自体骨髓间充质干细胞和同种异体肋软骨细胞共培养修复五指山小型猪膝关节软骨缺损的实验研究

发布时间：2018-07-06 09:59

本文选题：组织工程 + 骨髓间充质干细胞　；参考：《中南大学学报(医学版)》2017年08期

【摘要】：目的:探讨骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)和肋软骨细胞(costal chondrocytes,CCs)共培养构建组织工程软骨及共培养细胞修复五指山小型猪膝关节软骨缺损的可行性,为临床修复关节软骨缺损提供理论依据和奠定实验基础。方法:密度梯度离心法分离五指山小型猪BMSCs,双酶消化法分离CCs。取第3代BMSCs和第2代CCs,随机分为3组:BMSCs:CCs为1:2的共培养组为A组,单纯CCs为B组,单纯BMSCs为C组。绘制细胞生长曲线图,测定软骨细胞外分泌基质糖胺多糖(glycosaminoglycan,GAG)的能力。将12只五指山小型猪随机分为共培养细胞/胶原膜实验组、胶原膜对照组和空白组。共培养细胞/胶原膜实验组髁间窝软骨缺损处植入共培养细胞/胶原膜,胶原膜对照组单纯植入胶原膜,空白组不做任何植入,于术后第8和16周分别处死6只动物,每组2只。取材行大体观察、软骨组织学评分及病理组织学检测。结果:密度梯度离心法分离的BMSCs生长良好,双酶消化法分离的CCs生物活性良好,共培养细胞生长良好。术后16周共培养细胞/胶原膜实验组修复组织呈透明软骨样,表面光滑平坦,周围软骨及软骨下骨整合良好,而胶原膜对照组和空白组为纤维性修复和无修复。共培养细胞/胶原膜实验组修复组织大体评分明显优于胶原膜对照组和空白组(均P0.05),胶原膜对照组和空白组之间差异无统计学意义(P0.05)。结论:BMSCs,CCs和共培养细胞均适合作为软骨组织工程的种子细胞,其中共培养细胞(BMSCs:CCs为1:2)更具优势;共培养细胞复合胶原膜修复软骨缺损近期效果满意。
[Abstract]:Objective: to investigate the feasibility of co-culture of bone marrow mesenchymal stem cells (bone marrow mesenchymal stem cells) and costal chondrocytes (costal chondrocytes) to construct tissue engineered cartilage and co-culture cells to repair articular cartilage defects in Wuzhishan miniature pig. To provide theoretical basis and experimental basis for clinical repair of articular cartilage defects. Methods: BMSCs were isolated from Wuzhishan miniature pig by density gradient centrifugation and CCs were isolated by double enzyme digestion. The third generation of BMSCs and the second generation of CCS were randomly divided into three groups: group A, group B, group C, group B: BMSCs: 1. Cell growth curves were plotted to determine the ability of chondrocytes to secrete glycosaminoglycan polysaccharide (GAG). Twelve Wuzhishan miniature pigs were randomly divided into co-cultured cells / collagen membrane experimental group, collagen membrane control group and blank group. The co-cultured cells / collagen membrane was implanted into the cartilage defect of the intercondylar fossa in the experimental group, the collagen membrane was implanted only in the control group, and the blank group was not implanted at all. Six animals were killed at the 8th and 16th week after operation. 2 rats in each group. Gross observation, cartilage histological score and histopathological examination were performed. Results: BMSCs isolated by density gradient centrifugation grew well, CCs isolated by double enzyme digestion had good bioactivity and co-cultured cells grew well. 16 weeks after operation, the cultured cells / collagen membrane in the experimental group showed hyaline cartilage, smooth surface and good integration of the surrounding cartilage and subchondral bone, while the collagen membrane control group and the blank group were fibrous repair and no repair. The gross score of repair tissue in co-cultured cells / collagen membrane group was significantly better than that in collagen membrane control group and blank group (P0.05), but there was no significant difference between collagen membrane control group and blank group (P0.05). Conclusion both the CCs and the co-cultured cells are suitable for the seed cells of cartilage tissue engineering, among which the co-cultured cells (BMSCs: CCs is 1:2) have more advantages, and the short-term effect of the co-cultured cells combined with collagen membrane on the repair of cartilage defects is satisfactory.
【作者单位】：中南大学湘雅二医院骨科;中南大学湘雅医学院附属海口医院骨科中心;
【基金】：海南省自然科学基金(814379)~~
【分类号】：R684

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