Combination of Chitosan Nanoparticles, EDTA, and Irrigation Activation Enhances TGF-Β1 Release From Dentin: A Laboratory Study

Abstract

Introduction: This study aimed to investigate the effects of sonic (EDDY), ultrasonic (PUI), and laser (shock wave-enhanced emission photoacoustic streaming [SWEEPS]) activation of chitosan-nanoparticles enriched ethylenediaminetetraacetic acid (EDTA) irrigation on the release of transforming growth factor beta 1 (TGF-(31) from dentin in regenerative endodontic treatments (RETs). Methods: One hundred twenty extracted human teeth were prepared up to size #100 to mimic the open apex. Root canals were irrigated with 20 mL 1.5% NaOCl according to the RETs protocol. Teeth were divided into 3 main groups according to the irrigation solution (17% EDTA, 0.2% chitosan nanoparticles, and the combination of EDTA 1 chitosan nanoparticles) and 5 subgroups based on the activation methods (syringe-needle irrigation, Irriflex, PUI, EDDY, and SWEEPS). TGF-(31 release was measured on days 1 and 7 using ELISA. Data were analyzed with the significance level set at P < .05. Results: TGF-(31 release increased progressively from syringe-needle irrigation to Irriflex, PUI, EDDY, and SWEEPS, with SWEEPS achieving the highest release at both time points (P < .05). The combination of EDTA and chitosan nanoparticles yielded significantly higher TGF-(31 levels compared to EDTA or chitosan nanoparticles alone, particularly with SWEEPS and EDDY (P < .05). The percentage increase in TGF-(31 release from day 1 to day 7 was significantly higher in all activation groups (P < .05). Conclusion: The combination of EDTA and chitosan nanoparticles, particularly when used with advanced irrigation activation methods such as SWEEPS and EDDY, significantly increased TGF-(31 release from dentin. These results suggest that both the choice of irrigant and activation method can influence growth factor release in RETs. (J Endod 2025;51:1081-1087.)