Euppayo Thippaporn, Siengdee Puntita, Limlenglert Pakorn, Nganvongpanit Korakot, Watanabe Gen, Kasashima Yoshinori, Arai Katsuhiko
Faculty of Veterinary Medicine, Maejo University, Chiang Mai, 50300, Thailand.
Program in Applied Biological Sciences: Environmental Health, Chulabhorn Graduate Institute, Kamphaeng Phet 6 Road, Laksi, Bangkok, 10210, Thailand.
In Vitro Cell Dev Biol Anim. 2025 May 27. doi: 10.1007/s11626-025-01049-8.
The self-renewal capacity of chondrocytes in osteoarthritis (OA) joints is limited, and mesenchymal stem cells (MSCs) are crucial in disease treatment. This study established an OA model from equine bone marrow-derived mesenchymal stem cells (eBMSCs). The eBMSCs were cultured and differentiated into chondrocytes to generate cartilage pellets, which were induced for 7 d with inflammatory cytokines, interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) to mimic OA conditions. Treated culture medium was collected to estimate enzyme activity (MMP-2, MMP-3, and MMP-9) using zymography, and the cartilage pellets were collected to estimate both anabolic gene (COL2A1) and catabolic gene expression (MMP2, MMP3, and MMP9) using qRT-PCR. Cartilage degradation was observed when induced with IL-1β + TNF-α on cartilage pellets. IL-1β + TNF-α decreased the expression levels of COL2A1 and MMP2 genes, and enhanced their enzymatic activities, while Alcian blue-positive glycosaminoglycan in cartilage pellets induced by IL-1β + TNF-α groups decreased. These results suggested that IL-1β + TNF-α induced on cartilage pellets from eBMSCs could be used as an in vitro OA model in horses.
骨关节炎（OA）关节中软骨细胞的自我更新能力有限，间充质干细胞（MSCs）在疾病治疗中至关重要。本研究从马骨髓来源的间充质干细胞（eBMSCs）建立了OA模型。将eBMSCs培养并分化为软骨细胞以生成软骨微珠，用炎性细胞因子白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）诱导7天以模拟OA情况。收集处理后的培养基，使用酶谱法估计酶活性（MMP-2、MMP-3和MMP-9），收集软骨微珠，使用qRT-PCR估计合成代谢基因（COL2A1）和分解代谢基因表达（MMP2、MMP3和MMP9）。当用IL-1β + TNF-α诱导软骨微珠时，观察到软骨降解。IL-1β + TNF-α降低了COL2A1和MMP2基因的表达水平，并增强了它们的酶活性，而IL-1β + TNF-α组诱导的软骨微珠中阿尔新蓝阳性糖胺聚糖减少。这些结果表明，在eBMSCs来源的软骨微珠上诱导的IL-1β + TNF-α可作为马的体外OA模型。
