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Abstract Aim of the study: This study intended to examine the influence of Acetyl-11-keto-β-boswellic acid(AKBA)on Dental pulp Stem cells(DPSCs)viability and proliferation and its influence on the differentiating activity of the DPSCs to be used as a prospective root canal medicament. Materials and Methods :. DPSCs were isolated from human third molars, Flow cytometry analysis was employed to determine the phenotypic characterization of DPSCs, The cytotoxicity assay was applied using the Methyl-Thiazoltetrazolium (MTT) assay Cells were medicated with different concentration of Triple antibiotic paste (TAP) and Calcium Hydroxide Ca(OH2)(5,2.5,1,0.5,and0.25 mg/ml),AKBA(10,5,1,0.1,and0.01µM). All experiments were done in separate triplicate experiments The differentiation of DPSCs and calcium deposition was then examined by ALP activity,Alizarin red and Von Kossa stains, Also Real Time quantitative PCR analysis of gene expression for(DMP-1), (DSPP), (COLLA1),(RUNX2),(FN),(OPN), (GLAP), (BSP(102), and (MEPE) of DPSCs was executed after treatment of cells with TAP, Ca(OH)2 in concentration 0.5mg/ml and AKBA in concentrations(0 01, 0.1, and 1µM). Results:DPSCs were characterized by flow cytometry.Cells medicated with Ca(OH)2(1mg/ml,2.5mg/ml,and 5mg/ml)showed significant DROP in viability relative to the control(p < 0.05).DPSCs medicated with(1,2.5, and 5mg/ml)TAP resulted in a significant reduction in viability(p<0.05),Cells medicatedwithAKBAin(1,0.1and,0.01µM)concentrations resulted in more viability than the control(p <0.05).Concerning cell density assay,AKBA resulted in increase in cell density after 5 and 7days in comparison to cells, medicated by TAP and Ca(OH)2.After 14 days,calcified nodules were more enlarged and homogenously distributed in DPSCs cultured with(0.01,0.1,and1µM AKBA.Cells medicated with different AKBA concentrations resulted in significantly higher(p < 0.05)ALP activity than the other tested groups.Calcium clusters of variable densities were evident by the use of alizarin red staining, which also revealed other mineral deposits of varying densities.Formation of mineralized dentin-like calcified nodules was highly observed in DPSCs treated with different concentration of AKBA especially cells treated with 0.1 µM AKBA with Von Kossa staining.In Gene expression analysis treatment with AKBA showed higher levels of COL-1,OPN,BSB and MEPE than NC and TAP groups(p<0.05) specially with 0.1 µM AKBA While the other tested genes didn’t show any significant difference with the different AKBA concentrations,where DMP1,COLLA 1,FN,RUNX ,and MEPE showed significant increase with Ca(OH)2. Conclusion and Recommendations : AKBA in lower concentrations(0.01,0.1, and 1µM) resulted in superior viability than TAP and Ca(OH)2,and it may possess the potential to be an intracanal medicament in regenerative endodontics. Also AKBA may promote the osteoblastic differentiating ability of DPSCs.It is recommended to be tested clinically. Key Words (not more than 10) :Intracanal medications , Cytotoxicity , DPSCs, MTT , AKBA,TAP, Ca(OH)2 ,PCR, Gene expression. |