人胎盘提取物对成骨细胞增殖、分化、矿化及骨愈合的实验研究

摘要

目的 研究人胎盘提取物(human placenta extract,HPE)对体外成骨细胞的增殖、分化、矿化的影响,进一步了解HPE促进骨折愈合的机理.方法 用成骨细胞系(hFob1.19细胞),通过CCK-8、流式细胞术、Real-Time PCR法分别检测不同剂量的HPE(0、18 μl、100μl、180μl、360μl)在不同时间对成骨细胞增殖的影响;通过检测碱性磷酸酶(alkaline phosphatase,ALP)观察不同剂量的HPE和Rhbmp-2(0、18 μl HPE、100 μl HPE、100 μlRhbmp-2、100 μl HPE+ 100 μl Rhbmp-2)在不同时间对成骨细胞分化的影响;通过茜素红染色检测不同剂量的HPE(0、50 μl、100μl、200μl)在不同时间对成骨细胞矿化的影响,系统研究体外HPE对成骨细胞增殖、分化、矿化的影响及其促进骨愈合的可能机制.结果 空白对照组和加入不同剂量的HPE各组对成骨细胞的增殖、分化未观察到明显的影响(P＞0.05),而加入不同剂量的HPE(50μl、100μl、200μl)各组对成骨细胞的矿化比空白对照组有明显促进作用(P＜0.01).结论 体外HPE对成骨细胞的增殖、分化无明显影响,对成骨细胞的矿化有明显促进作用;HPE促进成骨细胞矿化可能是促进骨折愈合机制.%Objective To evaluate the effect of human placenta extract (HPE) on the proliferation,differentiation and mineralization of primary osteoblasts,and to further explore the possible mechanism for improving fracture healing.Methods Osteoblast cell line,hFob1.19,was used for the overall study.Osteoblasts were treated with HPE of different doses (0,18 μl,100 μl,180 μl,360 μl),and the cell proliferation was determined by CCK-8,FACS or Real-Time PCR.Alkaline Phosphatase production was assessed to evaluate the cell differentiation level.The effect of HPE (0,50 μl,100 μl,200 μl) on the mineralization of osteoblasts was detected by Alizarin Red S Staining.Results Compared with the control group,the mineralization of osteoblast increased after treatment with higher doses of HPE (P ＜ 0.01),but no significant enhancement of cell proliferation and cell differentiation were observed after cultured with different doses of HPE (P ＞ 0.05).Conclusion Our study suggests that HPE may contribute to fracture healing not through improvement of osteoblast proliferation or differentiation,but through enhancement of bone mineralization.