纳米微孔聚乙酸内酯联合骨髓间充质干细胞对兔损伤关节软骨结缔组织生长因子蛋白的影响

摘要

目的：研究纳米微孔聚乙酸内酯（NSP-PCL）联合骨髓间充质干细胞（BMSCs）对兔损伤关节软骨中结缔组织生长因子（CTGF）蛋白的影响。方法：将50只新西兰白兔随机分为空白组、模型组、NSP-PCL 组、BMSCs 组、NSP-PCL ＋ BMSCs 组。建立新西兰白兔双膝软骨损伤模型，术后1周将纳米微孔聚乙酸内酯（NSP-PCL）、骨髓间充质干细胞（BMSCs）、NSP-PCL ＋ BMSCs 分别注射到 NSP-PCL 组、BMSCs 组和 NSP-PCL＋ BMSCs 联合治疗组的兔软骨损伤处，连续注射3周，每周注射1次。末次给药后24 h，切取缺损软骨组织。通过 HE 染色法，对双膝关节软骨进行病理学检查；Hoeehst33342标记检测移植1周和3周后兔关节软骨中 BMSCs 的存活细胞数；免疫组化法检测软骨中 CTGF 蛋白的表达。免疫蛋白印记（Western blotting）法检测软骨中 CTGF 的蛋白水平。结果：模型组软骨层变薄，有纤维原细胞增殖，软骨表面凹陷较明显。各治疗组的损伤软骨表面凹陷处均有新生软骨细胞，软骨层较模型组厚。 BMSCs 移植3周后 BMSCs 组和 NSP-PCL ＋BMSCs联合治疗组的 BMSCs 细胞数量较移植1周后的细胞数量明显增加。免疫组化法和免疫蛋白印记法的结果显示，与模型组相比，各治疗组软骨中的 CTGF 的蛋白水平均有提高，差异有显著意义（P ＜0．05）。结论：纳米微孔聚乙酸内酯联合骨髓间充质干细胞通过提高损伤软骨中 CTGF 的蛋白水平修复损伤的软骨组织。%Objective To study nano-structured porous polycaprolactone combined with bone marrow stromal cells on CTGF expression in rabbit impaired cartilage. Methods 50 rabbits were randomly divided into normal group, model group, NSP-PCL group, BMSCs group, or NSP-PCL + BMSCs group. A model of rabbit impaired cartilage was surgically established. Then NSP-PCL, BMSCs, and NSP-PCL + BMSCs were separately administered to the latter three groups once a week from the 2nd week to the 5th week after the procedure. The impaired cartilage tissues were collected 24 h after the last administration. The cartilage tissues were pathologically examined by H & E staining. Number of surviving BMSCs was detected by Hoeehst33342 Markers 1 week and 3 weeks after transplantation. Levels of CTGF protein in cartilage were determined by immunohistochemistry and Western blotting. Results In the model group, cartilage layer became thinner, with proliferation of fibroblast cells and a obvious cartilage surface hollow; New cartilage cells appeared in the surface hollow of the impaired cartilage in each treatemnt group, with a thicker layer. The number of transplanted BMSCs after 3 weeks was significantly increased in BMSCs group and NSP-PCL + BMSCs group. As compared with the model group, levels of CTGF protein were increased in each treatment group, with significant differences (P < 0.05). Conclusions Nano-porous polycaprolactone combined with bone marrow mesenchymal stem cells can repair cartilage damage by enhancing the expression of CTGF protein.