In this prospective randomized clinical trial, we aimed to evaluate the safety and efficacy of endourethrotomy with holmium:yttrium-aluminium-garnet (HO:YAG) laser and compare the outcomes with the conventional cold-knife urethrotomy. Fifty-one male patients with single, iatrogenic, annular strictures of the urethra were randomly divided into two groups; 21 patients who underwent direct-vision endoscopic urethrotomy with Ho:YAG laser (15 W; 1,200-1,400 mJ; 8-12 Hz) at 12 o'clock position (laser group) and 30 patients who underwent direct-vision endoscopic urethrotomy with cold-knife incision at 12 o'clock position (cold-knife group). The results obtained were analyzed and compared at 3 months, 6 months, 9 months, and 12 months postoperatively by clinical evaluation, uroflowmetry, and retrograde urethrographies. Variables were compared among groups using Fisher's exact and Mann Whitney U tests. There were no differences between two groups in terms of patient age, preoperative Qmax value, stricture location, and length. Operative time was shorter in laser group (16.4 ± 8.04 minutes) when compared with cold-knife group (23.8 ± 5.47 minutes) (p<0.001). Recurrence-free rate at 3 months was similar between two groups (p=0.122). However, recurrence-free rates at 6 months, 9 months, and 12 months were significantly higher in laser group when compared with cold-knife group (p values were 0.045, 0.027, and 0.04, respectively). No intra- or postoperative complications were encountered. Use of Ho:YAG laser in the management of urethral stricture disease is a safe and effective method. In addition, it provides shorter operative time and lower recurrence rate when compared with the conventional technique.