The aim of this study was to evaluate the cytotoxicity of the root canal sealers RoekoSeal Automix (RSA) and AH Plus in vitro on human cervical carcinoma (HeLa) cells and mouse skin fibroblasts (L929). The sealers were covered with a 3 x 10(4) cell suspension 1 h, 24 h, 48 h, 7 days and 1 month after mixing. Four samples and respective controls without sealer were prepared. After 5 days of incubation, the number of cells was determined using an electronic counter; the number of viable cells was determined under light microscopy following the addition of nigrosin dye. The experiment was repeated twice for each sealer and each period. AH Plus was significantly more cytotoxic after 1 h, 24 h, and 48 h, compared to the 7 day and 1 month setting period on both cell lines. RoekoSeal had no cytotoxic effect on either cell line at any setting time.