胰岛素样生长因子1基因转染成肌细胞促小鼠胫骨骨折愈合的作用  

Effect of myoblast carrying insulin-like growth factor 1 on healing of tibial fracture in mice

作　　者：郑浩[1] 黄婷婷[2] 孔抗美[3] 王梅[3] 齐伟力[3] 梅海龙[4] 

机构地区：[1]九江学院附属医院骨科,江西省九江市332000 [2]九江学院附属医院放射科,江西省九江市332000 [3]汕头大学医学院第二附属医院骨科,广东省汕头市515000 [4]荆门市第一人民医院骨科,湖北省荆门市448000

出　　处：《中国组织工程研究与临床康复》2009年第24期4637-4642,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:随着基因治疗的研究发展,将转有胰岛素样生长因子1基因的细胞移植入损伤区,达到该基因在体内的持续表达已可行,并有可能解决骨折患者骨不连及骨延迟愈合等难题。目的:观察携带人胰岛素样生长因子1基因的成肌细胞体内局部多点注射对C57BL/6J小鼠胫骨骨折愈合的影响。设计、时间及地点:随机对照动物实验,于2007-01/2008-04在汕头大学医学院完成。材料:8周龄雄性C57BL/6J小鼠42只,体质量22～25g,建立左胫骨骨折的动物模型。方法:42只小鼠随机数字表法分为2组,每组21只。实验组于骨折处周围肌肉中多点注射0.3mL转人胰岛素样生长因子1基因成肌细胞悬液;对照组注射0.3mL生理盐水。分别在造模后2,3,4周每组各随机取7只小鼠处死,取材,行苏木精-伊红染色。主要观察指标:各组动物骨折部位形成骨痂面积的大小和组成成分的变化;外源性细胞在体内存活及表达人胰岛素样生长因子1情况;各组动物骨密度及骨痂细胞生长情况。结果:①实验组转基因成肌细胞BrdU及人胰岛素样生长因子1免疫组织化学染色均阳性,4周实验组平均灰度值虽稍高于2周实验组,但差异无显著性意义(P>0.05)。②携人胰岛素样生长因子1基因的成肌细胞移植入C57BL/6J小鼠胫骨骨折处周围肌肉中可存活4周以上,并能稳定地分泌人胰岛素样生长因子1。③实验组造模后各时间点外骨痂中小梁骨占骨痂总面积的百分比均高于对照组,其中第3,4周经F检验差异有显著性意义(P<0.05);4周实验组骨痂骨密度高于对照组(P<0.05)。④电镜观察可见实验组骨生成细胞的数量、活跃程度和胶原纤维的数量、分布均优于对照组。结论:局部注射转人胰岛素样生长因子1基因的成肌细胞可以促进C57BL/6J小鼠胫骨骨折的愈合。BACKGROUND： With the development of gene therapy, it is feasible to transplant cells carrying insulin-like growth factor 1 （IGF-1） gene into the injury area in order to continuously express this gene in vivo and to treat bone nonunion and delayed healing in the fracture patients. OBJECTIVE： To observe the effects of myob/asts carrying IGF-1 gene injected into the mice on the healing of tibiat fracture. DESIGN, TIME AND SETTING： Randomized control animal experiment was performed in the Shantou University Medical College between January 2007 and April 2008. MATERIALS： A total of 42 male C57BL/6J mice of 8 weeks old, weighing 22 25 g, were used to establish fracture in left tibial bone. METHODS： Forty-two model mice were divided into two groups by random with 21 mice in each. Experimental animals were injected with 0.3 mL myoblast suspension carrying IGF-1 gene into the muscle surrounding fracture site, while control animals were injected with 0.3 mL saline solution. Seven mice in each group were executed at 2, 3, 4 weeks after operation, the bone callus were stained by hematoxylin-eosin. MAIN OUTCOME MEASURES： Quantitative analysis of the formed callus area and the relative amounts of components; exogenous cells＇ survival and the expression of IGF-1 ; bone mineral density and bony callus cellular growth. RESULTS： （1）In the experiment group, the BrdU staining and IGF-1 immunohistochemistry in mice transplanted with myoblasts were positive. The average grey value at 4 weeks was slightly higher than that at 2 weeks in the experiment group, but no significant difference was found （P 〉 0.05）. （2）The transplanted myoblasts carrying IGF-1 gene could survive for more than 4 weeks in the muscle surrounding fracture site, they were shown to stably express human IGF-1. （3）The percent of trabecular bone on the total area of bony callus in the experimental group was greater than that in the control group at each period following modeling, and the difference at 3 and 4 weeks post-surgery dem

关 键 词：人胰岛素样生长因子1 成肌细胞 骨折小鼠 

分 类 号：R318[医药卫生—生物医学工程]