A basic study on Hypericin-PDT in vitro

Abstract

The effect of photo dynamic therapy (PDT) using hypericin as a photosensitiser and the effect of PDT on intracellular ATP levels using different lamps in a human leukemic monocyte lymphoma cell line (U937) were studied. The time required for hypericin to penetrate into the cancer cells was 1h, and incubation for more than 3h post-irradiation with hypericin-PDT was required to observe effects. Thus, if cancer cell death does not occur immediately following irradiation, it is unnecessary to perform additional irradiation, as most of the cells die via apoptosis during the incubation period post-irradiation. When hypericin-PDT was performed using a Na-Li lamp as a light source, the cell viability decreased approximately 55% immediately following irradiation for 5 min; however, after a 5-h post-irradiation incubation, the cell viability approached 0%. Concurrently, intracellular ATP levels increased markedly; thus, irradiation (0.225 J/cm(2)) for 5 min provided the best results in terms of the highest degree of cancer cell apoptosis. Similar experiments were performed using three different LED lamps respectively. When cells were treated with the LED lamps, with maximum peaks of 599 nm and 595 nm, the cell viability approached 0% after incubation for 5h following 15 min of irradiation (0.04 J/cm(2) and 0.099 J/cm(2), respectively). We confirmed that incubating the cells for more than 3h in a 100 × diluted hypericin solution was the most effective for PDT and that a LED lamp of low light intensity led to the highest apoptosis rate in the U937 cells.