玻璃化冷冻法保存关节软骨  

Vitrification method for cryopreservation of the articular cartilage

作　　者：宋洪强[1] 玄燕华[2] 吴雅迪[1] 亓建洪[1] 

机构地区：[1]泰山医学院运动医学研究所,山东省泰安市271000 [2]泰山医学院附属医院,山东省泰安市271000

出　　处：《中国组织工程研究》2012年第46期8615-8619,共5页Chinese Journal of Tissue Engineering Research

基　　金：山东省医药卫生科技发展计划项目(2011HW082);项目名称:同种异体骨软骨组织培养保存方法的改良设计及其延时机制研究~~

摘　　要：背景:同种异体骨软骨移植技术是治疗关节软骨缺损有效的方法之一,但由于移植物保存方法不理想,明显制约着该技术的临床应用。目的:探讨玻璃化冷冻法保存关节软骨组织的可行性和优越性。方法:切取成年猪骨软骨,制成约5mm×6mm(直径×长度)大小的圆柱形骨软骨块。以新鲜软骨组为对照,分别采用0.5mol/L甘油、1mol/L二甲基亚砜、1mol/L玻璃化溶液3种方法预处理软骨块,再行冷冻法保存软骨块8周,采用组织化学染色、免疫荧光染色观察并比较软骨细胞活性的变化。结果与结论:玻璃化溶液预处理组的关节软骨细胞存活率达到74.5%,明显高于甘油和二甲基亚砜预处理组,软骨基质成分仅少量丢失。3种方法相比较,玻璃化溶液预处理后慢速梯度降温冷冻保存法可以明显提高冻存关节软骨组织的活性。BACKGROUND：Articular cartilage allograft transplantation is one of effective methods for treatment of articular cartilage defects.However,the short preservation time of osteochondral allograft in vitro limits its clinical application OBJECTIVE：To discuss the feasibility and superiority of vitrification method on the viability of chondrocytes.METHODS：Osteochondral tissues were isolated from adult pigs,and cut into approximately 5mm×6mm（diameter×length） cylindrical osteochondral blocks.Fresh cartilage was taken as control.The osteochondral blocks was pretreated with 0.5mol/L glycerol,1mol/L dimethyl sulfoxide,and 1 mol/L vitrification solution,respectively,and then cryopreserved for 8 weeks.Histochemistry staining and immunofluorescence staining were performed to observe and compare the changes in chondrocyte viability.RESULTS AND CONCLUSION：The viability of chondrocytes was 74.5% in the vitrification pretreatment group,significantly higher than the glycerol and dimethyl sulfoxide pretreatment groups.In addition,there was a small loss of cartilage matrix in the vitrification pretreatment group.In summary,the vitrification-cryopreservation method can greatly improve the viability of frozen chondrocytes.

关 键 词：玻璃化 梯度降温 关节软骨 冷冻保存 细胞活性 

分 类 号：R318[医药卫生—生物医学工程]