Comparison of photodynamic therapy with methylene blue associated with ceftriaxone in gram-negative bacteria; an in vitro study.

The resistance of microorganisms increases the need for new antimicrobial therapies. The aim of this study was to evaluate the in vitro action of photodynamic therapy and its combination with ceftriaxone in third generation cephalosporin resistant gram-negative bacteria. Clinical strains of Klebsiella pneumoniae, Enterobacter aerogenes and Escherichia coli were obtained, incubated with MB for 15 min combined or not with ceftriaxone and irradiated with fluence of 10 and 25 J/cm². MB internalization was evaluated by confocal microscopy. Cell viability was assessed by counting colony forming units and bacterian metabolism by the resazurin test. MB has been observed within cells, although not in all bacteria. PDT-MB alone and combined with Ceftriaxone reduced bacterial growth by approximately 1 log at 10 J/cm² of fluence and 4 logs by 25 J/cm², with a significant difference from the control group. The reduction in bacterial growth between the treated groups was similar, without significant difference between them. The Resazurin test showed lower bacterial metabolic activity in the treated groups, but it did not allow to observe difference between fluences. It was concluded with this study that the internalization of MB was not observed in all cells of K. pneumoniae, E. aerogenes and E. coli strains. There was less bacterial metabolic activity in the treated groups, with no variation between different fluences. PDT-MB 25 J/cm² alone and combined with Ceftriaxone showed antimicrobial action, but the PDT-MB/Ceftriaxone combination had no potentiating effect.

Photodynamic therapy mediated by chlorin-type photosensitizers against Streptococcus mutans biofilms.

Photodynamic therapy (PDT) can be used for the control of oral pathogens and different photosensitizers (PS) have been investigated. This study evaluated the efficacy of PDT against Streptococcus mutans biofilms using two second-generation PS derived from chlorin: Photoditazine® (PDZ) and Fotoenticine® (FTC). These PS were compared to methylene blue (MB), a dye with proven antimicrobial activity against S. mutans. Suspensions of S. mutans were cultured in contact with bovine tooth disks for biofilm formation. After 48 h, the biofilms were treated with PDZ (0.6 mg/mL), FTC (0.6 mg/mL) or MB (1 mg/mL) and submitted to laser irradiation (660 nm, 50 mW/cm2). The biofilms were quantified by the determination of CFU/mL count and analyzed by scanning electron microscopy (SEM). All PS used for PDT reduced the number of S. mutans, with a statistically significant difference compared to the untreated groups. PDT achieved microbial reductions of 4 log with MB and 6 log with PDZ, while the use of FTC resulted in the complete elimination of S. mutans biofilms. SEM analysis confirmed the CFU/mL results, showing that all PS, particularly FTC, were able to detach the biofilms and to eliminate the bacteria. In conclusion, PDT mediated by chlorin-type PS exhibited greater antimicrobial activity against S. mutans than MB-mediated PDT, indicating that these PS can be useful for the control of dental caries.