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Optimizing photodynamic therapy by liposomal formulation of the photosensitizer pyropheophorbide-a methyl ester: In vitro and ex vivo comparative biophysical investigations in a colon carcinoma cell line

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Abstract

Photodynamic therapy (PDT), induced by a photosensitizer (PS) encapsulated in a nanostructure, has emerged as an appropriate treatment to cure a multitude of oncological and non-oncological diseases. Pyropheophorbide-a methyl ester (PPME) is a second-generation PS tested in PDT, and is a potential candidate for future clinical applications. The present study, carried out in a human colon carcinoma cell line (HCT-116), evaluates the improvement resulting from a liposomal formulation of PPME versus free-PPME. Absorption and fluorescence spectroscopies, fluorescence lifetime measurements, subcellular imaging and co-localization analysis have been performed in order to analyze the properties of PPME for each delivery mode. The benefit of drug encapsulation in DMPC-liposomes is clear from our experiments, with a 5-fold higher intracellular drug delivery than that observed with free-PPME at similar concentrations. The reactive oxygen species (ROSs) produced after PPME-mediated photosensitization have been identified and quantified by using electron spin resonance spectroscopy. Our results demonstrate that PPME-PDT-mediated ROSs are composed of singlet oxygen and a hydroxyl radical. The small amounts of PPME inside mitochondria, as revealed by fluorescence co-localization analysis, could maybe explain the very low apoptotic cell death measured in HCT-116 cells.

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Authors and Affiliations

  1. Laboratory of Biomedical Spectroscopy, Institute of Physics, B5a, University of Liège, Sart-Tilman, B-4000, Liège, Belgium

    Pierre-Henri Guelluy, Angeliki Grammenos & Maryse Hoebeke

  2. Institut des Sciences Moleculaires d’Orsay, FRE 3363, University of Paris-Sud 91405 Orsay, France - CNRS, 91405, Orsay, France

    Marie-Pierre Fontaine-Aupart

  3. CPBM/LUMAT, FR 2764, University of Paris-Sud, 91405, Orsay, France

    Sandrine Lécart

  4. GIGA-R: Virology and Immunology, B34, University of Liège, B-4000, Liège, Belgium

    Jacques Piette

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  1. Pierre-Henri Guelluy
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Correspondence to Pierre-Henri Guelluy.

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Guelluy, PH., Fontaine-Aupart, MP., Grammenos, A. et al. Optimizing photodynamic therapy by liposomal formulation of the photosensitizer pyropheophorbide-a methyl ester: In vitro and ex vivo comparative biophysical investigations in a colon carcinoma cell line. Photochem Photobiol Sci 9, 1252–1260 (2010). https://doi.org/10.1039/c0pp00100g

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