肝细胞生长因子基因修饰的骨髓间充质干细胞移植治疗兔股骨头坏死(英文)  

Transplantation of hepatocyte growth factor gene-modified mesenchymal stem cells for treatment of femoral head necrosis in rabbits

作　　者：宫恩年 商丰元 郭子宽[2] 楼晓[3] 

机构地区：[1]解放军66400部队骨病医院骨科,北京市100143 [2]北京放射与辐射医学研究所,北京市100850 [3]解放军第三0七医院,北京市100071

出　　处：《中国组织工程研究与临床康复》2009年第28期5411-5415,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家重点基础发展规划项目(2006CB504105);国家高技术研究发展计划(2007AA02Z454)~~

摘　　要：背景：课题利用实验室自主研制、已获国家SFDA批准临床试验的重组肝细胞生长因子腺病毒感染骨髓间充质干细胞，结合肝细胞生长因子的促血管新生和抗细胞凋亡以及间充质干细胞的成骨功能，通过细胞治疗的手段进行骨损伤修复。目的：探讨肝细胞生长因子基因修饰的骨髓间充质干细胞移植对兔股骨头坏死骨缺损的治疗效果。设计、时间及地点：细胞-材料学体内实验，于2007—09／12在北京放射与辐射医学研究所和解放军66400部队骨病专科医院完成。材料：清洁级26-28周龄新西兰大白兔18只，由北京开源兔业养殖场提供。骨基质明胶为上海骁博科技发展有限公司产品。方法：采用贴壁法分离培养兔自体骨髓间充质干细胞，体外鉴定成骨和成脂肪能力。18只兔均建立双侧股骨头坏死骨缺损模型，随机分为3组，6只，组，单纯材料组缺损区仅填塞骨基质明胶，细胞一材料组填塞复合骨髓间充质干细胞的骨基质明胶，基因修饰细胞一材料组填塞复合Ad—HGF感染的骨髓间充质干细胞的骨基质明胶，各组细胞用量为10^7个，缺损，骨基质明胶用量为27mm3，缺损，修复3个月后取材进行组织学检查。骨髓间充质干细胞和Ad-HGF感染的骨髓间充质干细胞经荧光染料二醋酸琥珀酰酯羟基荧光素标记后，氯化钴处理72h，流式细胞学分析细胞增殖情况。主要观察指标：骨髓间充质干细胞的诱导分化，骨髓间充质干细胞对兔股骨头坏死的修复效果，Ad-HGF感染的骨髓间充质干细胞抗低氧损伤能力。结果：培养的细胞在适当诱导条件下可分化为成骨和成脂肪细胞。修复后3个月组织学分析显示，单纯材料组仅见疏松的纤维肉芽组织填充：细胞一材料组可见致密的纤维肉芽组织填充，其间有较多的新生觑管，但未见新生骨组织：基因修饰细胞一材料组多为软骨样组�BACKGROUND： Using bone marrow mesenchymal stem cells （MSCs） infected by an adenoviral vector carrying human hepatocyte growth factor （HGF） gene, which was independently developed by our laboratory and has been approved by SFDA in clinical trial to repair bone defects via cell therapy. OBJECTIVE： To investigate the bone restoration effect of mesenchymal stem cell and hepatocyte growth factor in the pathophysiological course of femoral head osteonecrosis defects. DESIGN, TIME AND SETTING： An in vivo experiment of cellular-material science. The experiment was performed at the Beijing Institute of Radiation Medicine and the 66400 Orthopaedic Hospital of Chinese PLA from September to December 2007. MATERIALS： Eighteen New Zealand rabbits with clean grade, aged 26-28 weeks, were supplied by Beijing Kaiyuan rabbit livestock farm. Bone matrix gelatin was purchased from Shanghai Xiaobo Technological Development Limited Co., Ltd. METHODS： MSCs from New Zealand white rabbit were isolated and culture-expanded by adhesion method and their in vitro osteogenesis and adipogenesis were identified. Eighteen New Zealand white rabbits were created femoral heads defect models and randomly divided into 3 groups, with 6 animals in each group. Animals in each group were treated with scaffold of bone matrix gelatin （27 mm3 per defect） only, scaffolds seeded with MSCs （1×10^7 per defect） or MSCs infected by an adenoviral vector carrying human hepatocyte growth factor gene （MSC/HGF, 1×10^7 per defect）, respectively. Histological examination was conducted at 3 months post operation. MSCs or MSC/HGF were labeled with carboxyfluorescein diacetate succinmidyl ester dye, treated with cobalt chloride for 72 hours and their proliferative status was evaluated and compared by flow cytometdc techniques. MAIN OUTCOME MEASURES： The differentiation of MSCs, therapeutic effect of MSCs in treating femoral head necrosis, and potent ability of MSCs/HGF against hypoxia. RESULTS： The adherent cells could differentiate into

关 键 词：骨髓间充质干细胞 肝细胞生长因子 股骨头坏死 

分 类 号：R318[医药卫生—生物医学工程]