Abstract

Background and Aim: Evaluating the alkaline phosphatase expression is known as one of the diagnostic markers to confirm the stem cells' differentiation into osteoblasts. Several methods, including immunocytochemistry, are used to measure alkaline phosphatase in research. Due to some limitations of using this method, this study aimed to optimize the calorimetric method as an alternative, low cost and efficient method for quantitative assessment of alkaline phosphatase. Materials and Methods: Mesenchymal stem cells derived from human adipose tissue were used. Cells were differentiated into osteoblasts using an induction medium. Alkaline phosphatase levels were assessed on days 0 and 21 of differentiation using both immunocytochemistry and calorimetric methods. Results: Immunocytochemistry findings demonstrated a significant increase (approximately 5-fold) in alkaline phosphatase expression level on the 21st day of differentiation compared to day 0 (p <0.0001). Also, the results of statistical analysis of the calorimetric test showed a significant increase (approximately 9-fold) in alkaline phosphatase expression level on the 21st day of differentiation compared to day 0 (p<0.0001). Conclusion: The comparative data of both calorimetric and immunocytochemistry showed similar results. These findings suggest the use of colorimetric assay as an alternative, quantitative, fast, and low cost for determining the levels of alkaline phosphatase in osteoblasts differentiated from mesenchymal stem cells. © 2018 the Author (s).