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To quantify alkaline phosphatase (ALP) activity of MG-63 cells after 7, 14, and 21 days, the Alkaline phosphatase assay was done based on the manufacturer's guidance (Pars Azmoon, Iran). Briefly, the supernatants were obtained, and a p-nitrophenyl phosphate transformed to a p-nitrophenol. The reaction was terminated by adding 100 μl NaOH solution (1 N). The absorbance was read by an ELISA reader (Microplate Reader Model 1680, Bio-Rad, USA) at 405 nm.
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