细胞支架与骨形态发生蛋白及血管内皮生长因子复合修复大鼠股骨缺损  

Vascular endothelial growth factor and bone morphogenic protein combined with cell scaffold for the repair of rat femoral bone defects

作　　者：徐晓峰[1] 徐成振[2] 马鹏 李阳[2] 钱栋[2] 崔学文[1] 

机构地区：[1]江苏大学附属医院骨科,江苏省镇江市212001 [2]江苏大学临床医学院,江苏省镇江市212001

出　　处：《中国组织工程研究与临床康复》2009年第16期3052-3057,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：镇江市社会发展基金资助项目(SH2002019)~~

摘　　要：背景:随着骨组织工程学研究的深入,细胞因子对干细胞向成骨细胞转化和对骨愈合的影响越来越受到重视。目的:观察骨形态发生蛋白(bone morphogenic protein,BMP)、血管内皮生长因子(vascular endothelial growth factor,VEGF)复合骨髓间充质干细胞(marrowmesenchymal stem cells,MSCs)和纳米晶胶原基骨(nano-Hydroxyapatite/collagen,nHAC)复合物修复大鼠股骨骨缺损的效果。设计、时间及地点:完全随机分组设计,对比观察实验,于2007-12/2008-08在江苏大学临床医学院实验室完成。材料:将培养的第3代大鼠MSCs密度调整为5×109L-1,接种到nHAC材料表面,制成MSCs/nHAC复合支架。取复合培养的支架分别与BMP混悬液、聚乙烯吡咯烷酮、VEGF溶液混匀,负压下真空抽吸,冻干24h备用(每鼠所植入支架含BMP15mg,VEGF0.8μg)。方法:将30只SD大鼠按照随机数字表法分3组制成股骨缺损模型:单纯nHAC组骨缺损处单纯植入nHAC;MSCs/nHAC组植入MSCs/nHAC复合物;VEGF/BMP/MSCs/nHAC组植入VEGF/BMP/MSCs/nHAC复合物。主要观察指标:术后2,4,8,12周行影像学和组织学观察;术后12周行环境扫描电镜观察。结果:①各时间点动物均未出现排斥及炎症反应,创口愈合良好。②术后各时间点VEGF/BMP/MSCs/nHAC组放射学评分高于与其余2组(P<0.05)。③组织学检查结果显示MSCs/nHAC组、VEGF/BMP/MSCs/nHAC组较单纯nHAC组能更快的促进大鼠股骨缺损处骨再生;VEGF/BMP/MSCs/nHAC组的促进作用更明显。④术后12周扫描电镜观察单纯nHAC组骨纤维排列不规则,可见大量骨陷窝;MSCs/nHAC组可见大量成骨细胞,中量骨小梁结构,但仍可见间隙;VEGF/BMP/MSCs/nHAC组可见哈佛系统,大量骨小梁排列规则。结论:VEGF/BMP/MSCs/nHAC复合物较单纯nHAC或MSCs/nHAC复合物有更好的成骨能力。BACKGROUND： With the further development of bone tissue engineering, cytokines have been aroused more and more attention to affect the differentiation from stem cells into osteoblasts and bone healing. OBJECTIVE： To observe the effect of composite material, bone morphogenic protein （BMP）, vascular endothelial growth factor （VEGF）, marrow mesenchymal stem cells （MSCs） and nano-hydroxyapatite/collagen （nHAC）, on the treatment of rat femoral bone defects. DESIGN, TIME AND SETTING： A randomized grouping and contrast observational study was performed at the Laboratory of Clinical Medicine School of Jiangsu University between December 2007 and August 2008. MATERIALS： After adjusted the density of the third MSCs generation to 5× 10^9/L-1, MSCs were inoculated onto the surface of nHAC obtaining the MSCs/nHAC compound. The MSCs/nHAC compound was respectively mixed with the BMP suspension, polyvinylpyrrolidone （PVP） and VEGF solution completely, and aspirated the final compound under pressure. The final compound was froze to dry for 24 hours （The implant compound of every rat included 15 mg BMP and 0.8μg VEGF）. METHODS： Animal models of femoral bone defect were established in 30 Sprague-Dawley rats. They were randomly divided into 3 groups： nHAC group, defect was filled with nHAC; MSCs/nHAC group, with MSCs/nHAC; VEGF/BMP/MSCs/nHAC group, with VEGF/BMP/MSCs/nHAC. MAIN OUTCOME MEASURES： Imaging and histological test were made 2, 4, 8, 12 weeks after surgery. Environment scanning electron microscope （ESEM） observation was made 12 weeks after surgery. RESULTS： （1） No inflammation or rejection response was detectable during the experiment, and the wound healed well. （2） Radiograph score of VEGF/BMP/MSCs/nHAC group was significantly higher than other groups at different time points after surgery （P 〈 0.05）. （3） Histological examination showed that VEGF/BMP/MSCs/nHAC and MSCs/nHAC were superior to nHAC to promote regeneration of defected femoral bone; in particular,

关 键 词：骨形态发生蛋白:血管内皮生长因子 骨髓间充质干细胞 纳米晶胶原基骨 骨缺损 组织工程骨 

分 类 号：R318[医药卫生—生物医学工程]