过表达BMP-4基因的BMSCs复合生物陶瓷骨在同种异体骨缺损模型中的研究  

The composite bioceramic bone of BMSCs overexpressing BMP-4 gene in a homogeneous rabbit bone defect model

作　　者：白宇 徐立[1] 张旭然[1] BAI Yui;XU Li;ZHANG Xu-ran(Ward 3 of Orthopedic Surgery,Fuxin Central Hospital,Fuxin 123099,China)

机构地区：[1]阜新市中心医院骨外科三病房,辽宁阜新123099

出　　处：《解剖科学进展》2022年第2期185-188,196,共5页Progress of Anatomical Sciences

基　　金：辽宁省自然科学基金(20180550528)。

摘　　要：目的 研究骨形态发生蛋白BMP-4转染骨髓间充质干细胞后与生物陶瓷骨形成复合材料对兔桡骨缺损的修复作用和机制。方法 离体分离培养鉴定兔BMSCs,构建过表达BMP-4基因的慢病毒载体并转染BMSCs细胞,并通过流式细胞术和Western blot鉴定;制备复合生物陶瓷骨,ALP试剂盒检测桡骨组织ALP活性;HE染色观察桡骨组织形态变化;依据Lane-Sandhu法评价受体兔挠骨改善情况;采用免疫荧光检测桡骨组织Col I、OC和OPN表达;Western blot检测桡骨骨膜VEGF和EGF水平。结果 成功分离兔BMSCs细胞,并获得BMP-4基因过表达的BMSCs细胞。BMP-4过表达的BMSCs复合生物陶瓷骨能够显著修复缺损的兔桡骨组织形态,提高组织的Lane-Sandhu评分,并上调挠骨的I型胶原、骨钙素和骨桥素的表达水平和骨膜中血管内皮生长因子(VEGF)和表皮生长因子(EGF)的表达。结论 BMP-4转染BMSCs与生物陶瓷骨复合材料能显著修复兔骨缺损,并增加骨膜组织中VEGF和EGF的表达。Objective To study the effect and mechanism of bone morphogenetic protein BMP-4 transfected bone marrow mesenchymal stem cells(BMSCS) and bioceramics bone composite for repairing rabbit radial defect.Methods BMSCs were isolated and cultured in vitro. Lentiviral vectors overexpressing BMP-4 gene were constructed and transfected into BMSCs. BMSCs were identified by flow cytometry and Western blot. Composite bioceramic bone was prepared and ALP kit was used to detect ALP activity in radial tissue. HE staining was used to observe the morphological changes of radial bone. Lane-sandhu method was used to evaluate the improvement of the recipient rabbit’s flexural bone.The expressions of Col I, OC and OPN in radial tissue were detected by immunofluorescence. Western blot was used to detect VEGF and EGF levels in radial periosteum. Results Rabbit BMSCs were successfully isolated and BMSCs with BMP-4overexpression were obtained. BMSCs overexpressed by BMP-4 can significantly repair the tissue morphology of rabbit radius defect, improve lan-Sandhu score, and up-regulate the expression of type I collagen, osteocalcin and osteopontin, as well as the expression of vascular endothelial growth factor(VEGF) and epidermal growth factor(EGF) in the periosteum.Conclusion BMSCs and bioceramic bone composite transfected with BMP-4 could significantly repair bone defects and increase the expression of VEGF and EGF in the periosteum.

关 键 词：BMP-4 BMSCS 骨缺损 生物陶瓷骨 

分 类 号：R683[医药卫生—骨科学]