Effect of Interleukin 1 Receptor Antagonist Gene on Stable Expression Bone Marrow Mesenchymal Stem Cells and Early Aseptic Loosening of Hip Prosthesis of Mouse.

Abstract

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 293T 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 72h 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.