Antimicrobial effect on Candida albicans biofilm by application of different wavelengths and dyes and the synthetic killer decapeptide KP.

Abstract

The aim of this study was to test the application in vitro of different laser wavelengths at a low fluence in combination or not with proper photosensitizing dyes on Candida albicans biofilm with or without a synthetic killer decapeptide (KP). Candida albicans SC5314 was grown on Sabouraud dextrose agar plates at 37°C for 24 h. Cells were suspended in RPMI 1640 buffered with MOPS and cultured directly on the flat bottom of 96-wells plates. The previously described killer decapeptide KP was used in this study. Three different combinations of wavelengths and dyes were applied, laser irradiation has been performed at a fluence of 10 J/cm². The effect on C. albicans biofilm was evaluated by the XTT assay. Microscopic observations were realized by fluorescence optic microscopy with calcofluor white and propidium iodide. Compared with control, no inhibition of C. albicans biofilm viability was obtained with application of red, blue and green lasers alone or with any combination of red diode laser, toluidine blue and KP. The combined application of blue diode laser with curcumin and/or KP showed always a very significant inhibition, as curcumin alone and the combination of curcumin and KP did, while combination of blue diode laser and KP gave a less significant inhibition, the same obtained with KP alone. The combined application of green diode laser with erythrosine and/or KP showed always a very significant inhibition, as the combination of erythrosine and KP did, but no difference was observed with respect to the treatment with erythrosine alone. Again, combination of green diode laser and KP gave a significant inhibition, although paradoxically lower than the one obtained with KP alone. Treatment with KP alone, while reducing biofilm viability did not cause C. albicans death in the adopted experimental conditions. On the contrary, combined treatment with blue laser, curcumin and KP, as well as green laser, erythrosine and KP led to death most C. albicans cells. The combination of laser light at a fluence of 10 J/cm² and the appropriate photosensitizing agent, together with the use of KP, proved to exert differential effects on C. albicans biofilm.