| Background:Fracture is a common disease but occur to a more frequently unfortunate consequence of nonunion or delayed union because of more high-energy trauma caused by industry and traffic, aging population and other factors. Therefore, it has been being one of the most important studies for orthopedics to explore an effective treatment of this medical problem. Extracorporeal Shock Waves(ESW) is a new doveloping technology in the latest twenty years with the advantages of avoidance of surgery and few complications, and had been widely applied in medicine now. Extracorporeal shock wave treatment was first applied to cure urinary stones. It was found in the later study that shock waves could activate osteoblasts to accelerate the healing of fractures. So it was more widely used in the treatment of Musculoskeletal Diseases. It was documented that extracorporeal shock waves therapy in nonunion or delayed union of fracture had a success rate of 62% -83%, and achieved good clinical efficacy. Thus, it was considered as an effective treatment of nonunion or delayed union of fractures. However, the mechanisms of the effect were still unclear currently. Many scientists have undertaked considerable studies, while the effect of shock wave on cell adhesion and migration of osteoblasts has not been reported now. The present study, therefore, is intended to investigate the effect of shock wave on on cell behaviors of osteoblasts and its cellular mechanism in response to shock wave.Objective:Primary rat osteoblasts were treated by shock wave in order to determine the appropriate energy of shock wave ,and then the effects of shock wave on cell proliferation, differentiation, adhesion and migration were observed. The expression of integrin ,acted as an important molecule of cell adhesion, were detected and the related cell signal transduction pathways were analyzed to reveal undiscovered mechanism of shock wave.Methods:Primary ostsoblasts were isolated from newborn rat calvaria and seeded in plates,and An optimal dose of ESW was selected according to survival and proliferation of osteoblasts treated by different energies of ESW. The optimal energy of ESW treating osteoblasts was 10 kV (500), which had the best promotion effect on the osteoblasts survival and proliferation. The 4th subculture osteoblasts treated by optimal energy of ESW were cultured and harvested for the analysis of cell growth by CCK-8 Kit, alkaline phosphatase activity by ALP kit, mineralized nodules by Alizarin red staining,cell adhesion and migration by wound healing assay andβ1 integrin expression by Flow CytoMeter and RT-PCR.Results:It was found that cell growth, alkaline phosphatase activity and cell mineralization was increased as compared with that in control group. Higher rates of cell adhesion and further distance of cell migration were found in ESW group, and mRNA expression ofβ1 integrin was also deteched in ESW group.Conclusion:First, primary osteoblasts collected using planting pieces method could be high purity through a repetitive adhesion method, and also maintain the biological characteristics better because of not damaging cell membrane, so they were suitable for cell experiments. Second, ESW promoted the proliferation,differentiation,adhersion and migration of osteoblast in vitro,andβ1 integrin may play an important role in this process. |
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