软骨细胞迁徙能力与自体细胞软骨移植中整合的相关性  

Correlation between chondrocyte migration and integration capability during autologous chondrocyte transplantation

作　　者：陆一鸣[1] 桂鉴超[1] 徐扬[1] 尹昭伟[1] 杨晓斐[1] 蒋逸秋[1] 

机构地区：[1]南京医科大学附属南京医院骨科,江苏省南京市210006

出　　处：《中国组织工程研究》2013年第44期7721-7728,共8页Chinese Journal of Tissue Engineering Research

基　　金：江苏省"科教兴卫"工程资助项目(YZK07119)~~

摘　　要：背景:在关节外科中常用自体细胞软骨移植技术修复软骨缺损,整合不良是导致修复失败的原因之一。软骨细胞迁徙能力被证明和整合有相关性,某些通路如Src-磷酸化磷脂酶Cγ1-细胞外调节蛋白激酶1/2已经被证实和软骨细胞的迁徙能力相关,但是否和整合相关还是个未知数。目的:阐明软骨细胞迁徙通路在自体细胞软骨移植中和整合之间的相关性。方法:将来源于猪膝关节的软骨细胞分成4组,为Src、磷酸化磷脂酶Cγ1、细胞外调节蛋白激酶1/2抑制组以及正常组,通过博伊登小室来量化4组软骨细胞的迁徙能力。将处理后的软骨细胞与软骨环建立共培养模型培养28 d后,进行组织学、生物化学、生物力学、免疫蛋白印迹以及细胞示踪等分析来观察正常组与抑制组之间的差异。结果与结论:当用抑制剂处理软骨细胞后,软骨细胞的迁徙能力明显下降。在软骨细胞软骨环共培养28 d后,免疫印迹蛋白分析表明通路抑制剂一直存在于整个培养周期中。同时正常组迁徙到整合区域中的软骨细胞数量和距离以及分泌的胶原、基质和力学强度明显高于其他3个抑制组。说明可以通过Src-磷酸化磷脂酶Cγ1-细胞外调节蛋白激酶1/2通路调节软骨细胞的迁徙能力从而影响软骨的整合能力。BACKGROUND：In joint surgery, the commonly used autologous chondrocyte transplantation often used to repair cartilage defects, and poor integration is one of the reasons that leading to failure repairing. Chondrocytes migration capability is proven to have correlation with integration and some pathways, such as Src-phosphorylated phospholipase Cγ1-extracellular regulated kinase 1/2 has been confirmed to have correlation with the migration ability of chondrocytes, but the correlation with the integration is stil unknown. OBJECTIVE：To determine the chondrocyte signaling pathways involved in autologous chondrocyte migration and their effects on cartilage integration in autologous chondrocyte implantation. METHODS：Articular chondrocytes were isolated from immature pig knee joints. The cells were divided into four groups：Src group, phosphorylated phospholipase Cγ1 group, extracellular regulated kinase 1/2 group and control group, then the Boyden chambers were used to quantify the chondrocyte migration. The chondrocytes/cartilage ring integration model was developed and cultured for 28 days, and then histology, biochemistry, biomechanics, western blot analysis and celltracking analysis were performed to observe the differences between the control group and the suppression groups. RESULTS AND CONCLUSION：The migration ability of chondrocytes was significantly decreased after pretreated with inhibitors. After the chondrocytes/cartilage ring co-cultured for 28 days, Western blot analysis showed that the pathway inhibitors has been presented in the entire culture cycle. The number and length of chondrocytes migrated into the integration area, col agen secretion level, matrix and mechanical strength in the control group were higher than those in three suppression groups. The results suggest that chondrocyte migration ability can affect the cartilage integration capability through Src-phosphorylated phospholipase Cγ1-extracellular regulating kinase 1/2 signal transduction pathway.

关 键 词：器官移植 细胞移植 SRC 磷酸化磷脂酶Cγ1 细胞外调节蛋白激酶1/2 软骨细胞 迁徙 整合 自体软骨细胞移植 省级基金 

分 类 号：R617[医药卫生—外科学]