论文部分内容阅读
背景:体外冲击波是治疗骨折延迟愈合或不愈合的一种有效方法,成骨细胞在此过程中发挥着重要的作用。目的:研究成骨细胞在体外冲击波促进骨折愈合过程中的作用,为提高冲击波治疗效果提供理论支持。方法:将原代培养的SD大鼠成骨细胞随机分成冲击波组和对照组,应用不同能量的冲击波进行处理后接种于96孔培养板,根据细胞存活率和细胞增殖情况确定适宜的冲击波能量值。钙钴法染色观察成骨细胞碱性磷酸酶,CCK-8试剂盒检测细胞存活,AKP试剂盒检测AKP表达,茜素红染色观察矿化结节,流式细胞仪检测整合素β1及RT-PCR检测整合素β1mRNA表达,伤口愈合试验观察成骨细胞的迁移率。结果与结论:冲击波处理体外原代培养成骨细胞的适宜能量为10kV(500脉冲),冲击波组细胞增殖速度快,细胞分泌碱性磷酸酶水平高,矿化结节面积大,细胞黏附率高,整合素β1及其mRNA的表达均高于对照组(P<0.01),且冲击波组细胞迁移的平均距离大于对照组(P<0.05),提示适宜能量冲击波可促进成骨细胞的增殖、分化、黏附及迁移,同时,整合素β1在细胞黏附及迁移过程中可能扮演了重要的角色。
BACKGROUND: Extracorporeal shock waves are an effective method for the treatment of delayed union or non-union fracture. Osteoblasts play an important role in this process. OBJECTIVE: To study the role of osteoblasts in shock wave promotion of fracture healing in vitro and to provide theoretical support for improving the therapeutic effect of shock waves. Methods: Primary cultured SD rat osteoblasts were randomly divided into shock wave group and control group, treated with shock wave of different energy and then inoculated into 96-well culture plate, and the appropriate shock wave energy value was determined according to cell survival rate and cell proliferation . Calcium and cobalt staining for osteoblast alkaline phosphatase, CCK-8 kit for cell survival, AKP kit for AKP expression, alizarin red staining for mineralized nodules, flow cytometry for the detection of integrin beta1 and RT- PCR was used to detect integrin β1 mRNA expression. Osteoblast migration was observed by wound healing test. RESULTS AND CONCLUSION: The optimal energy of shock wave treatment for primary cultured osteoblasts was 10 kV (500 pulses). The shock wave group showed rapid proliferation, high level of alkaline phosphatase secreted by cells, large mineralized nodules and high cell adhesion rate , And the expression of integrin β1 and its mRNA were higher than that of the control group (P <0.01). The average distance of migration in shock wave group was larger than that in control group (P <0.05), suggesting that appropriate energy shock waves could promote the proliferation and differentiation of osteoblasts , Adhesion and migration, at the same time, integrin β1 may play an important role in cell adhesion and migration.