2.3. Tartrate resistant acid phosphatase (TRAP) assay

After seven days of culture, the cells were histochemically stained to detect TRAP by using an Acid Phosphatase staining kit (387 A, Sigma‐Aldrich) according to the manufacturer's instructions. Briefly, the media was removed and the wells were gently rinsed with PBS. BMMs were fixed with 4% PFA and then incubated at 37°C with a solution containing deionized water, Fast Garnet GBC, Napthol phosphate, Acetate, and Tartrate for 1 h. TRAP positive cells with three or more nuclei were counted as osteoclast using light microscopy.