骨髓干细胞聚乳酸羟基乙酸修复大鼠软骨缺损△

Bone marrow stem cells composited with polylactic glycolic acid frame for repairing cartilage defect in rats

摘要:

[目的] 探索骨髓间充质干细胞 （bone marrow mesenchymal stem cells, BMSCs） 与纳米聚乳酸羟基乙酸 （polyaiticgly- colic Acid, PLGA）复合物修复大鼠股骨软骨缺损的效果。[方法]从大鼠胫骨和股骨骨髓腔内获取 BMSCs 并培养传代，之后培养到纳米 PLGA 支架材料上，进行电镜下观察。将 SD 大鼠的双后肢进行手术制造股骨远端软骨缺损，右侧缺损填充 BMSCPLGA 复合材料（移植组），而左侧缺损不再做任何处理（空白对照组）。于术后 1 个月和 3 个月进行大体观察评分和组织学评分，测定并比较两组甲苯胺蓝染色和免疫组化染色光密度值。[结果]BMSCs 呈特有的螺旋状生长，PLGA 电镜下呈纤维交织网状结构，BMSCs 在材料上生长良好。植入体内后 1、3 个月移植组大体观察评分 [(8.1±0.8) vs (1.7±0.8), P＜0.001; (10.3±1.2) vs (3.8± 1.5), P＜0.001]、组织学评分 [(11.6±1.0) vs (4.0±0.9), P＜0.001; (15.5±1.0) vs (4.8±0.8), P＜0.001]、甲苯胺蓝染色 OD 值 [(0.2±0.0) vs(0.1± 0.0), P＜0.001; (0.4±0.0) vs (0.1±0.0), P＜0.001]，术后 3 个月免疫组化 II 胶原检测 OD 值 [(0.4±0.0) vs (0.1±0.0), P＜0.001 均显著优于空白组。[结论]LGA 结合 BMSCs 有效修复了大鼠软骨缺损，为软骨缺损治疗展示了可能的组织工程修改方法。

Abstract:

[Objective] To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) composied with nano-polyaiticglycolic acid (PLGA) frame on repairing femoral cartilage defect in rats. [Methods] BMSCs were obtained from the bone marrow cavity of the tibia and femur of rats, cultured and passed through, and then cultured on the nano-PLGA scaffold frame, and the graft was observed under electron microscope. Eighteen SD rats were operated on both hind limbs to create cartilage defect on the distal femur, and the defect on the right side was filled with BMSC-PLGA composited material (the graft group), whereas the defect on the left side left not further treated [the blank control (BC) group]. Gross and histological observation and scoring, as well as optical density values measured in toluidine blue staining and immunohistochemical staining were compared between the two groups. [Results] BMSCs grew in a unique spiral shape, the PLGA frame showed fiber interwoven network structure under electron microscope, and BMSCs grew well on the PLGA frame. At 1 and 3 months after implantation, the graft group proved significantly superior to the BC group in terms of gross observation scores [(8.1±0.8) vs (1.7±0.8), P＜ 0.001; (10.3±1.2) vs (3.8±1.5), P＜0.001], histological score [(11.6±1.0) vs (4.0±0.9), P＜0.001; (15.5±1.0) vs (4.8±0.8), P＜0.001], toluidine blue staining OD value [(0.2±0.0) vs (0.1±0.0), P＜0.001; (0.4±0.0) vs (0.1± 0.0), P＜0.001], in addition, immunohistochemical II collagen detection OD value [(0.4±0.0) vs (0.1±0.0), P＜0.001] 3 months postoperatively. [Conclusion] BMSCs -PLGA composition does effectively repair cartilage defects in rats in this study, demonstrating a potential tissue engineering modification method for the treatment of cartilage defects.