同种异体软骨脱细胞基质在关节软骨缺损中的应用研究  

Repair of joint cartilage defect by allogenic decellularized cartilage matrix

作　　者：金诺 王璐 张洲铭 同瑾 李德超[1,2] JIN Nuo;WANG Lu;ZHANG Zhouming;TONG Jin;LI Dechao(Jiamusi University AffiliatedStomatological Hospital,Department of Oral Implantology,154002,China;United Jia Medical Co.,Ltd.,Xi'an)

机构地区：[1]佳木斯大学附属口腔医院口腔种植科 [2]青岛市口腔医院 [3]联合佳医医疗有限公司

出　　处：《实用口腔医学杂志》2019年第4期495-500,共6页Journal of Practical Stomatology

基　　金：黑龙江省卫生计生委科研课题(编号:2018037);佳木斯大学协同创新计划(编号:2011XTCX2016-02)

摘　　要：目的:探索使用同种异体软骨脱细胞基质( DCM)修复关节软骨缺损的效果及可能机制。方法:原代培养新西兰大白兔耳软骨细胞和骨髓间充质干细胞( BMMSCs),软骨细胞扩增后诱导形成软骨细胞膜片,将细胞膜片脱细胞处理后制作为DCM,冷冻干燥备用。将BMMSCs 附和在DCM 表面共培养,SEM 观察材料微观形态以及BMMSCs 与材料的附和情况。qPCR检测DCM 对BMMSCs 软骨向分化的影响。使用同种异体DCM 修复兔膝关节软骨缺损,3个月后使用HE 染色和免疫组织化学染色检测缺损修复效果。结果:成功培养出复层软骨细胞膜片并制作出DCM;在SEM 下观察可见BMMSCs 与DCM结合良好;并发现DCM 可诱导BMMSCs 分泌细胞外基质。qPCR检测发现DCM 促进BMMSCs 表达COL-II 和SOX-9 表达,抑制COL-I 和COL-X 表达,不影响Aggrecan 表达。动物实验显示DCM + BMMSCs 组关节软骨缺损的修复程度优于对照组。结论:同种异体DCM 可能是通过SOX-9 通路诱导BMMSCs 的软骨向分化,促进软骨缺损的修复。Objective: To explore the effects of allogenic decellularized cartilage matrix( DCM) in the repair the articular cartilage defect and the mechanism. Methods: Chondrocytes from the auricular cartilage of rabbits were isolated and expanded,cultured to form cell sheet and then the sheet was dellularized to be DCM. The bone marrow mesenchymal stem cells( BMMSCs) was co-cultured with DCM,the micromorphology of the DCM and the BMMSCs on DCM were observed by SEM. The expression of differentiation-related genes of BMMSCs was detected by qPCR. The DCM allografts were applied to repair the articular cartilage defect of rabbits,the effect was evaluated with HE staining and immunohistochemical staining( IHC). Results: SEM observation showed that the DCM scaffold had good combination ability with BMMSCs and induced the secretion of extracellular matrix. qPCR revealed that DCM may increase COL-Ⅱ and SOX-9 expression,decrease COL-I and COL-X expression of BMMSCs. The DCM allograft had a good histocompatibility in vivo and exhibited prominent effect in repairing articular cartilage defect. Conclusion: Allogenic DCM may promote the differentiation of BMMSCs into cartilage cells through SOX-9 pathway and favoure chondrogenesis for the repair of cartilage defect.

关 键 词：软骨脱细胞基质( DCM) 关节软骨缺损 软骨再生 

分 类 号：Q813[生物学—生物工程]