Micro-CT Analysis of Bone Mineralization

Micro-CT analysis of mineralized bone mass in tibiae was performed using the SkyScan 1272 system (Bruker) at a pixel size of 5 µm with 50 kV tube voltage, 200 µA current and 0.5 mm aluminium filter^{39 (link),41 (link)}. Projection data were reconstructed using the NRecon software (Bruker) and trabecular and cortical volumes of interest were selected manually. For trabecular bone, the region between 0.5 and 3.5 mm distal from the growth plate was selected, and for cortical bone, the region between 2 and 2.5 mm was selected. 3D morphometric parameters were calculated using the CT Analyzer software (Bruker) at a threshold index of 100 to 255 (global Otsu thresholding). Data were presented according to the guidelines of the American Society for Bone and Mineral Research^{42 (link)}. 3D image rendering was performed using the 3D visualization software (Bruker).