The Effect And Mechanism Of IRoot BP Plus On Proliferation And Differentiation Of Human Dental Pulp Stem Cells

Objectives This study aims to investigate the effect of iRoot BP Plus on the proliferation,osteogenesis of human dental pulp stem cells(hDPSCs),as well as the mechanism involved in.This study will provide basic research basis for the application of new bioactive material,iRoot BP Plus,in the preservation of vital pulp.Methods 1 The dental pulp tissues were isolated and cultured by the tissue block enzymolytic method,Flowcytometry was used to analyze the surface markers of hDPSCs.2 Preparation iRoot BP Plus and MTA extracts with concentration of 5.56 mg/ml,11.11 mg/ml and 33.33 mg/ml respecttively.The hDPSCs were treated with the extracts at different concentrations for 48 hours,the relative proliferation rate of hDPSCs was detected by CCK-8.3 The hDPSCs were treated with iRoot BP Plus and MTA at the optimal concentration(33.33 mg/ml)for 14 days,respectively,the mineralized nodules of iRoot BP Plus group,MTA group and negative control(NC)were stained by alizarin red.4 The relative mRNA levels of OC,BSP and COL-I genes in each detected by qRTPCR.5 The relative protein levels of ERK,JNK,phosphorylated-ERK(p-ERK)and p-JNK in MAPK signaling pathway in DPSCs were detected by Western Blot.Results 1 The derived hDPSCs were positive expression of CD44,STRO-1and negative expression of CD34,these consistent with the characteristics of pulp stem cells.2 The proliferation activitites of hDPSCs were significantly increased by iRoot BP Plus and MTA with each concentration(P< 0.01).The effect of iRoot BP Plus on the proliferation ability of hDPSCs was higher than that of the same concentration of MTA(P< 0.05).3 The number of calcium nodules formed in iRoot BP Plus group was large quantity and high density than that in MTA group,and the IOD value in iRoot BP Plus group was significantly higher than that in MTA group(P< 0.01).4 The relative mRNA of osteogenic genes OC,BSP and COL-I in hDPSCs in both iRoot BP Plus group and MTA group were significantly higher than those in the control group(P< 0.01).The relative mRNA levels in the iRoot BP Plus group was significantly higher than that in the MTA group(P< 0.01),while the relative mRNA level of BSP in MTA group was markedly higher than that in iRoot BP Plus group(P< 0.01).5 The relative protein levels of p-ERK and p-JNK in both iRoot BP Plus group and MTA group were significantly higher than those in the control group(P< 0.01),and the relative protein level of p-ERK in the iRoot BP Plus group was higher than that in the MTA group(P< 0.05).Conclusions 1 The effect of iRoot BP Plus on promoting the proliferation ability of hDPSCs is better than MTA at the same concentration.2 The effect of iRoot BP Plus on promoting the mineralization ability of hDPSCs is better than MTA.3 The effect of iRoot BP Plus on promoting the osteogenic differentiation of hDPSCs is may mediated by activating the MAPK signaling pathway and followlled by the osteogenic genes OC,BSP and COL-I.iRoot BP Plus may be better than MTA in regulating MAPK signaling pathway and downstream genes.