Interleukin-4 and Prednisolone affect the matrix metalloproteinase-13 releasing from degenerative bovine nasal cartilage in vitro

Background: Proteolysis of the extracellular matrix (ECM) of articular cartilage by matrix metalloproteinases (MMPs) is a key characteristic of joint diseases. Thus, identification of MMPs expression mechanism in chondrocytes may lead to the development of new treatment for arthritis.

Objectives: Investigate the combination effects of interleukin-4 (IL-4) and Prednisolone on ECM alterations and production of matrix metalloproteinase-13 (MMP-13) induced by interleukin-1α (IL-1α) in bovine nasal cartilage (BNC) explants.

Materials and Methods: Ninety mg of BNC explants (10 slices in each well) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with IL-1α (10 ng/ml), IL-4 (50 ng/ml) and Prednisolone (1nM, 1000 nM) at the same time for 28 days. At days 3, 6,7,14, 21, and 28 media were removed and replaced with fresh media and the removed media were stored in -20°C. The alterations of ECM were assessed by using histology techniques. The combination effects of IL-4 and Prednisolone on MMP-13 production were analyzed by Western immunoblots. The cell viability was evaluated by using lactate dehydrogenase (LDH) assay kit.

Results: The results showed that after 28 days treated with IL-1α, matrix blue staining significantly disappeared. In addition, a clear band of MMP-13 was detected. In the presence of IL-1α and Prednisolone (1000 nM), matrix breakdown was partially decreased and MMP-13 production was significantly reduced. In combination of IL-4 with both low and high dose Prednisolone, proteogycan loss significantly reduced and synergistic inhibition of MMP-13 production were observed. LDH level was not significantly increased in the presence of cytokines and Prednisolone.

Conclusion: The combination of IL-4 and Prednisolone showed profound protective effects on cartilage matrix alterations. A better understanding of the mechanisms by this combination affects MMPs expression in human chondrocytes could provide valuable insight into new therapeutic strategies of cartilage destruction.