髂骨来源的“双相”BMG结合软骨细胞修复兔关节软骨缺损的实验研究

摘要

Objective:To study the effectiveness and feasibility of the “bipolar”BMG which come from ilium combined with chondrocytes transplanted to repair articular cartilage defects.Methods:Culture the primary rabbit cartilage cells with the enzyme diges-tion method in vitro.Construct “bipolar”BMG scaffold material with the autogenous iliac.Construct cells-BMG composites with the cartilage cells and BMG scaffold material.The 27 cartilage defect animal models were randomly divided into three groups.The cells-BMG composite was transplanted to repair cartilage defect model as experimental group.The BMG scaffold material transplanted to re-pair cartilage defect model as materials control group.The defect model was without any treatment as blank control group.The repair effect was evaluated by gross morphology (according to the ICRS rating scale),pathological histology staining and immunohistochemical staining method at 4 weeks,8 weeks and 12 weeks respectively.Results:Safranin O,toluidine blue and collagen II immunohistochemical staining demonstrate that regenerate tissue of experimental group was similar with normal cartilage,and the surface of regenerate tissue and normal cartilage tissue were almost flat at 12 weeks after surgery.However,the defects in materials control group had been repaired partly and the repair effect of blank control group was very poor.According to the ICRS rating scale,the repair effect scores of gross morphology and pathologic histology showed that the scores of experimental group were significant higher than the other two groups at 12 weeks after surgery (P <0.05).Conclusion:“Bipolar”BMG combined with cartilage cells can form cartilage tissue in vivo which pos-sess a certain structure function.The tissue engineering cartilage can be used in regenerative repair of cartilage defects.%目的：探讨应用髂骨来源的“双相”骨基质明胶（bone matrix gelatin，BMG）结合软骨细胞移植修复兔关节软骨缺损的有效性和可行性。方法：使用酶消化法体外分离培养并扩增兔肋软骨细胞；使用自体来源髂骨构建“双相”BMG 支架材料；将软骨细胞种植与 BMG 支架材料上构建成为细胞-BMG 复合物；将制作好的27只软骨缺损动物模型随机分为3组。将细胞-BMG 复合物移植于软骨缺损模型进行修复治疗作为实验组。将 BMG 支架移植入软骨缺损模型作为材料组；空白对照组不做处理。分别在术后4周、8周和12周通过大体形态学（依据国际软骨修复协会评分量表）、病理组织学染色及免疫组织化学染色方法对各组修复效果进行评价。结果：番红 O 染色，甲苯胺蓝染色和 Collagen II 免疫组化染色分别提示术后12周时实验组的修复组织非常类似于正常关节软骨，组织表面与正常软骨组织平面几乎持平，材料组软骨缺损处得到了部分修复，而缺损组的修复效果较差。按照 ICRS 量表，大体形态学和病理组织学评分结果提示：术后12周实验组的修复效果与其他两组相比有明显统计学差异（P ＜0.05）。结论：自体髂骨来源的“双相”BMG 结合软骨细胞在体内可形成具有一定结构功能的软骨组织，能够应用于再生修复软骨的缺损。