CAJ | 학술논문

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 mL，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2×10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。7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。
배경：대량래원우동물실험혹실험실적증거표명골수간충질간세포구유강유력적면역억제、항염증、연골재생능력。단시상대우동물실험，골관절염환자적관절미배경경복잡，섭급대량적생물활성인자급면역궤제。목적：분석경피천자관절강주사자체골수간충질간세포치료경중도골성관절염환자적료효，탐토기염증억제、면역조절、완해연골강해등궤제。방법：수집26례경중도골성관절염환자골수15 mL，재실험실분리、배양、획득족량적골수간충질간세포，병이류식세포의검측기표면표지물，분석기체외유도성골、성지분화능력。26례환자쌍측슬관절분별입조실험조급대조조，실험조슬주사자체래원적체외배양적골수간충질간세포(2×107결과여결론：안조표준화적세포분리、배양확증방안，능획득족량적용우관절강주사적골수간충질간세포(초과2×10개)，대조조칙주사등량불함세포안위제。채용WOMAC평분평개치료전후슬관절공능변화。재치료전、치료후2주，이용ELISA검측관절액중백세포개소1、종류배사인자α、백세포개소10、연골과취기질단백수평。7제3대세포)。종환자분리출래적골수간충질간세포균양성표체CD105、CD29，음성표체CD45、CD34，체외구유량호적성골、성지능력。치료후환자적슬관절공능명현개선，이차상대우대조조，실험조관절액중백세포개소1、종류배사인자α、연골과취기질단백수평명현하강，이백세포개소10수평상승。이상결과표명，관절강주사자체골수간충질간세포치료경중도골성관절염구유량호적료효，진일보추측주사도관절강내적골수간충질간세포유가능통과염증억제급역전연골강해등궤제발휘작용，종이유망성위치료경중도골성관절염유효、가행적방법。
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 (2×107 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 2×107 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 capacities. The function of the knee joints was improved obviously after treatment. In the experimental group, the secretion of both interleukin-1, tumor necrosis factor-αand cartilage oligomeric matrix protein in the joint fluid was suppressed, while the interleukin-10 level was up-regulated compared to the control group. The results suggest that intra-articular injection of autologous bone marrow mesenchymal stem cel s can exert good effect in mild-to-moderate osteoporosis patients. Furthermore, bone marrow mesenchymal stem cel s via intra-articular injection can function through anti-inflammation and reversion of degradation of the articular cartilage, which is a new promising approach for treating mild-to-moderate osteoarthritis.