CAJ | 학술논문

目的研究髋关节骨关节炎(osteoarthritis,OA)及股骨颈骨折(femoral neck fracture,FNF)患者软骨细胞血管内皮生长因子(vascular endothelial growth factor,VEGF)表达,同时对比分析OA滑膜液及创伤后滑膜液对去分化软骨细胞分泌VEGF的影响,从而探索炎性因子致去分化软骨细胞异常的VEGF表达变化.方法取12例髋关节OA与8例FNF患者的软骨进行HE和番红O-固绿染色后行Mankin评分.将OA组及FNF组软骨组织分别粉碎后富集软骨细胞进行单层培养,取细胞换液时遗弃的上清液作为检测标本,以cathepsin B作为检测OA组软骨细胞去分化程度的指标,并且动态检测VEGF水平,并各培养基于传代培养时分别加入OA滑膜液、DMEM及创伤后滑膜液以观察其对软骨细胞VEGF分泌的影响.数据统计分析使用SPSS 11.0统计软件.结果 OA组软骨Mankin评分明显较骨折组升高,随着培养时间延长软骨细胞逐渐丧失其原有细胞形态并伴显著的cathepsin B上调和VEGF下凋;OA滑膜液可促使软骨细胞特别是OA组软骨细胞上调分泌VEGF,但刺激所产生的VEGF较原代培养初期明显减少,而FNF滑膜液对VEGF上调影响不大.结论 OA软骨损害Mankin评分与原代培养初期VEGF成正相关;软骨细胞体外原代培养后迅速表现为去分化特性;OA较FNF滑膜液可以促使软骨细胞特别是OA组细胞表达更多的VEGF,提示OA滑膜液参与病程进展,并且OA组软骨细胞病理表型更倾向于表达较多VEGF;而FNF滑膜液则相对有利于维持软骨细胞的原有表型.
목적 연구관관절골관절염(osteoarthritis,OA)급고골경골절(femoral neck fracture,FNF)환자연골세포혈관내피생장인자(vascular endothelial growth factor,VEGF)표체,동시대비분석OA활막액급창상후활막액대거분화연골세포분비VEGF적영향,종이탐색염성인자치거분화연골세포이상적VEGF표체변화.방법 취12례관관절OA여8례FNF환자적연골진행HE화번홍O-고록염색후행Mankin평분.장OA조급FNF조연골조직분별분쇄후부집연골세포진행단층배양,취세포환액시유기적상청액작위검측표본,이cathepsin B작위검측OA조연골세포거분화정도적지표,병차동태검측VEGF수평,병각배양기우전대배양시분별가입OA활막액、DMEM급창상후활막액이관찰기대연골세포VEGF분비적영향.수거통계분석사용SPSS 11.0통계연건.결과 OA조연골Mankin평분명현교골절조승고,수착배양시간연장연골세포축점상실기원유세포형태병반현저적cathepsin B상조화VEGF하조;OA활막액가촉사연골세포특별시OA조연골세포상조분비VEGF,단자격소산생적VEGF교원대배양초기명현감소,이FNF활막액대VEGF상조영향불대.결론 OA연골손해Mankin평분여원대배양초기VEGF성정상관;연골세포체외원대배양후신속표현위거분화특성;OA교FNF활막액가이촉사연골세포특별시OA조세포표체경다적VEGF,제시OA활막액삼여병정진전,병차OA조연골세포병리표형경경향우표체교다VEGF;이FNF활막액칙상대유리우유지연골세포적원유표형.
Objective To investigate the vascular endothelial growth factor (VEGF) expression level by chondrocytes isolated from patients with osteoarthritis (OA) in hip or femoral neck fracture (FNF) and explore the effect of synovial fluid from OA or FNF on secretion of VEGF. Methods The cartilage tissues were collected from 12 patients with OA in hip and 8 patients with FNF. Cartilage was stained with HIM and Safranin O/Fast Green (S/F) method. The damage of cartilage was evaluated using Mankin scores.Cathepsin B which was selected for cell dedifferentiation monitoring marker and VEGF level was detected in the supernatant fluid. The synovial fluid from OA, FNF and DMEM were respectively added to the culture medium to explore their effects on regulating VEGF. Results Cartilage the Mankin scores of OA group were higher than that of FNF group. Chondrocytes gradually lost their original spherical appearance, with Cathepsin B upregulated while VEGF downregulated. The OA synovial fluid can stimulate chongdrocytes to secrete more VEGF than the one from patients with FNF. However, chondrocytes gradually produced less VEGF after passaging. Conclusion Mankin scores had good correlation with chondrocytes' VEGF production in the early stage of primary culture. Chondrocytes showed quick dedifferentiation characteristics in vitro. OA synovial fluid showed abig ger capability in stimulating chondrocytes to express more VEGF, which might indicate that OA synovial fluid participated in the pathological process of OA.