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Influences of IL-6R antibody on PMMA bone cement-mediated expression of OPG and RANKL in synovial fibroblasts
Tao, Ke; Zeng, Hui; Xiao, De-ming; Xiong, Ao; Weng, Jian; Kang, Bin
关键词interleukin-6 receptor polymethyl methacrylate bone cement synovial fibroblasts osteoprotegerin receptor activator of nuclear factor-kappaB ligand
刊名JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY-MEDICAL SCIENCES
2014-04-01
DOI10.1007/s11596-014-1265-5
34期:2页:241-246
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Biochemistry & Molecular Biology
研究领域[WOS]Biochemistry & Molecular Biology
关键词[WOS]INTERLEUKIN-6 RECEPTOR INHIBITION ; FACTOR-KAPPA-B ; RHEUMATOID-ARTHRITIS ; PERIPROSTHETIC OSTEOLYSIS ; TOTAL HIP ; IN-VITRO ; TOCILIZUMAB ; ARTHROPLASTY ; TISSUE ; SYSTEM
英文摘要

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 mu g/mL PMMA bone cement particles), IL-6R antibody group (10 ng/mL IL-6R antibody+75 mu g/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 (P < 0.01), but there was no statistically significant difference in A value at 450 nm between the control group and PMMA group (P > 0.05). Results of ELISA indicated that the expression of OPG was significantly higher in IL-6R antibody group than in PMMA group and control group (P < 0.01). The expression of RANKL was inhibited (P < 0.05), and the ratio of OPG/RANKL was significantly increased in IL-6R antibody group as compared with PMMA group and control group. There was no significant difference in the expression of OPG between control group and PMMA group (P > 0.05), but the expression of RANKL was higher in PMMA group than in control group (P < 0.05), and there was a significant difference in the ratio of OPG/RANKL between them (P < 0.05). Results of FQ-PCR revealed the expression of RANKL mRNA was significantly inhibited (P < 0.01) and the expression of OPG mRNA was significantly increased (P < 0.01) in IL-6R antibody group as compared with PMMA group and control group. The expression of RANKL mRNA was higher in PMMA group than in control group (P < 0.05), but the expression of OPG mRNA had no significant difference between them (P > 0.05). 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.

语种英语
WOS记录号WOS:000334426900017
项目编号JCYJ20130402114702130 ; 201302064
资助机构Research and Development Projects of Shenzhen of P. R. China ; Healthcare and Medical Research Fund of Shenzhen of P. R. China
引用统计
被引频次:2[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/58484
专题北京大学深圳医院_骨关节外科
作者单位Peking Univ, Shenzhen Hosp, Dept Orthoped, Shenzhen 518036, Peoples R China
推荐引用方式
GB/T 7714
Tao, Ke,Zeng, Hui,Xiao, De-ming,et al. Influences of IL-6R antibody on PMMA bone cement-mediated expression of OPG and RANKL in synovial fibroblasts[J]. JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY-MEDICAL SCIENCES,2014,34(2):241-246.
APA Tao, Ke,Zeng, Hui,Xiao, De-ming,Xiong, Ao,Weng, Jian,&Kang, Bin.(2014).Influences of IL-6R antibody on PMMA bone cement-mediated expression of OPG and RANKL in synovial fibroblasts.JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY-MEDICAL SCIENCES,34(2),241-246.
MLA Tao, Ke,et al."Influences of IL-6R antibody on PMMA bone cement-mediated expression of OPG and RANKL in synovial fibroblasts".JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY-MEDICAL SCIENCES 34.2(2014):241-246.
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