Synergistic effect of methylene blue and biogenic gold nanoparticles against Enterococcus faecalis

This study reports successful photodynamic inactivation of planktonic and biofilm cells of Enterococcus faecalis using Methylene Blue (MB) in combination with gold nanoparticles synthesized using the cell-free filtrate obtained from 3-day biomass of Trichoderma asperellum strain. Monodispersed colloidal gold nanoparticles were characterized by UV-vis absorption, TEM and DLS to be 13 ± 3 nm spheres. Diode lasers with the peak-power wavelength ʎ = 660 nm (output power of 21, 41 and 68 mW; power density of 55, 108 and 179 mW∙cm^-2, respectively, were used as a light source to study the effects of MB alone, the gold nanoparticles alone (AuNPs) and the MB + AuNPs mixture on the viability of E. faecalis cells. The lethal effect of planktonic cells was achieved for MB after 30 min of laser irradiation with energy fluence of 322 J∙cm^-2. When MB + AuNPs mixture was used as photosensitizer, the lethal effect was achieved with energy fluence of 292 J∙cm^-2. The biofilm culture was more resistant to photo-inactivation and the best bactericidal effect of MB as photosensitizer was found after light dose of 483 J∙cm^-2. The bacterial cell viability was reduced by 99.92%. It was proved that MB + AuNPs mixture synergistically enhances the kill of the studied microorganism as the same light dose resulted in 99.991% kill.