Introduction: EDTA has been considered the gold standard in regenerative endodontic treatments. The aim of this study was to evaluate the effects of different dentin conditioning agents other than EDTA on released growth factors, mesenchymal stem cell attachment, and morphology. Methods: Transforming growth factor beta 1, vascular endothelial growth factor, bone morphogenetic protein 2, and fibroblast growth factor 2 release from prepared dentin discs conditioned with 17% EDTA, 10% citric acid, 1% phytic acid (IP6), or 37% phosphoric acid were quantified using the enzyme-linked immunosorbent assay after final irrigation and after 3 days of adipose-derived mesenchymal stem cell (adMSC) seeding. Forty root fragments were prepared from extracted Single-rooted teeth. The morphology and attachment of adMSCs on the conditioned root fragments were observed using a scanning electron microscope. Data for growth factor quantification were analyzed using 1-way analysis. Results: The highest transforming growth factor beta 1 release was observed after citric acid treatment followed by phosphoric acid; there was no significant difference between them, but compared with EDTA and 1% IP6, there were significant differences observed. The enzyme-linked immunosorbent assay detected a very minor exposure of vascular endothelial growth factor and fibroblast growth factor 2 after dentin conditioning, but there were no significant differences between the groups. The greatest bone morphogenetic protein 2 release was observed in the 1% IP6 group, but there were no significant differences between the groups. Three days of adMSC seeding after dentin conditioning has made a dramatic increase in all of the growth factors, and phosphoric acid appeared to be the most effective agent with significant differences compared with the remaining groups. Scanning electron microscopic observations showed that none of the conditioning solutions had an adverse effect on stem cell proliferation and attachment to root dentin. Different cell morphologies like round, oblong, flat, and well-attached cells with developed filopodia were observed in the dentin-conditioned groups. Conclusions: Phosphoric acid conditioning could be useful and may have beneficial effects in regenerative endodontic treatments.