Mechanical properties of 12 tibiae per group were measured as previously described. After removing soft tissues, tibia samples were wrapped in wet tissue and frozen at −20°C. Sixteen hours before testing, the samples were transferred to 4°C and then to RT an hour before testing. The four-point bending mode of ElectroForce 3220 (TA Instruments, New Castle, DE) instrument with 225-N load cell was used. Samples were aligned on the fixtures in a manner that the load was applied perpendicular to the principal axis of tibia. The span length of the bottom support was 9.2 mm, while the top span length was 2.8 mm. Bending test was performed in displacement control mode at a rate of 0.025 mm/s. Load-displacement data were recorded at a data acquisition rate of 10 Hz. Displacement was tared at the first data point at which the load equaled or exceeded 1 N. Maximum load is the highest load (newtons) before the sample fractures. Bending stiffness (newtons per millimeter) was calculated as the slope of load versus displacement between 30 and 70% of maximum load to failure in the linear region.

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