髁突软骨诱导外源性骨髓间充质干细胞成软骨分化的实验研究  

Chondrogenic differentiation of bone marrow mesenchymal stem cells induced by condylar cartilage

作　　者：张赵一淳 鹿蕾 于世宾 杨鸿旭 叶涛 冯剑颖[1] ZHANG Zhaoyichun;LU Lei;YU Shibin;YANG Hongxu;YE Tao;FENG Jianying(School of Stomatology,Zhejiang Chinese Medicine University,Hangzhou 310054,China;State Key Laboratory of Military Stomatology&National Clinical Research Center for Oral Diseases&Shaanxi Key Laboratory of Stomatology,Department of Oral Anatomy and Physiology,School of Stomatology,The Fourth Military Medical University,Xi’an 710032,China)

机构地区：[1]浙江中医药大学口腔医学院,杭州310054 [2]军事口腔医学国家重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔疾病国际联合研究中心,第四军医大学口腔医院口腔解剖生理学教研室

出　　处：《实用口腔医学杂志》2020年第3期417-421,共5页Journal of Practical Stomatology

基　　金：国家自然科学基金资助项目(编号:81500875);陕西省自然科学基础研究计划(编号:2016JQ8037).

摘　　要：目的:研究正常和骨关节炎髁突软骨诱导外源性骨髓间充质干细胞(BMSCs)成软骨分化能力的差别及调节机制。方法:采用单侧前牙反[牙合](UAC)诱导的颞下颌关节骨关节炎动物模型。BMSCs分别与正常髁突软骨(对照组)和UAC髁突软骨(实验组)在transwell系统中共培养48 h,用realtime-PCR检测BMSCs中Col2a1基因表达情况,用甲苯胺蓝染色及免疫荧光染色观察BMSCs软骨基质表达情况和c-myc和ERK1/2表达情况。结果:BMSCs与对照组和UAC组细胞培养后,甲苯胺蓝染色均为阳性,Col2a1的mRNA表达水平均升高,UAC组BMSCs比对照组共培养的BMSCs表达低。共培养的BMSCs的c-myc和ERK1/2均为阳性,但与UAC组共培养的BMSCs的c-myc和ERK1/2表达水平较对照组共培养的低。结论:骨关节炎软骨较正常软骨诱导BMSCs成软骨分化能力弱,c-myc和ERK1/2可能参与了髁突软骨诱导的BMSCs成软骨分化。Objective:To investigate the difference of chondrogenesis of exogenous bone marrow mesenchymal stem cells(BMSCs)respectively induced by normal and osteoarthritic condylar cartilage.Methods:The unilateral anterior crossbite(UAC)induced temporomandibular joint osteoarthritis model was used.BMSCs were co-cultured in the transwell system with either normal(control)and UAC condylar cartilage for 48 h respectively.The expression of the Col2a1 gene was detected by realtime-PCR.Cartilage matrix and c-myc and ERK1/2 of BMSCs were detected with toluidine blue staining and immunofluoresent staining,respectively.Results:After co-culture,the BMSCs was positive for toluidine blue staining.The expression level of Col2a1 mRNA of co-cultured BMSCs was higher than that of BMSCs without co-culture,the level of BMSCs co-cultured with UAC group was lower than that of control group.The co-cultured BMSCs were c-myc and ERK1/2 positve,the expression level of UAC group was lower than that of the control group.Conclusion:Osteoarthritic condylar cartilage has weaker effect in the induction of chondrogenesis of BMSCs,c-myc and ERK1/2 might be involved in the BMSCs chondrogenic differentiation induced by condylar cartilage.

关 键 词：骨髓间充质干细胞(BMSCs) 成软骨分化 髁突软骨 Ⅱ型胶原(Col2a1) c-myc 细胞外信号调节激酶1和2(ERK1/2) 

分 类 号：R782.6[医药卫生—口腔医学]