猪腹膜脱细胞基质联合微骨折技术修复兔膝关节软骨缺损

Application of Combined Pig Peritoneum-derived Acellular Matrix and Microfracture Technique for Repairing Articular Cartilage Defect in Rabbit

目的:检测猪腹膜脱细胞基质(pig peritoneum-derived acellular matrix,PPAM)支架材料联合微骨折修复兔膝关节软骨缺损的效果。方法:采用CCK-8 Kit测定SD大鼠骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMMSCs)在PPAM支架上的生长状态,Live/Dead染色观察细胞在PPAM支架上的活性。用直径4mm环钻造成深度约1.5 mm的新西兰大白兔膝关节软骨缺损,微骨折术后将PPAM支架植入软骨缺损处,单纯微骨折技术(Microfracture,MF)作为对照组,分别在第6周、12周和24周取材。大体观察、HE染色、甲苯胺蓝染色和II型胶原(COL II)免疫组化染色检测软骨缺损修复的效果。结果:CCK-8 Kit和Live/Dead染色结果显示PPAM支架支持细胞生长和增殖,无细胞毒性。大体观察和组织学检测显示PPAM联合MF修复软骨缺损能促进软骨缺损填充组织的质量,组织学评分优于单纯MF组。结论:PPAM支架是一种有良好生物相容性的天然材料来源组织工程材料,联合MF技术能促进关节软骨缺损的修复。

Objective To evaluate the outcome of articular cartilage repair using pig peritoneum-derived acellular peritoneum matrix（PPAM）in combination with microfracture（MF）technique in rabbit. Methods The growth of bone marrow-derived mesenchymal stem cells（BMMSCs）and the activity of BMMSCs on the PPAM was respectively observed by CCK-8 Kit and Live/Dead staining. The cartilage defect（4mm in diameter and1.5mm in depth）in knee trochlea of rabbit was made by a trephine. A part of the defects was implanted with PPAM in treatment group,and the remaining defects were served as MF group. Gross observation,HE staining,toluidineblue staining and immunohistochemical staining of collagen type II were used to evaluate the outcome of articular cartilage repair at 6,12 and 24 weeks after the operation. Results The PPAM supported the growth and proliferation of BMMSCs well enough without any cytotoxicity. The histological score in treatment group was significantly higher than in the MF group. Conclusions The combination of PPAM and microfracture technique was beneficial for articular cartilage repair due to its good biocompatibility.