Basic research for the molecular mechanism of regeneration in bone and cartilage by the fibroblast-like synovial cells

A.研究目的：成纤维样滑膜细胞（FLS）与间充质干细胞具有相似性，细胞因子调节间充质干细胞的成脂作用。我们在这里研究了FLS的多系分化潜力，以及细胞因子在FLS分化过程中的作用。B.方法：根据我们机构的人体实验指南，在骨科手术时从滑液组织中分离FLS。FLS经PPARγ配体曲格列酮诱导向脂肪样细胞分化。还通过每种分化培养基诱导来自FLS的软骨细胞样细胞和成骨细胞样细胞的分化。通过油红O染色（脂肪细胞样细胞）、Alucian蓝/番红O染色（软骨细胞样细胞）或von Kossa染色（成骨细胞样细胞）定量IFN-γ、TNF-α或IL-1β在此过程中的作用。 ...更多信息 曲格列酮诱导的脂细胞样细胞。与FLS相比，脂肪细胞样细胞中的NP-κB活性降低，并且脂肪细胞样细胞的MMP-3、IL-6和IL-8的产生也受到抑制。IFN-γ、TNF-α和IL-β可抑制FLS中PPARy和C/EBP的表达，从而抑制FLS向脂肪样细胞的分化。除了脂肪细胞样细胞分化之外，FLS通过使用每种分化培养基成功地分化成软骨细胞样细胞和成骨细胞样细胞。与FLS相比，软骨细胞样细胞或成骨细胞样细胞产生的MMP-3、IL-6和IL-8也趋于减少。D.讨论：FLS具有多向分化潜能，被认为是滑膜组织中的局部干细胞。这些结果提示FLS可能是再生治疗的候选靶细胞。FLS易于从患者体内分离，避免了接种过程中的排斥反应。此外，“。由细胞因子形成的“炎性类风湿性滑膜微环境”抑制FLS的分化过程，从而可能抑制类风湿性关节炎患者的“关节组织修复”。少

A. Purpose of Study: Fibroblast-like synovial cells (FLS) has similarity with mesenchymal stem cells, and cytokines regulate adipogenesis of mesenchymal stem cells. We examined here the multilineage differentiation potential of FLS, and the effect of cytokines during differentiation process of FLS.B.Methods : FLS were isolated from the synovial tissues at the time of orthopedic surgery in accordance with human experimental guidelines of our institution. Adipocyte-like cell differentiation from FLS was induced by PPARγ ligand, troglitazone. Differentiation of chondrocyte-like cells and osteoblast-like cells from FLS was also induced by each differentiation medium. Effect of IFN-γ, TNF-α, or IL-1β during the process was quantified by Oil red O staining (adipocyte-like cells), Alucian blue staining/Safranin O staining (chondrocyte-like cells), or von Kossa staining (osteoblast-like cells).C.Results : FLS expressed PPARy and C/EBP, key molecules for adipogenesis, and differentiated into ad … More ipocyte-like cells by troglitazone. NP-κB activity in adipocyte-like cells was diminished as compared with FLS, and the production of MMP-3, IL-6, and IL-8 from adipocyte-like cells was also inhibited. IFN-γ, TNF-α, and IL-β suppressed the expression of PPARy and C/EBP, resulting in inhibition of adipocyte-like cell differentiation from FLS. In addition to adipocyte-like cell differentiation, FLS succeeded in differentiation into chondrocyte-like cells and osteoblast-like cells by the use of each differentiation medium. The production of MMP-3, IL-6, and IL-8 from chondrocyte-like cells or osteoblast-like cells also tended to be diminished as compared with FLS.D.Discussion : FLS have the multilineage differentiation potential, and thought to be the local stem cells in synovial tissues. These results may indicate that FLS could be the candidate target cells of regeneration therapy. FLS are easy to be isolated from the patients, thus, the rejection during the inoculation is evaded. In addition, "the. inflammatory rheumatoid synovial microenviroments formed by cytokines" inhibits the differentiation process of FLS, and thus may suppress "the tissue repair of joints" in patients with rheumatoid arthritis. Less