巨噬细胞移动抑制因子促进骨髓间充质干细胞归巢修复急性膝关节软骨损伤  

Macrophage migration inhibition factor promotes bone marrow mesenchymal stem cells homing to repair acute knee cartilage injury

作　　者：陆定贵[1] 姚顺晗 唐乾利[1] 唐毓金[1] Lu Dinggui;Yao Shunhan;Tang Qianli;Tang Yujin(Affiliated Hospital of Youjiang Medical College for Nationalities,Baise 533000,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]右江民族医学院附属医院,广西壮族自治区百色市533000

出　　处：《中国组织工程研究》2021年第25期4013-4018,共6页Chinese Journal of Tissue Engineering Research

基　　金：广西高校中青年教师基础能力提升项目(2018KY0449),项目负责人:陆定贵;广西壮族自治区卫生和计划生育委员会自筹经费科研项目(Z20170243),项目负责人:陆定贵;百色市科学研究与技术开发计划项目(20170512),项目负责人:陆定贵;广西重点研发项目(桂科A18050008),项目负责人:唐毓金。

摘　　要：背景:软骨缺乏血管、神经,自我修复能力有限。青年人的软骨修复主要方法有软骨下骨钻孔术、微骨折术,其目的是打通软骨下骨板,使骨髓腔受到炎症刺激从而动员骨髓间充质干细胞归巢到损伤区域分化为软骨细胞、成纤维细胞,从而发挥修复作用。微骨折术后的软骨生长取决于损伤区归巢干细胞的数量,因此,提高干细胞归巢数量可提高手术成功的机会。目的:探讨巨噬细胞移动抑制因子对骨髓间充质干细胞归巢治疗软骨急性损伤的影响。方法:由人股骨骨髓中分离培养骨髓间充质干细胞,并检测其分化为软骨细胞的能力。采用免疫组化方法测定骨髓间充质干细胞和成软骨细胞的CXCR2表达,采用细胞划痕实验探讨巨噬细胞移动抑制因子对骨髓间充质干细胞迁移动力学的影响。制作大鼠急性膝关节软骨损伤模型,造模后第1,2,3天关节内注射巨噬细胞移动抑制因子,造模后第7天采用免疫化学荧光染色和流式细胞术检测损伤区域PECAM-1阳性细胞(血管内皮细胞)数量,采用DAPI染色检测损伤区域单核细胞数量。结果与结论:①骨髓间充质干细胞表面标记物CD166、CD29阳性,CD34阴性;②骨髓间充质干细胞诱导分化为软骨细胞后CXCR2抗体表型退化、消失,而Ⅱ型胶原抗体表型呈阳性;③巨噬细胞移动抑制因子可促进骨髓间充质干细胞迁移,阻断CXCR2受体后巨噬细胞移动抑制因子促进骨髓间充质干细胞迁移的作用被抑制;④在动物模型中,DAPI染色结果提示巨噬细胞移动抑制因子组损伤区有核细胞数明显高于对照组,免疫化学荧光染色及流式细胞术显示巨噬细胞移动抑制因子组修复损伤区域PECAM-1阳性细胞明显高于对照组;⑤以上结果表明,巨噬细胞移动抑制因子可促进骨髓间充质干细胞归巢修复急性软骨损伤。BACKGROUND:Cartilage lacks blood vessels and nerves,so its ability to repair itself is limited.The main method of young people’s cartilage repair is subchondral bone drilling and microfracture surgery.The purpose is to open up the subchondral bone plate and make bone marrow mesenchymal stem cells home to the injured area.Bone marrow mesenchymal stem cells differentiate into chondrocytes and fibroblasts to play a repair role.The growth rate of cartilage after microfracture surgery depends on the number of stem cells in the injured area.Therefore,increasing the number of stem cells homing can increase the chance of successful surgery.OBJECTIVE:To explore the effect of macrophage migration inhibition factor on bone marrow mesenchymal stem cells homing treatment of acute cartilage injury.METHODS:Bone marrow mesenchymal stem cells were obtained from the bone marrow of the human femur,and then cultured.The ability of bone marrow mesenchymal stem cells to differentiate into chondrocytes was tested.The CXCR2 receptor expression of bone marrow mesenchymal stem cells and chondrocytes was measured by immunohistochemical method.The effect of macrophage migration inhibition factor on the migration kinetics of bone marrow mesenchymal stem cells was investigated by cell scratch test.A rat model of acute knee articular cartilage injury was made,and macrophage migration inhibition factor was injected intra-articularly at 1,2,and 3 days after model establishment.Immunofluorescence staining and flow cytometry were used to detect the number of PECAM-1 positive cells(vascular endothelial cells)in the injured area,and DAPI staining was used to detect the number of monocytes in the damaged area.RESULTS AND CONCLUSION:(1)The surface markers of bone marrow mesenchymal stem cells were positive for CD166,CD29,and negative for CD34.(2)The CXCR2 antibody phenotype degenerated and disappeared after bone marrow mesenchymal stem cells differentiated into chondrocytes,while the phenotype expression of type 2 collagen antibody was positive.(3)M

关 键 词：干细胞 骨髓间充质干细胞 巨噬细胞移动抑制因子 膝关节 软骨损伤 细胞归巢 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]