滑液干细胞对关节盘软骨细胞的作用研究  

Study the role of synovial fluid mesenchyme stem cellson articular disc cells

作　　者：邢超 孙养鹏 张志光 XING Chao;SUAN Yang-peng;ZHANG Zhi-guang(People’s Hospital of Shenzhen Baoan District,Shenzhen 518101,China)

机构地区：[1]深圳市宝安区人民医院,广东518101 [2]中山大学光华口腔医学院·附属口腔医院广东省口腔医学重点实验室,广东510055

出　　处：《中华老年口腔医学杂志》2020年第1期38-42,共5页Chinese Journal of Geriatric Dentistry

基　　金：国家自然科学基金(项目编号:81271115)。

摘　　要：目的:初步探讨人颞下颌关节滑液来源的间充质干细胞条件培养基对大鼠关节盘软骨细胞的作用。方法:临床收取颞下颌关节紊乱病患者的滑液干细胞进行细胞培养,三向分化验证其细胞干性。激光共聚焦显微镜下观察获取的大鼠关节盘软骨细胞Ⅰ型胶原、Ⅱ型胶原免疫荧光表达,确认细胞属性。收集滑液干细胞培养24h的上清液制作条件培养基,设置空白对照组分别培养大鼠关节盘软骨细胞,CCK-8检测大鼠关节盘软骨细胞增殖情况,实时荧光定量PCR(RT-PCR)检测培养7天、14天大鼠关节盘软骨细胞相关基因Ⅰ、Ⅱ、Ⅹ型胶原表达变化。结果:获取的滑液干细胞具有成脂、成骨、成软骨三向分化潜能;大鼠关节盘软骨细胞表达Ⅰ型胶原、Ⅱ型胶原;CCK-8结果显示条件培养基组48h后细胞增殖速度较对照组明显加快(P<0.05);第7天、14天实时荧光定量PCR显示条件培养基组Ⅹ型胶原表达明显低于对照组。结论:人颞下颌关节滑液来源的间充质干细胞条件培养基促进大鼠关节盘软骨细胞增殖,抑制关节盘软骨细胞Ⅹ型胶原表达。Objective:Research the role of synovial fluid mesenchyme stem cells from patients with temporomandibular joint disorders on articular disc cells.Methods:Synovial fluid-derived adherent cells were collected during TMJ cavity irrigation arthrocentesis in patients with temporomandibular joint disorders,these cells were induced into osteoblasts,chondrocytes,adipocytes to prove the potential of multilineage differentiation.The Col 1a1,Col 2a1 of rat disc cells immunofluorescence assay in Laser Scanning Confocal Microscope.Prepare Synovial fluid Mesenchyme stem cells conditioned medium(SFMSC-CM)to culture rat TMJ disc cells,the effect of conditioned medium on cell proliferation was evaluated by Cell Counting Kit-8(CCK-8)analysis,the changes in transcription genes of Col 1a1,Col 2a1,Col 10a1 at 7 day and 14 day were assayed by real-time PCR.Results:Synovial fluid-derived cells were successfully induced into adipocytes,osteoblasts,chondrocytes.Col 1a1,Col 2a1 of rat disc cells immunofluorescence were detected in Laser Scanning Confocal Microscope.CCK-8 showed after culture 48 h the disc cells proliferation increase statistically significant in condition medium.RT-PCR revealed that in condition medium group Col 10a1 was decreased compared with control group.Conclusion:Synovial fluid mesenchyme stem cells promote the disc cells proliferation and decrease the Col 10a1 expression.

关 键 词：颞下颌关节紊乱病 颞下颌关节盘 滑液 间充质干细胞 

分 类 号：R783[医药卫生—口腔医学]