应用GFP基因转染示踪BMSCs在组织工程下颌骨内的分化  

A pilot study of using GFP labelling to monitor tissue engineered bone formation in repairing of canine segmental mandibular defects

作　　者：袁捷[1,2] 韦敏[1,2] 祁佐良[1,2] 陈瑾君[1,2] 崔磊[1,2] 曹谊林[1,2] 

机构地区：[1]上海交通大学医学院附属第九人民医院整形外科,上海市组织工程实验室,上海200011 [2]上海市组织工程研发中心,上海200235

出　　处：《中国口腔颌面外科杂志》2010年第2期165-170,共6页China Journal of Oral and Maxillofacial Surgery

基　　金：国家"973"组织工程基本科学问题项目(2005CB522700);国家"863"组织工程化骨构建技术研究与产品开发(2002AA205011);上海市教育委员会自然科学类科研创新项目(08YZ49)~~

摘　　要：目的:应用绿色荧光蛋白(GFP)标记技术,观察组织工程化下颌骨形成过程中种子细胞的变化与转归。方法:用GFP反转录病毒载体与疱疹性口炎病毒糖蛋白G基因(VSV-G)蛋白重组形成假病毒,高效转染犬骨髓间质干细胞(bone marrow stromal cells,BMSCs),将其接种于珊瑚,形成细胞、材料复合物,植入修复犬下颌骨标准缺损。术后32周取材,进行大体观察,组织学观察组织结构,免疫组化检测GFP蛋白表达情况。结果:32周后,大体可见组织工程化下颌骨形成,Van Gieson染色示新生骨大部分为成熟骨质,免疫组化显示新骨表面有GFP阳性成骨细胞,连接处无GFP阳性细胞,,珊瑚大部分降解。结论:组织工程下颌骨结构与正常皮质骨类似,新生组织表达GFP,证实组织工程下颌骨组织的形成来源于植入细胞。PURPOSE：To reconstruct tissue engineered mandible with cultured bone marrow stromal cells（BMSCs） and trace it by green fluorescent protein（GFP） gene transfer.METHODS：The pseudotyped RV-GFP expression vector was used to infect canine BMSCs directly.GPF-labeled BMSCs were seeded within coral to form a cell-scaffold construct,then implanted to repair canine 3cm segmental mandibular defects.New bone was harvested at 32 weeks post-operation and evaluated by gross view,Van Gieson staining and GFP immunohistochemistry.RESULTS：The engineered mandible was formed after 32 weeks.Newly formed mature bone occurred and coral was mostly degraded.Immunohistochemistry of GFP revealed that the labeled cells existed in the surface of the new bone and differentiated into osteoblast-like cells,while there were no GFP-labeled cells in the connection region.CONCLUSION：The results show that engineered mandible is similar to cortical bone and the composed cells originate from implanted BMSCs.

关 键 词：疱疹性口炎病毒糖蛋白G基因 绿色荧光蛋白 组织工程 下颌骨 

分 类 号：R318.0[医药卫生—生物医学工程]