Enhancement of the photodynamic activity of tri-cationic porphyrins towards proliferating keratinocytes by conjugation to poly-S-lysine

Abstract

A fast uptake of the tri-cationic 5-(4-carboxyphenyl)-10,15,20-tris(4-methylpyridinium-4-yl)porphyrin tri-iodide (P-H), independent of the presence or absence of proteins in the culture medium, occurs during incubation of NCTC 2544 human keratinocytes with this porphyrin. By contrast, the uptake of the poly-S-lysine conjugate (P-(Lys)n) is faster in serum-free medium than in the supplemented medium suggesting that P-(Lys)n interacts with serum proteins. The P-(Lys)n uptake is almost an order of magnitude greater than that of P-H in serum-free or supplemented culture medium. With histidine as a specific probe of type II photodynamic reactions, the relative photosensitizing effectiveness of the conjugate is only one fourth that of P-H. Nevertheless, the photocytotoxicity of the conjugate is strongly enhanced as compared to that of P-H as a result of its larger uptake. Thus, the doses achieving 50% of photocytotoxicity after incubation with 5 μM of the conjugate and its parent cationic porphyrin are about 20 min and 1 h, respectively. Similarly, the initial rate of the cell lipid peroxidation induced by photosensitization with P-(Lys)n is about 8 times higher than that obtained with P-H. Fluorescence microscopy reveals that P-H is more diffusely located in the cytoplasm than P-(Lys)n which seems to accumulate in lysosome-like structures. Little if any staining of the nucleus is observed with both photosensitizers. © 2006 The Royal Society of Chemistry and Owner Societies.