CAJ | 학술논문

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β1 mRNA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10 kV(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.
배경:체외충격파시치료골절연지유합혹불유합적일충유효방법,성골세포재차과정중발휘착중요적작용.목적:연구성골세포재체외충격파촉진골절유합과정중적작용,위제고충격파치료효과제공이론지지.방법:장원대배양적SD대서성골세포수궤분성충격파조화대조조,응용불동능량적충격파진행처리후접충우96공배양판,근거세포존활솔화세포증식정황학정괄의적충격파능량치.개고법염색관찰성골세포감성린산매,CCK-8시제합검측세포존활,AKP시제합검측AKP표체,천소홍염색관찰광화결절,류식세포의검측정합소β1급RT-PCR검측정합소β1 mRNA표체,상구유합시험관찰성골세포적천이솔.결과여결론:충격파처리체외원대배양성골세포적괄의능량위10 kV(500맥충),충격파조세포증식속도쾌,세포분비감성린산매수평고,광화결절면적대,세포점부솔고,정합소β1급기mRNA적표체균고우대조조(P<0.01),차충격파조세포천이적평균거리대우대조조(P<0.05),제시괄의능량충격파가촉진성골세포적증식、분화、점부급천이,동시,정합소β1재세포점부급천이과정중가능분연료중요적각색.
BACKGROUND: Extracorporeal shock wave therapy is indicated as an effective method for treatment of delayed fracture healing or nonunion. Osteoblasts plays an important role in this process.OBJECTIVE: To investigate the function of osteoblasts in the process of extracorporeal shock wave promoting fractures healing, and to provide theoretical support for improving shock wave therapy on fracture healing.METHODS: Primary cultured ostsoblasts were isolated from newbon SD rat calvaria and randomly divided into two groups, shock wave and control. Treated by different energies of extracorporeal shock wave, cells were incubated onto 96-well culture plate. An optimal dose of extracorporeai shock wave was selected according to survival and proliferation of osteoblasts. The osteoblasts treated by optimal energy of extracorporeal shock wave were cultured and harvested for the analysis of alkaline phosphatase by calcium cobolt stain, cell survival by CCK-8 Kit, alkaline phosphatase expression by AKP kit, mineralized nodules by Alizarin red staining, integrin β1 and β1 mRNA expressions by flow cytometry and RT-PCR, cell migration by wound healing assay.RESULTS AND CONCLUSION: The optimal energy of extracorporeal shock wave treating primary cultured osteoblasts was 10 kV (500 impulses). Following extracorporeal shock wave therapy, the cell proliferation, alkaline phosphatase activity, cell mineralization, rates of cell adhesion, as well as β1 integrin and its mRNA expressions were increased as compared with those in control group (P ＜ 0.01). Further distance of cell migration was found in extracorporeal shock wave group (P ＜ 0.05). The results showed that the optimal energy of extracorporeal shock wave could promote the proliferation, differentiation, adhesion and migration of osteoblasts in vitro, and β1 integrin may play an important role in the process of cell adhesion and migration.