The purpose of this study was to investigate the evaluation of laser-assisted irrigation (LAI) on the removal of the smear layer as compared to passive ultrasonic irrigation (PUI). Forty-eight single-rooted, upper-central incisor teeth were selected and prepared with ProTaper rotary instruments up to size #40 (F4) at the working lengths. Specimens were divided into four groups, as follows: (a) PUI with 5 mL of 2.5% NaOCl for 60 s; (b) PUI with 2.5 mL of 17% EDTA and 2.5 mL of 2.5% NaOCl each for 30 s; (c) LAI with 5 mL of 2.5% NaOCl for 60 s; and (d) LAI with 2.5 mL of 17% EDTA and 2.5 mL of 2.5% NaOCl each for 30 s. In the PUI groups, the ultrasonically activated file was inserted 1 mm short of the working length, but in the LAI groups, the fiber tip was applied into the pulp chamber. LAI in the pulp chamber with the combination of 17% EDTA and 2.5% NaOCl removed more of the smear layer than the other groups (p < 0.018). LAI in the pulp chamber with the combination of 2.5% NaOCl and 17% EDTA better removed the smear layer than LAI applied similarly but without EDTA or PUI with the same NaOCl and EDTA combinations using an ultrasonically activated file inserted 1 mm short of the working length.