Pigmentation of gingiva is an aesthetic problem. Until now, various methods have been introduced for removal of gingival pigmentation. The purpose of this study was to compare bur abrasion and CO2 laser methods in removing gingival pigmentation. Twelve patients aged 20-40 years old with the chief complaint of physiologic gingival pigmentation participated in this study. For these patients, gingival depigmentation was performed using two treatment modalities including bur abrasion and CO2 laser in a split-mouth design. Gingival depigmentation was performed in the right half of the anterior maxillary and mandibular sextants using bur abrasion method by means of diamond bur on a high-speed headpiece with vigorous water irrigation and the left half of the anterior maxillary and mandibular sextants using a CO2 laser. CO2 laser was set at: peak power 252 watts, repeat time 20 ms and pulse duration 200 microsecond which was used in a non-contact position. The area of pigmentation was calculated via gridlines in the Microsoft Paint software 1 and 6 months after the procedure. Gingival recession was also determined before, and at 1 and 6 months after the procedure. The area of gingival pigmentation was not statistically significantly different between the two treatment modalities before the procedure (p = 0.452), 1 month (p = 0.443) and 6 months after the procedure (p = 0.202). There was no statistically significant difference in the variance of pigmented area at different times in the two methods. In both CO2 laser and bur abrasion methods, the mean area of pigmentation was statistically significantly different in the follow-up period (p < 0/001), in a way that the change in the area of pigmentation was statistically significant 1 month after treatment (p <0.001) and also 6 months after treatment (p < 0.001) compared to before. The change in the area of pigmentation between 1 and 6 months after treatment was not statistically significant in both CO2 laser (p = 0.157) and bur abrasion method (p = 0.150). No increase in gingival recession was observed in any of the patients. Both treatment modalities can effectively treat gingival pigmentation. No increase in gingival recession was observed. Conventional method and CO2 laser were not statistically significantly different during a follow-up period of 1 and 6 months.