Tartrate-resistant acid phosphatase (TRAP) staining for callus was performed, as previously described29 (link),30 (link). In brief, decalcified sections were first deparaffinized and preincubated. Subsequently, the slides were incubated for 45–60 min at 37 °C with TRAP staining solution, based on the manufacturer’s protocol (387A-1KT, Sigma-Aldrich, MO, USA). The slides were also counterstained with hematoxylin solution. For quantification, the OC surface was calculated using an image processing program (ImageJ software, National Institutes of Health, Bethesda, MD, USA).
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