The Osteogenic Differentiation of Mice Bone Marrow Mesenchymal Stem Cells with Fisetin Phytoestrogen

Abstract

Objective(s): The aim of this study was to evaluate the effects of fisetin to promote osteogenic differentiation in mice bone marrow mesenchymal stem cells (BMSCs).Materials and Methods: In this study cytotoxicity and viability of fisetin was measured by MTT assay. The differentiation effects of fisetin on BMSCs into osteoblast was assessed with alkaline phosphatase (ALP) activity measurement. Alizarin red staining and Real time PCR for osteoblast specific marker, Osteocalcin (OCN) ,Osteopontin (OPN), Runt-related transcription factor 2 (RUNX2), ERK and MAPK were investigatedResults: The results showed that fisetin does not have toxicity effect on BMSCs and it causes cell proliferation; hence 200, 400 and 800 µg/ml of fisetin was selected for the assessment of differentiation progress. Alizarin red staining (ARS) showed that fisetin promotes osteogenic differentiation on BMSCs at 21st day; dependently also higher alkaline phosphates activity was observed in the treatment groups of 10 days culture, compared to the control groups. The evaluation of Real time -PCR result evaluated showed that OCN OPN, RUNX2,ERK and MAPK genes expressions were increased.Conclusion: The results of this method, showed that differentiation in bone marrow stem cells took place through p38 MAPK, and ERK1 gene activation.