Effective Inactivation of Enterococcus faecalis Biofilms via Singlet Oxygen Induced by Calcination Betaine Hydrochloride Carbon Dots Under Light Irradiation

This study synthesizes novel photosensitizer calcination betaine hydrochloride carbon dots (CBCDs) to address the critical challenge of Enterococcus faecalis (E. faecalis) biofilms, a major cause of root canal treatment failure. To this end, this study investigates the effective elimination via reactive oxygen species (ROS) mediated by these CBCDs. CBCDs were prepared by calcining betaine hydrochloride and rigorously characterized for their structural and chemical properties using transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS). Their optical characteristics were also thoroughly analyzed through UV-Vis and fluorescence spectroscopy. The RNO-ID assay was performed to explicitly confirm ROS production, particularly verifying significant singlet oxygen (1O2) generation. Bactericidal efficacy of the CBCDs was comprehensively evaluated against planktonic E. faecalis and its formed biofilms. Live/dead staining was subsequently performed to observe their state after treatment. As a result, TEM confirmed nanosized CBCDs, and FTIR/XPS analyses identified crucial functional groups. Colony Forming Unit (CFU) assays revealed a dose-dependent reduction in E. faecalis viability, achieving complete eradication at 200 mg/L under light irradiation. Complete cell death and inactivation of the formed biofilms with increasing CBCD concentrations were also strongly evidenced by red fluorescence. The obtained results underscore CBCDs as highly effective photodynamic agents for the robust elimination of E. faecalis biofilms, offering a promising new strategy to combat persistent oral infections.