Background:​ Because embalming has been demonstrated to decrease the mechanical integrity of bone, most investigators favor fresh-frozen specimens for biomechanical evaluation. However, little is known about how the integrity of fresh-frozen specimens may change during biomechanical testing or may be affected by standard practices in testing.

Objective:​ The purpose of this study was to evaluate how the time after removal from a freezer may affect the mechanical properties of fresh-frozen diaphyseal bone.

Methods:​ Matched pairs of nonosteoporotic fresh-frozen human cadaveric femora were thawed before instrumentation with bicortical screws. Matched femora were reserved for either control or delayed use. Each specimen received standard diaphyseal bicortical screws (six or more in each group). At specified time points, screws were axially pulled out following the guidelines of ASTM F543-07. Test groups were stored in air (21 ± 0.5°C) for 16, 50, or 90 hours. In the control group, screws were pulled out at 16 hours, which corresponds to the minimum elapsed time for specimen thawing, instrumentation, potting, and biomechanical test initiation. This represents the baseline mechanical properties of the fresh-frozen bone at the inception of any biomechanical test. The 90-hour group corresponds to the time needed to cycle a construct 300,000 times at a physiological test frequency of 1 Hz. This corresponds approximately to 2 to 4 months of in vivo loading. A midpoint of 50 hours was also tested, representing approximately 180,000 cycles.

Results:​ Failure for all specimens occurred as a result of bone failure at the screw-to-bone interface. There was a decrease in screw pullout strength as exposure time in air increased. The 50-hour test group showed a 9% decrease in screw pullout strength as compared with the 16-hour control group (P = 0.81). However, the 90-hour test group showed a 30% decrease in screw pullout strength as compared with the 16-hour control group (P = 0.04).

Conclusion:​ This study indicates that when using fresh-frozen cadaveric bone in biomechanical tests to simulate the orthopaedic clinical setting, specimen exposure time should be considered. The timing of testing should be kept constant between specimens to allow for a proper comparison. Furthermore, for fresh-frozen cadavers, the physical properties of bone may be detrimentally affected in biomechanical testing that exceeds the 50-hour time point after removal from the freezer.