The initial fixation strength of pedicle screws is commonly tested using a standard pull-out test with load applied at a constant rate. This method overlooks the cyclic nature of in situ loading responsible for clinical failure. This study was undertaken to determine the effects of stress relaxation properties at the bone-screw interface on screw fixation strength. Pedicle screws were inserted into calf lumbar vertebrae using a paired testing array. After embedding and mounting in a custom fixture, axial pull-out tests were performed at the rates of 1, 5, and 25 mm/min. For each vertebra, one screw was pulled at a continuous rate. The other screw was pulled at increments of 0.5 mm, at the same rate, with 1000 s pause between increments. Peak load, energy-to-failure, displacement-to-failure, and stiffness were calculated for each screw pull-out test. Two-way ANOVA showed that the standard pull-out method yielded significantly higher peak loads (p < 0.05) at faster pull-out rates and higher stiffnesses (p < 0.05) at all rates compared to the stress relaxation pull-out protocol. These results suggest that the stress relaxation properties of bone significantly affect the pull-out behavior of pedicle screws, reducing the peak load and stiffness values observed during testing. This mode of testing may provide a better biomechanical model of screw pull-out failure and a more accurate estimate of initial fixation strength.