Osteogenic gene expression in osteoblastic differentiation of human mesenchymal stem cells

Background and ObjectivesDuring differentiation of mesenchymal stem cells (MSCs) into various cells, the expression of a variety of genes undergoes some changes; in this study we decided to investigate the expression rate of some genes like osteopontin (OPN) and osteocalcin (OCN) during this process in order to find a better and faster way for these cells to be differentiated into osteoblasts. Material and MethodsIn this experimental study, the mononuclear cells of bone marrow were separated and then cultured in DMEM-LG culture media with 10% FBS. During some definite days, the RNA of differentiating cells was extracted. Then, the effective genes in osteogenesis like OPN and OCN were amplified by speciefic primers. The mesenchymal cells were cultured on 3D calcium phosphate scaffolds, and finally the activity rate of the alkaline phosphatase was examined. ResultsThis research has demonstrated that in the process of differentiation, the expression of the two genes of OPN and OCN changed orderly with the maximum expression of OPN in the 6th day and the maximum expression of OCN in the 7th and 8th days of differentiation. The osteogenic differentiation of MSCs was not confirmed by the coloration of mineral sediments. The activity rate of alkaline phosphatase revealed the preference of 3D calcium phosphate scaffold to 2D environment in this differentiation. Conclusion The calcium phosphate scaffold positively affects the differentiation process. The expression of OPN and OCN genes changes during differentiation and can be used as away to a better and faster differentiation of these cells into osteoblast.