人重组骨形成蛋白-2对骨髓成骨细胞增殖和碱性磷酸酶活性的影响

The effect of rhBMP-2 on the alkaline phosphatase activity and proliferation of osteoblast cells from human mononuclear bone marrow in vitro

目的探讨人重组骨形成蛋白-2(rhBMP-2)对人骨髓成骨细胞增殖和分化的影响以及剂量效应关系。方法以改良方法培养的骨髓成骨细胞为作用底物,加入梯度浓度的rhBMP-2溶液,应用MTT法和碱性磷酸酶(ALP)染色法检测其对细胞增殖和ALP活性的影响。结果rhBMP-2浓度低于1μg/ml时对人骨髓成骨细胞的增殖无明显影响,高于此浓度时对细胞的增殖有明显的促进作用,但无显著的剂量效应关系。从0.1～10μg/ml,ALP与rhBMP-2具有剂量依赖性促进关系,rhBMP-2浓度为10μg/ml时,其ALP活性是对照组的4倍,在高于10μg/ml时ALP活性并没有进一步显著提高。结论rhBMP-2对骨髓成骨细胞的增殖和碱性磷酸酶活性具有促进作用,其最佳效应浓度为10μg/ml。

Objective To investigate the effect of rhBMP-2to the differentiation and proliferation of osteoblast cells from human mononu-clear bone marrow cultures in vitro.Methods The bone marrow was obtained by aspir ation from a healthy male volunteer,mononuclear cells were then isolated,plating,incubated and passaged.The fourth passage cells were subcultured in culture me dium containing gradient con-centration rhBMP-2,respectively.After 72hours,cells were used to measure ALP(alkaline phosphatase)activity with ALP staining and analy ze the degree of cell proliferation with MTT-test.Results The rhBMP-2had no effect to cell proliferation when the concentration of rhBMP-2was under 1μg /ml,and it had a more significant pr omotion to cell proliferation when t he concentration of rhBMP-2was betw een 0.1～10μg /ml.The ALP activity of cells increased with the concentration of rhBMP-2increasing,the ALP activity of cells wa s four times when the concentration of rhBMP-2was 10μg /ml.Conclusion RhBMP-2could promote the prolifera tion and increase the ALP activity of osteoblast cells,and the optimal concentration of rhBMP-2wa s 10μg /ml ?