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该研究探讨从颞下颌关节骨关节病患者的关节冲洗液中获取滑膜间充质干细胞(synovial mesenchymal stem cells,SMSCs)并进行体外培养,研究其间充质干细胞特性。采用离心、贴壁培养法从颞下颌关节骨关节病患者的关节冲洗液中提取并分离细胞,进行细胞形态学观察、细胞生长及周期的分析,细胞表面物标志物流式细胞术检测,间充质干细胞标志物免疫荧光检测,成骨、成脂、成软骨多向分化能力检测,促血管分泌因子检测。结果显示,通过离心、贴壁培养法可稳定地从患者颞下颌关节冲洗液中获取滑膜间充质干细胞,体外培养呈对数生长。SMSCs表达间质源性细胞的特异性标记物CD73、CD90、CD105、波形蛋白(Vimentin)以及α平滑肌肌动蛋白的特异性标记物α-SMA。经诱导培养基诱导,SMSCs可分化为骨、软骨和脂肪细胞,细胞分泌液中可检测到如成纤维细胞生长因子(fibroblast growth factor,FGF)、angiopoietin、转化生长因子-β(transforming growth factor-β,TGF-β)等10余种促血管生长因子。以上结果表明,该研究成功建立了以微创的方式获取颞下颌关节骨关节病患者的滑膜间充质干细胞,有望为组织工程应用于骨软骨修复提供良好种子细胞的来源。
This study investigated synovial mesenchymal stem cells (SMSCs) from synovial fluid of patients with temporomandibular joint osteoarthrosis and cultured them in vitro to study the characteristics of mesenchymal stem cells. Centrifugal and adherent culture methods were used to extract and separate the cells from the joint rinse fluid of patients with temporomandibular joint osteoarthrosis. Cell morphological observation, cell growth and cycle analysis, cell surface marker flow cytometry were performed, Immunofluorescence detection of mesenchymal stem cell markers, osteogenic, adipogenic, multi-directional differentiation into cartilage, and detection of pro-angiogenic factors. The results showed that synovial MSCs could be stably obtained from temporomandibular joint rinse solution by centrifugation and adherent culture, and the growth in vitro was logarithmic growth. SMSCs express stromal-derived markers CD73, CD90, CD105, vimentin and α-SMA, a specific marker of α-smooth muscle actin. After induced by induction medium, SMSCs can differentiate into bone, cartilage and adipose cells, detectable in cell exudates such as fibroblast growth factor (FGF), angiopoietin, transforming growth factor- β, TGF-β) and more than 10 kinds of pro-angiogenic growth factor. The above results show that the successful establishment of a minimally invasive manner to obtain temporomandibular joint osteoarthritis patients with synovial mesenchymal stem cells is expected to tissue engineering for osteochondral repair to provide a good source of seed cells.