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Combinatory approach for developing silk fibroin scaffolds for cartilage regeneration
Several processing technologies and engineering strategies have been combined to create scaffolds with superior performance for efficient tissue regeneration. Cartilage tissue is a good example of that, presenting limited self-healing capacity together with a high elasticity and load-bearing properties. In this work, novel porous silk fibroin (SF) scaffolds derived from horseradish peroxidase (HRP)-mediated crosslinking of highly concentrated aqueous SF solution (16 wt.%) in combination with salt-leaching and freeze-drying methodologies were developed for articular cartilage tissue engineering (TE) applications. The HRP-crosslinked SF scaffolds presented high porosity (89.3 ± 0.6%), wide pore distribution and high interconnectivity (95.9 ± 0.8%). Moreover, a large swelling capacity and favorable degradation rate were observed up to 30 days, maintaining the porous-like structure and β-sheet conformational integrity obtained with salt-leaching and freeze-drying processing. The in vitro studies supported human adipose-derived stem cells (hASCs) adhesion, proliferation, and high glycosaminoglycans (GAGs) synthesis under chondrogenic culture conditions. Furthermore, the chondrogenic differentiation of hASCs was assessed by the expression of chondrogenic-related markers (collagen type II, Sox-9 and Aggrecan) and deposition of cartilage-specific extracellular matrix for up to 28 days. The cartilage engineered constructs also presented structural integrity as their mechanical properties were improved after chondrogenic culturing. Subcutaneous implantation of the scaffolds in CD-1 mice demonstrated no necrosis or calcification, and deeply tissue ingrowth. Collectively, the structural properties and biological performance of these porous HRP-crosslinked SF scaffolds make them promising candidates for cartilage regeneration. ; This study was funded by the Portuguese Foundation for Science and Technology (FCT) projects HierarchiTech and PEst (PEst- C/SAU/LA0026/2013). The project FROnTHERA (NORTE-01-0145-FEDER-000023), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), is also greatly acknowledge. The authors are also grateful for the FCT distinctions attributed to J. M. Oliveira (IF/00423/2012 and IF/01285/2015), A. L. Oliveira (IF/00411/2013) and F. R. Maia (SFRH/BPD/117492/2016). V. P. Ribeiro (PD/BD/113806/2015) and J. Costa (PD/BD/113803/2015) were awarded PhD scholarships under the financial support from FCT/MCTES and FSE/POCH, PD/59/2013. R. Canadas was also awarded a FCT PhD scholarship (SFRH/BD/92565/2013). A. da Silva Morais acknowledge ERC-2012-ADG 20120216–321266 (ComplexiTE) for his Post-doc scholarship. The authors also thank to Teresa Oliveira for the assistance with the histological preparation and to Le-Ping Yan for the valuable help with silk fibroin purification and hydrogels processing. ; info:eu-repo/semantics/publishedVersion
Combinatory approach for developing silk fibroin scaffolds for cartilage regeneration
Several processing technologies and engineering strategies have been combined to create scaffolds with superior performance for efficient tissue regeneration. Cartilage tissue is a good example of that, presenting limited self-healing capacity together with a high elasticity and load-bearing properties. In this work, novel porous silk fibroin (SF) scaffolds derived from horseradish peroxidase (HRP)-mediated crosslinking of highly concentrated aqueous SF solution (16 wt.%) in combination with salt-leaching and freeze-drying methodologies were developed for articular cartilage tissue engineering (TE) applications. The HRP-crosslinked SF scaffolds presented high porosity (89.3 ± 0.6%), wide pore distribution and high interconnectivity (95.9 ± 0.8%). Moreover, a large swelling capacity and favorable degradation rate were observed up to 30 days, maintaining the porous-like structure and β-sheet conformational integrity obtained with salt-leaching and freeze-drying processing. The in vitro studies supported human adipose-derived stem cells (hASCs) adhesion, proliferation, and high glycosaminoglycans (GAGs) synthesis under chondrogenic culture conditions. Furthermore, the chondrogenic differentiation of hASCs was assessed by the expression of chondrogenic-related markers (collagen type II, Sox-9 and Aggrecan) and deposition of cartilage-specific extracellular matrix for up to 28 days. The cartilage engineered constructs also presented structural integrity as their mechanical properties were improved after chondrogenic culturing. Subcutaneous implantation of the scaffolds in CD-1 mice demonstrated no necrosis or calcification, and deeply tissue ingrowth. Collectively, the structural properties and biological performance of these porous HRP-crosslinked SF scaffolds make them promising candidates for cartilage regeneration. ; This study was funded by the Portuguese Foundation for Science and Technology (FCT) projects HierarchiTech and PEst (PEst- C/SAU/LA0026/2013). The project FROnTHERA (NORTE-01-0145-FEDER-000023), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), is also greatly acknowledge. The authors are also grateful for the FCT distinctions attributed to J. M. Oliveira (IF/00423/2012 and IF/01285/2015), A. L. Oliveira (IF/00411/2013) and F. R. Maia (SFRH/BPD/117492/2016). V. P. Ribeiro (PD/BD/113806/2015) and J. Costa (PD/BD/113803/2015) were awarded PhD scholarships under the financial support from FCT/MCTES and FSE/POCH, PD/59/2013. R. Canadas was also awarded a FCT PhD scholarship (SFRH/BD/92565/2013). A. da Silva Morais acknowledge ERC-2012-ADG 20120216–321266 (ComplexiTE) for his Post-doc scholarship. The authors also thank to Teresa Oliveira for the assistance with the histological preparation and to Le-Ping Yan for the valuable help with silk fibroin purification and hydrogels processing. ; info:eu-repo/semantics/publishedVersion
Combinatory approach for developing silk fibroin scaffolds for cartilage regeneration
Ribeiro, V. P. (author) / da Silva Morais, A. (author) / Maia, F. R. (author) / Canadas, R. F. (author) / Costa, J. B. (author) / Oliveira, A. L. (author) / Oliveira, J. M. (author) / Reis, R. L. (author)
2018-04-01
29626700
Article (Journal)
Electronic Resource
English
DDC:
690
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