Cultispher微载体构建组织工程软骨修复关节软骨缺损

Tissue-engineered cartilage construction by Cultispher microcarrier for articular cartilage repair

背景:纤维蛋白胶和微载体均可作为软骨组织工程的良好载体,但因力学性能差和可塑性差等缺点限制其广泛应用。目的:以负载软骨细胞的Cultispher微载体为基础,复合纤维蛋白胶,构建Cultispher微载体/纤维蛋白胶复合支架,观察其用于修复兔膝关节软骨缺损的效果。方法:将兔软骨细胞与Cultispher微载体置于搅拌式生物反应器中三维悬浮共培养,待细胞帖附至微载体表面并大量扩增后,将负载有软骨细胞的Cultispher微载体与纤维蛋白胶复合,构建Cultispher微载体/纤维蛋白胶复合支架,并用于修复兔膝关节股骨滑车软骨缺损。实验按不同的软骨缺损处植入物分为3组:MCF组以负载软骨细胞的Cultispher微载体/纤维蛋白胶复合支架修复软骨缺损;MF组以单纯Cultispher微载体/纤维蛋白胶复合支架修复软骨缺损;空白对照组旷置软骨缺损,不作任何处理。术后3,6个月取材。检测并记录大体观、大体观评分、病理学染色、病理学评分、Micro-CT扫描等指标,评估软骨修复效果。结果与结论:①大体观显示MCF组的软骨修复效果明显优于MF组和空白对照组;②番红"O"、天狼猩红病理染色结果显示MCF组的修复组织主要以透明软骨为主,而MF组和空白对照组的修复组织主要以纤维组织为主;③Micro-CT扫描结果显示,MCF组较MF组和空白对照组获得更好的软骨下骨重建;④MCF组的大体观评分与病理学评分均明显高于MF组和空白对照组;⑤结果证实,负载软骨细胞的Cultispher微载体/纤维蛋白胶复合支架能成功修复兔股骨滑车软骨缺损。

BACKGROUND:Both Cultispher microcarrier and fibrin can act as carriers in cartilage tissue engineering,but their application is limited by poor mechanical properties and poor plasticity.OBJECTIVE:To combine Cultispher microcarrier carrying chondrocytes with fibrin glue to construct Gultispher/fibrin composite scaffold,and to investigate the effect of this composite scaffold in the articular cartilage repair in a rabbit model.METHODS:Rabbit chondrocyte and Cultispher microcarriers were co-cultured in a stirred bioreactor until the chondrocytes adhered to and proliferated quickly on the microcarrier surface.Chondrocytes-seeded microcarries were then combined with fibrin glue to construct microcarrier/fibrin glue composite scaffolds,to repair trochlear cartilage defects of the knee joint in the rabbit model.In the experiment,three different treatments were respectively done for repair of cartilage defects,including implantation of chondrocytes-seeded microcarries/fibrin glue composite scaffold(MCF group),implantation of Gultispher/fibrin composite scaffold(MF group),and no treatment(blank control group).At3and6months after surgery,gross observation,histological evaluation,pathological evaluation and Micro-CT scanning were conducted to evaluate the cartilage repair effects.RESULTS AND CONCLUSION:Gross observation showed that the MCF group achieved better effect on cartilage repair,compared to the other two groups.Histopathological evaluation revealed hyaline-like cartilage tissues in the MCF group while fibrocartilage tissues were seen in the other two groups,shown by safranin O staining and sirius red staining.Micro-CT scanning results showed better subchondral bone remodeling was found in the MCF group than the other two groups.Gross observation and pathological observation showed better outcomes in the MCF group than the MF and blank control groups.To conclude,the chondrocyte-seeded Cultispher microcarrier/fibrin glue composite scaffold succeeds in the articular cartilage repair.