Internalization of Aggregated Photosensitizers by Tumor Cells: Subcellular Time-resolved Fluorescence Spectroscopy on Derivatives of Pyropheophorbide-a Ethers and Chlorin e6 under Femtosecond One- and Two-photon Excitation¶
- 1 May 2007
- journal article
- Published by American Society for Photobiology in Photochemistry and Photobiology
- Vol. 76 (6), 686-694
- https://doi.org/10.1562/0031-8655(2002)0760686ioapbt2.0.co2
Abstract
Amphiphilic sensitizers self‐associate in aqueous environments and form aggregated species that exhibit no or only negligible photodynamic activity. However, amphiphilic photosensitizers number among the most potent agents of photodynamic therapy. The processes by which these sensitizers are internalized into tumor cells have yet to be fully elucidated and thus remain the subject of debate. In this study the uptake of photosensitizer aggregates into tumor cells was examined directly using subcellular time‐resolved fluorescence spectroscopy with a high temporal resolution (20–30 ps) and high sensitivity (time‐correlated single‐photon counting). The investigations were performed on selected sensitizers that exhibit short fluorescence decay times (<50 ps) in aggregated form. Derivatives of pyropheophorbide‐a ether and chlorin e6 with varying lipophilicity were used for the study. The characteristic fluorescence decay times and spectroscopic features of the sensitizer aggregates measured in aqueous solution also could be observed in A431 human endothelial carcinoma cells administered with these photosensitizers. This shows that tumor cells can internalize sensitizers in aggregated form. Uptake of aggregates and their monomerization inside cells were demonstrated directly for the first time by means of fluorescence lifetime imaging with a high temporal resolution. Internalization of the aggregates seems to be endocytosis mediated. The degree of their monomerization in tumor cells is strongly influenced by the lipophilicity of the compounds.Keywords
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