骨生长相关因子对兔骨膜细胞生长分化的量效作用

目的　研究地塞米松 (dexamethasone)、重组人碱性成纤维细胞生长因子 (recombinanthumanbasicfibroblastgrowthfator,rhFGF)、重组人骨形态发生蛋白 -2 (recombinanthumanbonemorpho geneticprotein -2 ,rhBMP -2 )三种骨生长相关因子对兔骨膜细胞生长及分化的量效作用 ,为其在骨组织工程中的应用提供实验基础。　方法　体外分离培养兔骨膜细胞 ,分别与终浓度为 10 - 8,10 - 7,10 - 6 mol/L的地塞米松 ;终浓度为 50 ,2 0 0 ,50 0ng/ml的rhFGF ;终浓度为 50 ,50 0 ,10 0 0ng/ml的rhBMP -2共培养。分别于 4,7d后终止培养并测定细胞生长及分化指标。　结果 10 - 6 mol/L地塞米松显著抑制细胞蛋白合成 ,且对骨膜细胞碱性磷酸酶 (alkalinephosphatase ,ALP)表达无明显影响。各浓度组rhFGF显著促进了细胞蛋白合成量 ,而表现出明显的ALP活性抑制。各浓度组rhBMP -2对细胞增殖无明显影响 ;50ng/ml的rhBMP -2不影响骨膜细胞的ALP活性 ;当浓度增加到 50 0ng/ml与 10 0 0ng/ml时 ,ALP活性显著增加 (P <0 .0 1)。　结论　rhBMP -2在不影响骨膜细胞增殖的前提下 ,适当的浓度能够显著促进骨膜细胞向成骨细胞转化 ,在骨组织工程的研究中有重要的应用价值

Dose-effect effects of relating bone growth factors on proliferation and differentiation of periosteal cells in vitro

Objective To investigate the dose-effect effects of th ree relating bone growth factors, dexamethasone, recombinant human basic fibro blastic growth factor (rhFGF) and recombinant human bone morphogenetic protein- 2 (rhBMP-2), on proliferation and differentiation of periosteal cells so as to provide experimental basis for their further application in bone tissue engineer ing. Methods Periosteal cells were isolated and cultured in vitro and then exposed to dexamethasone (10 -8 mol/L, 10 -7 mol/L and 10 -6 mol/L), rhFGF (50 ng/ml, 200 ng/ml and 500 ng/ml) and rhBMP-2 (50 n g/ml, 500 ng/ml and 1 000 ng/ml) respectively. At the 4th and 7th days respectiv ely, the culture stopped and the total protein and alkaline phosphatase (ALP) ac tivities were measured. ResultsDexamethasone at concentratio n of 10 -6 mol/L significantly inhibited protein synthesis without obvious effects on ALP expression. The rhFGF at various concentrations significantly pro moted cell proliferation but inhibited ALP activity. The rhBMP-2 at various con centrations exerted insignificant effect on cell proliferation. In comparison, A LP expression was significantly enhanced by treatment of rhBMP-2 at concentrati on of 500 ng/ml and 1 000 ng/ml ( P <0.01). ConclusionThe rhBMP-2 at appropriate concentration promotes transformation of periostea l cells to osteogenic cells without reverse effects on cell proliferation and ha s great potential in bone tissue engineering.