Bone mechanical testing by 3-point bending test

Bone mechanical testing of femurs was performed as our previously described protocol ¹. Bone femurs were fixed in 4% PFA for overnight and stored in 70% ethanol at -80 °C until usage. A 3-point bending test was performed to study bone mechanical properties of cortical strength. All femur bones were tested under a load applied at a constant rate of 20 mm/min. This test predominantly measured the cortical bone strength parameters like maximum load (ultimate strength) and stiffness (elastic deformation) by using Bone Strength Tester TK-252C (Muromachi Kikai Co. Ltd., Tokyo, Japan).