1 March 2006 Photochemical damage from chronic 458-nm laser exposures in an artificially pigmented hTERT-RPE1 cell line
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Artificially pigmented hTERT-RPE1 cells were exposed to a mode-locked or continuous wave (CW) laser at 458 nm for one hour in a modified culture incubator. Exposure conditions were selected to give greatest likelihood of damage due to a photochemical mechanism, with interest in possible differences between CW and mode-locked damage thresholds. After post-exposure-recovery (PER) for either 1-hour or 24-hour, cells were concurrently stained with annexin V and 6-CFDA to determine if they had undergone necrosis or apoptosis. Alternatively, cells were stained with Ethidium Homodimer (EthD-1) and Calcein AM to determine if they had undergone necrosis following 1-hour and 24-hours PER. Preliminary results indicate that laser exposure induced some apoptosis following 1-hour PER, with irradiance required for apoptosis being lower than that for necrosis with mode-locked exposure conditions. Probit analysis yielded necrosis thresholds for cell culture following 1-hour PER using data compiled from both dye sets. CW exposures resulted in a lower threshold than mode-locked exposures for necrosis following 1-hour PER. A thermal model provided the predicted temperature rise in cell culture due to laser exposure. The thermal model validates our choice of laser parameters to obtain photochemical damage. Data following 24-hours PER were inconclusive. Considerations of cell migration are included in the interpretation of data and further improvements to methods when using live cell assays are recommended.
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Michael S. Foltz, Michael S. Foltz, Norris A. Whitlock, Norris A. Whitlock, Larry E. Estlack, Larry E. Estlack, Manuel A. Figueroa, Manuel A. Figueroa, Robert J. Thomas, Robert J. Thomas, Benjamin A. Rockwell, Benjamin A. Rockwell, Michael L. Denton, Michael L. Denton, } "Photochemical damage from chronic 458-nm laser exposures in an artificially pigmented hTERT-RPE1 cell line", Proc. SPIE 6084, Optical Interactions with Tissue and Cells XVII, 608415 (1 March 2006); doi: 10.1117/12.644690; https://doi.org/10.1117/12.644690

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