Photodynamic inactivation of Candida albicans sensitized by tri- and tetra-cationic porphyrin derivatives

Abstract

The photodynamic action of 5-(4-trifluorophenyl)-10,15,20-tris(4-trimethylammoniumphenyl)porphyrin iodide (TFAP3+) and 5,10,15,20-tetra(4-N,N,N-trimethylammonium phenyl)porphyrin p-tosylate (TMAP4+) has been studied in vitro on Candida albicans. The results of these cationic porphyrins were compared with those of 5,10,15,20-tetra(4-sulphonatophenyl)porphyrin (TPPS4-), which characterizes an anionic sensitizer. In vitro investigations show that these cationic porphyrins are rapidly bound to C. albicans cells, reaching a value of ∼1.4 nmol/106 cells, when the cellular suspensions were incubated with 5 μM sensitizer for 30 min. In contrast, TPPS4- is poorly uptaken by yeast cells. The fluorescence spectra of these sensitizers into the cells confirm this behaviour. The amount of porphyrin binds to cells is dependent on both sensitizer concentrations (1-5 μM) and cells densities (106-108 cells/mL). Photosensitized inactivation of C. albicans cellular suspensions increases with sensitizer concentration, causing a ∼5 log decrease of cell survival, when the cultures are treated with 5 μM of cationic porphyrin and irradiated for 30 min. However, the photocytotoxicity decreases with an increase in the cell density, according to its low binding to cells. Under these conditions, the photodynamic activity of TFAP3+ is quite similar to that produced by TMAP4+, whereas no important inactivation effect was found for TPPS4-. The high photodynamic activity of cationic porphyrins was confirmed by growth delay experiments. Thus, C. albicans cell growth was not detected in the presence of 5 μM TFAP3+. Photodynamic inactivation capacities of these sensitizers were also evaluated on C. albicans cells growing in colonies on agar surfaces. Cationic porphyrins produce a growth delay of C. albicans colonies and viability of cells was not observed after 3 h irradiation, indicating a complete inactivation of yeast cells. Therefore, these results indicate that these cationic porphyrins are interesting sensitizers for photodynamic inactivation of yeasts in liquid suspensions or in localized foci of infection. © 2008 Elsevier Masson SAS. All rights reserved.