关节腔注射自体骨髓间充质干细胞治疗轻中度骨性关节炎  

Intraarticular injection of autologous bone marrow mesenchymal stem cells for mild-to-moderate osteoarthritis

作　　者：梁建基 何智勇[1] 刘康[1] 李晓玲[1] 程伟民[1] 余新平[1] 陈尔东[1] 

机构地区：[1]中山市人民医院,广东省中山市528400

出　　处：《中国组织工程研究》2015年第14期2216-2223,共8页Chinese Journal of Tissue Engineering Research

摘　　要：背景:大量来源于动物实验或实验室的证据表明骨髓间充质干细胞具有强有力的免疫抑制、抗炎症、软骨再生能力。但是相对于动物实验,骨关节炎患者的关节微环境更复杂,涉及大量的生物活性因子及免疫机制。目的:分析经皮穿刺关节腔注射自体骨髓间充质干细胞治疗轻中度骨性关节炎患者的疗效,探讨其炎症抑制、免疫调节、缓解软骨降解等机制。方法:收集26例轻中度骨性关节炎患者骨髓15 m L,在实验室分离、培养、获得足量的骨髓间充质干细胞,并以流式细胞仪检测其表面标志物,分析其体外诱导成骨、成脂分化能力。26例患者双侧膝关节分别入组实验组及对照组,实验组膝注射自体来源的体外培养的骨髓间充质干细胞(2×107个),对照组则注射等量不含细胞安慰剂。采用WOMAC评分评价治疗前后膝关节功能变化。在治疗前、治疗后2周,利用ELISA检测关节液中白细胞介素1、肿瘤坏死因子α、白细胞介素10、软骨寡聚基质蛋白水平。结果与结论:按照标准化的细胞分离、培养扩增方案,能获得足量的用于关节腔注射的骨髓间充质干细胞(超过2×107第3代细胞)。从患者分离出来的骨髓间充质干细胞均阳性表达CD105、CD29,阴性表达CD45、CD34,体外具有良好的成骨、成脂能力。治疗后患者的膝关节功能明显改善,而且相对于对照组,实验组关节液中白细胞介素1、肿瘤坏死因子α、软骨寡聚基质蛋白水平明显下降,而白细胞介素10水平上升。以上结果表明,关节腔注射自体骨髓间充质干细胞治疗轻中度骨性关节炎具有良好的疗效,进一步推测注射到关节腔内的骨髓间充质干细胞有可能通过炎症抑制及逆转软骨降解等机制发挥作用,从而有望成为治疗轻中度骨性关节炎有效、可行的方法。BACKGROUND：Increasing evidence col ected from animal experiments or laboratories shows that bone mesenchymal stem cel s possess potent immunosuppression and anti-inflammation effects and cartilage regenerative capability. The microenvironment in human knee joint of osteoporosis is more complex and involves lots of bioactive factors and immunologic mechanisms. OBJECTIVE：The analyze the therapeutic effects of intra-articular injection of autologous bone marrow mesenchymal stem cel s on mild-to-moderate osteoporosis, and to investigate the mechanisms for anti-inflammation, immunoregulation and reversion of cartilage degradation. METHODS：About 15 mL bone marrow was aspired from 26 patients with mild-to-moderate osteoporosis and taken to the laboratory where bone marrow mesenchymal stem cel s were isolated and characterized in terms of some surface markers by a flow cytometer, and the ability of osteogenic and adipogenic differentiation was analyzed. The bilateral knees of each patient were divided into two groups at random. The experiment group were treated with autologous bone marrow mesenchymal stem cel s by intra-articular injection （2＆#215;107 cel s）, while the control group injected the same volume of control medium without bone marrow mesenchymal stem cel s. Western Ontario and McMaster Universities Osteoarthritis Index was used to estimate the function of the knee joints before and after the treatment. We col ected the joint fluid before, 2 and 4 weeks post treatment, and then measured the production of interleukin-1, interleukin-10, tumor necrosis factor-α, cartilage oligomeric matrix protein using ELISA. RESULTS AND CONCLUSION：According to the standardized culture-expansion protocol, a sufficient number of bone marrow mesenchymal stem cel s （more than 2＆#215;107 passage 3 cel s） were obtained for intra-articular injection. The bone marrow mesenchymal stem cel s isolated from patients were positive for CD105, CD29 and negative for CD45, CD34, and had the fair osteogenic and adipogenic cap

关 键 词：干细胞 移植 骨髓间充质干细胞 骨性关节炎 白细胞介素1 肿瘤坏死因子α 白细胞介素10 软骨寡聚基质蛋白 炎症抑制 软骨降解 

分 类 号：R394.2[医药卫生—医学遗传学]