慢病毒介导人骨形态发生蛋白2/骨髓间充质干细胞/脱钙骨基质修复股骨大段缺损  

Repair of large-segmental femoral defects using lentivirus-mediated bone morphogenetic protein 2/bone marrow mesenchymal stem cells/demineralized bone matrix

作　　者：陶旋[1] 李强[1] 李诗鹏 石正松[1] 周桢杰 Tao Xuan;Li Qiang;Li Shi-peng;Shi Zheng-song;Zhou Zhen-jie(Department of Emergency Traumatic Surgery,the Affiliated Hospital of Guilin Medical University,Guilin 541001,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]桂林医学院附属医院急诊创伤外科,广西壮族自治区桂林市541001

出　　处：《中国组织工程研究》2018年第9期1338-1343,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(31160199);广西壮族自治区自然科学基金资助项目(2014GXNSFAA118263);桂林医学院科研项目(LX2014254)~~

摘　　要：背景:课题组前期研究已证实,慢病毒载体介导EGFP/骨形态发生蛋白2基因转染兔骨髓间充质干细胞在表达持续时间上具有一定的优势。但慢病毒介导人骨形态发生蛋白基因转染兔骨髓间充质干细胞复合脱钙骨基质能否在体内促进骨缺损修复的问题有待探究。目的:观察兔股骨大段缺损中植入慢病毒介导人骨形态发生蛋白2转染骨髓间充质干细胞复合脱钙骨基质后的修复效果,探寻治疗股骨大段缺损的新途径。方法:选取48只新西兰大白兔,股骨中段截骨钢板内固定法制备兔左侧股骨大段缺损模型。造模成功后,随机分成A,B,C,D组,每组12只。A组无修复材料植入,B,C,D组分别植入脱钙骨基质、脱钙骨基质/骨髓间充质干细胞、慢病毒载体介导人骨形态发生蛋白2/骨髓间充质干细胞/脱钙骨基质。造模后2,4,8及12周分别处死每组3只兔子,通过苏木精-伊红染色、生物力学分析,X射线检查评价股骨大段缺损修复的情况。结果与结论:(1)X射线示A,B,C,D组均有不同的骨痂修复形成,且Lane-Sandhu X射线评分A<B<C<D组(P<0.05);(2)三点弯曲实验结果表明造模后8,12周时D组最大负荷显著高于C组,但显著低于正常股骨(P<0.05);(3)苏木精-伊红染色显示A组可见少量排列紊乱的骨小梁和大量的纤维组织;B组脱钙骨基质支架已降解,可见部分排列紊乱的骨小梁和大量的纤维组织;C组骨小梁已基本连接成线,开始塑形为皮质骨,髓腔再通不明显;D组已基本形成新的骨皮质,有髓腔再通;(4)结果提示,骨缺损区域植入慢病毒载体介导人骨形态发生蛋白2/骨髓间充质干细胞/脱钙骨基质后有大量的骨痂形成,良好的生物力学性能,完整的骨皮质,说明支架促进了骨传导、骨诱导和骨生成,对兔股骨大段缺损有较好的修复效果。BACKGROUND:Our research team has confirmed that compared to the adenoviral vector,transfection by lentiviral vector to rabbit bone marrow mesenchymal stem cells(BMSCs)is more effective that the expression of enhanced green fluorescent protein(EGFP)/bone morphogenetic protein 2(BMP-2)can be persistent for a longer term.But further investigations are needed to explore whether BMSCs transfected with hBMP-2 through lentivirus combined with demineralized bone matrix(DBM)can promote bone defect repair.OBJECTIVE:To evaluate the effect of lentivirus-mediated hBMP-2/BMSCs/DBM(LV-hBMP-2/BMSCs/DBM)on the repair of large-segmental femoral defects and to explore the new treatments for large-segmental femoral defects.METHODS:Large-segmental bone defect models were made in the right femur of 48 New Zealand white rabbits by cutting the middle femoral bone and steel plate fixation.Then,animal models were randomly divided into four groups(n=12 per group)and implanted with nothing(control),DBM,hBMP-2/DBM,and LV-hBMP-2/BMSCs/DBM.Three rabbits from each group were sacrificed at 2,4,8 and 12 weeks after surgery to evaluate the repairing effect of femoral defects through hematoxylin-eosin staining,biomechanical analysis and radiological examination.RESULTS AND CONCLUSION:X-ray results revealed that osteotylus formed in all the four groups to different extents,and Lane-Sandhu X-ray scores were ranked as follows:control group<DBM group<hBMP-2/DBM group<LV-hBMP-2/BMSCs/DBM group(P<0.05).Findings from the three-point bending test showed that the maximum load of the LV-hBMP-2/BMSCs/DBM group was significantly higher than that of the hBMP-2/DBM group,but is still lower than that of the normal femur at 8 and 12 weeks after modeling(P<0.05).Hematoxylin-eosin staining results revealed that a few trabecular bones arranged disorderedly and a large amount of fibrous tissues in the control group;the DBM scaffold was basically degraded in the DBM group presenting with partially disordered trabecular bones and a large amount of fibrous tissues;the tr

关 键 词：慢病毒属 骨形态发生蛋白类 间质干细胞 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]