MTA和三种改良盖髓剂对人牙髓干细胞增殖及分化为成牙本质细胞的促进作用观察OACSTPCD

Objective To observe the promoting effects of mineral trioxide aggregate(MTA)and three modified pulp-capping agents(namely nRoot,Vitapex,and iRoot BP Plus)on the proliferation and differentiation of human dental pulp stem cells(hDPSCs)into odontoblasts.Methods ① Observation on the promoting effects of four different concen-trations of pulp-capping agents on proliferation of hDPSCs:In this study,the fifth-passage hDPSCs were divided into the MTA,iRoot BP Plus,nRoot,Vitapex,and control groups.The MTA,iRoot BP Plus,nRoot,and Vitapex groups were treated with various concentrations(0.02,0.2,1,and 2 mg/mL)of MTA,iRoot BP Plus,nRoot,and Vitapex culture media,respectively;the control group was treated with DMEM/F12 culture medium containing 10%fetal bovine serum(FBS).Cell proliferation activity was assessed using the CCK-8 method at 24 and 48 h after incubation.② Observation on the promoting effects of four pulp-capping agents on differentiation of hDPSCs into odontoblasts:The optimal inducing con-centrations of the four materials for hDPSCs were screened through alkaline phosphatase(ALP)activity detection.The fourth-passage hDPSCs were divided into five groups:control,MTA,iRoot BP Plus,nRoot,and Vitapex groups.The cells in the MTA,iRoot BP Plus,nRoot,and Vitapex groups were treated with osteogenic media containing 0.2 mg/mL of MTA,iRoot BP Plus,nRoot,and Vitapex,respectively,while the control group was treated with normal osteogenic me-dia.On day 21 of culture,alizarin red staining and semi-quantitative analysis were used to observe the formation of miner-alized nodules in each group.On day 7 of culture,Western blotting was employed to detect the expression levels of runt-re-lated transcription factor 2(RUNX-2),osteocalcin(OCN),dentin sialophosphoprotein(DSPP),and dentin matrix pro-tein 1(DMP-1).Results On the second day of culture,compared with the control group,the cell proliferation activity was significantly higher in the MTA group,iRoot BP Plus group and nRoot group at concentrations of 0.02,0.2,and 1 mg/mL(all P<0.05).Additionally,compared with that of the first day of culture,the cell proliferation activity decreased in the MTA group and Vitapex group at 2 mg/mL on the second day(all P<0.05).Consequently,0.2 mg/mL was select-ed as the subsequent test concentration.In comparison to the nRoot and Vitapex groups,the iRoot BP Plus and MTA groups exhibited higher cellular calcium deposition(all P<0.05).Furthermore,compared with the control group,the rela-tive expression levels of OCN,RUNX-2,DSPP,and DMP-1 were up-regulated in the MTA,iRoot BP Plus,nRoot,and Vitapex groups(all P<0.05).Notably,the iRoot BP Plus group demonstrated higher relative expression levels of OCN,RUNX-2,DSPP,and DMP-1 as compared with both the nRoot and Vitapex groups(all P<0.05).Conclusion Com-pared with MTA and Vitapex,nRoot and iRoot BP Plus are more effective in promoting the proliferation of hDPSCs and their differentiation into odontoblasts.