IGF1 诱导的滑膜间充质干细胞条件培养基增加软骨细胞炎症中的软骨生成和软骨保护标志物。
Conditioned medium of IGF1-induced synovial membrane mesenchymal stem cells increases chondrogenic and chondroprotective markers in chondrocyte inflammation.
机构信息
Faculty of Pharmacy, Andalas University, Jl. Limau Manis, Padang 25166, West Sumatera, Indonesia.
Faculty of Medicine, Andalas University, Jl. Limau Manis, Padang 25166, West Sumatera, Indonesia.
出版信息
Biosci Rep. 2021 Jul 30;41(7). doi: 10.1042/BSR20202038.
Recently, mesenchymal stem cells (MSCs) have been the most explored cells for cell therapy for osteoarthritis (OA) that can be obtained from various sources. Synovial membrane MSCs (SMMSCs) provide best potential for OA therapy, however they are not widely explored. Conditioned medium of SMMSCs (SMMSCs-CM) rich in growth factors and cytokines can inhibit apoptosis and increase chondrocytes cell proliferation. The aim of the present study was to determine growth factors content in SMMSCs-CM as well as the chondrogenic and chondroprotective markers expression in OA model after insulin-like growth factor (IGF)1-induced and non-induced SMMSCs-CM treatments. Chondrocyte cell line (CHON002) was induced by IL1β as OA model (CHON002 with IL1β (IL1β-CHON002)) and treated with SMMSCs-CM with or without IGF1 induction to determine its effectiveness in repairing OA cells model. ELISA was used to assay BMP2, fibroblast growth factor 18 (FGF18) and transforming growth factor (TGF) β1 (TGFβ1) levels in SMMSCs-CM, matrix metalloproteinase (MMP) 13 (MMP13) and a disintegrin and metalloproteinase with thrombospondin motif 4 (ADAMTS4) levels in OA cells model treated with SMMSCs-CM. RT-qPCR analyses were used to investigate the gene expression of SOX9, COL2, and COL10. CM from SMMSCs cultured and induced by IGF1 150 ng/mL was the most effective concentration for increasing the content of growth factor markers of SMMSCs-CM, which had successfully increased negative cartilage hypertrophy markers (SOX9 and COL2) and reduced hypertrophy markers (COL10, MMP13, and ADAMTS4). Preconditioning with IGF1 has better and very significant results in lowering MMP13 and ADAMTS4 levels. The present study supports IGF1 pre-conditioned SMMSCs-CM to develop a new therapeutic approach in OA improvement through its chondrogenic and chondroprotective roles.
最近,间充质干细胞(MSCs)已成为关节炎(OA)细胞治疗中最受关注的细胞,可以从多种来源获得。滑膜间充质干细胞(SMMSCs)为 OA 治疗提供了最佳的潜力,但尚未广泛探索。富含生长因子和细胞因子的 SMMSCs 条件培养基(SMMSCs-CM)可以抑制细胞凋亡并增加软骨细胞的增殖。本研究旨在确定 SMMSCs-CM 中的生长因子含量,以及 IGF1 诱导和非诱导 SMMSCs-CM 处理后 OA 模型中的软骨生成和软骨保护标志物的表达。通过 IL1β 诱导 CHON002 细胞系(CHON002)作为 OA 模型(用 IL1β 诱导的 CHON002(IL1β-CHON002)),并用 SMMSCs-CM 处理,无论是否有 IGF1 诱导,以确定其在修复 OA 细胞模型中的效果。ELISA 用于测定 SMMSCs-CM 中的 BMP2、成纤维细胞生长因子 18(FGF18)和转化生长因子(TGF)β1(TGFβ1)水平,OA 细胞模型中 SMMSCs-CM 处理后的基质金属蛋白酶(MMP)13 和 a 分解素和金属蛋白酶与血小板反应蛋白基序 4(ADAMTS4)水平。RT-qPCR 分析用于研究 SOX9、COL2 和 COL10 的基因表达。培养和 IGF1 诱导的 SMMSCs 培养的 CM,浓度为 150ng/ml,是增加 SMMSCs-CM 中生长因子标志物含量的最有效浓度,成功增加了负性软骨肥大标志物(SOX9 和 COL2),降低了肥大标志物(COL10、MMP13 和 ADAMTS4)。IGF1 预处理具有更好和非常显著的降低 MMP13 和 ADAMTS4 水平的效果。本研究支持 IGF1 预处理的 SMMSCs-CM 通过其软骨生成和软骨保护作用,为改善 OA 提供新的治疗方法。