This study evaluated the mechanisms involved and the influence of photosensitizer solvent in the killing of Enterococcus faecalis using photodynamic therapy (PDT). Enterococcus faecalis cells incubated with 100 μm methylene blue dissolved in water and in MIX (a mixture of glycerol:ethanol:water) were irradiated with 664 nm diode laser (63.69 J cm−2). The effect of PDT on the viability of bacteria, and the functional integrity of cell wall, chromosomal DNA and membrane proteins were analyzed. The bactericidal action of PDT was significantly higher when a MIX-based photosensitizer solvent was used (P < 0.001). Fluorimetric and fluorescence microscopy-based analysis showed the functional impairment of E. faecalis cell wall which was significantly higher when a MIX-based photosensitizer solvent was used (P < 0.001). PDT with MIX-based photosensitizer solvent showed extensive damage to chromosomal DNA. However, both PDT conditions showed similar trend in the degradation of membrane proteins, although cross-linked proteins were evident only in PDT conducted with MIX-based photosensitizer solvent. The findings from our study showed that PDT destroyed the functional integrity of cell wall, DNA and membrane proteins of E. faecalis. The degrees of damage on these targets were influenced by the photosensitizer solvent used during PDT.
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