Micro-computed tomography (micro-CT) assay

Right femora bones of mice were taken and the soft tissue around the bone was removed. These bones were fixed in 4% polyformaldehyde (PFA) solution and kept in 4° C for 48 hours. To estimate the change of bone microarchitecture, 1176 micro-computed tomography scanner (Skyscan, Belgium) with 9-mm voxel size, 59 KVp, 127 μA, and 0.48 rotation step, was used to measure the distal region of the femur from 60 mm approximately to the end of the distal growth plate over 1.7 mm toward the diaphysis. The following trabecular parameters were analyzed: bone mineral density (BMD), percent bone volume (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N) and trabecular separation (Tb.Sp).