The aim of this in vitro study was to investigate the effect of the smear layer on apical microleakage in teeth obturated with mineral trioxide aggregate (MTA). Fifty single-rooted central maxillary teeth were used in this study. All teeth were instrumented to size 60 by using the step-back technique. The selected teeth were randomly divided into 2 groups (n = 20). In the first group (smear [+]), the teeth were irrigated with only NaOCl (5.25%). In the second group (smear [-]), the teeth were irrigated with ethylenediaminetetraacetic acid (EDTA) (17%) and NaOCl (5.25%) to remove the smear layer. The teeth were then filled with MTA. The computerized fluid filtration method was used for evaluation of apical microleakage. The quantitative apical leakage of each tooth was measured after 2, 30, and 180 days. Statistical analysis was done with the Mann-Whitney U and Wilcoxon signed ranks tests. At the end of this study there was no difference in 2 days between the groups, but removal of the smear layer caused significantly more apical microleakage than when the smear layer was left intact for 30 and 180 days (P < .005). Within the constraints of this study it can be concluded that the apical microleakage of MTA is less when the smear layer is present than when it is absent.