Repetition-rate-dependent oxygen consumption modifies cytotoxicity in photodynamic therapy using pulsed light

We studied the repetition-rate dependence of PDT cytotoxicity and relation between PDT cytotoxicity and both oxygen consumption and photobleaching during PDT in vitro. Mice renal carcinoma cells were incubated with second-generation photosensitizer, PAD-S31, and were irradiated with 670-nm nanoseconds pulsed light from YAG-OPO system. Four repetition rates of 30, 15, 5 and 3 Hz were investigated, provided that the incident peak intensity and the total light dose adjusted to 1.2 MW/cm² and 40 J/cm², respectively. We found limited cytotoxicity about 40% at 30 and 15 Hz and sufficient cytotoxicity about 80% at 5 and 3 Hz. The oxygen measurement during irradiation revealed that the 5- and 3-Hz irradiation caused slow oxygen consumption, while rapid oxygen consumption followed by a rapid recovery of oxygenation at 30 and 15 Hz. Interestingly, the fluorescence measurement during irradiation also demonstrated that photobleaching discontinued in the same period of oxygen recovery at 30 and 15 Hz. These discontinued oxygen consumptiona and photobleaching at 30 and 15 Hz might have limited effective total fluence and resulted in suppressed cytotoxicity. These results suggest that the PDT efficacy using a pulsed laser significantly depends on the pulse repetition rate probably due to different oxygen consumption process during PDT.