1,25-二羟维生素D3和地塞米松促进人颞下颌关节滑液间充质干细胞的体外成骨分化  

Combined use of 1,25-dihydroxyvitamin D3 and dexamethasone promotes osteogenic differentiation of synovial fluid mesenchymal stem cells from human temporomandibular joint

作　　者：刘文静 孙养鹏[2,3] 郑有华 张志光[2,3] Liu Wen-jing;Sun Yang-peng;Zheng You-hua;Zhang Zhi-guang(Department of Prosthodontics,Stomatological Hospital,Southern Medical University,Guangzhou 510280,Guangdong Province,China;Department of Oral and Maxillofacial Surgery,Hospital of Stomatology,Guanghua School of Stomatology,Sun Yat-sen University,Guangzhou 510055,Guangdong Province,China;Guangdong Key Laboratory of Stomatology,Guangzhou 510070,Guangdong Province,China)

机构地区：[1]南方医科大学口腔医院修复科,广东省广州市510280 [2]中山大学光华口腔医学院·附属口腔医院口腔颌面外科,广东省广州市510055 [3]广东省口腔医学重点实验室,广东省广州市510070

出　　处：《中国组织工程研究》2018年第9期1443-1449,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81271115)

摘　　要：背景:人颞下颌关节滑液间充质干细胞获取微创简便且体外扩增能力强,成为组织工程修复一重要的种子细胞来源,优化其成骨分化能力有利于在关节骨组织再生中的应用。目的:分析1,25-二羟维生素D3和地塞米松对关节滑液间充质干细胞成骨能力的影响,获得人颞下颌关节滑液间充质干细胞成骨分化的理想条件。方法:将获得的人颞下颌关节滑液标本进行体外培养扩增,分别加入不同组分的成骨诱导液进行成骨诱导,通过茜素红和Von kossa染色比较各组矿化形成情况,通过RT-PCR定量比较成骨相关标志ALP、RUNX2和OCN的表达。结果与结论:(1)体外培养的人颞下颌关节滑液间充质干细胞可以向成骨细胞分化,其中含有100 nmol/L地塞米松和10 nmol/L 1,25-维生素D3的成骨诱导液其矿化形成明显增强;(2)含有1,25-二羟维生素D3的成骨诱导液诱导后ALP、RUNX2和OCN的基因表达明显增加;(3)适当浓度的地塞米松和1,25-二羟维生素D3可以促进人颞下颌关节滑液间充质干细胞的成骨分化,从而可为其自体应用修复颞下颌关节组织提供依据。BACKGROUND:Synovial fluid mesenchymal stem cells can be amplified rapidly in vitro and collected by a minimally invasive method.Recent studies have suggested that synovial fluid mesenchymal stem cells have become an important kind of seed cells for bone tissue engineering.Osteogenic differentiation is required to be optimized prior to the application of synovial fluid mesenchymal stem cells in the bone regeneration.OBJECTIVE:To describe a protocol to generate osteoblast-lineage cells from human synovial fluid mesenchymal stem cells of the temporomandibular joint using a cocktail that includes glutamax,dexamethasone,β-glycerophosphate,vitamin C,1,25-dihydroxyvitamin D3,and to investigate the effect of 1,25-dihydroxyvitamin D3 and dexamethasone on the osteogenic capacity of synovial fluid mesenchymal stem cells.METHODS:Synovial fluid mesenchymal stem cells from the human temporomandibular joint were expanded in vitro and cultured in different osteogenic induction media.The mineralization capacity of osteogenic differentiation was evaluated by alizarin red and Von kossa staining.And the osteogenic markers,including ALP,RUNX2 and OCN,were assessed by reverse transcription-PCR.RESULTS AND CONCLUSION:The mineralization formation increased greatly in the medium with 100 nmol/L dexamethasone and 10 nmol/L 1,25-dihydroxyvitamin D3.The expression of ALP,RUNX2 and OCN was enhanced distinctly in the 1,25-dihydroxyvitamin D3-induced differentiated cells.These findings reveal that appropriate concentration of 1,25-dihydroxyvitamin D3 and dexamethasone can be ideal ingredients to induce the osteogenic differentiation of human synovial fluid mesenchymal stem cells of the temporomandibular joint.Thus,this effective condition can be used to induce the osteogenic differentiation of synovial fluid mesenchymal stem cells for the bone regeneration in the temporomandibular joint.

关 键 词：滑液 间质干细胞 细胞分化 骨化三醇 地塞米松 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]