旋转反应器胶原支架的制备及复合软骨细胞修复骨软骨损伤的初步研究

目的以胶原为原料开发一种小型胶原支架材料,尝试是否适合旋转反应器来复合软骨细胞,并探讨复合细胞后修复软骨损伤的效果。方法成年猪跟腱中提取胶原,制备胶原支架,进行细胞毒性检测和生物相容性分析。将其切成1 mm~3小块,置于旋转培养器中与软骨细胞共培养,倒置相差显微镜观察细胞贴附效果。建立兔关节软骨缺损模型,编号后,将14只新西兰大白兔随机分为2组:支架材料组(n=8),兔膝关节软骨缺损区植入胶原支架材料和软骨细胞;空白对照组(n=6),不进行任何植入处理。进行HE染色后观察。结果胶原支架呈瓷白色,表面空隙均匀,无细胞毒性,生物相容性良好,但在体内降解较快。在旋转反应器中,支架可以与软骨细胞良好结合。胶原支架植入动物体内12周,虽未完全修复缺损,但已有少量软骨细胞在缺损处出现,修复效果优于对照组。结论制备的胶原支架复合软骨细胞短期内有一定的修复软骨缺损的能力,长期效果欠佳,可能与胶原支架在体内过快降解有关,尚需对制备方法进行改进。

Preparation of collagen scaffold for rotary cell culture system and effect on repairing cartilage damages of it with adhension of chondrocytes

Objective Repair of focal articular cartilage injury may be solved by tissue engineering.The preparation of proper scaffold is one of the most important problems to be solved.The aim of the study was to construct a kind of collagen scaffold,co-culture with L929 cells in rotary cell culture system,and implant into animal models with cartilage damage. Methods Collagen scaffold was prepared from achilles tendon of pigs.Collagen scaffold was cut into small pieces with the size of 1 mm~3 and co-cultured with L929 cells in rotary cell culture system,and the cytotoxicity and biocompatibility were tested.Rabbit articular cartilage damage models were prepared and divided into 2 groups:the experimental group treated by scaffold with chondrocytes(n = 8) and control group(n = 6) without treatment.After 4 or 12 weeks,the specimens were extracted after surgery. The effects on repairing cartilage injury were examined with haematoxylin and erosin staining. Results The collagen scaffold exhibited good biocompatibilities,and had no cell toxicity.It was proper to co-culture with chondrocytes in rotary cell culture system.The collagen scaffold with adhension of chondrocytes showed effect of repairing cartilage damage,but still could not completely reconstruct the damage. Conclusion The collagen scaffold with adhension of chondrocytes can repair cartilage damage in a short period,but it shows less potent effect in a long period,which may be correlated with the rapid degradation of the scaffold.The method of collagen scaffold preparation should be improved in our future work.