CAJ | 학술논문

目的：探讨结缔组织生长因子（CTGF）对纤维软骨细胞外基质分泌、VEGF表达及促进半月板无血管区的损伤修复中的作用。方法：将新西兰大白兔半月板白区，经胶原酶消化、离心分离后，提取半月板纤维软骨细胞，培养至第2代。用流式细胞仪鉴定该细胞表面CD31，CD44，CD45和CD105标志物，并用Ⅱ型胶原抗体做免疫细胞化学鉴定，以证明所培养、传代的细胞是纤维软骨细胞。分别将细胞培养在浓度100 ng/ml的CTGF培养基中3、14 d后，通过实时定量聚合酶链反应，检测Ⅰ型胶原、Ⅱ型胶原和VEGF基因的表达变化情况。造模，在兔半月板中央区，制作长3 mm的纵行撕裂。将45只大白兔随机分为3组，处理方式为：半月板单纯缝合术，缝合术加填充PBS-纤维蛋白胶，缝合术加填充1．5滋g CTGF-纤维蛋白胶。术后第1、4、10周用荧光免疫组织化学分析法显示Ⅰ型、Ⅱ型胶原和VEGF在损伤处的表达与分布情况，直观撕裂处的愈合情况。结果：体外实验第14天，定量RT-PCR结果显示，100 ng/ml CTGF组中的Ⅰ型、Ⅱ型胶原和VEGF mRNA表达比PBS对照组，明显增加。体内实验的荧光免疫组织化学结果显示，在术后第10周，CTGF治疗组中的Ⅰ型、Ⅱ型胶原和VEGF已完全填充撕裂损伤处。 PBS-蛋白胶组中，损伤处仍有明显裂隙。结论：CTGF可促进半月板无血管区重要的细胞外基质（Ⅰ型、Ⅱ型胶原）的合成，同时损伤处VEGF的表达活性明显增强，有利于促进无血管区半月板撕裂损伤的愈合。
목적：탐토결체조직생장인자（CTGF）대섬유연골세포외기질분비、VEGF표체급촉진반월판무혈관구적손상수복중적작용。방법：장신서란대백토반월판백구，경효원매소화、리심분리후，제취반월판섬유연골세포，배양지제2대。용류식세포의감정해세포표면CD31，CD44，CD45화CD105표지물，병용Ⅱ형효원항체주면역세포화학감정，이증명소배양、전대적세포시섬유연골세포。분별장세포배양재농도100 ng/ml적CTGF배양기중3、14 d후，통과실시정량취합매련반응，검측Ⅰ형효원、Ⅱ형효원화VEGF기인적표체변화정황。조모，재토반월판중앙구，제작장3 mm적종행시렬。장45지대백토수궤분위3조，처리방식위：반월판단순봉합술，봉합술가전충PBS-섬유단백효，봉합술가전충1．5자g CTGF-섬유단백효。술후제1、4、10주용형광면역조직화학분석법현시Ⅰ형、Ⅱ형효원화VEGF재손상처적표체여분포정황，직관시렬처적유합정황。결과：체외실험제14천，정량RT-PCR결과현시，100 ng/ml CTGF조중적Ⅰ형、Ⅱ형효원화VEGF mRNA표체비PBS대조조，명현증가。체내실험적형광면역조직화학결과현시，재술후제10주，CTGF치료조중적Ⅰ형、Ⅱ형효원화VEGF이완전전충시렬손상처。 PBS-단백효조중，손상처잉유명현렬극。결론：CTGF가촉진반월판무혈관구중요적세포외기질（Ⅰ형、Ⅱ형효원）적합성，동시손상처VEGF적표체활성명현증강，유리우촉진무혈관구반월판시렬손상적유합。
Objective:To investigate effect of connective tissue growth factors (CTGF) on secretion of extracellular matrix synthesis of meniscal fibrochondrocytes,expression of vascular endothelial growth factors (VEGF),and angiogenesis during the repair of meniscal tearing damage. Methods:Meniscal fibrochondrocytes were isolated from the inner-1/2 of rabbits'meniscus by collagenase enzymatic digestion,centrifugal separation,and treated with 100 ng/ml CTGF in vitro. Characteriza-tion of fibrochondrocytes was identified by flow cytometry analysising CD31 ,CD44,CD45 and CD105,and was further tested by typeⅡcollagen immunocytochemistry. Changes in gene expression of meniscal fibrochondrocytes were monitored by quanti-tative real-time polymerase chain reaction. In vitro ,the sections of the 3 mm of the longitudinal teared in the middle of the rab-bit's meniscus,and then the defects were dealed with simple suture,suture and implanting with PBS-fibrin glue,sutured and implanting with 1.5μg CTGF respectively. Expression and distribution of typeⅠandⅡcollagen and VEGF,the tearing healing were observed by fluorescence-immunohistochemisty analysis on the 1st week ,the 4th week and the 10th week. Results:Quantitative RT-PCR assays showed that typeⅠand typeⅡcollagen,and VEGF mRNA expression in the 100 ng/ml CTGF group had been remarkably enhanced than in the PBS group on the 14th day. Consistent with these effects in vitro ,fluores-cence-immunohistochemical analysis revealed that in the group implanted with CTGF-fibrin glue ,typeⅠcollagen,typeⅡcol-lagen and capillaries completely filled the defect on the 10th week postoperatively. In contrast ,only soft tissue repair occurred after the PBS-fibrin glue was implanted. Conclusion:CTGF can significantly promote extracellular matrix (Ⅰcollagen,Ⅱcol-lagen) of the meniscal avascular zone synthesis,and CTGF can greatly heighten the expression of VEGF activity at the same time in vitro,so that it can further enhance the repair of meniscal tearing damage in the avascular zone.