Todd; Anastasia L. Elias
      Abstract: Publication date: Available online 19 July 2017
      Source:Acta Biomaterialia
      Author(s): Sangita Sridar, Matthew A. Churchward, Vivian K. Mushahwar, Kathryn G. Todd, Anastasia L. Elias
      The goal of this study is to improve the integration of implanted microdevices with tissue in the central nervous system (CNS). The long-term utility of neuroprosthetic devices implanted in the CNS is affected by the formation of a scar by resident glial cells (astrocytes and microglia), limiting the viability and functional stability of the devices. Reduction in the proliferation of glial cells is expected to enhance the biocompatibility of devices. We demonstrate the modification of polyimide-insulated microelectrodes with a bioactive peptide KHIFSDDSSE. Microelectrode wires were functionalized with (3-aminopropyl) triethoxy silane (APTES); the peptide was then covalently bonded to the APTES. The soluble peptide was tested in 2D mixed cultures of astrocytes and microglia, and reduced the proliferation of both cell types. The interactions of glial cells with the peptide-modified wires was then examined in 3D cell-laden hydrogels by immunofluorescence microscopy. As expected for uncoated wires, the microglia were first attracted to the wire (7 days) followed by astrocyte recruitment and hypertrophy (14 days). For the peptide-treated wires, astrocytes coated the wires directly (24 hours), and formed a thin, stable coating without evidence of hypertrophy, and the attraction of microglia to the wire was significantly reduced. The results suggest a mechanism to improve tissue integration by promoting uniform coating of astrocytes on a foreign body while lessening the reactive response of microglia. We conclude that the bioactive peptide KHIFSDDSSE may be effective in improving the biocompatibility of neural interfaces by both reducing acute glial reactivity and generating stable integration with tissue. Statement of Significance The peptide KHIFSDDSSE has previously been shown in vitro to both reduce the proliferation of astrocytes, and to increase the adhesion of astrocyte to glass substrates. Here, we demonstrate a method to apply uniform coatings of peptides to microwires, which could readily be generalized to other peptides and surfaces. We then show that when peptide-modified wires are inserted into 3D cell-laden hydrogels, the normal cellular reaction (microglial activation followed by astrocyte recruitment and hypertrophy) does not occur, rather astrocytes are attracted directly to the surface of the wire, forming a relatively thin and uniform coating. This suggests a method to improve tissue integration of implanted devices to reduce glial scarring and ultimately reduce failure of neural interfaces.
      Graphical abstract image

      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.026
  • Regulated viral BDNF delivery in combination with Schwann cells promotes
           axonal regeneration through capillary alginate hydrogels after spinal cord
    • Authors: Shengwen Liu; Beatrice Sandner; Thomas Schackel; LaShae Nicholson; Abdelwahed Chtarto; Liliane Tenenbaum; Radhika Puttagunta; Rainer Müller; Norbert Weidner; Armin Blesch
      Abstract: Publication date: Available online 19 July 2017
      Source:Acta Biomaterialia
      Author(s): Shengwen Liu, Beatrice Sandner, Thomas Schackel, LaShae Nicholson, Abdelwahed Chtarto, Liliane Tenenbaum, Radhika Puttagunta, Rainer Müller, Norbert Weidner, Armin Blesch
      Grafting of cell-seeded alginate capillary hydrogels into a spinal cord lesion site provides an axonal bridge while physically directing regenerating axonal growth in a linear pattern. However, without an additional growth stimulus, bridging axons fail to extend into the distal host spinal cord. Here we examined whether a combinatory strategy would support regeneration of descending axons across a cervical (C5) lateral hemisection lesion in the rat spinal cord. Following spinal cord transections, Schwann cell (SC)-seeded alginate hydrogels were grafted to the lesion site and AAV5 expressing brain-derived neurotrophic factor (BDNF) under control of a tetracycline-regulated promoter was injected caudally. In addition, we examined whether SC injection into the caudal spinal parenchyma would further enhance regeneration of descending axons to re-enter the host spinal cord. Our data show that both serotonergic and descending axons traced by biotinylated dextran amine (BDA) extend throughout the scaffolds. The number of regenerating axons is significantly increased when caudal BDNF expression is activated and transient BDNF delivery is able to sustain axons after gene expression is switched off. Descending axons are confined to the caudal graft/host interface even with continuous BDNF expression for 8weeks. Only with a caudal injection of SCs, a pathway facilitating axonal regeneration through the host/graft interface is generated allowing axons to successfully re-enter the caudal spinal cord. Statement of Significance Recovery from spinal cord injury is poor due to the limited regeneration observed in the adult mammalian central nervous system. Biomaterials, cell transplantation and growth factors that can guide axons across a lesion site, provide a cellular substrate, stimulate axon growth and have shown some promise in increasing the growth distance of regenerating axons. In the present study, we combined an alginate biomaterial with linear channels with transplantation of Schwann cells within and beyond the lesion site and injection of a regulatable vector for the transient expression of brain-derived neurotrophic factor (BDNF). Our data show that only with the full combination axons extend across the lesion site and that expression of BDNF beyond 4weeks does not further increase the number of regenerating axons.
      Graphical abstract image

      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.024
  • Nanofiber-Structured Hydrogel Yarns with pH-Response Capacity and
           Cardiomyocyte-Drivability for Bio-Microactuator Application
    • Authors: Shaohua Wu; Bin Duan; Xiaohong Qin; Jonathan T. Butcher
      Abstract: Publication date: Available online 18 July 2017
      Source:Acta Biomaterialia
      Author(s): Shaohua Wu, Bin Duan, Xiaohong Qin, Jonathan T. Butcher
      Polymeric hydrogels have great potential in soft biological micro-actuator applications. However, inappropriate micro-architecture, non-anisotropy, weak biomechanics, and inferior response behaviors limit their development. In this study, we designed and manufactured novel polyacrylonitrile (PAN)-based hydrogel yarns composed with uniaxially aligned nanofibers. The nanofibrous hydrogel yarns possessed anisotropic architecture and robust mechanical properties with flexibility, and could be assembled into defined scaffold structures by subsequent processes. The as-prepared hydrogel yarns showed excellent pH response behaviors, with around 100% maximum length and 900% maximum diameter changes, and the pH response was completed within several seconds. Moreover, the hydrogel yarns displayed unique cell-responsive abilities to promote the cell adhesion, proliferation, and smooth muscle differentiation of human adipose derived mesenchymal stem cells (HADMSC). Chicken cardiomyocytes were further seeded onto our nanofibrous hydrogel yarns to engineer living cell-based microactuators. Our results demonstrated that the uniaxially aligned nanofibrous networks within the hydrogel yarns were the key characteristics leading to the anisotropic organization of cardiac cells, and improved sarcomere organization, mimicking the cardiomyocyte bundles in the native myocardium. The construct is capable of sustaining spontaneous cardiomyocyte pumping behaviors for 7 days. Our PAN-based nanofibrous hydrogel yarns are attractive for creating linear microactuators with pH-response capacity and biological microactuators with cardiomyocyte-drivability. Statement of Significance A mechanically robust polyacrylonitrile-based nanofibrous hydrogel yarn is fabricated by using a modified electrospinning setup in combination with chemical modification processes. The as-prepared hydrogel yarn possesses a uniaxially aligned nanofiber microarchitecture and supports a rapid, pH-dependent expansion/contraction response within a few seconds. Embryonic cardiomyocytes-seeded hydrogel yarn improves the sarcomere organization and mimics the cardiomyocyte bundles in the native myocardium, which sustains spontaneous cardiomyocyte pumping behaviors. The nanofibrous hydrogel yarn has several advantages over traditional bulk hydrogel scaffolds in terms of robust biomechanics, anisotropic aligned architecture, and superior pH response behaviors. Our nanofibrous hydrogel yarn holds the potential to be developed into novel linear and biological microactuators for various biomedical applications.
      Graphical abstract image

      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.023
  • A nanoporous titanium surface promotes the maturation of focal adhesions
           and formation of filopodia with distinctive nanoscale protrusions by
           osteogenic cells
    • Authors: Dainelys Guadarrama Bello; Aurélien Fouillen; Antonella Badia; Antonio Nanci
      Abstract: Publication date: Available online 17 July 2017
      Source:Acta Biomaterialia
      Author(s): Dainelys Guadarrama Bello, Aurélien Fouillen, Antonella Badia, Antonio Nanci
      While topography is a key determinant of the cellular response to biomaterials, the mechanisms implicated in the cell-surface interactions are complex and still not fully elucidated. In this context, we have examined the effect of nanoscale topography on the formation of filopodia, focal adhesions, and gene expression of proteins associated with cell adhesion and sensing. Commercially pure titanium discs were treated by oxidative nanopatterning with a solution of H2SO4/H2O2 50:50 (v/v). Scanning electron microscopy and atomic force microscopy characterizations showed that this facile chemical treatment efficiently creates a unique nanoporous surface with a root-mean-square roughness of 11.5nm and pore diameter of 20±5nm. Osteogenic cells were cultured on polished (control) and nanotextured discs for periods of 6, 24, and 72h. Immunofluorescence analysis revealed increases in the adhesion formation per cell area, focal adhesion length, and maturity on the nanoporous surface. Gene expression for various focal adhesion markers, including paxillin and talin, and different integrins (e.g. α1, β1, and α5) was also significantly increased. Scanning electron microscopy revealed the presence of more filopodia on cells grown on the nanoporous surface. These cell extensions displayed abundant and distinctive nanoscale lateral protrusions of 10–15nm diameter that molded the nanopore walls. Together the increase in the focal adhesions and abundance of filopodia and associated protrusions could contribute to strengthening the adhesive interaction of cells with the surface, and thereby, alter the nanoscale biomechanical relationships that trigger cellular cascades that regulate cell behavior. Statement of Significance Oxidative patterning was exploited to create a unique three-dimensional network of nanopores on titanium surfaces. Our study illustrates how a facile chemical treatment can be advantageously used to modulate cellular behavior. The nanoscale lateral protrusions on filopodia elicited by this surface are novel adhesive structures. Altogether, the increases in focal adhesion, length, maturity, and filopodia with distinctive lateral protrusions could substantially increase the contact area and adhesion strength of cells, thereby promoting the activation of cellular signaling cascades that may explain the positive osteogenic outcomes previously achieved with this surface. Such physicochemical cueing offers a simple attractive alternative to the use of bioactive agents for guiding tissue repair/regeneration around implantable metals.
      Graphical abstract image

      PubDate: 2017-07-24T19:30:21Z
      DOI: 10.1016/j.actbio.2017.07.022
  • On the relationship between indentation hardness and modulus, and the
           damage resistance of biological materials
    • Authors: David Labonte; Anne-Kristin Lenz Michelle Oyen
      Abstract: Publication date: 15 July 2017
      Source:Acta Biomaterialia, Volume 57
      Author(s): David Labonte, Anne-Kristin Lenz, Michelle L. Oyen
      The remarkable mechanical performance of biological materials is based on intricate structure–function relationships. Nanoindentation has become the primary tool for characterising biological materials, as it allows to relate structural changes to variations in mechanical properties on small scales. However, the respective theoretical background and associated interpretation of the parameters measured via indentation derives largely from research on ‘traditional’ engineering materials such as metals or ceramics. Here, we discuss the functional relevance of indentation hardness in biological materials by presenting a meta-analysis of its relationship with indentation modulus. Across seven orders of magnitude, indentation hardness was directly proportional to indentation modulus. Using a lumped parameter model to deconvolute indentation hardness into components arising from reversible and irreversible deformation, we establish criteria which allow to interpret differences in indentation hardness across or within biological materials. The ratio between hardness and modulus arises as a key parameter, which is related to the ratio between irreversible and reversible deformation during indentation, the material’s yield strength, and the resistance to irreversible deformation, a material property which represents the energy required to create a unit volume of purely irreversible deformation. Indentation hardness generally increases upon material dehydration, however to a larger extent than expected from accompanying changes in indentation modulus, indicating that water acts as a ‘plasticiser’. A detailed discussion of the role of indentation hardness, modulus and toughness in damage control during sharp or blunt indentation yields comprehensive guidelines for a performance-based ranking of biological materials, and suggests that quasi-plastic deformation is a frequent yet poorly understood damage mode, highlighting an important area of future research. Statement of Significance Instrumented indentation is a widespread tool for characterising the mechanical properties of biological materials. Here, we show that the ratio between indentation hardness and modulus is approximately constant in biological materials. A simple elastic-plastic series deformation model is employed to rationalise part of this correlation, and criteria for a meaningful comparison of indentation hardness across biological materials are proposed. The ratio between indentation hardness and modulus emerges as the key parameter characterising the relative amount of irreversible deformation during indentation. Despite their comparatively high hardness to modulus ratio, biological materials are susceptible to quasiplastic deformation, due to their high toughness: quasi-plastic deformation is hence hypothesised to be a frequent yet poorly understood phenomenon, highlighting an important area of future research.
      Graphical abstract image

      PubDate: 2017-06-24T05:51:15Z
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