Dentin conditioning protocol for regenerative endodontic procedures.

Abstract Aim This study focused on (1) the optimization of NaOCl-EDTA irrigation in terms of the viability and morphology of dental pulp stem cells (DPSCs), (2) the effects of optimized EDTA protocol alone or prepared with nanobubble water (NBs) on cell behavior. Materials and Methods In the first part, human dentin discs were conditioned with the following protocols; (1)NaOCl, followed by PBS (NP); (2)NP, followed by EDTA (NPE); (3)NPE, followed by PBS (NPEP); (4)without any PBS rinse (NE) or (5)only final PBS rinse (NEP). DPSC viability and morphology were determined. In the second part, dentin discs were conditioned with (1) the optimized protocol in the first part (EDTA); (2) EDTA prepared using NBs (EN); (3) ultrasonic-activated EDTA (EU) or (4) EN irrigation (ENU). Transforming growth factor (TGF)-β release and DPSC viability & morphology, and migration were determined using ELISA, WST-1 cell viability assay & live-dead assay, and transwell migration, assay, respectively. Data were analyzed using Kruskal Wallis or one-way ANOVA and post-hoc tests. Results The highest cell viability was observed in the NPEP group (p 0.05). Conclusion Removing the residual EDTA using PBS improved the cell viability on the dentin surface. Ultrasonic activation enhanced the growth factor release and biological properties while the preparation of EDTA with NBs showed a similar effect to regular EDTA without compromising cellular effect.