羊胎素对体外成骨细胞增殖、分化及矿化功能的影响

目的 观察羊胎素在体外对新生大鼠颅骨成骨细胞增殖、分化及矿化功能的影响。方法将提取之羊胎素稀释加入体外培养的SD新生大鼠颅骨成骨细胞体系中,终浓度分别为0.625％、1.25％、2.5％、5％和10％;加药后24、48和96 h用MTT法检测细胞的增殖并绘制生长曲线:培养5 d时用PNPP法测定细胞碱性磷酸酶(ALP)活性;培养31 d时用茜素红染色2.5％羊胎素组形成之矿化骨结节,用图像分析仪计算骨结节的面积。结果 所有含羊胎素浓度组,其MTT法测得的A值都显著高于空白对照组(P<0.01),生长曲线表现为含羊胎素组的细胞增殖加快,以2.5％浓度组最为显著;ALP结果显示,羊胎素各浓度组的碱性磷酸酶活性都显著高于空白对照组(P<0.01);茜素红染色后显示,2.5％羊胎素组所形成的骨结节面积显著高于空白对照组(P<0.01)。结论羊胎素对体外培养的SD新生大鼠颅骨成骨细胞有显著的促进增殖、分化和矿化的作用。

Effects of placentin on osteoblast proliferation and differentiation in vitro

Objective To investigate the effects of placentin extracted from lamb placenta on rat calvarium osteoblast proliferation and differentiation in vitro. Methods Different dilutions of placentin were added to the cultured osteoblasts from calvarium of newborn SD rats. The final concentrations of placentin were 0.625% , 1.25% , 2.5% , 5% ,and 10% . The MTT method was employed to detect the osteoblast proliferation after culture with placentin for 24 h, 48 h, and 96 h separately. The osteoblast differentiation was evaluated by detecting activities of alkaline phosphatase (ALP) using PNPP method after culture with placentin for 5 days. Mineralized nodules were stained with alizarin after culture with placentin for 31 days, and the areas of nodules were measured and computed by digitized image analyses. Results The OD values by MTT method in all placentin groups were much higher than that in control vehicle group(P < 0.01) and were in dose-dependent manner; placentin in concentrations of 2.5 % and 5 % showed better effects. ALP activities in all placentin groups were significantly higher than that in control group (P < 0.01) . The summed areas of mineralized nodules in 2.5% placentin group were significantly higher than that in control group (P < 0.01). Conclusions Placentin extracted from lamb placenta stimulates the proliferation, differentiation and mineralization of cultured osteoblasts from SD rat calvarium in vitro.