C60 fullerene accumulation in human leukemic cells and perspectives of LED-mediated photodynamic therapy

Abstract

Recent progress in nanobiotechnology has attracted interest to a biomedical application of the carbon nanostructure C₆₀ fullerene since it possesses a unique structure and versatile biological activity. C₆₀ fullerene potential application in the frame of cancer photodynamic therapy (PDT) relies on rapid development of new light sources as well as on better understanding of the fullerene interaction with cells. The aim of this study was to analyze C₆₀ fullerene effects on human leukemic cells (CCRF-CEM) in combination with high power single chip light-emitting diodes (LEDs) light irradiation of different wavelengths: ultraviolet (UV, 365 nm), violet (405 nm), green (515 nm) and red (632 nm). The time-dependent accumulation of fullerene C₆₀ in CCRF-CEM cells up to 250 ng/10⁶ cells at 24 h with predominant localization within mitochondria was demonstrated with immunocytochemical staining and liquid chromatography mass spectrometry. In a cell viability assay we studied photoexcitation of the accumulated C₆₀ nanostructures with ultraviolet or violet LEDs and could prove that significant phototoxic effects did arise. A less pronounced C₆₀ fullerene phototoxic effect was observed after irradiation with green, and no effect was detected with red light. A C₆₀ fullerene photoactivation with violet light induced substantial ROS generation and apoptotic cell death, confirmed by caspase3/7 activation and plasma membrane phosphatidylserine externalization. Our work proved C₆₀ fullerene ability to induce apoptosis of leukemic cells after photoexcitation with high power single chip 405 nm LED as a light source. This underlined the potential for application of C₆₀ nanostructure as a photosensitizer for anticancer therapy.

Keywords: Apoptosis; C(60) fullerene; HPLC-ESI-MS; Immunocytochemistry; LEDs; Leukemic cells; Photodanamic therapy.