Recently, we reported induced anoxia as a limiting factor for photodynamic tumor therapy (PDT). This effect occurs in vivo if the amount of generated singlet oxygen that undergoes chemical reactions with cellular components exceeds the local oxygen supply. The amount of generated singlet oxygen depends mainly on photosensitizer (PS) accumulation, efficiency, and illumination intensity. With illumination intensities above a certain threshold, singlet oxygen is limited to the blood vessel and the nearest vicinity; lower intensities allow singlet oxygen generation also in tissue which is a few cell layers away from the vessels. While all experiments so far were limited to light intensities above this threshold, we report experimental results for intensities at both sides of the threshold for the first time, giving proof for the described model. Using time-resolved optical detection in NIR, we demonstrate characteristic, illumination intensity-dependent changes in signal kinetics of singlet oxygen and photosensitizer phosphorescence in vivo. The described analysis allows for better optimization and coordination of PDT drugs and treatment, as well as new diagnostic methods based on gated PS phosphorescence, for which we report a first in vivo feasibility test.

Fujian Provincial Key Laboratory of Photonics Technology Fujian Normal University Fuzhou 350007 China

Institute of Macromolecular Chemistry Czech Academy of Sciences Heyrovského nám 2 16200 Prague Czech Republic

Institute of Physics Photobiophysics Humboldt University of Berlin Newtonstr 15 12489 Berlin Germany

Laboratory of Microbiology and Oncology Faculty of Pharmaceutical Sciences Sojo University Kumamoto 860 0082 Japan

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