In this study, we aimed to assess the long-term effects of the mucosal layer plastic surgery of the vocal folds performed on the voices of male-to-female transgenders. This retrospective cohort study enrolled 22 patients who were operated during 2004-2010 by a combined technique of transmucosal anterior suturing and stiffening of the vocal folds through a longitudinal cordotomy incision using CO2 laser. Long-term effects were assessed based on completed questionnaires on four different categories including subjective evaluation of patient's own voice perception and a standardized voice inventory (VHI), as well as an acoustic assessment and videolaryngoscopic examination. Out of the 22 patients contacted, 16 returned the questionnaires, and 13 of them participated in the subsequent acoustic analysis and videolaryngoscopic assessment. Results of the study were as follows: The total VHI score, after the procedure, was 32. F0 increased significantly for both the vowel/a/ and text from 157 to 207 Hz and 139 to 162 Hz, respectively. Perturbation measures did not show a significant change. F0 did not correlate with the VHI score. One patient had symptomatic anterior perforation that needed re-operation. The combination of transmucosal anterior suturing and stiffening of the vocal folds by laser cordectomy results in significant elevation of the F0 in the long term and in acceptable levels of VHI score compared to other reports. In conclusion, the results of the procedures show that it is enough to get the mucosal edges sutured together without the need to suture either the ligaments or the muscle of the vocal folds. The need for revision is minimal with this procedure.