核结合因子a1基因修饰的皮肤成纤维细胞修复骨缺损的实验研究  

Experimental study of core binding,factor al gene-modified rabbit skin fibroblasts enhance bone defect repair

作　　者：肖德常[1] 邓廉夫[1] 杨庆铭[1] 谭延斌[1] 吕学敏[1] 张伟[1] 冯伟[1] 何亚峰[1] 梁静[1] 朱雅萍[1] 齐进[1] 周琦[1] 王君[1] 

机构地区：[1]上海市伤骨科研究所 上海交通大学医学院附属瑞金医院骨科,山东省青岛市市立医院骨科200025

出　　处：《中华外科杂志》2007年第22期1565-1568,共4页Chinese Journal of Surgery

基　　金：国家自然科学基金(30070759);上海市青年科技启明星(后)计划资助项目(01QMH1409);上海市科学技术委员会科研计划资助项目(06DZ22020)

摘　　要：目的探讨核结合因子 al(Cbfal)基因修饰的兔皮肤成纤维细胞与煅烧骨构建的移植材料对长骨大段骨缺损的修复作用。方法分离、纯化、培养兔皮肤成纤维细胞(RSF),应用pSG5-cbfal 及 pSG5质粒分别转染 RSF,获得 pSG5-Cbfal/RSF 及 pSG5/RSF,培养生长48 h 后,分别接种于长2 cm、直径4 mm 的圆柱状牛煅烧骨(TBC)上,制成 pSG5-Cbfal/RSF/TBC 及 pSG5/RSF/TBC复合物材料。制备兔桡骨中部长2 cm 的骨缺损模型,随机分组后,植入 pSG5-Cbfal/RSF/TBC 材料作为实验组(Ⅰ组),以 pSGS/RSF/TBC 材料植入组(Ⅱ组)、单纯 TBC 材料植入组(Ⅲ组)和无植入材料的自然修复组(Ⅳ组)作为对照,每组6只(12个桡骨)动物。分别于术后第4周及第12周摄 X 线片检查并最终获取骨折处标本,对植骨处进行三点抗折弯测试、脱钙骨组织切片的 HE 染色和 Masson染色及新生骨小梁图像计量分析。结果Ⅰ组标本骨质生长活跃,有大量新生骨小梁,生物力学性能良好,骨缺损处已完成早期愈合及修复;Ⅱ组、Ⅲ组及Ⅳ组无明显新生骨形成,未显示骨修复现象。结论 Cbfa1 基因修饰的兔皮肤成纤维细胞经与煅烧骨复合植入兔体内后,能完成对大段骨缺损的修复。Objective To investigate bone defect healing by true bone ceramic complex carrying core binding factor al （Cbfal） gene modified rabbit skin fibroblasts. Methods Transfect rabbit skin fibroblasts （RSF） with both eukaryotic expression vector pSG5 which could express Cbfal gene and pSG5. After being cultured for 48 h, the transfected RSF were seeded into true bone ceramic （ TBC ） of 2 cm in length and 4 mm in diameter to construct pSG5-Cbfal/RSF/TBC complex and pSG5/RSF/TBC complex. Forty-eight bone defect model rabbits were randomized into four groups, each has 6 rabbits （ 12 radiuses ） , due to different treatment, group I ： with pSG5-Cbfal/RSF/TBC complex, group II ： with pSG5/RSF/TBC complex, group III ： with TBC, GrouplV ： empty control. After being seeded and cultured for about 24 h the complexes were implanted into 2 cm long bone defects in the middle of bilateral radiuses of rabbits. The radiuses were inspected by X-ray and then the specimens were collected at the end of the fourth and twelfth weeks after operation. Then, the specimens were decalcified and histologically investigated with Hematoxylin eosin staining and Masson staining methods. Newly synthesized trabecular bone was inspected by image analysis system and the strength of bone defect area treated with graft-implantation was tested with biomechanical method-three point bending test. Results In group I , trabecular bone was actively synthesized to generate a great amount of trabecular bone and osteon. Preliminary union and bone defect healing were completed with good biomechanical characteristics. There were no newly synthesized trabecular in the other three groups, and bone defect healing were not discovered. The amount of newly synthesized trabecular bone and the results of biomechanical testing differed significantly between group I and the other three（P 〈 0.01 ）. The efficacy of group I was significantly better than that of the other three groups. Conclusion True bone ceramic complex composed with Cbfal gene modif

关 键 词：成纤维细胞 转染 核结合因子a1基因 骨缺损 骨修复 

分 类 号：R687[医药卫生—骨科学]