Melanin and the cellular effects of ultrashort-pulse near-infrared laser radiation

Randolph D. Glickman; Neeru Kumar; Benjamin A. Rockwell; Gary D. Noojin; Michael L. Denton; David J. Stolarski

doi:10.1117/12.477653

27 August 2003 Melanin and the cellular effects of ultrashort-pulse, near-infrared laser radiation

Randolph D. Glickman, Neeru Kumar, Benjamin A. Rockwell, Gary D. Noojin, Michael L. Denton, David J. Stolarski

Proceedings Volume 4961, Laser-Tissue Interaction XIV; (2003) https://doi.org/10.1117/12.477653
Event: Biomedical Optics, 2003, San Jose, CA, United States

Abstract

Our research into laser bioeffects has increasingly focused on cytotoxic mechanisms affecting genomic expression and programmed cellular stress responses. In the context of DNA damage, we previously reported that more DNA strand breaks were produced in cultured retinal pigment epithelium (RPE) cells exposed to ultrashort pulse, than to CW, near-infrared (NIR) laser radiation. To test the hypothesis that RPE melanin was the cellular chromophore responsible for mediating this damage, the experiments were repeated with a line of human-derived RPE cells that could be grown in culture expressing varying levels of pigmentation. Lightly-pigmented cells were either unexposed, or exposed to the output of a Ti:Sapphire laser producing 810 nm light in mode-locked pulses (48-fsec at 80 MHz), or as CW radiation. Cells were irradiated at 160 W/cm² or 80 W/cm² (the estimated ED50 or half-ED50 for a retinal lesion). Immediately following the laser exposure, cells were processed for the comet assay. Longer "comet" tails and larger "comet" areas indicated more DNA strand breaks. In lightly-pigmented RPE cells, the overall comet assay differences among the laser-exposed groups were smaller than those observed in our earlier experiments which utilized highly pigmented primary cells. The comet tail lengths of cells exposed to the mode-locked pulses at the ED50, however, were significantly longer than those of the controls or the CW-exposed cells. The other comet assay parameters examined (tail moment, comet area) did not show consistent differences among the groups. While these results support the involvement of melanin in the ultrashort pulse laser-induced damage to DNA, they do not exclude the involvement of other cellular chromophores. Some preliminary experiments describing other measures of cellular stress responses to laser-induced oxidative stress are described.

Citation Download Citation

Randolph D. Glickman, Neeru Kumar, Benjamin A. Rockwell, Gary D. Noojin, Michael L. Denton, and David J. Stolarski "Melanin and the cellular effects of ultrashort-pulse, near-infrared laser radiation", Proc. SPIE 4961, Laser-Tissue Interaction XIV, (27 August 2003); https://doi.org/10.1117/12.477653

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