[Effects of Different Concentrations of Glucose on the Osteogenic Differentiation of Orofacial Bone Mesenchymal Stem Cells].

OBJECTIVES

To determine the effects of different concentrations of glucose on the proliferation and osteogenic differentiation of orofacial bonemesenchymal stem cells (OFMSCs).

METHODS

OFMSCs were primarily cultured and identified to undergo osteogenic/adipogenic/chondrogenic differentiation. The cells were exposed to osteogenic medium containing different levels of glucose: 5.5,11,16.5,25,44 mmol/L. The cell activity and proliferation index were measured using a cell counting kit (CCK)-8 and flow cytometry. The alkaline phosphatase (ALP) activity of the cells was measured at the 4 and 7 day. Alizarin red staining was carried out at the 21 day. RT-PCR detecting osteogenesis-related gene and Osterix mRNA expression was performed at the 4, 7, 14 and 21 day.

RESULTS

Osteogenesis induced calcium nodes was observed with Alizarin red staining at the 21 day. Adipogenic induced red lipid droplets was observed with Oil Red O staining at the 14 day. Chondrogenic induced blue cytoplasm was observed with Alcian blue staining at the 14 day. With 5.5 to 25 mmol/L glucose,OFMSCs proliferation was promoted.But when the concentration of glucose continued to increase (from 25 to 44 mmol/L),OFMSCs proliferation significantly reduced.The ALP activity decreased with glucose in a concentration-dependent manner ( <0.05). Osteogenesis induced Alizarin red staining and mineralization showed at the 21 day. The calcium nodes and mineralization quantity decreased with glucose in a concentration-dependent manner ( <0.05).The cells exposed to 5.5 mmol/L glucose had a higher level of expression of and Osterix mRNA than the others (<0.05).The experssion of and Osterix mRNA in all groups showed a trend of rising first, followed by an obvious down regulation.

CONCLUSIONS

With certain limits,OFMSC proliferation is promoted by glucose. Osteogenic differentiation is inhibited by glucose in a concentration-dependent manner.