中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (5): 668-672.doi: 10.3969/j.issn.2095-4344.1606

• 干细胞移植 stem cell transplantation • 上一篇    下一篇

骨髓间充质干细胞移植联合川芎嗪膝关节腔注射治疗兔膝关节骨关节炎

李 巍1，冼 呈2，赵劲民1，劳 山1   

  1. 1广西医科大学第一附属医院骨关节外科，广西壮族自治区南宁市 530021；2广西医科大学附属埌东医院，广西壮族自治区南宁市 530021
  • 修回日期:2018-11-15 出版日期:2019-02-18 发布日期:2019-02-18
  • 通讯作者: 劳山，主任医师，广西医科大学第一附属医院骨关节外科，广西壮族自治区南宁市 530021
  • 作者简介:李巍，男，1984年生，河北省饶阳县人，汉族，2011年广西医科大学毕业，硕士，主治医师，主要从事关节与运动医学研究。
  • 基金资助:

    中医药自筹课题(GZZC15-33)，项目负责人：李巍；广西医科大学青年基金项目(GXMUYSF201618)，项目负责人：李巍

Intra-articular injection of bone marrow mesenchymal stem cells combined with ligustrazine in a rabbit knee osteoarthritis model

Li Wei1, Xian Cheng2, Zhao Jinmin1, Lao Shan1   

  1. 1Department of Osteoarthrosis, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China; 2Langdong Hospital, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Revised:2018-11-15 Online:2019-02-18 Published:2019-02-18
  • Contact: Lao Shan, Chief physician, Department of Osteoarthrosis, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • About author:Li Wei, Master, Attending physician, Department of Osteoarthrosis, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Supported by:

    the Self-Financing Project of Traditional Chinese Medicine, No. GZZC15-33 (to LW); the Youth Foundation of Guangxi Medical University, No. GXMUYSF201618 (to LW)

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摘要：

文章快速阅读：

文题释义：
骨髓间充质干细胞在骨关节炎中的应用：骨关节炎的最终病理发展为关节软骨损伤，因此，治疗骨关节炎的关键是阻止这一过程的进展，使受损软骨细胞及时得到修复。骨髓间充质干细胞是一种多能成体干细胞，具有多向分化及自我更新的潜能，还具有明显的可塑性，易于体外培养、扩增。研究表明，骨髓间充质干细胞被证明具有较强的软骨分化能力，体内可迁移到损伤处，参与损伤组织的修复。
川芎嗪：是从川芎生物碱中分离得到的有效单体，能干扰血流、红细胞变形、白细胞黏附、血小板聚集和血栓溶解等血液流变学事件，对改善血液循环有极好的作用。软骨作为微循环较差的组织之一，应用改善微循环的药物，或许可以增强其自我修复的能力。

 

摘要
背景：单纯应用骨髓间充质干细胞或单纯应用川芎嗪治疗骨关节炎疗效有限。骨髓间充质干细胞移植复合川芎嗪是否能起到更好的效果未见报道。
目的：了解川芎嗪对骨髓间充质干细胞成软骨分化及软骨再生的作用。
方法：分离出新西兰白兔(1周龄，广西医科大学动物实验中心提供)骨髓间充质干细胞，培养传至第3代。将健康新西兰大白兔(五六个月龄，广西医科大学动物实验中心提供)40只随机分成4组进行实验：除空白对照组外，其余3组应用伸直位石膏固定法建立兔膝关节骨关节炎模型，所有建模部位均为右侧后肢。造模成功后第2天，空白对照组兔膝关节腔注射生理盐水0.3 mL，每周1次，共4次；单纯川芎嗪组兔膝关节腔注射川芎嗪0.3 mL，每周1次，共4次；单纯干细胞移植组兔膝关节腔内一次性注射骨髓间充质干细胞0.3 mL(细胞浓度为3×108 L-1)，同时膝关节腔内注射生理盐水0.3 mL，每周1次；干细胞移植联合川芎嗪组兔膝关节腔内一次性注射骨髓间充质干细胞0.3 mL(细胞浓度为3×108 L-1)，同时膝关节腔内注射川芎嗪0.3 mL，每周1次，共4次；干预后第6，12周采集各组兔膝关节液。
结果与结论：①空白对照组膝关节软骨无明显磨损，周围滑膜无明显增生及炎性改变；单纯川芎嗪组膝关节磨损较为严重，软骨破坏达到3级，但并未暴露软骨下骨，周围滑膜增生明显；单纯干细胞移植组、干细胞移植联合川芎嗪组膝关节轻度磨损，1级或2级，周围滑膜增生及炎性改变不明显，两组在大体观下无法辨别磨损严重程度的差异；②单纯川芎嗪组、干细胞移植联合川芎嗪组兔关节液中白细胞介素1β及白细胞介素6水平明显低于单纯干细胞移植组(P < 0.05)。在第6周时，也就是兔膝关节骨关节炎初期，软骨破坏不明显的时候，单纯川芎嗪组兔膝关节液炎症因子水平最低；在第12周时，干细胞移植联合川芎嗪组兔关节液炎症因子水平最低；③结果表明，川芎嗪对骨髓间充质干细胞成软骨分化及软骨再生的促进作用不明显，但是川芎嗪能明显降低膝关节骨关节炎的炎症水平，可以用来缓解膝关节骨关节炎患者的临床症状。

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
ORCID: 0000-0002-1649-5787(李巍)

关键词: 膝关节, 骨关节炎, 骨髓间充质干细胞, 骨髓间充质干细胞移植, 川芎嗪, 软骨分化, 软骨再生

Abstract:

BACKGROUND: Mesenchymal stem cells or ligustrazine alone has limited efficacy in the treatment of osteoarthritis. Whether bone marrow mesenchymal stem cell transplantation combined with ligustrazine can achieve better results has yet to be reported.
OBJECTIVE: To define whether ligustrazine has the function to promote the chondrogenic differentiation and regeneration of bone marrow mesenchymal stem cells.
METHODS: Bone marrow mesenchymal stem cells were isolated from New Zealand white rabbits (1 week old, provided by the Animal Experimental Center of Guangxi Medical University in China) and cultured to the third generation. Forty healthy New Zealand white rabbits (5-6 months old, provided by the Animal Experimental Center of Guangxi Medical University) were randomly divided into four groups (n=10 per group). Except for the blank control group, models of knee osteoarthritis were made in the right hind limbs of rabbits in the extension position using the plaster fixation method in the other three groups. On the 2nd day after the successful modeling, the rabbits in the blank control group were injected with 0.3 mL of normal saline once a week for 4 weeks. In the pure ligustrazine group, the rabbit knee joint cavity was injected with 0.3 mL of ligustrazine once a week for 4 weeks. In the cell transplantation group, the rabbit knee joint cavity was injected with one-off bone marrow mesenchymal stem cells (0.3 mL, 3×108 cells/L), and the knee joint was injected with 0.3 mL of normal saline once a week for 4 weeks. In the combination group, one-off bone marrow mesenchymal stem cells (0.3 mL, 3×108 cells/L) were injected into the rabbit knee joint, while intra-articular injection of ligustrazine, 0.3 mL, once a week, for 4 weeks, were simultaneously performed. The knee joint fluid of each group was collected at 6 and 12 weeks after the intervention.
RESULTS AND CONCLUSION: (1) The knee joint cartilage of the blank control group had no obvious wear as well as no obvious hyperplasia and inflammatory changes in the surrounding synovial membrane. The knee joint of the ligustrazine group was seriously worn, and the cartilage damage was defined as grade 3, but the subchondral bone was not exposed, accompanied with obvious surrounding synovial hyperplasia. In the cell transplantation and combination groups, the knee joint was slightly worn, the cartilage damage was confirmed as grade 1 or 2, and the surrounding synovial hyperplasia and inflammatory changes were not obvious. We could not distinguish the severity of wear between the cell transplantation and combination groups under the general view. (2) Compared with the cell transplantation group, the levels of interleukin-1β and interleukin-6 in the knee joint fluid of rabbits were significantly lower in the ligustrazine group and combination group (P < 0.05). At the 6th week, when the cartilage destruction was not obvious in the early stage of rabbit knee osteoarthritis, the levels of inflammatory factors in the knee joint fluid were the lowest in the ligustrazine group. At the 12th week, the levels of inflammatory factors in the joint fluid were the lowest in the combination group. To conclude, ligustrazine cannot obviously promote the chondrogenic differentiation and regeneration of bone marrow mesenchymal stem cells, but it can significantly reduce the level of inflammatory factors and relieve clinical symptoms in patients with knee 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

Key words: Osteoarthritis, Knee, Bone Marrow, Mesenchymal Stem Cells, Drugs, Chinese Herbal, Tissue Engineering

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