Aggrecan过表达质粒修饰BMSC-外泌体联合丝素蛋白支架对软骨缺损的修复作用  

Repair effect of modifying BMSC exosomes+Aggrecan overexpression plasmid+fibroin scaffold on cartilage defects in New Zealand white rabbits

作　　者：谷铭勇 郑润泉 GU Ming-yong;ZHENG Run-quan(Bone Trauma Surgery,the 960th Hospital of the PLA Joint Logistics Support Force,Jinan 250031,China)

机构地区：[1]中国人民解放军联勤保障部队第九六〇医院创伤外科,山东济南250031

出　　处：《解剖科学进展》2024年第3期305-309,313,共6页Progress of Anatomical Sciences

基　　金：中国人民解放军联勤保障部队第九六〇医院院长基金(2018ZD01)。

摘　　要：目的探究Aggrecan过表达质粒修饰BMSC-外泌体联合丝素蛋白支架对软骨缺损的修复作用。方法以健康清洁级新西兰大白兔为研究对象,提取大白兔后腿股骨内骨髓组织进行BMSCs的培养,利用流式细胞仪进行鉴定。利用Cas-9技术构建多聚蛋白多糖(aggrecan)的过表达载体,利用慢病毒介导转染BMSCs,然后提取外泌体(exosome),联合丝素蛋白支架,构建软骨缺损治疗的联合体系,利用番红染色和Micro-CT等方法检测联合体系的修复效果,利用免疫组织化学染色和RT-PCR定量分析修复软骨组织中的Aggrecan和Ⅱ型胶原蛋白的含量。结果BMSCs-exo+丝素蛋白支架组膝关节软骨缺损处组织修复较好,软骨缺损愈合完全,组织严密,多为软骨组织,软骨缺损处填充物中Ⅱ型胶原蛋白表达较多,兔膝关节软骨缺损修复组织的Collagen Ⅱ和Aggrecan mRNA表达水平明显高于BMSCs+丝素蛋白支架组和Aggrecan过表达载体修饰BMSCs组。结论Aggrecan过表达载体转染BMSCs来源的外泌体联合丝素蛋白支架系统可以发挥更有效的软骨修复作用。Objective To investigate the repairing effect of Aggrecan overexpression plasmid modified BMSC exosomes combined with fibroin scaffold on cartilage defect.Methods BMSCs were taken from the femur of healthy and clean New Zealand white rabbits,cultured and identified by flow cytometry.The overexpression vector of Aggrecan was constructed by Cas-9 technology,BMSCs were transfected by lentivirus,exosomes from BMSCs were extracted,silk fibroin scaffolds were combined to construct a combined system for cartilage defect treatment.The repair effect of the combined system was detected by crocin staining and micro CT,and the content of Aggrecan and type Ⅱ collagen in repaired cartilage was determined by immunohistochemistry and RT-PCR.Results BMSCs-exo+fibroin scaffold group had better tissue repair,with complete healed cartilage defects and more type Ⅱ collagen expression level in the cartilage defects.The expression levels of collagen Ⅱ and Aggrecan mRNA in the repair tissues of knee joint cartilage defects were significantly higher in BMSCs-exo+silk fibroin scaffold group than in BMSCs+silk fibroin scaffold group or in Aggrecan overexpression vector modified BMSCs group.Conclusion The exosomes derived from BMSCs with Aggrecan overexpression vector play more effective role in cartilage repair.

关 键 词：多聚蛋白多糖 过表达载体 外泌体 骨髓间充质干细胞 丝素蛋白支架 新西兰大白兔 

分 类 号：R681.3[医药卫生—骨科学]