透明质酸钠复合外周血间充质干细胞修复软骨缺损

背景：近几年外周血来源的间充质干细胞被重视,但并未见应用其修复软骨缺损的报道。目的：将动员后外周血来源的间充质干细胞与透明质酸钠复合后注入关节腔内,探讨修复关节软骨缺损的可行性。方法：联合应用干细胞因子及粒细胞集落刺激因子动员2月龄兔,取外周血后分离间充质干细胞,进行原代及传代细胞培养。以透明质酸钠为细胞载体材料,将它与第3代间充质干细胞混合制成细胞混悬液。于3月龄兔双膝关节股骨内外髁负重区制造软骨缺损区,随机选取10只以左膝为实验组,术后1周关节腔内注射含透明质酸钠的细胞悬液0.5mL,右膝为透明质酸钠对照组,注射透明质酸钠0.5mL,其余5只作为生理盐水对照组,每周1次,连续3次。治疗后12周时处死取材,观察缺损处修复情况、新生组织类型及有无免疫反应。结果与结论：①兔外周血中分离培养的间充质干细胞与骨髓来源的形态相似,能够进行原代培养。②膝关节内注射外周血间充质干细胞与透明质酸钠的混悬液后未见红肿等现象发生,关节活动良好。③治疗后12周时,实验组部分缺损区基本被软骨样组织填满,表面较平整光滑。但部分边界尚可见,浅层纤维化较明显。实验组的修复效果明显优于透明质酸钠对照组及生理盐水对照组。说明动员后外周血间充质干细胞与透明质酸钠的混悬液短期内对关节软骨缺损具有一定的修复作用。

Hyaluronate sodium and peripheral blood mesenchymal stem cells compound for repairing cartilage defects

BACKGROUND：In recent years,peripheral blood mesenchymal stem cells（MSCs） have been greatly investigated,but no studyreports its application for the repair of cartilage defects.OBJECTIVE：To inject the compound of mobilized peripheral blood derived MSCs and hyaluronate sodium into articular cavity,and to investigate the feasibility of applying the compound to repair articular cartilage defect.METHODS：Two-month aged rabbits were used to isolate MSCs from peripheral blood after granulocyte colony-stimulating factorand stem cell factor mobilization.Primary and passage culture was to be continued.Taking hyaluronate sodium as the carrier materials,cell suspension was prepared by mixing with passage 3 MSCs.The full-thickness cartilage defects were created on both medial and lateral femoral condyles on every knee joint of three-month aged rabbits.Left knees were taken as the experimental group,injecting cell suspension（containing 0.5 mL hyaluronate sodium） into articular cavity,while right knees as control group,injecting hyaluronate sodium 0.5 mL once a week,for continuous three times.Twelve weeks later,rabbits were killed to observe the state of repairing the defects,the type of new tissues and the articular immune response.RESULTS AND CONCLUSION：①The appearance of peripheral blood derived MSCs was similar with bone marrow derived MSCs.When they were isolated from peripheral blood,primary cells could be cultured.②After injection,no inflammatory reaction and infection were seen,joint motion was well.③After injection for 12 weeks,the defects in experiment group were partially stuffed by cartilage-like tissues.The surface in defects was smooth,but parts of the boundary between repair tissue and surrounding could be seen.Superficial layer＇s fibrosis was more obviously.Effect of articular cartilage repair in the experimental group was better than the control groups.Peripheral blood derived MSCs mixed with hyaluronate sodium possess the capability of repairing articular cartilage defect.