Cytotoxic response of three cell lines exposed in vitro to dental endodontic sealers

Abstract

The in vitro cytotoxicity of five endodontic sealers was measured >8-12 weeks using L929 mouse fibroblasts, osteoblastic cells (ROS) 17/2.8 rat osteoblasts, and MC3T3-E1 mouse osteoblasts. Discs (n = 6) of AH-plus Jet (AHP), two versions of Endo Rez (ER, ERx), Epiphany (EPH), and Pulp Canal Sealer (PCS) were prepared. The sealers and Teflon (Tf, negative control) were placed in direct contact with cells after immersion in phosphate-buffered saline for 1-12 wk. Cellular succinate dehydrogenase (SDH) activity was estimated using the MTT method (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a yellow tetrazole), and activities were normalized to Teflon® controls. The cellular responses to the materials were compared using analysis of variance with Tukey posthoc analyses (α = 0.05). Initially, all sealers suppressed normalized SDH activity of L929 fibroblasts by >90%. After 12 weeks of immersion in saline, AHP exhibited the SDH activity above Tf (120%), followed by ERx (78%), ER (58%), PCS (38%), and EPH (28%), all statistically distinct (p < 0.05). In general, the three cell lines responded similarly to the sealers. However, AHP caused unique responses: ROS cells were significantly (p < 0.05) less sensitive initially, and AHP was severely cytotoxic to MC3T3 cells (<35% of Tf) through 8 weeks. The data suggest that with "aging" in saline, current endodontic sealers decrease in in vitro cytotoxicity at different rates.