The objective of this study was to evaluate the anti-biofilm efficacy of photodynamic therapy by conjugating a photosensitizer (TBO) with silver nanoparticles (AgNP). Streptococcus mutans was exposed to laser light (630 nm) for 70 s (9.1 J cm(-2)) in the presence of a toluidine blue O-silver nanoparticle conjugate (TBO-AgNP). The results showed a reduction in the viability of bacterial cells by 4 log10. The crystal violet assay, confocal laser scanning microscopy and scanning electron microscopy revealed that the TBO-AgNP conjugates inhibited biofilm formation, increased the uptake of propidium iodide and leakage of the cellular constituents, respectively. Fluorescence spectroscopic studies confirmed the generation of OH(*) as a major reactive oxygen species, indicating type I phototoxicity. Both the conjugates down-regulated the expression of biofilm related genes compared to TBO alone. Hence TBO-AgNP conjugates were found to be more phototoxic against S. mutans biofilm than TBO alone.
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