Influence of 17β-E2 on biological behavior of HDPCs cultured in vitro

doi:10.3969/j.issn.1002-7386.2016.13.002

Abstract：Objective To investigate the effects of 17 beta estradiol ( 17β-E2 ) on proliferation activity , alkaline phosphates’(ALP) activity, the cell cycle of human dental pulp cells (HDPCs) in vitro.Methods The dental pulp tissue was taken from healthy young people'molars or premolar ,then the dental pulp cells were obtained by tissue explant-collagenase digestion method and cultured in vitro .The fourth generation dental pulp cells were identified by vimentin and keratin immunohistochemisty staining for cell origin .The fourth generation dental pulp cells at logarithmic phase were digested and made into cell suspension with 0.25%trypsin,and the cell suspension was placed into 96-well microplates, then the HDPCs at logarithmic phase were randomly divided into 5 experimental groups ,1 control group and 1 blank control group ,with 6 holes in each group.The experimental groups: 17 beta estradiol 150μl with terminal concentration being 10 -9 ,10 -8 ,10 -7 ,10 -6 , 10 -5 mol/L,respectively was added into corresponding cell holes ,and 17 beta estradiol was placed into phenol red free high sugar DMEM medium containing 2% activated carbon adsorption fetal calf serum .The control group: only DMEM nutrient solution was added into corresponding cell holes . The blank control group: DMEM nutrient solution was added into corresponding holes without cells.The effects of 17β-E2 on the proliferation of HDPCs were detected by MTT at 48h,72h, respectively .The effects of 17β-E2 on the cell cycle of HDPCs were detected by flow cytometry .Results In contrast with control group, 17β-E2 could promote proliferation of HDPCs and cell cycle process within a certain range of concentration . The effects of 17β-E2 in concentration of 10 -8 mol/L were the most significant ( P <0.05).Conclusion The 17β-E2 can induce proliferation of HDPCs at some extent in vitro .