The aim of this study was to evaluate the relationship between the tongue volume and lower incisor irregularity, using cone-beam computed tomography (CBCT), and to identify the possible gender differences. CBCT images of 60 patients between 16 and 36 years of age were selected from 1400 sets of images in the database. Tomography was carried out using iCAT (R) (Imaging Sciences International, Hatfield, Pennsylvania, USA) and segmentation was carried out by using Mimics 10.1 software (Materialise NV, Leuven, Belgium). The tongue volume was calculated by using the volume of the voxels from the scan and the number of voxels selected for a given mask. Lower incisor crowding was measured with the Little's irregularity index and divided into three groups: mild, moderate, and severe. Independent samples t-test, analysis of variance (ANOVA), and Tukey test were used at P < 0.05 level. Pearson correlation coefficients and linear regression model were calculated to determine the correlation between tongue volume and incisor irregularity. No significant gender dimorphism was found for the tongue volume (females: 28.13 +/- 8.54 cm(3) and males: 31.02 +/- 9.75 cm(3)). According to ANOVA, there was statistically significant difference in the tongue volume measurements among subjects with different levels of irregularity. Tukey analysis indicated that mild irregularity group (33.97 cm(3)) showed higher values for tongue volume than severe irregularity group (26.60 cm(3); P = 0.025). The relationship between incisor irregularity and tongue volume was evaluated for both genders and significant inverse correlation (r = -0.429; P = 0.029) was determined between lower incisor irregularity and tongue volume in males. In female group, no signifi cant correlation was determined between tongue volume and incisor irregularity.