Regulation of IL-6R antibody on PMMA bone cement mediated mRNA expression of MMPs and vascular factors by synovial fibroblasts

Objective To investigate the PMMA bone cement-mediated expression of MMP-1,MMP-3,VEGF,and PDGF mRNA in synovial fibroblasts regulated by IL-6R antibody.Methods The synovial tissue obtained from total hip arthroplasty was digested and subcultured.Phase-contract microscope was employed to observe the synovial cells and immunocytochemistry(SABC method) staining was used to identify synovial fibroblasts.The experiment was divided into 3 groups according to the different supplement in cultures: PMMA group(75μg/ml PMMA bone cement particles);IL-6R antibody group(10 ng/ml IL-6R antibody and 75 μg/ml PMMA bone cement particles);and the control group.Cell proliferation activity regulated by IL-6R antibody or PMMA was measured by using a cell counting kit-8(CCK-8).FQ-PCR was used to detect the mRNA expressions of MMP-1,MMP-3,VEGF,and PDGF.Results The primary synovial cells showed a short spindle shape after adhesion.After 3 consecutive passages,more than 95% of the cells became fibroblast-like cells with a long spindle shape.SABC staining showed that the anti-CD68 antibody was negative and anti-vimentin antibody was positive in fibroblast-like cells.CCK-8 test demonstrated that the absorbance value(A) was significantly lower in IL-6R antibody group than that in PMMA group and control group(P0.01).The A value was not statistically different between the control group and PMMA group(P0.05).FQ-PCR measurement showed that mRNA expressions of MMP-1,MMP-3,VEGF,and PDGF in IL-6R antibody group were significantly inhibited than those in PMMA group and control group(P0.01).The mRNA expressions of MMP-1,MMP-3,VEGF,and PDGF in PMMA group were higher than those in control group(P0.05).Conclusion IL-6R antibody could significantly inhibit the mRNA expressions of MMP-1,MMP-3,VEGF,and PDGF in synovial fibroblasts.This may provide theoretical basis of molecular biology for the prevention and treatment of aseptic loosening of the prosthesis.