Chondrogenic potential of human dermal fibroblasts in a contractile, soft, self-assembling, peptide hydrogel

The present paper describes a simple approach to obtain three‐dimensional (3D) cartilage constructs using human normal dermal fibroblasts (hNDFs) cultured in a self‐assembling peptide nanofibre scaffold. During the first days of culture, the 3D constructs underwent morphological changes consisting o...

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Veröffentlicht in:Journal of tissue engineering and regenerative medicine 2016-02, Vol.10 (2), p.E54-E62
Hauptverfasser: Bussmann, Bianca M., Reiche, Sven, Marí-Buyé, Núria, Castells-Sala, Cristina, Meisel, Hans Jörg, Semino, Carlos E.
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container_issue 2
container_start_page E54
container_title Journal of tissue engineering and regenerative medicine
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creator Bussmann, Bianca M.
Reiche, Sven
Marí-Buyé, Núria
Castells-Sala, Cristina
Meisel, Hans Jörg
Semino, Carlos E.
description The present paper describes a simple approach to obtain three‐dimensional (3D) cartilage constructs using human normal dermal fibroblasts (hNDFs) cultured in a self‐assembling peptide nanofibre scaffold. During the first days of culture, the 3D constructs underwent morphological changes consisting of a substantial contraction process that ended in a small compact structure. During this process the system became sensitive to induction with standard chondrogenic medium, evidenced by the expression of specific markers of mature cartilage. First, it was detected that the samples become highly stained with toluidine blue dye over time (40–50 days), indicating that the system produced significantly high amounts of glycosaminoglycans. By quantitative PCR, it was confirmed that the system significantly upregulated the expression of the proteoglycan aggrecan, a good indicator of cartilage commitment. Moreover, collagen type II was upregulated at protein level, confirming that the system differentiated to a chondrocyte‐like construct. Additionally, during the first days of culture in control medium analysed hNDFs proliferation capacity in this 3D system was analysed. This platform could be used in the future to obtain an autologous source of cells from a simple patient skin biopsy, which could be easily translated into a low‐cost and effective regenerative therapy. Copyright © 2013 John Wiley & Sons, Ltd.
doi_str_mv 10.1002/term.1766
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During the first days of culture, the 3D constructs underwent morphological changes consisting of a substantial contraction process that ended in a small compact structure. During this process the system became sensitive to induction with standard chondrogenic medium, evidenced by the expression of specific markers of mature cartilage. First, it was detected that the samples become highly stained with toluidine blue dye over time (40–50 days), indicating that the system produced significantly high amounts of glycosaminoglycans. By quantitative PCR, it was confirmed that the system significantly upregulated the expression of the proteoglycan aggrecan, a good indicator of cartilage commitment. Moreover, collagen type II was upregulated at protein level, confirming that the system differentiated to a chondrocyte‐like construct. Additionally, during the first days of culture in control medium analysed hNDFs proliferation capacity in this 3D system was analysed. This platform could be used in the future to obtain an autologous source of cells from a simple patient skin biopsy, which could be easily translated into a low‐cost and effective regenerative therapy. 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During the first days of culture, the 3D constructs underwent morphological changes consisting of a substantial contraction process that ended in a small compact structure. During this process the system became sensitive to induction with standard chondrogenic medium, evidenced by the expression of specific markers of mature cartilage. First, it was detected that the samples become highly stained with toluidine blue dye over time (40–50 days), indicating that the system produced significantly high amounts of glycosaminoglycans. By quantitative PCR, it was confirmed that the system significantly upregulated the expression of the proteoglycan aggrecan, a good indicator of cartilage commitment. Moreover, collagen type II was upregulated at protein level, confirming that the system differentiated to a chondrocyte‐like construct. Additionally, during the first days of culture in control medium analysed hNDFs proliferation capacity in this 3D system was analysed. This platform could be used in the future to obtain an autologous source of cells from a simple patient skin biopsy, which could be easily translated into a low‐cost and effective regenerative therapy. 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subjects Ascorbic Acid - pharmacology
Biomarkers - metabolism
Calcification, Physiologic - drug effects
Cell Differentiation - drug effects
Cells, Cultured
chondrogenesis
Chondrogenesis - drug effects
Collagen Type II - metabolism
dermal fibroblasts
Dermis - cytology
Fibroblasts - cytology
Fibroblasts - drug effects
Glycosaminoglycans - metabolism
Humans
Hydrogel, Polyethylene Glycol Dimethacrylate - pharmacology
in vitro
Peptides - pharmacology
Regenerative medicine
self-assembling peptide
three-dimensional
Tissue engineering
Transforming Growth Factor beta3 - pharmacology
title Chondrogenic potential of human dermal fibroblasts in a contractile, soft, self-assembling, peptide hydrogel
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