The aim of this study was to propose the use of red light-emitting diode (LED) as an alternative light source for methylene blue (MB) photosensitizing effect in photodynamic therapy (PDT). Its effectiveness was tested against Staphylococcus aureus (ATCC 26923), Escherichia coli (ATCC 26922), Candida albicans (ATCC 90028) and Artemia salina. The maximum absorption of the LED lamps was at a wavelength of 663 nm, at intensities of 2,4,6 and 12 J.cm-2 for 10, 20, 30 and 60 min of exposure, respectively. Assays with and without LED exposure were carried out in plates containing MB at concentrations of 7 to 140.8 (micro) M for microorganisms and 13.35 to 668.5 (micro) M for microorganisms or microcrustaceans. The LED exposure induced more than 93.05%, 93.7% and 93.33% of growth inhibition for concentrations of 42.2 (micro)M for S.aureus (D-value=12.05 min) and 35.2 (micro)M for E.coli (D-value=11.51 min) and C.albicans (D-value=12.18 min), respectively after 20 min of exposure. LED exposure for 1 h increased the cytotoxic effect of MB against A.salina from 27% to 75%. Red LED is a promising light device for PDT that can effectively inhibit bacteria, yeast and microcrustacean growth.
No presente estudo, o papel de diferentes centros metálicos (magnésio, zinco e cobre) sobre o caráter hidrofílico de metaloclorofilas foi avaliado. O solvatocromismo, assim como o processo de agregação para esses compostos, em misturas água/etanol foram avaliados usando medidas de fluorescência e de espalhamento de luz ressonante (ELR), visando caracterizar o comportamento desses compostos. Independente da metaloclorofila estudada, a presença de ao menos 60% de água resulta em incremento considerável na emissão de fluorescência apresentada por esses compostos. Isso, provavelmente, é uma conseqüência direta de uma menor agregação, o que é confirmado pelos resultados de ELR. Adicionalmente, os resultados sugerem que as clorofilas de magnésio e de zinco devem ser promissores agentes fototerapêuticos para emprego em Terapia Fotodinâmica.In this study the role of different metal centers (magnesium, zinc and copper) on the enhancement of the hydrophilic character of metallochlorophylls, was evaluated. The solvatochromism as well as the aggregation process for these compounds in water/ethanol mixtures at different volume ratios were evaluated using Fluorescence, and Resonant Light Scattering (RLS) measurements, aiming to characterize the behavior of these compounds. Independently on the studied metallochlorophyll, the presence of at least 60% of water results in a considerable increase in the fluorescence emission, probably a direct consequence of a lower aggregation of these compounds, which is confirmed by the results from RLS measurements. Additionally, the results suggest that magnesium and zinc chlorophyll should be promising phototherapeutic agents for Photodynamic Therapy.
Recebido em 12/12/08; aceito em 24/7/09; publicado na web em 8/1/10 INFLUENCE OF DIFFERENT WATER-ETHANOL SOLVENT SYSTEMS ON THE SPECTROSCOPIC AND PHYSICO-CHEMICAL PROPERTIES OF THE MACROCYCLIC COMPOUNDS PHEOPHYTIN AND CHLOROPHYLL a. This work focus on the influence of solvent on the photophysical properties of chlorophyll a and pheophytin. Both compounds are related to the photosynthesis process and are considered prototypes of photosensitizers in Photodynamic Therapy. Fluorescence measurements were developed using water/ethanol mixtures at different compositions, since both solvents could be employed in biological applications. The spectroscopic properties of these compounds undergo profound changes depending on water content in the ethanol due to autoaggregation processes. The major hydrophobicity and the lower dielectric constant of ethanol when compared with water precluded significantly the auto-aggregation process of these compounds.
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