The pullout strength of the pedicle screws after direct vertebral rotation (DVR) maneuver is not known. This biomechanical study was performed to quantitatively analyze the pullout strength of a pedicle screw after DVR maneuver using human cadaveric vertebrae. Thoracic vertebral bodies from three cadavers were harvested and stripped of soft tissues. Thirty pedicles of 15 vertebrae were separated into two groups after bone mineral density measurements. Polyaxial 5.5 mm pedicle screws with appropriate length were inserted with a freehand technique for each pedicle. One Kirschner wire was inserted to the anterior part of each vertebral corpus the half depth of each corpus was embedded into PVC pipes using polyester paste. In the DVR group, each screw was pulled horizontally with 2 kg (similar to 20 N) load over a screwdriver rigidly attached to the screw, and a DVR maneuver was simulated. The control group did not load with a DVR maneuver. Samples were placed on a universal testing machine and pullout loads were measured. The Mann-Whitney U test was utilized, and the P value <0.05 was considered as statistically significant. In the DVR group, the mean pullout strength was 183.35 N (SD +/- 100.12), and in the control group, the mean pullout strength was 279.95 N (SD +/- 76.26). Intergroup comparisons revealed that DVR maneuver significantly decreases the pullout strength (P = 0.012). The results of this study confirm that the pullout strength of pedicle screw significantly decreases by approximately 35% when DVR maneuver is applied.