The right femurs of the rats were wrapped with saline saturated gauze to maintain moisture

For this section, dilution buffer without antibody was used. After thawing at room temperature, the femurs were moisturized by soaking in saline solution, the residual muscle removed and the length of femur measured with a ruler. The femur bone mineral density between the midpoint and the distal end of the femur was scanned with a SD-1000 single-photon bone mineral density instrument to measure the bone mineral content and bone width. The bone mineral density measurements were performed at the midpoint of the femur and 2 cm proximal and the BMD was calculated as BMC/BW. Bone mechanical properties determination After measuring BMD, the femur was used to determine the bone mechanical properties through three-point bending using Bose ElectroForce Testing system. Bone samples were tested with a 1 mm indenter, at speed of 0.01 mm/s with a 15 mm span. Force and deflection that automatically recorded. The output parameters include elastic force, maximum force, fracture force and the maximum deflection. The stiffness coefficient WZ8040 EGFR/HER2 inhibitor was also calculated based on the above parameters. Bone Histomorphometry XAV939 Wnt/beta-catenin inhibitor The right proximal tibial growth plate and metaphyses were processed for cartilaginous longitudinal growth rate and cancellous bone histomorphometric analyses.