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1 June 2003 Encapsulation of p-THPP into Nanoparticles: Cellular Uptake, Subcellular Localization and Effect of Serum on Photodynamic Activity
Y. N. Konan, J. Chevallier, R. Gurny, E. Allémann
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The cellular uptake, localization and efflux of meso-tetra-(4-hydroxyphenyl)porphyrin (p-THPP)–loaded nanoparticles have been studied in EMT-6 tumor cells. The effect of blood serum on photocytotoxicity has also been evaluated. Sub–130 nm nanoparticles based on poly(d,l-lactide-co-glycolide) (PLGA) (50:50 PLGA and 75:25 PLGA) and poly(d,l-lactide) (PLA) have been examined in comparison with free p-THPP. For all formulations tested, uptake of photosensitizer into cells was dependent on concentration, time and temperature. All nanoparticulate formulations accumulated within the cells to a greater extent relative to free drug. Indeed, the fluorescence intensities measured on EMT-6 cells treated with p-THPP–loaded nanoparticulate formulations were at least two-fold higher than those obtained with free dye. Furthermore, the highest accumulation level was found with PLGA nanoparticles. Fluorescence microscopy revealed that endocytosis is a major intracellular sequestration mechanism of these p-THPP formulations and that these were localized into early and late endosomes. The efflux study performed on both nonirradiated and irradiated cells indicated that free and p-THPP–loaded nanoparticles gradually escaped from EMT-6 cells as a function of time. This was more pronounced when cells were treated with nanoparticles and irradiated, reflecting important photodamage. It was also found that regardless of the nanoparticulate formulations tested, p-THPP photocytotoxicity was influenced by the concentration of the serum.

Y. N. Konan, J. Chevallier, R. Gurny, and E. Allémann "Encapsulation of p-THPP into Nanoparticles: Cellular Uptake, Subcellular Localization and Effect of Serum on Photodynamic Activity," Photochemistry and Photobiology 77(6), 638-644, (1 June 2003).<0638:EOPINC>2.0.CO;2
Received: 22 October 2002; Accepted: 1 March 2003; Published: 1 June 2003

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