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学科主题: 临床医学
题名:
Influences of IL-6R antibody on PMMA bone cement-mediated expression of OPG and RANKL in synovial fibroblasts
作者: 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
发表日期: 2014-04-01
DOI: 10.1007/s11596-014-1265-5
卷: 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
所属项目编号: JCYJ20130402114702130 ; 201302064
项目资助者: Research and Development Projects of Shenzhen of P. R. China ; Healthcare and Medical Research Fund of Shenzhen of P. R. China
WOS记录号: WOS:000334426900017
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/58484
Appears in Collections:北京大学深圳医院_骨关节外科_期刊论文

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作者单位: Peking Univ, Shenzhen Hosp, Dept Orthoped, Shenzhen 518036, Peoples R China

Recommended Citation:
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.
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