HE staining was conducted according to routine protocols [17 (link)]. Briefly, after deparaffinization and rehydration, 5 μm longitudinal sections were stained with hematoxylin solution for 5 min followed by 5 dips in 1% acid ethanol (1% HCl in 70% ethanol) and then rinsed in distilled water. Then the sections were stained with eosin solution for 3 min and followed by dehydration with graded alcohol and clearing in xylene. The mounted slides were then examined and photographed using an Olympus BX53 fluorescence microscope (Tokyo, Japan). The staining intensity of the trabecular bone was analyzed by Image-Pro Plus 6.0 software and expressed as IOD value.
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