CAJ | 학술논문

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。
배경：재관절외과중상용자체세포연골이식기술수복연골결손，정합불량시도치수복실패적원인지일。연골세포천사능력피증명화정합유상관성，모사통로여Src-린산화린지매Cγ1-세포외조절단백격매1/2이경피증실화연골세포적천사능력상관，단시부화정합상관환시개미지수。목적：천명연골세포천사통로재자체세포연골이식중화정합지간적상관성。방법：장래원우저슬관절적연골세포분성4조，위Src、린산화린지매Cγ1、세포외조절단백격매1/2억제조이급정상조，통과박이등소실래양화4조연골세포적천사능력。장처리후적연골세포여연골배건립공배양모형배양28 d후，진행조직학、생물화학、생물역학、면역단백인적이급세포시종등분석래관찰정상조여억제조지간적차이。결과여결론：당용억제제처리연골세포후，연골세포적천사능력명현하강。재연골세포연골배공배양28 d후，면역인적단백분석표명통로억제제일직존재우정개배양주기중。동시정상조천사도정합구역중적연골세포수량화거리이급분비적효원、기질화역학강도명현고우기타3개억제조。설명가이통과Src-린산화린지매Cγ1-세포외조절단백격매1/2통로조절연골세포적천사능력종이영향연골적정합능력。
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.