Background Mandibular sagittal split osteotomy (MSSO) may incur unfavorable split and sensorineural injuries. Knowledge of the anatomic location of the mandibular canal (MC) and bone thickness in the region of interest for MSSO, and the possible variations by age and gender can assist in avoiding such complications. Purpose: To study the location of the MC and bone thickness in the region of MSSO by cone-beam computed tomography (CBCT) radiographs and to evaluate the possible variations by age and gender in a Jordanian population. Material and Methods This retrospective radio-anatomical study examined all CBCT radiographs for patients treated over three years at the University of Jordan Hospital, Amman, Jordan. Distances from the MC to the cortical external surfaces and MC diameter (MCD) were measured by a reliable observer at three predetermined regions for MSSO: region (A) [mandibular foramen area], region (B) [mandibular angle area] and region (C) [directly mesial to the second molar]. Gender and age differences in all measurements were then compared using non-parametric Mann-Whitney U test. Results The final study radiographs comprised a total of 202 CBCT belonged to a cohort of 202 subjects; 91 males (45.1%) and 111 (54.9%) females, with mean age (± SD) of 42.94 ± 18.54 years (range 18–90 years). Whereas only the bone thickness superior, buccal and inferior to MC in regions (B) and (C), and MCD in the three regions exhibited significant ( p < 0.05) gender differences, all measured distances exhibited statistically significant ( p < 0.05) differences between young and adult patients. Conclusions The location of MC and bone thickness in the region of MSSO were significantly variable according to age, but exhibited sexual diamorphism only in regions (B) and (C). This fundamental knowledge should be considered during MSSO planning. Key words: Cone-Beam Computed Tomography, mandibular canal, mandibular sagittal split osteotomy.