Influences of IL-6R Antibody on PMMA Bone Cement-mediated Expression of OPG and RANKL in Synovial Fibroblasts

摘要

Effect of interleukin-6 receptor(IL-6R) antibody on polymethyl methacrylate(PMMA) bone cement-mediated expression of osteoprotegerin(OPG) and receptor activator of nuclear factor-kappaB ligand(RANKL) in synovial fibroblasts was investigated. Synovial tissue obtained from total knee arthroplasty was digested and cultured. Inverted microscope was employed to observe the synovial cells and immunocytochemistry(SABC method) staining was used to identify synovial fibroblasts. This experiment was divided into three groups according to different culture media: PMMA group(75 μg/mL PMMA bone cement particles), IL-6R antibody group(10 ng/mL IL-6R antibody+75 μg/mL PMMA bone cement particles), and control group(no IL-6R antibody or PMMA bone cement particles). Influence of IL-6R antibody and PMMA on proliferation of synovial fibroblasts was measured by cell counting kit-8(CCK-8). ELISA method was used to measure OPG and RANKL levels in culture solution. Fluorescence quantitative real-time PCR(FQ-PCR) was used to detect the expression of OPG and RANKL mRNA. After three consecutive passages, more than 95% of the primary synovial cells became long spindle fibroblast-like cells. SABC staining results showed that the fibroblast-like cells were negative for anti-CD68 antibody and positive for anti-vimentin antibody, with brown madder stained. CCK-8 test demonstrated that the absorbance(A) value at 450 nm was significantly lower in IL-6R antibody group than in PMMA group and control group(P0.05). Results of ELISA indicated that the expression of OPG was significantly higher in IL-6R antibody group than in PMMA group and control group(P0.05), but the expression of RANKL was higher in PMMA group than in control group(P0.05). IL-6R antibody could significantly increase the expression of OPG, but inhibit the expression of RANKL, which might provide a theoretical basis of molecular biology for the prevention and treatment of aseptic loosening of prosthesis.