Induction of heme oxygenase-1 expression by root canal sealers in human gingival fibroblasts is augmented by oxidative stress

Heme oxygenase-1 (HO-1) is known as a stress-inducible protein and functions as an antioxidant enzyme. It has been shown that HO-1 is induced rapidly by a variety of chemical and physical stimuli. However, little is known about the induction of cellular signaling events after cell exposure to root canal sealers. The aim of this study was to investigate the effects of zinc oxide-eugenol-based root canal sealer N2 and epoxy resin-based root canal sealer Topseal on the expression of HO-1 protein in cultured human gingival fibroblasts (HGFs). The results showed that both N2 and Topseal were cytotoxic to HGFs in a concentration-dependent manner (p < 0.05). The exposure of quiescent HGFs to N2 and Topseal resulted in the induction of HO-1 protein expression in a time-dependent manner (p < 0.05). Furthermore, to determine whether oxidative stress could modulate HO-1 expression in HGFs by root canal sealers, HGFs were pretreated with glutathione (GSH) synthesis precursor 2-oxothiazolidine-4-carboxylic acid (OTZ) and GSH synthesis inhibitor buthionine sulfoximine (BSO) for 24 h. The pretreatment with OTZ decreased the N2-induced HO-1 protein level by ∼32% (p < 0.05). However, BSO enhanced the N2-induced HO-1 protein level by up to twofold (p < 0.05). Similar results were found by Topseal. The pretreatment with OTZ decreased the Topseal-induced HO-1 protein level by ∼12% (p < 0.05). However, BSO enhanced the Topseal-induced HO-1 protein level by up to 1.7-fold (p < 0.05). Taken together, HO-1 expression might be one signal transduction pathway linked to the induction of stress-inducible protein by root canal sealers. In addition, the activation of HO-1 is augmented by oxidative stress. Factors that induce GSH synthesis may appear useful in preventing oxidative damage mediated by root canal sealers.