2.19. Immunohistochemical staining

The decalcified samples were fixed, dehydrated, made xylene-transparent, wax-dipped and embedded in paraffin. For H&E staining, the histological sections (5 μm) were dewaxed and immersed in hematoxylin staining solution for 3 min and eosin staining solution for 45 s. Finally, the histological sections were mounted in mounting media. For PDGF-B, TRAP, Col Ⅰ, and OCN immunohistochemical staining, the sections were treated with primary antibodies and peroxidase-conjugated secondary antibodies. The signals were detected with a DAB kit. Immunohistochemical staining was analyzed by using Image J software.