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There is a complex interaction between biomaterials placed as a coronal barrier with stem cells and dentin in regenerative procedures. In this study, the effect of Biodentine (BD; Septodont, Saint-Maurdes-Fossés, France), Endosequence BC Root Repair Material-Putty (ES; Brasseler, Savannah, GA), Endosequence BC Root Repair Material-Putty Fast set (ES-fast, Brasseler), and ProRoot (Dentsply Tulsa Dental Specialties, Johnson City, TN) mineral trioxide aggregate (MTA) on the viability and differentiation of stem cells of the apical papilla (SCAP) was evaluated using an ex vivo dentin disk model.Standardized human dentin disks were treated using an established protocol. Disk lumens were filled with BD, ES, ES-fast, or MTA, and SCAP were cultured directly onto the samples. Cell viability was measured at 7 days, whereas differentiation into a mineralizing phenotype was evaluated by real-time reverse-transcription polymerase chain reaction and alizarin red staining at 21 days in culture. Results were analyzed using 1-way analysis of variance with the Bonferroni post hoc test or the Mann-Whitney U test (P ≤ .05).All materials promoted SCAP viability and proliferation with a greater response in the BD and ES groups. Also, a greater expression of alkaline phosphatase messenger RNA and dentin sialophosphoprotein was noted in the BD and ES groups, whereas MTA promoted a greater expression of the osteoblastic marker IBSP. Interestingly, no difference in alizarin red staining was observed with MTA, BD, or ES, which were significantly greater than ES-fast.These data suggest that BD and ES promoted greater survival and differentiation of SCAP and the increase of the odontoblastic marker DSPP, whereas MTA appeared to promote greater osteoblastic differentiation. Thus, BD and ES can be considered for regenerative and vital pulp therapies.Endosequence root repair material and Biodentine promoted greater viability of stem cells of the apical papilla.Differentiation into a mineralizing phenotype is increased by all bioceramic materials.ProRoot mineral trioxide aggregate promoted up-regulation of osteogenic markers, whereas Endosequence Root Repair Material and Biodentine promoted a greater expression of the odontoblastic markers.The fast set formulation of Endosequence Root Repair Material resulted in reduced viability and differentiation potential.