The regulation of cytotoxicity and cyclooxygenase‐2 expression by 2‐hydroxy‐ethyl methacrylate in human osteoblasts are related to intracellular glutathione levels

Abstract

To investigate the effects of 2-hydroxy-ethyl methacrylate (HEMA) on cytotoxicity and cyclooxygenase-2 (COX-2) protein expression in human osteoblasts. Cytotoxicity was judged using an Alamar Blue reduction assay on human osteoblast cell line U2OS. Western blot was used to evaluate the expression of COX-2 protein by HEMA. To determine whether glutathione (GSH) levels were important in cytotoxicity and COX-2 expression of HEMA, cells were pre-treated with the GSH precursor, 2-oxothiazolidine-4-carboxylic acid (OTZ), to boost thiol levels, or buthionine sulfoximine (BSO) to deplete GSH. Paired Student's t-tests were applied for the statistical analysis of the results. HEMA demonstrated a cytotoxic effect to U2OS cells in a dose-dependent manner (P<0.05). The 50% inhibition concentration of HEMA was approximately 3mmol L(-1) . HEMA was found to induce COX-2 protein expression in U2OS cells (P<0.05). The addition of OTZ acted as a protective effect on HEMA-induced cytotoxicity and COX-2 expression (P<0.05). In contrast, the addition of BSO enhanced HEMA-induced cytotoxicity and COX-2 expression (P<0.05). Taken together, the levels of HEMA that were tested inhibited cell growth on U2OS cells. HEMA has a significant potential for periapical toxicity. The activation of COX-2 protein expression may be one of the mechanisms of HEMA-induced periapical inflammation. These inhibitory effects were associated with intracellular GSH levels.