Development of an in-vitro rheumatoid arthritis (RA) model and its application in anti-rheumatic drug testing
Tan, Kian Siong
Date of Issue2016-05-16
School of Chemical and Biomedical Engineering
This study describe the evaluation of Celecoxib on a newly established in vitro three-dimensional Rheumatoid Arthritis (RA) model, by co-culturing LhCG with effector cells of RA namely, RA Synovial Fibroblast, Macrophage and Chondrocytes, this model was able to mimic the cartilage degradation condition seen in RA. The validation of this model was based on several assays which includes qPCR expression level of inflammatory mediator (IKBKB), MMPs (MMP-1, MMP-3 and MMP-13) and cartilage ECM proteins content (Col-2 and AGG), biochemical analysis of cartilage ECM proteins content (Col-2 and GAG) and histology and immunochemistry analysis of GAG, calcium deposition, Col-1 and Col-2. As expected, celecoxib had a protective effect on LhCG degradation via reduction in the MMP-1 and MMP-3 expression, particularly with the concentration at T10 (10 μg/mL) and T20 (20 μg/mL), which corroborate well with a higher expression of cartilage ECM proteins (Col-2 and AGG) relative to control. The model treated with T40 (40 μg/mL) of celecoxib was observed to be most effective in reducing the inflammatory mediator gene IKBKB. Other qualitative and quantitative analysis on cartilage ECM content indicate a mixed effectiveness of the different celecoxib concentrations at various time point of the experiment. The low dosage of celecoxib (T10 and T20) seems to have a stimulatory effect on MMP-13 and ECM content while T40 maintain the ECM content level quite consistently throughout the experiment by preventing an upsurge in MMP-13 and keeping a low IKBKB expression level.
Final Year Project (FYP)
Nanyang Technological University