Fibroblast-Like Synovial Cells in Rheumatoid Arthritis- Impact of Infliximab on Hexosoaminidase Activity

Abstract

Background. The effect of multiple infusions of infliximab (INF), a chimeric anti-tumor necrosis factor alpha antibody, on the concentration of hexosaminidase (HEX) activity in a synovial cell culture derived from human synovial inflamed fluid obtained from patients suffering from rheumatoid arthritis (RA) has been evaluated. Objectives. The aim of this study was to prove INF efficacy in RA. Material and Methods. Inflamed synovial fluid was taken from RA patients (a study group) and patients who had undergone knee trauma within 7 days (a control group). The following solutions of infliximab were used: 40, 60 and 140 µg/mL. Determination of the concentration of HEX activity in cell cultures was performed after 24, 48, 72 and 96 h of infliximab administration. To identify synoviocytes in cell culture immunohistochemical staining with vimentin and pancytokeratin was performed. Results. A predominance of fibroblast-like synovial cells has been observed in the study group. In the control group the concentration of HEX activity without adding infliximab to the cell culture was 283.00 nkat/mL. After 96 h of incubation with infliximab, the concentrations of HEX activity in cultured synoviocytes according to infliximab doses of 40, 60 and 140 µg/mL were respectively: 280.00, 271.50 and 293.50 nkat/mL. In the study group, the concentration of HEX activity without adding infliximab to the cell culture was 542.27 nkat/mL. The final concentrations of HEX activity of cultured fibroblast-like synovial cells measured after 96 h of incubation with infliximab were: 471.72, 498.27 and 556.72 nkat/mL, according to infliximab doses of 40, 60 and 140 µg/mL. In all groups (besides the infliximab concentration of 140 µg/mL after 96 h of incubation), the level of concentration of HEX activity was significantly higher in the study group compared to the control group, irrespective of infliximab concentration and time of infliximab incubation. Conclusions. Infliximab changes the concentration of HEX activity depending on the drug dose and time of administration (Adv Clin Exp Med 2015, 24, 5, 807-813).

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“…Following isolation, the cells were cultured in Dulbecco's modified Eagle's medium (DMEM) (containing 10% fetal bovine serum, 1% penicillin, streptomycin, amphotericcin B) in 5% CO 2 at 37.5°C, with replacement of the mediun every 2–3 days. HVFs were identified by anti-vimentin antibody staining and subsequently stored in liquid nitrogen for further study ( 19 , 20 ).…”

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