定向诱导骨关节炎患者骨髓间充质干细胞向软骨细胞的分化

摘要

BACKGROUND:In recent years, many reports have focused on inflammatory cytokines, growth factors and mechanical loads affecting the cartilage and subchondral regeneration, but there is a lack of comprehensive understanding about the mechanism of osteoarthritis. OBJECTIVE: To explore the correlation between function status of bone marrow mesenchymal stem cels and disease progression in patients with osteoarthritis. METHODS:Femoral bone marrow was extracted from patients with femoral neck fractures (control group), mild (mild group) and severe (severe group) osteoarthritis to isolate and culture bone marrow mesenchymal stem cels. Cel counting kit-8 was used to detect the proliferative ability of bone marrow mesenchymal stem cels from different patient groups, and passage 3 bone marrow mesenchymal stem cels were subject to 2-week chondrogenic induction folowed by toluidine blue staining. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cels were isolated and cultured from the femoral bone marrow of different groups. The proliferative ability of cels in the control group was significantly higher than that in the mild and severe groups. After chondrogenic induction, bone marrow mesenchymal stem cels varied obviously in the morphology that was from fusiform to qusi-circular or polygon, the percentage of nucleoplasm became smaler, and cels were positive for toluidine blue staining. The number of chondrocytes generated in the severe group was less than that in the control group, but there was no great difference in cel morphology. These findings indicate that the occurrence of osteoarthritis is negatively correlated with the functional status of autologous bone marrow mesenchymal stem cels.%背景：近年来有关影响软骨和软骨下骨组织再生的炎症因子、生长因子以及力学负荷已有许多报道，但对骨关节炎整体发生机制还欠缺全面地了解。目的：探讨骨关节炎患者骨髓间充质干细胞功能状态与病情进展的相关性。方法：抽取股骨颈骨折患者、轻度及重度骨性关节患者股骨骨髓，贴壁法分离培养出骨髓间充质干细胞，采用CCK-8法检测各组患者骨髓间充质干细胞的增殖能力，取第3代骨髓间充质干细胞成软骨诱导培养2周后进行甲苯胺蓝染色。结果与结论：从各组患者骨髓中均培养出骨髓间充质干细胞，对照组细胞的增殖能力明显高于轻度骨性关节炎组、重度骨性关节炎组；成软骨诱导分化后骨髓间充质干细胞形态发生明显改变，由长梭形逐渐变为类圆形或多角形，核浆比例明显减小，诱导细胞甲苯胺蓝染色阳性。重度骨性关节炎组诱导出的软骨细胞的数量少于对照组，但细胞形态差异不大。结果表明骨性关节炎的发生与自身的骨髓间质干细胞的功能形态呈现负相关趋势。