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.
Keywords: Ceftriaxone; Enterobacter aerogenes and Escherichia coli; Klebsiella pneumoniae; Methylene blue; Photodynamic therapy.
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