中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (24): 4547-4552.doi: 10.3969/j.issn.1673-8225.2010.24.041

• 骨组织构建 bone tissue construction • 上一篇    下一篇

体外冲击波对大鼠成骨细胞增殖、分化、黏附及迁移的影响

黄钟炼，胡  军，于萌蕾，卢志军   

  1. 汕头大学医学院第一附属医院骨科，广东省汕头市  515041
  • 出版日期:2010-06-11 发布日期:2010-06-11
  • 通讯作者: 胡 军，博士，博士后，主任医师。汕头大学医学院第一附属医院骨科，广东省汕头市 515041
  • 作者简介:黄钟炼★，男，1981年生，广东省普宁市人，汉族，2004年汕头大学医学院毕业，在读硕士，医师，主要从事骨组织工程的研究。

Effect of extracorporeal shock wave on proliferation, differentiation, adhesion and migration of rat osteoblasts in vitro

Huang Zhong-lian, Hu Jun, Yu Meng-lei, Lu Zhi-jun   

  1. Department of Orthopaedics, the First Affiliated Hospital, Shantou University Medical College, Shantou   515041, Guangdong Province, China
  • Online:2010-06-11 Published:2010-06-11
  • Contact: Hu Jun, Doctor, Chief physician, Department of Orthopaedics, the First Affiliated Hospital, Shantou University Medical College, Shantou 515041, Guangdong Province, China hjzkm@yahoo.com.cn
  • About author:Huang Zhong-lian★, Studying for master’s degree, Physician, Department of Orthopaedics, the First Affiliated Hospital, Shantou University Medical College, Shantou 515041, Guangdong Province, China stuhzl@yahoo.com.cn

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摘要：

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。

关键词: 冲击波, 成骨细胞, 增殖, 分化, 黏附, 迁移, 整合素, 骨组织工程

Abstract:

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 newborn 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 extracorporeal 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.

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