以转染CDMP1基因的BMSCs修复软骨缺损  

Repairing larynx cartilage defects with rabbit bone marrow mesenchymal stem cells transfected by CDMP1

作　　者：崔颖[1] 王月田[1] 姚梅[1] 胡晶[1] 李俊霞[1] 王宇[1] 张本[1] 

机构地区：[1]辽宁医学院附属第一医院耳鼻咽喉科,辽宁锦州121000

出　　处：《基础医学与临床》2013年第4期450-457,共8页Basic and Clinical Medicine

基　　金：辽宁省教育厅创新团队项目(LT2010063);辽宁省教育厅基金项目(L2010290)

摘　　要：目的探讨CDMP1基因转染的骨髓间充质干细胞(BMSCs)负载于聚乳酸-羟基乙酸(PLGA)支架上修复喉软骨缺损的能力,并对其修复效果做出初步评估。方法用反转录聚合酶链式反应(RT-PCR)和免疫印迹法(Western blot)检测hCDMP1 mRNA和蛋白的表达;用免疫组织化学方法检测Ⅱ型胶原蛋白(ColⅡ)以及糖胺聚糖(GAG)的表达;将转染前后的细胞支架培养体系移植入兔甲状软骨全层缺损处,从大体、组织学方面观察其对软骨缺损的修复作用。结果腺病毒感染方法可以将外源hCDMP1基因成功转入BMSCs,并使其获得稳定表达;和对照组比较,转染hCDMP1基因的BMSCs分泌ColⅡ、GAG等软骨特异性基质的能力增强,有促进软骨分化趋势;转染细胞支架复合物可更加有效地修复喉软骨缺损。结论转染hCDMP1基因的BMSCs/PLGA三维生物支架复合物移植动物体内可修复喉软骨缺损。Objective To explore the clinical application of BMSCs in PLGA scaffold transfected by CDMP and to make a preliminary assessment on the repairing effect. Methods Using RT-PCR and Western blot for detection of hCDMP1 mRNA and protein expression ; using immunohistochemistry for detection of collagen type Ⅱ（ ColⅡ ） as well as glycosaminoglycan （GAG） expression. The transfected and untransfected cell scaffold culture systems were implanted into the rabbit thyroid cartilage defects and the culturing systems were analyzed at the gross level as well as at the histological level to observe the ability to repair cartilage defects. Results Adenovirus infection was suc- cessfully transfected into BMSCs of exogenous hCDMPI gene with stable expression as compared with the control group, the BMSCs transfected by hCDMP1 gene enhanced the secretion of type Ⅱ collagen, GAG and cartilage specific matrix ability; transfected cell scaffold complex could effectively repair laryngeal cartilage defects. Conclu- sions BMSCs / PLGA 3D scaffold composite transfected by hCDMP1 gene can repair laryngeal cartilage defects ef- fectively.

关 键 词：软骨组织工程 软骨源性形态发生蛋白1 骨髓间充质干细胞 腺病毒转染 

分 类 号：R318.19[医药卫生—生物医学工程]