Photodynamic therapy using methylene blue, combined or not with gentamicin, against Staphylococcus aureus and Pseudomonas aeruginosa.

Antimicrobial photodynamic therapy (a-PDT), combined or not with antibiotics, constitutes a promising therapy for superficial infections caused by bacteria implicated in multidrug resistance processes. We compared the efficacy of aPDT using the photosensitizer methylene blue (MB), combined or not with the antibiotic gentamicin (GN), against Staphylococcus aureus and Pseudomonas aeruginosa. Different concentrations of MB (0.03-7000 µg/mL), with or without GN (1-20 µg/mL), were added to planktonic cultures or biofilms and the samples irradiated with a LED lamp (λ 625 nm, 7 mW/cm2, 18 J/cm2). The number of viable bacteria in the samples and in corresponding nonirradiated controls was quantified by counting colony-forming units to evaluate the individual effects of MB, GN, and irradiation. MB-aPDT resulted in significant bacterial photoinactivation. The combination of GN and MB-aPDT exerted a synergistic bactericidal effect against planktonic cultures of S. aureus and P. aeruginosa. This combination did not significantly alter the photoinactivating effect of MB against S. aureus biofilms, but exerted a positive bactericidal effect against P. aeruginosa biofilms. These results underscore the need for further clinical studies of this therapeutic combination for the management of difficult-to-treat skin and mucous infections, especially those caused by P. aeruginosa.

Antimicrobial photodynamic activity of Rose Bengal, alone or in combination with Gentamicin, against planktonic and biofilm Staphylococcus aureus.

Antimicrobial photodynamic therapy (aPDT) could constitute an alternative therapy to antibiotics especially against superficial infections caused by bacteria involved in multidrug resistance processes. The aim of this study is to compare the efficacy of aPDT using the photosensitizer rose bengal (RB), combined or uncombined with gentamicin (GN), against Staphylococcus aureus. Different concentrations of RB (ranging from 0.03 to 64 µg/ml) were added to S. aureus in water suspensions or forming biofilms in the absence or presence of GN (1-40 µg/ml) and the samples were irradiated (18 or 37 J/cm2). The number of viable bacteria was quantified by counting colony-forming units. RB-aPDT shows significant photoactivity. The combination of GN and RB-aPDT exerts a synergistic bactericidal effect against planktonic S. aureus. On the other hand, a synergistic effect is observed only when the maximum concentration tested of RB and GN was used in biofilm. According to these result the use of RB-aPDT alone or in combination with GN could be implemented against S. aureus.

Bactericidal Effect of Photodynamic Therapy, Alone or in Combination with Mupirocin or Linezolid, on Staphylococcus aureus.

Antibiotic treatments frequently fail due to the development of antibiotic resistance, underscoring the need for new treatment strategies. Antimicrobial photodynamic therapy (aPDT) could constitute an alternative therapy. In bacterial suspensions of Staphylococcus aureus, which is commonly implicated in cutaneous and mucosal infections, we evaluated the in vitro efficacy of aPDT, using the photosensitizing agents rose bengal (RB) or methylene blue (MB), alone or combined with the antibiotics mupirocin (MU) or linezolid (LN). RB or MB, at concentrations ranging from 0.03 to 10 µg/ml, were added to S. aureus ATCC 29213 suspensions containing >108 cells/ml, in the absence or presence of MU or LN (1 or 10 µg/ml). Suspensions were irradiated with a white metal halide (λ 420-700 nm) or light-emitting diode lamp (λ 515 and λ 625 nm), and the number of viable bacteria quantified by counting colony-forming units (CFU) on blood agar. Addition of either antibiotic had no significant effect on the number of CFU/ml. By contrast, RB-aPDT and MB-aPDT effectively inactivated S. aureus, as evidenced by a 6 log10 reduction in bacterial growth. In the presence of MU or LN, the same 6 log10 reduction was observed in response to aPDT, but was achieved using significantly lower concentrations of the photosensitizers RB or MB. In conclusion, the combination of MU or LN and RB/MB-aPDT appears to exert a synergistic bactericidal effect against S. aureus in vitro.

Comparative effect of photodynamic therapy on separated or mixed cultures of Streptococcus mutans and Streptococcus sanguinis.

Antimicrobial photodynamic therapy (aPDT) has shown to exert a bactericidal effect against Streptococcus sanguinis and Streptococcus mutans. However, this efficacy has been reported for either type of bacteria separately. Bacterial suspensions of both strains, separately or together, were treated with concentrations of methylene blue (MB) and rose bengal (RB). Suspensions were irradiated with a light-emitting diode lamp (λ center at 625nm for MB and λ center at 515nm for RB) using a fluence of 18J/cm2. RB-aPDT at concentrations of 0.16-0.62 and 0.16-0.31µg/mL, and MB-aPDT at concentrations of 0.62-1.25 and 0.31-1.25µg/mL inhibited the growth of S. mutans and S. sanguinis respectively by 6 log10. In suspensions of both strains together, the same 6 log10 reduction in bacterial growth was achieved using the same concentrations of each photosensiziser. In conclusion, RB-aPDT and MB-aPDT appear to exert the same bactericidal effect against suspensions of S. sanguinis and S. mutans either for single strain treatment or for samples constituted by both bacteria mixed together. RB shows to be slightly more efficient than MB.