Effects of gradient glucose solution glucose on cultured osteoblasts in vitro

Objective:To explore the effect of high gradient glucose on the proliferation and differentiation of osteoblastic cells(OB) cultured in vitro.Methods:OB was isolated from the skull of 1～2 day newly born SD rats by means of Trypsin-collagenase digestion,and the osteoblasts were identified by morphologic analysis,V-G collagen staining,ALPase staining,calcification nod staining etc.Different concentrations of glucose were added to the cultured OB.The effects of glucose on the proliferation and differentiation of osteoblasts was monitored by MTT analysis,ALPase activity analysis and formation of mineralized nodulus.Results:Different concentrations of glucose promoted the proliferation of osteoblastic cells for 48 hours culture,but inhibited for 72 hours when it was beyond 15 mmol/L.Among these different concentrations glucose,the glucose of less than 15 mmol/L could promote osteoblastic cells proliferation on the early period of cultivation (P0.01,P0.05) and has no suppression during a relative long time,and the glucose of more than 30 mmol/L inhibited the secretion of ALPase for 7 days(P0.05),and the glucose of beyond 10 mmol/L could delay formation of mineralized cell nodulus for 18 days,below 10 mmol/L glucoses did not inhibit the secretion of ALPase and formation of mineralized nodulus.Conclusion:High gradient glucose could influence on the proliferation,differentiation and mineralization of osteoblastic cells.The tolerance concentrations of glucose were no more than (15 mmol/L) on the proliferation of OB,and the differentiation and mineralization of OB were below 10 mmol/L.