Objectives:The objective of this study was to evaluate the efficacy of erbium-doped yttrium aluminum garnet (Er:YAG) laser irradiation in different energy outputs versus ultrasonic in gutta-percha removal during the endodontic retreatment.Materials and Methods:A total of 21 extracted human lower premolars were divided into three groups (n = 7). Following the standardized preparation of the root canals with Wave One Rotary system and obturation with gutta-percha: Group I was treated with ultrasonic, Group II by Er:YAG laser with 40 mJ/Pulse, and Group III by Er:YAG laser with 50 mJ/Pulse for the removal of gutta-percha from the canals. Two extra teeth were treated by Er:YAG laser with 135 mJ/Pulse as control group. For all groups, time for gutta-percha removal was recorded. Samples were then splited into two halves and tested by scanning electron microscope and stereomicroscopic evaluation under different magnification power to observe the efficacy of each method used in the removal of gutta-percha.Results:Statistical analysis of Kruskal–Wallis suggested that there are significant difference between the groups in relation to removal time (P < 0.05) and 2 × 2 Mann–Whitney U-test among the groups revealed that there is no significant difference between 40 and 50 mJ laser outputs (P > 0.05), but ultrasonic versus 40 and/or 50 mJ laser outputs were significantly different (P < 0.05).Conclusions:Er:YAG laser beam was not so efficient when compared to ultrasonic to reach the deeper parts of the canals as it was asserted, thermal side effects and burning damages were observed on the root canal dentinal walls. Moreover, the delivery system was not flexible enough to compensate the curvature of the canal system even though we used more straight canals as the sample ones as well as more time-consuming than the ultrasonic and more clinical time, rendering it to be less efficient in the removal of the obturation material during endodontic retreatment procedures.