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

doi:10.3969/j.issn.2095-4344.2015.14.015

Abstract

Abstract:

BACKGROUND: Increasing evidence collected from animal experiments or laboratories shows that bone mesenchymal stem cells 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 cells 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 cells 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 cells by intra-articular injection (2×10⁷ cells), while the control group injected the same volume of control medium without bone marrow mesenchymal stem cells. Western Ontario and McMaster Universities Osteoarthritis Index was used to estimate the function of the knee joints before and after the treatment. We collected 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 cells (more than 2×10⁷ passage 3 cells) were obtained for intra-articular injection. The bone marrow mesenchymal stem cells 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-α and 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 cells can exert good effect in mild-to-moderate osteoporosis patients. Furthermore, bone marrow mesenchymal stem cells 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.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: Bone Marrow, Mesenchymal Stem Cell Transplantation, Osteoarthritis

CLC Number:

R394.2

Liang Jian-ji, He Zhi-yong, Liu Kang, Li Xiao-ling, Cheng Wei-min, Yu Xin-ping, Chen Er-dong. Intraarticular injection of autologous bone marrow mesenchymal stem cells for mild-to-moderate osteoarthritis [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(14): 2216-2223.

Figures/Tables 7

2.5 参与者数量分析 26例膝关节骨性关节炎患者均进入结果分析。 2.6 治疗前后患者膝关节功能变化的结果分析治疗前、治疗后2周、治疗后4周膝关节的WOMAC评分数据的分布符合正态(正态性检验采用Shapiro-Wilk检验，设定检验水准α=0.1)，采用x(_)±s进行数据描述，以配对t 检验进行比较。治疗后2，4周的膝关节功能评分[(39.62±15.27)分，(33.65±14.07)分]明显低于治疗前(82.27±12.76)分，差异有显著性意义(P < 0.05)，见图7。治疗后2周膝关节功能评分与治疗后4周比较，差异无显著性意义，结合临床表现可见，相对于治疗后2周，治疗后4周患者的临床症状改善不大。 2.7 实验组及对照组治疗前后关节液中白细胞介素1、白细胞介素10、软骨寡聚基质蛋白、肿瘤坏死因子α水平变化治疗前，对实验组及对照组白细胞介素1、肿瘤坏死因子α、白细胞介素10、软骨寡聚基质蛋白资料基线水平进行了比较，因各独立样品资料不符合正态分布，故分别采用两独立样本的Mann-Whitney U检验，设定检验水准 P=0.05，得出各组资料的统计结果差异无显著性意义(P > 0.05)，见表1，治疗前实验组及对照基线水平一致，故具有可比性。统计分析显示治疗前后白细胞介素1、肿瘤坏死因子α、白细胞介素10、软骨寡聚基质蛋白的差值分布不符合正态分布，采用配对资料的秩和检验。实验组经皮膝关节注射自体骨髓间充质干细胞2周后，关节液中白细胞介素1、肿瘤坏死因子α和软骨寡聚基质蛋白明显下降，白细胞介素10明显升高，实验组治疗前后的白细胞介素1、肿瘤坏死因子α、白细胞介素10、软骨寡聚基质蛋白资料经统计，差异均有显著性意义(P < 0.05)；对照组治疗前后白细胞介素1、肿瘤坏死因子α资料经统计发现差异无显著性意义(P > 0.05)，而白细胞介素10及软骨寡聚基质蛋白资料经统计发现差异有显著性意义(P < 0.05)，见图8和表2。通过比较正负平均秩和得出对照组治疗后白细胞介素10平均水平降低，软骨寡聚基质蛋白平均水平升高，见表3。说明骨髓间充质干细胞的植入能很大程度上抑制炎症因子白细胞介素1、肿瘤坏死因子α的产生，促进抗炎症因子白细胞介素10的产生，使软骨降解产物软骨寡聚基质蛋白下降，从而缓解软骨的降解。 2.8 不良反应注射骨髓间充质干细胞过程中未发生发热、感染、软骨软化等不良反应。"

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