In Vitro Mitigation of Arsenic-Induced Toxicity by Reduced Glutathione in Rat Pulp Cells

Objective: Despite the controversial results regarding the amount of arsenic (As) in mineral trioxide aggregate (MTA) and MTA-like cements, it is prudent to assess the effect of this heavy metal on pulpal cells and search for methods to attenuate its toxicity. This study investigated the toxic effect of As on pulpal-like cells and evaluated the influence of reduced glutathione (GSH) on As-induced toxicity. Methods: The cytotoxicity of 50 µm As, 50 µm As+50 µM GSH, 50µm As+500 µM GSH or 50 µm As+5000 µM GSH on rat pulpal cells (RPC-C2A) was evaluated at 24 hours and 72 hours. Cell culture in fresh medium without experimental solution served as the control. Cell viability was measured by means of 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) assay and the optical density was measured with microplate reader. The morphology of the cultured cells was observed under phase contrast microscope. Cytotoxicity data were analyzed by two-way ANOVA and Tukey post hoc tests (P<0.05). Results: There were statistically significant differences in cell viability amongst the tested groups (P<0.05). As elicited remarkable toxic effect on pulpal cells, while 5000 µM GSH protected the cells from As-induced damage at 24-hour exposure time. The cultured control cells were polygonal-shaped; however, As-treated cells exhibited contracted and spherical morphology with increased intercellular spaces indicative of cellular death and decreased proliferation. Conclusion: As negatively affected the viability of pulpal cells; however, controlled concentration of GSH had a short-term protective effect against As-induced toxicity. Future research is warranted on the clinical use of GSH with MTA and MTA-like cements to minimize initial inflammation resulting from As release during the setting of the aforementioned cements thus enhancing the success of procedures where these cements are placed in direct contact with vital pulp tissues.