Mice were subcutaneously administered with calcein (Sigma) at a dose of 15 mg/kg at day 10 and day 3 before sacrifice. After fixation and embedding in polymethyl acrylate, femurs were serially cut into 50-μm-thick sections, and the double calcein labeling was observed with a fluorescence microscope (Nikon). The bone formation rate (BFR) was calculated as the inter-label width/labeling period with Image-Pro software.
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