不同磨损颗粒对体外培养人外周血单核细胞的影响  

Influence of different wear particles on human peripheral blood mononuclear cells in vitro

作　　者：李朋[1] 杨淑野[1] 张锴[1] 贾龙[1] 杜刚强[1] 刘栋[1] 张新军[1] 张德刚[1] 王志刚[1] Li Peng;Yang Shuye;Zhang Kai;Jia Long;Du Gangqiang;Liu Dong;Zhang Xinjun;Zhang Degang;Wang Zhigang(Department of Trauma Orthopedics, Binzhou Medical University Hospital, Binzhou 256600, Shandong Province, China)

机构地区：[1]滨州医学院附属医院创伤骨科,山东省滨州市256600

出　　处：《中国组织工程研究》2019年第6期894-900,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81241061);项目负责人:张锴;山东省医药卫生科技发展计划项目(2016WS0023);项目负责人:杨淑野~~

摘　　要：背景:人工关节置换后正常的行走活动会产生大量的钛铝钒合金(Titanium6Aluminium4Vanadium,Ti6Al4V)、聚甲基丙烯酸甲酯(Polymethylmethacrylate,PMMA)、超高分子量聚乙烯(Ultra-High-MolecularWeight-Polyethylene,UHMWPE)、钴铬合金(Chromium-Cobalt Alloy,Co-Cr)等磨损颗粒,激活单核细胞释放大量的炎性细胞因子和趋化因子,诱发骨溶解和形成假性肉芽肿等一系列生物学级联反应,最终导致假体发生无菌性松动。目的:探讨Ti6Al4V、PMMA、UHMWPE和Co-Cr颗粒对体外培养人外周血单核细胞分泌肿瘤坏死因子α、白细胞介素1和白细胞介素6的影响及诱导骨溶解引起无菌性松动的作用机制。方法:将人外周血单核细胞分别与不同浓度的Ti6Al4V、PMMA、UHMWPE和Co-Cr颗粒进行共培养,72 h后提取细胞上清液,用ELISA法检测肿瘤坏死因子α、白细胞介素1和白细胞介素6的表达水平,观察各种颗粒浓度和理化性质对外周血单核细胞活性的影响。结果与结论:(1)各组人外周血单核细胞存活率均大于95%,仅有Co-Cr组颗粒/细胞比值为1 000时单核细胞存活率降至87%;(2)当颗粒/细胞比值为10,100,1000时,活化的单核细胞分泌肿瘤坏死因子α、白细胞介素1和白细胞介素6的水平与原始幼稚单核细胞(颗粒/细胞比值为0时)相比显著增高,差异有显著性意义(P <0.05);(3)Ti6Al4V组、PMMA组和UHMWPE组中随颗粒浓度增加,单核细胞分泌肿瘤坏死因子α、白细胞介素1和白细胞介素6出现不同程度的增强。Co-Cr组中当颗粒/细胞比值达到1 000时却出现肿瘤坏死因子α、白细胞介素1和白细胞介素6的表达水平降低;(4)在骨溶解过程中Ti6Al4V颗粒的功能生物学活性最强,其次是Co-Cr和PMMA颗粒,UHMWPE颗粒最低弱;Ti6Al4V、PMMA、UHMWPE和Co-Cr颗粒在骨溶解过程中对外周血单核细胞的影响存在剂量依赖性。BACKGROUND:After artificial joint replacement,daily activities lead to the production of a large number of titanium6aluminium4vanadium(Ti6Al4V),polymethylmethacrylate(PMMA),ultra-high-molecular-weight-polyethylene(UHMWPE)and chromium-cobalt alloy(Co-Cr)particles,activating monocytes to release a large number of inflammatory cytokines and chemokines,including a series of biological cascade reactions,such as inducing bone dissolution and forming pseudo-granuloma,and eventually leading to aseptic loosening of the prosthesis.OBJECTIVE:To investigate the effect of Ti6Al4V,PMMA,UHMWPE and Co-Cr particles on tumor necrosis factor-α,interleukin-1and interleukin-6in the human peripheral blood monocytes cultured in vitro and to explore the underlying mechanism of aseptic loosening of the prosthesis by inducing bone dissolution.METHODS:Human peripheral blood mononuclear cells were cultured with Ti6Al4V,PMMA,UHMWPE and Co-Cr particles at different concentrations.After72hours of culture,ELISA technique was utilized to quantify the secretion of different cytokines includes tumor necrosis factor-α,interleukin-1and interleukin-6.We observed the influence of different wear particle concentrations and particle properties on the viability of human peripheral blood mononuclear cells.RESULTS AND CONCLUSION:Cell viability exceeded95%for all groups,except for87%in the Co-Cr group when the particle/cell ratio was1000.Compared with the naive mononuclear cells(particle/cell ratio of0),the expression of tumor necrosis factor-α,interleukin-1and interleukin-6in the activated mononuclear cells increased significantly when the particle/cell ratio was10,100and1000(P<0.05).Ti6Al4V,PMMA and UHMWPE particles stimulated human mononuclear cells to overexpress tumor necrosis factor-α,interleukin-1and interleukin-6,which were positively correlated with the concentration of particles.However,when the Co-Cr particle/cells ratio was1000,the expression levels of tumor necrosis factor-α,interleukin-1and interleukin-6were decreased.During the osteolysi

关 键 词：人工关节置换 无菌性松动 骨溶解 磨损颗粒 单核细胞 细胞因子 国家自然科学基金 关节成形术 置换 颗粒物 肿瘤坏死因子α 白细胞介素1 白细胞介素6 组织工程 

分 类 号：R459.9[医药卫生—治疗学]