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Objective To evaluate the biomechanical stability of anterior transarticular crossing screw (ATCS) and compare it with anterior transarticular screw (ATS) which may provide basic evidence for clinical application. Methods Eight human fresh cadaveric specimens (occiput-C4) were tested with 5 conditions including the intact status, the injury status (type II odontoid fracture), the injury+ATS fixation status (traditional bilateral ATS fixation); the injury+unilateral ATCS fixation status; and the injury+bilateral ATCS fixation status. Specimens were applied to a pure moment of 1.5 Nm in flexion-extension, lateral bending, and axial rotation, respectively. The range of motions (ROMs) and the neutral zones (NZs) of C1 to C2 segment were calculated and compared between 5 status. Results ATS and ATCS fixations significantly reduced the motions in all directions when compared with the intact and injury statues (p < 0.05). In flexion-extension, the ROMs of ATS, unilateral ATCS, and bilateral ATCS were 4.7° ±2.5°, 4.1° ±1.9°, and 3.2° ±1.2°, respectively. Bilateral ATCS resulted in a significant decrease in ROM in flexion-extension when compared with ATS and unilateral ATCS (p = 0.035 and p = 0.023). In lateral bending and axial rotation, there was no significant difference in ROM between the 3 fixations (p > 0.05). Three fixations resulted in similar NZs in all directions (p > 0.05). Conclusion ATCS is a biomechanically effective alternative or supplemental method for atlantoaxial instability.