Effects of a Bacterial Lipid Byproduct on Human Pulp Fibroblasts In Vitro

Butyrate, a short chain fatty acid, is a metabolic lipid byproduct of various root canal pathogens, such as Porphyromonas endodontalis. However, little is known about the effects of butyrate on cultured human pulp fibroblasts. H33258 fluorescence, flow cytometry, and protein synthesis assays were used to investigate the pathobiologic effects of butyrate on cultured human pulp fibroblasts. Butyrate exhibited cytotoxic effects on human pulp fibroblasts in a concentration-dependent manner (p < 0.05). The addition of butyrate resulted in G2/M phase arrest (p < 0.05). Butyrate also inhibited protein synthesis in a dose-dependent manner (p < 0.05). To determine whether glutathione (GSH) levels were important in the cytotoxicity of butyrate, we pretreated cells with the GSH precursor, 2-oxothiazolidine-4-carboxylic acid (OTZ), to boost thiol levels, or buthionine sulfoximine (BSO) to deplete GSH. The addition of OTZ acted as a protective effect on the butyrate-induced cytotoxicity (p < 0.05). In contrast, the addition of BSO enhanced the butyrate-induced cytotoxicity (p < 0.05). These results indicate that butyrate is cytotoxic to human pulp fibroblasts by inhibiting cell growth, cell-cycle kinetics, and protein synthesis. These inhibitory effects were associated with intracellular GSH levels.