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The purpose of this study was to investigate the relationships between the location of the mental foramen (MF) and the mandibular canal (MC) and the surgical access line (SAL) of the mandibular posterior teeth using cone-beam computed tomographic (CBCT) scans.CBCT scans of 204 subjects ranging in age from 18–76 years old were evaluated. The vertical and horizontal distances from the MF to the SAL of the mandibular premolars and first molars and the vertical distance from the MC to the SAL of the second premolars and first molars were measured via CBCT scans.The average vertical distance between the MF and the SALs showed significant increases sequentially from the first premolars to the distal roots of the first molars, and the shortest average distance of 2.74 mm was obtained for the first premolars. The SALs of the second premolars were the closest to the MF in the horizontal direction with an average distance of 1.5 mm. In 19.9% of the cases, the vertical and horizontal distances between the MF and the SALs of the second premolars were less than 2 mm. In addition, the MF was located superior to the root apices in 6.62% of the cases. The majority of the SALs were located at a vertical distance from the MC that was more than 2 mm. Men and women exhibited significant differences in both the horizontal distance from the MF to the SALs of the first premolars and the vertical distance from the MC to the SALs of the second premolars.To improve the success of endodontic microsurgery, adequate knowledge of the anatomic relationships between the location of the MF and MC and the SAL of the mandibular posterior teeth is indispensable to surgeons.