Combination of Chitosan Nanoparticles, EDTA, and Irrigation Activation Enhances TGF-β1 Release from Dentin: A Laboratory Study.

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-β1) 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 + chitosan nanoparticles) and 5 subgroups based on the activation methods (syringe-needle irrigation, Irriflex, PUI, EDDY, and SWEEPS). TGF-β1 release was measured on days 1 and 7 using ELISA. Data were analyzed with the significance level set at P < .05.

Results: TGF-β1 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-β1 levels compared to EDTA or chitosan nanoparticles alone, particularly with SWEEPS and EDDY (P < .05). The percentage increase in TGF-β1 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-β1 release from dentin. These results suggest that both the choice of irrigant and activation method can influence growth factor release in RETs.