Evaluation of antibiofilm activity of phytofabricated magnetic iron nanoparticles against Enterococcus faecalis biofilm

Objectives: This study aimed to evaluate the antibiofilm efficacy of phytofabricated magnetic iron nanoparticles against Enterococcus faecalis , a key pathogen implicated in persistent endodontic infections. Methods: Methanol-water solvent extraction of Murraya koenigii was performed and utilized for the phytofabrication of iron nanoparticles using iron (III) chloride under controlled conditions. Nanoparticle formation was confirmed using UV–Vis spectroscopy and further characterized by X-ray diffraction (XRD) to determine structural properties and particle size. Antibiofilm activity against mature E. faecalis biofilms was assessed using crystal violet assay and confocal laser scanning microscopy (CLSM). Statistical analysis was performed to validate the findings. Results: Successful phytofabrication of iron nanoparticles was indicated by the formation of a dark black-colored solution. UV–Vis spectroscopy confirmed nanoparticle synthesis, while XRD analysis revealed crystalline nanoparticles with an average particle size of 19 nm. Crystal violet assay demonstrated a concentration-dependent inhibition of biofilm formation, with a maximum suppression of 76% at 1000 μg/ml (p < 0.001). CLSM analysis showed a significant reduction in microbial load and biofilm thickness from 15 μm to 3.5 μm following nanoparticle treatment. Conclusions: Green-synthesized iron nanoparticles derived from Murraya koenigii exhibit significant antibiofilm activity against Enterococcus faecalis . Their eco-friendly synthesis and potent biofilm disruption highlight their potential as an adjunctive strategy for root canal disinfection and management of persistent endodontic infections. Further in vivo studies are warranted to establish clinical applicability.