人骨形态发生蛋白7基因转染对骨髓间充质干细胞增殖及成骨分化性能的影响(英文)  

Human bone morphogenetic protein 7 gene transfection for the proliferation and osteogenetic differentiation of the bone marrow mesenchymal stem cells

作　　者：金丹[1] 裴国献[1] 王珂 魏宽海[1] 陈滨[1] 

机构地区：[1]南方医科大学附属南方医院创伤骨科,广东省广州市510515 [2]解放军第一五三医院骨科,河南省郑州市450000

出　　处：《中国临床康复》2005年第14期206-208,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家重点基础研究发展规划九七三项目(G 1999054309);全军医药卫生"十五"科研基金重点项目(01Z045)资助~~

摘　　要：背景:骨组织工程中一个重要的研究内容就是如何保持并提高成骨细胞的体内外成骨能力,采用基因转移技术有可能为此问题的解决提供一种新的有效方法。目的:探讨逆转录病毒介导的人骨形态发生蛋白7基因转染对兔骨髓间充质干细胞增殖和向成骨细胞分化的影响。设计:以细胞为研究对象,分组对照,重复观察测量。对象:实验于2001-07/2003-07在南方医科大学附属南方医院创伤骨科实验室完成。新西兰大白兔4只由第一军医大学动物实验中心提供,雌雄不拘,体质量1.0～1.5kg。方法:构建人骨形态发生蛋白7逆转录病毒载体,使用含目的基因的病毒液感染骨髓间充质干细胞,免疫组织化学方法检测人骨形态发生蛋白7蛋白的表达。四甲基偶氮唑盐法检测细胞增殖能力,使用流式细胞仪检测细胞周期,NPP法检测碱性磷酸酶合成情况。主要观察指标:主要结局:①细胞增殖检测结果。②碱性磷酸酶检测结果。次要结局:①转染骨髓间充质干细胞中人骨形态发生蛋白7蛋白的表达。②细胞周期检测结果。结果:经骨形态发生蛋白7基因转染的兔骨髓间充质干细胞免疫组织化学检测显示有人骨形态发生蛋白7的阳性表达。经人骨形态发生蛋白7基因转染的兔骨髓间充质干细胞增殖能力无明显改变(P>0.05)。经转染的骨髓间充质干细胞碱性磷酸酶合成(294.592±86.567)BACKGROUND:The main aspect of the study in the bone histological engineering i s how to maintain and improve the osteogenesis of the osteoblasts in vivo and in vitro.The gene transference may provide a new effective method to deal with the problem. OBJECTIVE:To discuss the effect of the reverse transcription virus mediated hu man bone morphogenetic protein 7(hBMP 7) gene transfection on the proliferation and osteogenetic differentiation of the bone marrow mesenchymal stem cells (BMS Cs) of the rabbits. DESIGN:Cells taken as the study object, grouping control, repeat observation a nd measurement. SETTING:Traumatological and othopaedic lab of a medical university hospital. PARTICIPANTS:The study was completed in the Traumatological and Othopaedic Lab in the Affiliated Nanfang Hospital of the Southern Medical University from July 2001 to July 2003. Four New Zealand rabbits,whose weights varied from 1.0 to 1. 5 kg,were provided without sex limit by the Animal Experiment Center of the Firs t Military Medical University of Chinese PLA. METHODS:The reverse transcription virus carriers of the hBMP 7 were construct ed,and then the BMSCs were transfected by the virus containing target genes.The expression of the hBMP 7 protein was detected with the immunohistochemical meth od. The cell proliferation, cycle and ALP synthesis were respectively detected w ith the MTT method,flow cytometer and NPP method. MAIN OUTCOME MEASURES:Primary results:①the detection results of the cell prol iferation.②the detection results of the ALP. Secondary results:①the expression of the hBMP 7 protein in the transfected BMSCs.②the detection results of the cell cycle. RESULTS:After the BMP 7 gene transfection,there was hBMP 7 positive expressi on in the BMSCs of the rabbits,using the immunohistochemical detection.There was no significant change in the BMSCs proliferation of the rabbits after the hBMP 7 gene transfection (P >0.05).Compared with the ALP synthesis of the transfect ed BMSCs(294.592±86.567) nkat/L,there was significant differen

关 键 词：骨形态发生蛋白质类 干细胞 骨髓细胞 碱性磷酸酶 组织工程 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]