兔骨折血肿细胞分离回植对骨折愈合的影响及机制的研究

The Study on the Impact and Mechanisms of Fracture Healing by the Rabbit Fracture Hematoma Cells Separation and Replantation

目的:研究血肿分离细胞对骨折愈合的影响及其相关分子机制,为骨折血肿细胞的应用提供实验依据。方法:将新西兰兔随机分成对照组和实验组。制备股骨髁上骨折模型。对照组:骨折后常规状态下观察骨折愈合,不做任何特殊处理。实验组:将骨折血肿取出,分离、除去红细胞及血清,将制备后的血肿细胞回植于骨折处。分别于骨折后第10、20、30天,对实验兔进行X线摄影检查,骨折断端行病理组织学检查,应用免疫组织化学染色技术检测血小板衍生生长因子(PDGF)和转化生长因子-β(TGF-β)的表达。比较2组实验兔的骨折愈合情况。结果:(1)X线摄影检查,实验组兔在不同时间点与对照组进行比较,其骨折区域骨痂形成量多,出现时间早。(2)HE染色及免疫组织化学染色检测,实验组与对照组比较,其不同时间点骨细胞及成骨细胞数量多、出现早,PDGF、TGF-β的免疫表达强度也更高。结论:(1)骨折血肿细胞回植可促进骨折愈合,回植后骨折愈合无明显感染及排斥反应。(2)骨折血肿细胞回植促进骨折愈合的分子机制可能部分与其PDGF、TGF-β表达增强有关。

Purpose:To study the hematoma isolated cell on fracture healing and its associated molecular mechanism,so as to provide the experimental basis for the application of fracture hematoma cells.Methods:The New Zealand rabbits were randomly divided into two groups:group A(control group)and group B(experimental group).The fracture model of femoral supracondylar was prepared.In group A,the normal fracture healing was observed after fracture.In group B,the fracture hematoma cells were replanted after being prepared by separating from the fracture hematoma and isolating from the red blood cells and serum.X-ray examinations and histopathological examination were taken in the 10 th,20 th,and 30 thday,respectively after fracture.Immunohistochemistry was also used to detect the expression of platelet derived growth factor(PDGF)and transforming growth factor-β(TGF-β).The rabbit fracture healing were compared between the two groups.Results:(1)At different time points,the fracture callus formation of rabbits in group B appeared more and earlier than that in group A.(2)At different time points,through the HE staining and immunohistochemistry detection,the osteocytes and osteoblasts of rabbits in group B appeared more and earlier than that in group A,and the expression of PDGF and TGF-βin group B was higher than that in group A.Conclusion:(1)Fracture hematoma cells are rich in cytokines,and have no obvious fracture healing infection and rejection in replantation.(2)The molecular mechanisms of fracture cells replantation promoting fracture healing may be partially related to the higher secretion of PDGF and TGF-β.