Identification of OC precursors and mOC by fluorescent triple staining
TRAP-positive
Fluorescent triple staining of OC precursors and mOCs was performed as previously described (Using a confocal microscope (DMI4000B, Leica, Wetzlar, Germany), microscopic pictures were taken (5-10 images per bone slice). According to established criteria, the cells were classified into three groups: pro- (spindle-shaped with a single nucleus), pre- (round with one or two nuclei) and mOCs (TRAP-positive and with three or more nuclei) (