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目的探讨重组人胰岛素样生长因子-Ⅰ(rhIGF-Ⅰ)和重组人转化生长因子β1(rhTGF-β1)联合应用对成骨细胞增殖和分化的影响。方法①体外培养成骨细胞株MG-63,分组以不同浓度的IGF-Ⅰ(0.1、1.0、10.0和100.0ng/mL)或TGF-β1(0.01、0.1、1.0和10.0ng/mL)培养,MTT法检测细胞增殖,得到IGF-Ⅰ与TGF-β1单独作用的最适浓度。②选用最适浓度的IGF-Ⅰ,TGF-β1单独或联合培养,MTT法检测细胞增殖,ALP试剂盒检测细胞分化,分析两种细胞因子联合应用时,在成骨细胞增殖和分化过程中的作用。结果①IGF-Ⅰ与TGF-β1对成骨细胞MG-63均有一定的促增殖和促分化作用,促增殖作用以10ng/mL的IGF-Ⅰ在第3天,1ng/mL的TGF-β1在第5天最为显著。②在上述浓度联合刺激条件下,促增殖和促分化效果均优于单一的IGF-Ⅰ或TGF-β1单独应用组。结论IGF-Ⅰ与TGF-β1均有一定的促MG-63细胞增殖和分化效应,两者联合应用时具有良好的协同作用。
Objective To investigate the effect of recombinant human insulin-like growth factor-I (rhIGF-Ⅰ) and rhTGF-β1 on osteoblast proliferation and differentiation. Methods ① The osteoblast cell line MG-63 was cultured in vitro and cultured in different concentrations of IGF-Ⅰ (0.1, 1.0, 10.0 and 100.0ng / mL) or TGF-β1 (0.01,0.1,1.0 and 10.0ng / Cell proliferation was measured by MTT method and the optimal concentration of IGF-Ⅰ and TGF-β1 alone was obtained. ② The optimal concentration of IGF-Ⅰ, TGF-β1 alone or in combination culture, MTT assay of cell proliferation, ALP kit detection of cell differentiation, analysis of the two cytokines combined application in osteoblast proliferation and differentiation process effect. Results ①IGF-Ⅰ and TGF-β1 could promote proliferation and differentiation of osteoblasts MG-63, and promote proliferation. On the 3rd day, IGF-Ⅰ of 1ng / mL The fifth day is the most significant. ② Under the combined stimulation of above concentrations, the effects of promoting proliferation and promoting differentiation were better than single IGF-Ⅰ or TGF-β1 alone groups. Conclusion Both IGF-Ⅰ and TGF-β1 promote the proliferation and differentiation of MG-63 cells in a certain extent. The combination of IGF-Ⅰ and TGF-β1 has a good synergistic effect.