Phycocyanin: laser activation, cytotoxic effects, and uptake in human atherosclerotic plaque

Phycocyanin is a phycobiliprotein with peak absorption at 620 nm. The laser activation, cytotoxic effects, and uptake into atherosclerotic plaque of phycocyanin was studied. Optimal activation was produced by argon dye laser at 0.5 W and a total energy dose of 300 J/cm2 at 620 nm and 650 nm, irradiated through blood with a hematocrit of 8%. Activation was evidenced by reduction of optical density by 0.3 units at 340 nm caused by oxidation of the reduced nicotinamide adenine dinucleotide phosphate (NADPH) in a buffered reaction solution containing 0.1 mg/ml of phycocyanin. Cytotoxicity was evaluated by measuring viability of mouse myeloma cells in culture after incubation with phycocyanin (0.25 mg/ml) and irradiated by 300 J/cm2 at 514 nm. After 72 hours post-treatment the cells showed 15% viability compared to 69% and 71% for control cells exposed to laser only or phycocyanin only, respectively. Atherosclerotic artery segments obtained within 5 hours postmortem were perfused with 0.1 mg/ml phycocyanin in oxygenated Krebs Ringer solution at 30 mm Hg for 5 minutes followed by washout with phycocyanin-free Krebs for 10 minutes. Artery sections examined histologically by light and fluorescence microscopy showed specific fluorescence localization within the plaque particularly at the elastic laminae and to a larger extent at the internal elastic lamina but not in the medial muscle layer. In conclusion, phycocyanin is a cytotoxic photosensitizer that exhibits specific binding to plaque and is activated at a wavelength minimally absorbed by blood. These properties suggest potential therapeutic use for plaque localization and regression.