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: guangdongzhou : yilincao yahoo. : gaokongliuyun Use the link below to share a full-text version of this article with your friends and colleagues. Learn more. Functional reconstruction of large cartilage defects in subcutaneous sites remains clinically challenging because of limited donor cartilage. Tissue engineering is a promising and widely accepted strategy for cartilage regeneration. To date, however, this strategy has not achieved a ificant breakthrough in clinical translation owing to a lack of detailed preclinical data on cell yield and functionality of clinically applicable chondrocytes.
To address this issue, the current study investigated the initial cell yield, proliferative potential, chondrogenic capacity, and regenerated cartilage type of human chondrocytes derived from auricular, nasoseptal, and costal cartilage using a scaffold-free cartilage regeneration model cartilage sheet.
Chondrocytes from all sources exhibited high sensitivity to basic fibroblast growth factor within 8 passages. Nasoseptal chondrocytes presented the strongest proliferation rate, whereas auricular chondrocytes obtained the highest total cell amount using comparable cartilage sample weights. Importantly, all chondrocytes at fifth passage showed strong chondrogenic capacity both in vitro and in the subcutaneous environment of nude mice. Although some ificant differences in histological structure, cartilage matrix content and cartilage type specific proteins were observed between the in vitro engineered cartilage and original tissue; the in vivo regenerated cartilage showed mature cartilage features with high similarity to their original native tissue, except for minor matrix changes influenced by the in vivo environment.
The current study provides detailed preclinical data for choice of chondrocyte source and thus promotes the clinical translation of cartilage regeneration approach. Figure S1. Chondrocyte proliferation in different culture systems. Figure S2. Proliferation rates of different chondrocyte types.
Figure S3. Redifferentiated potential of chondrocytes from different sources. Under regular culture system, the expression of cartilage-specific genes decrease rapidly while the dedifferentiation-related genes increase dramatically with repeated passages in all types of chondrocytes A-D. The regenerated cartilage both in vitro and in vivo presented strong expression of cartilage-specific genes with very low level dedifferentiation-related genes A-D. The mechanical strength of in vitro regenerated cartilage sheets was too weak to be detected.
Figure S4. Immuno-fluorescent staining of auricular chondrocytes at different passages. Collagen II expression rapidly decreases with repeated passages A-E. After cultured in the chondrogenic medium for 5 days, the dedifferentiated chondrocytes P5 regained collagen II expression F. Figure S5. Comparison of histological characterization between the engineered cartilage and original tissue. All the in vitro engineered cartilage samples showed ificant differences in histological structure, cell density, and cell arrangement compared with their derived tissue.
After in vivo implantation, these differences were ificantly diminished and tissue resembling the derived cartilage was observed. Figure S6. Cartilage-specific staining of native cartilage. All native cartilage showed strong positive staining of Safranin-O and collagen II but negative staining of collagen I. AU cartilage showed strong positive expression of elastin after Victoria blue-Ponceau S staining, while both NS and CO cartilage showed negative staining of elastin.
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Corresponding Author guangdongzhou First published: 15 November Read the full text. Tools Request permission Export citation Add to favorites Track citation. Share Share Give access Share full text access. Share full-text access.
Please review our Terms and Conditions of Use and check box below to share full-text version of article. Shareable Link Use the link below to share a full-text version of this article with your friends and colleagues. Abstract Functional reconstruction of large cartilage defects in subcutaneous sites remains clinically challenging because of limited donor cartilage.
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