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标题:Silk-based anisotropical 3D biotextiles for bone regeneration
时间:2019-12-03 22:30:43
DOI:10.1016/j.biomaterials.2017.01.027
作者:Ribeiro, Viviana P.; Silva-Correia, Joana; Nascimento, Ana I.
关键词:Biotextile;Textile;Silk fibroin;Knitted spacer fabrics;Human adipose-derived stem cells;Craniofacial bone tissue engineering
出版源: 《Biomaterials》 ,123 :92-106
摘要:Bone loss in the craniofacial complex can been treated using several conventional therapeutic strategies that face many obstacles and limitations. In this work, novel three-dimensional (3D) biotextile architectures were developed as a possible strategy for flat bone regeneration applications. As a fully automated processing route, this strategy as potential to be easily industrialized. Silk fibroin (SF) yarns were processed into weft-knitted fabrics spaced by a monofilament of polyethylene terephthalate (PET). A comparative study with a similar 3D structure made entirely of PET was established. Highly porous scaffolds with homogeneous pore distribution were observed using micro-computed tomography analysis. The wet state dynamic mechanical analysis revealed a storage modulus In the frequency range tested, the storage modulus values obtained for SF-PET scaffolds were higher than for the PET scaffolds. Human adipose-derived stem cells (hASCs) cultured on the SF-PET spacer structures showed the typical pattern for ALP activity under osteogenic culture conditions. Osteogenic differentiation of hASCs on SF-PET and PET constructs was also observed by extracellular matrix mineralization and expression of osteogenic-related markers (osteocalcin, osteopontin and collagen type I) after 28 days of osteogenic culture, in comparison to the control basal medium. The quantification of convergent macroscopic blood vessels toward the scaffolds by a chick chorioallantoic membrane assay, showed higher angiogenic response induced by the SF-PET textile scaffolds than PET structures and gelatin sponge controls. Subcutaneous implantation in CD-1 mice revealed tissue ingrowth's accompanied by blood vessels infiltration in both spacer constructs. The structural adaptability of textile structures combined to the structural similarities of the 3D knitted spacer fabrics to craniofacial bone tissue and achieved biological performance, make these scaffolds a possible solution for tissue engineering approaches in this area.
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目录:
  • Silk-based anisotropical 3D biotextiles for bone regeneration
    • 1. Introduction
    • 2. Materials and methods
      • 2.1. Materials
      • 2.2. Production of the textile-based scaffolds
      • 2.3. Morphological characterization
        • 2.3.1. Scanning electron microscopy
        • 2.3.2. Micro-computed tomography
      • 2.4. Mechanical properties
        • 2.4.1. Compressive tests
        • 2.4.2. Dynamic mechanical analysis
      • 2.5. In vitro cell studies
        • 2.5.1. hASCs isolation and expansion
        • 2.5.2. Seeding and osteogenic differentiation of hASCs in the textile-based scaffolds
        • 2.5.3. Scanning electron microscopy and energy dispersive spectroscopy analysis
        • 2.5.4. DNA quantification
        • 2.5.5. Alkaline phosphatase activity quantification
        • 2.5.6. Alizarin red staining and calcium deposition quantification
        • 2.5.7. Immunodetection of bone-specific proteins
        • 2.5.8. RNA isolation and real-time reverse transcriptase-polymerase chain reaction
      • 2.6. In vivo angiogenesis evaluation
        • 2.6.1. Chick chorioallantoic membrane assay
        • 2.6.2. Analysis of blood vessel convergence
        • 2.6.3. Hematoxylin & eosin staining
        • 2.6.4. Immunohistochemical detection
      • 2.7. Subcutaneous implantation
      • 2.8. Statistical analysis
    • 3. Results
      • 3.1. Morphological characterization
      • 3.2. Mechanical properties
      • 3.3. In vitro assessment of the textile-based scaffolds
        • 3.3.1. Osteogenic differentiation in the textile-based scaffolds
        • 3.3.2. Extracellular matrix mineralization and elemental composition of the deposited matrix
        • 3.3.3. Genotypic and phenotypic expression of osteogenic-related markers
      • 3.4. Angiogenic potential of the textile-based scaffolds
      • 3.5. Subcutaneous implantation of the textile-based scaffolds
    • 4. Discussion
    • 5. Conclusions
    • Acknowledgments
    • Appendix A. Supplementary data
    • References

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