The research aimed to investigate the diagnostic value of Interleukin 1 receptor antagonist (IL-1Ra) in the early aseptic loosening of hip prosthesis and whether IL-1Ra can be expressed in bone marrow mesenchymal stem cells. In this study, the IL-1Ra gene was firstly connected to the lentiviral vector LV5, and the lentiviral vector LV5-home-IL1Ra was obtained by recombination. Then the recombinant LV5-home-IL1Ra was co-transfected with the virus-assisted plasmid into 293 T cells and packaged to produce lentivirus. Bone marrow-derived stem cells (BMSCs) were infected with packaged lentiviruses. The relative expression of IL-1Ra mRNA in BMSCs was detected by fluorescence quantitative PCR. The expression of IL-1Ra protein in BMSCs was detected by western blot transfer electrophoresis. Peripheral venous blood samples from 108 patients and healthy subjects underwent total hip replacement were collected to detect the levels of plasma biomarkers procollagen type I carboxy-terminal propeptide (PICP), N-telopeptide cross-links of type I collagen (NTX), osteoprotegerin (OPG), TNGα, receptor activator of NF-kappaB ligand (RANKL), and IL-1β. The recombinant lentivirus vector IL-1Ra was successfully constructed by 2% agarose gel electrophoresis. Lentivirus-mediated IL-1Ra gene could efficiently transfection bone marrow mesenchymal stem cells, and the cell growth density reached about 80% at 72 h after infection. The transfection rate was about 90%, and the fluorescence was enhanced. The relative mRNA and protein expression levels of IL-1Ra in the BMSCs-IL-1Ra group were significantly higher than those in the BMSCs group and the BMSCs-con group (P < 0.01). The late loosening group of IL-1β was significantly higher than the stable prosthesis group and the healthy group (P < 0.05). The ROC curve showed that IL-1 background had strong diagnostic sensitivity and specificity, which was similar to the X-ray score of osteolysis and had the most significant diagnostic significance. Lentivirus-transfected exogenous IL-1Ra can be expressed stably in mouse bone marrow mesenchymal stem cells, and IL-1β, an antagonist of IL-1Ra, plays an important role in the early aseptic loosening of hip prosthesis.

本研究旨在探讨白介素1受体拮抗剂（IL-1Ra）在髋关节假体早期无菌性松动中的诊断价值以及IL-1Ra是否可以在骨髓间充质干细胞中表达。本研究首先将IL-1Ra基因与慢病毒载体LV5连接，重组得到慢病毒载体LV5-home-IL1Ra。然后将重组的LV5-home-IL1Ra与病毒辅助质粒共转染293T细胞并包装生产慢病毒。骨髓来源的干细胞 (BMSCs) 被包装的慢病毒感染。荧光定量PCR检测BMSCs中IL-1Ra mRNA的相对表达。Western blot转移电泳检测BMSCs中IL-1Ra蛋白的表达。， I型胶原（NTX），骨保护素（OPG）的N-端肽交联， TNGα，NF-κB的配体的受体活化剂（ RANKL） ，和IL-1β。2%琼脂糖凝胶电泳成功构建重组慢病毒载体IL-1Ra。慢病毒介导的IL-1Ra基因可高效转染骨髓间充质干细胞，感染后72h细胞生长密度可达80%左右。转染率约90%，荧光增强。BMSCs-IL-1Ra 组 IL-1Ra mRNA 和蛋白相对表达水平显着高于 BMSCs 组和 BMSCs-con 组（P < 0.01)。IL-1β晚期松动组显着高于稳定假体组和健康组（P  < 0.05）。ROC曲线显示IL-1背景具有较强的诊断敏感性和特异性，与骨溶解的X线评分相似，具有最显着的诊断意义。慢病毒转染的外源性IL-1Ra可在小鼠骨髓间充质干细胞中稳定表达，IL-1Ra的拮抗剂IL-1β在髋关节假体早期无菌性松动中起重要作用。