After treatment with OM for 1, 3, 5 and 7 days, MC3T3-E1 cells were rinsed with PBS, solubilized in lysis buffer (50 mM Tris, 100 mM glycine) for 5 min and centrifuged (2,000 × g; room temperature; 5 min) to collect the supernatant. The ALP activity in the supernatant was then determined using ALP Assay kits (Nanjing Jiancheng Bioengineering Institute) according to the manufacturer's instructions. The ALP activity was normalized to total the protein concentration, which was determined using BCA Protein Assay kits (Thermo Fisher Scientific, Inc.).

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