Effects of different concentrations of type-I collagen hydrogel on the growth and differentiation of chondrocytes

The objective of this study was to analyze the effects of type-I collagen hydrogel of different concentrations on the growth and differentiation of rabbit chondrocytes. Articular cartilage from New Zealand white rabbits was harvested and cultured. Second-generation chondrocytes were collected for in vitro culture with 10, 7, and 5 mg/ml type-I collagen hydrogel, respectively (denoted as groups A, B, and C). After in vitro culture for 1 day, chondrocytes were stained with fluorescein diacetate (FDA)/propidium iodide (PI), and cell viability was observed by laser confocal microscopy. After in vitro culture for 14 days, the histological patterns were observed by H&E and toluidine blue staining. The expression of chondrocyte-related genes were measured by real-time quantitative RT-PCR. After in vitro culture for 1 day, FDA/PI staining showed that the cell density of group A was significantly higher than that of group B and C. After in vitro culture for 14 days, H&E staining showed that chondrocytes showed obvious aggregation in group A, partial proliferation and aggregation in group B, and uniform distribution in group C. Toluidine blue staining showed that chondrocytes in group A had aggregation areas and some were stained purple-red, fewer chondrocytes were aggregated with different staining around them in group B, and the aggregation of chondrocytes was not obvious. However, the distribution of chondrocytes was uniform with different staining in group C. After in vitro culture for 2 weeks, the levels of polymerized proteoglycan and type-II collagen mRNA were not significantly different between the three groups (P>0.05). The levels of type-I collagen, type-X collagen, and Sox9 mRNA in group A were significantly higher than those in group B and C (P<0.05). In conclusion, high concentration type-I collagen hydrogel can promote chondrocyte fibrosis and upregulation of type-I collagen, type-X collagen, and Sox9 mRNA.