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2022, Vol. 26 ›› Issue (17): 2673-2677Effect of connective tissue growth factor on the secretion of type I and III collagen in tenocytesWang Quanzhen1, Xiao Yingfeng1, Wan Shengxiang1, Zhang Jian1, Zhou Bo1, Meng Fanbin1, Yu Longbiao1, Yu Fei2, 3
Abstract: BACKGROUND: Tendon rupture is a common thorny problem in orthopedics, and collagen secretion in tenocytes affects the repair process of tendon rupture. Connective tissue growth factor affects the fibrosis process of tissues and organs, and plays an important role in the secretion of cell collagen. However, its effects on the secretion of tenocyte collagen have been rarely reported. OBJECTIVE: To investigate the expression of type I collagen and type III collagen in primary chicken tenocytes stimulated in vitro with different concentrations of connective tissue growth factors. METHODS: Six Leghorn chickens, aged 8 weeks, were killed, and deep digital flexor tendon was obtained to culture primary tenocytes. Immunofluorescence staining for type I and III collagen was used to detect the primary tenocytes. Then passage 4 tenocytes were stimulated with connective tissue growth factors at different concentrations (0, 5, 10, 20, 50, 100, 200, and 500 μg/L) for 4 days. Real-time fluorescence quantitative PCR and enzyme-linked immunosorbent assay were used to detect the mRNA and protein expression of type I collagen and type III collagen in different groups of primary tenocytes. RESULTS AND CONCLUSION: The primary cultured cells were identified as tenocytes by type I and III collagen immunofluorescence staining. Real-time fluorescence quantitative PCR detection showed that the mRNA expression of type I collagen and type III collagen was increased in tenocytes after connective tissue growth factor stimulation. Enzyme-linked immunosorbent assay detection showed that the protein expression of type I collagen and type III collagen was increased in tenocytes after connective tissue growth factor stimulation (P < 0.01), and both increased gradually with the increase of connective tissue growth factors concentration. Connective tissue growth factors at 20 and 50 μg/L might be the best concentrations to stimulate primary tenocytes to produce type I collagen and type III collagen in vitro. Therefore, connective tissue growth factors within a certain concentration range can promote the secretion of type I collagen and type III collagen in primary chicken tenocytes. Key words:connective tissue growth factor, tenocyte, type I collagen, type III collagen, cell experiment |