骨髓间充质干细胞对大鼠退变椎间盘的修复作用  

Effect of bone marrow mesenchymal stem cells on the repair of degenerative intervertebral disc in rats

作　　者：余英剑[1] 邱丽纯 廖丛娟 陈奕芝[1] 李桂涛[1] 曹瑞治[1] 张坤尧[1] 

机构地区：[1]深圳市龙岗中心医院骨科,广东深圳518116 [2]深圳市龙岗区第二人民医院,广东深圳518165 [3]深圳市龙岗区人民医院,广东深圳518172

出　　处：《中国临床研究》2017年第11期1473-1475,1480,共4页Chinese Journal of Clinical Research

基　　金：深圳市龙岗区科技创新局课题(20160606171351610)

摘　　要：目的研究骨髓间充质干细胞(MSCs)对大鼠退变椎间盘的修复作用。方法采用随机数字表法将2015年10月至2016年12月利用腰椎间盘可控轴向加压装置构建出来的40只退变椎间盘大鼠分为对照组(未接受任何治疗)、Sox9转染MSCs组(制备Sox9基因慢病毒载体转染MSCs用于注射)、RADA-16+MSCs组(制备荷载多肽粉末RADA-16的MSCs用于注射)、RADA-16+Sox9转染MSCs组(制备荷载RADA-16和Sox9转染的MSCs用于注射)各10只,分别向L3/4、L4/5、L5/6椎间隙注射制备的上述MSCs混合溶液。处死大鼠,取椎间盘制备病理切片,采用免疫组化染色法检测髓核组织中Ⅱ型胶原的表达水平(灰度值分级)。对治疗前后不同时间段影像学指标(MRI、X线)、Ⅱ型胶原表达水平结果进行比较和分析。结果术前四组髓核矢状径指数、根袖直径、神经根鞘袖和硬膜囊的夹角、病变间隙、邻近节段椎间隙高度变化、Ⅱ型胶原灰度值分级比较,差异无统计学意义(P均>0.05);术后1、2、3个月各指标比较表明,疗效由高至低分别为RADA-16+Sox9转染MSCs组、RADA-16+MSCs组、Sox9转染MSCs组、对照组,差异有统计学意义(P均<0.05)。结论 RADA-16+Sox9转染MSCs修复退变椎间盘效果理想。Objective To study the repair effect of bone marrow mesenchymal stem cells （ MSCs ） on degenerative intervertebral disc in rats. Methods Forty rats were selected to construct degenerative intervertebral disc model by pressurized device for lumbar intervertebral disc and were divided into four groups （ n = 10 each） ： control group （ no treatment was given） , MSCs transfected by Sox9 group （ MSCs transfected by Sox9 gene lentiviral vector ） , RADA-16 ＋ MSCs group （MSCs loaded peptide powder RADA-16） and RADA-16 ＋ MSCs transfected by Sox9 group （MSCs loaded peptide powder RADA-16 and transfeeted by Sox9 gene lentiviral vector）. After modeling, no treatment was given in control group,and the MSCs mixed solutions prepared by the above method were respectively injected into L3/4, L4/5, L5/6 intervertebral spaces in the remaining three grotlps. After being killed, the rats intervertebral disks were taken out to prepare pathological section,and immunohistochemical staining method was used to detect type II collagen expression level （ gray value grading） in nucleus pulposus tissues. The imaging index（ MRI, X-ray）and the type II collagen expression level before injection and at different time periods after injection were compared and analyzed. Results There were no significant differences in indexes of MRI （ sagittal diameter index of nucleus pulposus, the included angle of dural sac and sleeve of nerve root sheath,root sleeve diameter）, X-ray （lesion space, vertebral space height of adjacent segments） and the gray value grading of type II collagen before treatment of MSCs mixed solutions in four groups（ all P 〉 0.05 ）. The comparisons of aforementioned indexes at 1-,3-,5- months after surgery showed that the efficacies in the order from high to low were RADA-16 ＋ MSCs transfected by Sox9 group, RADA-16 ＋ MSCs group, MSCs transfected by Sox9 group and control group （P 〈 0.05 ）. Conclusion MSCs loaded RADA-16 and transfected by Sox9 have a ideal efficacy for the

关 键 词：骨髓间充质干细胞 退变 椎间盘 SOX9基因 RADA-16多肽 慢病毒载体 修复作用 

分 类 号：R681.5[医药卫生—骨科学]