保存方法对同种异体软骨移植物免疫原性的影响

背景：异体软骨移植是治疗关节软骨缺损主要手段,寻找软骨组织保存方法来降低移植软骨的免疫原性已成为临床上关键性的技术问题。目的：比较慢速梯度降温法、玻璃化法与60Co射线照射＋梯度降温法对同种异体骨软骨移植物免疫原性的影响。方法：将关节软骨块随机分为4组：新鲜组不采取任何措施;慢速梯度降温组给予程序降温法保存关节软骨;玻璃化组利用玻璃化溶液处理后保存;60Co射线照射＋梯度降温组给予60Co射线照射后,再按程序降温法保存关节软骨。保存8,15,30,60d后关节软骨块经细胞培养及扩增后制备成细胞悬液。流式细胞仪检测关节软骨细胞表面主要组织相容性复合体Ⅰ,Ⅱ抗原表达率。结果与结论：经过保存后,慢速梯度降温组、玻璃化组和60Co射线照射＋梯度降温组细胞表面的主要组织相容性复合体Ⅰ和Ⅱ抗原表达率有大幅下降,与新鲜组相比有显著性差异,但前3组间没有显著性差异。各保存组主要组织相容性复合体Ⅰ类抗原和主要组织相容性复合体Ⅱ类抗原表达率均随着时间的推移下降,并且在30d时3种不同保存方法保存的软骨细胞表面主要组织相容性复合体表达率就降低到最低。预示着经保存处理的关节软骨可能会提高同种异体骨软骨移植手术的成功率。

Effects of preservation methods on the immunogenicity of allograft cartilage

BACKGROUND：Allograft cartilage transplantation is the primary means of treatment of articular cartilage defects.It is a clinical key technical issue to reduce the immunogenicity of the cartilage transplant by looking for cartilage tissue preservation methods.OBJECTIVE：To compare the impact on the immunogenicity of allogeneic cartilage graft stored by slow cooling gradient store method,vitrification and 60Co-irradiation with cooling gradient store method.METHODS：The fresh cartilages were randomly divided into four groups：fresh group did not take any measures;slow cooling group were given procedural temperature gradient to preserve articular cartilage;vitrification group preserved articular cartilage by glass vitrification solution;60Co-irradiation with cooling gradient group were given 60Co-ray to irradiate,then we saved articular cartilage by procedural temperature gradient.Chondrocyte was separated and cultured in vitro and made cell suspension at 8,15,30,and 60 days after preservation.The expression rate of major histocompatibility complex （MHC）Ⅰand Ⅱwas measured by flow cytometry.RESULTS AND CONCLUSION：After preservation,the rate of MHCⅠ and MHCⅡantigen expression on chondrocyte surface was significantly decreased in slow cooling,vitrification,and 60Co-irradiation with cooling gradient groups.There were significant differences comparing with fresh group,but there were no significant differences among each experimental group.After preservation,the rate of MHC-Ⅰand MHC-Ⅱantigen expression on chondrocyte surface reduced to aminimum at 30 days after preservation.So,treatment by the preservation of articular cartilage may improve success rate of bone allograft cartilage transplantation.