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Cellular Imaging and Time-Domain FLIM Studies of Meso-Tetraphenylporphine Disulfonate as a Photosensitising Agent in 2D and 3D Models.
Balukova, Andrea; Bokea, Kalliopi; Barber, Paul R; Ameer-Beg, Simon M; MacRobert, Alexander J; Yaghini, Elnaz.
Afiliación
  • Balukova A; Department of Surgical Biotechnology, Division of Surgery and Interventional Science, University College London, London NW3 2QG, UK.
  • Bokea K; Department of Surgical Biotechnology, Division of Surgery and Interventional Science, University College London, London NW3 2QG, UK.
  • Barber PR; Department of Oncology, UCL Cancer Institute, University College London, London WC1E 6DD, UK.
  • Ameer-Beg SM; Comprehensive Cancer Centre, School of Cancer and Pharmaceutical Sciences, King's College London, London SE1 9RT, UK.
  • MacRobert AJ; Comprehensive Cancer Centre, School of Cancer and Pharmaceutical Sciences, King's College London, London SE1 9RT, UK.
  • Yaghini E; Department of Surgical Biotechnology, Division of Surgery and Interventional Science, University College London, London NW3 2QG, UK.
Int J Mol Sci ; 25(8)2024 Apr 11.
Article en En | MEDLINE | ID: mdl-38673807
ABSTRACT
Fluorescence lifetime imaging (FLIM) and confocal fluorescence studies of a porphyrin-based photosensitiser (meso-tetraphenylporphine disulfonate TPPS2a) were evaluated in 2D monolayer cultures and 3D compressed collagen constructs of a human ovarian cancer cell line (HEY). TPPS2a is known to be an effective model photosensitiser for both Photodynamic Therapy (PDT) and Photochemical Internalisation (PCI). This microspectrofluorimetric study aimed firstly to investigate the uptake and subcellular localisation of TPPS2a, and evaluate the photo-oxidative mechanism using reactive oxygen species (ROS) and lipid peroxidation probes combined with appropriate ROS scavengers. Light-induced intracellular redistribution of TPPS2a was observed, consistent with rupture of endolysosomes where the porphyrin localises. Using the same range of light doses, time-lapse confocal imaging permitted observation of PDT-induced generation of ROS in both 2D and 3D cancer models using fluorescence-based ROS together with specific ROS inhibitors. In addition, the use of red light excitation of the photosensitiser to minimise auto-oxidation of the probes was investigated. In the second part of the study, the photophysical properties of TPPS2a in cells were studied using a time-domain FLIM system with time-correlated single photon counting detection. Owing to the high sensitivity and spatial resolution of this system, we acquired FLIM images that enabled the fluorescence lifetime determination of the porphyrin within the endolysosomal vesicles. Changes in the lifetime dynamics upon prolonged illumination were revealed as the vesicles degraded within the cells.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Porfirinas / Especies Reactivas de Oxígeno / Fármacos Fotosensibilizantes Límite: Female / Humans Idioma: En Revista: Int J Mol Sci Año: 2024 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Suiza
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Porfirinas / Especies Reactivas de Oxígeno / Fármacos Fotosensibilizantes Límite: Female / Humans Idioma: En Revista: Int J Mol Sci Año: 2024 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Suiza