软骨素糖胺聚糖对二维培养和与 OA 骨软骨外植体共培养中人骨髓间充质干细胞向软骨分化的影响。

Influence of Kartogenin on Chondrogenic Differentiation of Human Bone Marrow-Derived MSCs in 2D Culture and in Co-Cultivation with OA Osteochondral Explant.

机构信息

Associated Tissue Bank of Faculty of Medicine of P. J. Safarik University and L. Pasteur University Hospital, Trieda SNP 1, 04011 Kosice, Slovakia.

Department of Orthopaedics and Traumatology of Faculty of Medicine of P. J. Safarik University and L. Pasteur University Hospital, Trieda SNP 1, 04011 Kosice, Slovakia.

出版信息

Molecules. 2018 Jan 16;23(1):181. doi: 10.3390/molecules23010181.

Articular cartilage has limited capacity for natural regeneration and repair. In the present study, we evaluated kartogenin (KGN), a bioactive small heterocyclic molecule, for its effect on in vitro proliferation and chondrogenic differentiation of human bone marrow-derived mesenchymal stromal cells (hBMSCs) in monolayer culture and in co-culture models in vitro. OA osteochondral cylinders and hBMSCs were collected during total knee replacement. The effect of KGN on hBMSCs during 21 days of culture was monitored by real-time proliferation assay, immunofluorescence staining, histological assay, scanning electron microscopy (SEM) (imaging and multiplex enzyme-linked immunosorbent assay) ELISA assay. The rate of proliferation of hBMSCs was significantly increased by treatment with 10 µM KGN during nine days of culture. Histological and SEM analyses showed the ability of hBMSCs in the presence of KGN to colonize the surface of OA cartilage and to produce glycosaminoglycans and proteoglycans after 21 days of co-culture. KGN treated hBMSCs secreted higher concentrations of TIMPs and the secretion of pro-inflammatory molecules (MMP 13, TNF-α) were significantly suppressed in comparison with control without hBMSCs. Our preliminary results support the concept that 10 µM KGN enhances proliferation and chondrogenic differentiation of hBMSCs and suggest that KGN is a potential promoter for cell-based therapeutic application for cartilage regeneration.

关节软骨的自然再生和修复能力有限。本研究评估了生物活性小分子 kartogenin（KGN）对单层培养和体外共培养模型中人骨髓间充质基质细胞（hBMSCs）体外增殖和软骨分化的影响。在全膝关节置换期间收集 OA 骨软骨圆柱和 hBMSCs。通过实时增殖测定、免疫荧光染色、组织学测定、扫描电子显微镜（SEM）（成像和多重酶联免疫吸附试验）ELISA 测定监测 KGN 在 21 天培养期间对 hBMSCs 的影响。在 9 天的培养过程中，用 10 μM KGN 处理可显著增加 hBMSCs 的增殖率。组织学和 SEM 分析表明，在 KGN 存在的情况下，hBMSCs 能够在 OA 软骨表面定植，并在共培养 21 天后产生糖胺聚糖和蛋白聚糖。与无 hBMSCs 的对照相比，用 KGN 处理的 hBMSCs 分泌更高浓度的 TIMPs，并且促炎分子（MMP13、TNF-α）的分泌明显受到抑制。我们的初步结果支持 10 μM KGN 增强 hBMSCs 增殖和软骨分化的概念，并表明 KGN 是一种潜在的促进细胞治疗软骨再生的应用的物质。