Laser Irradiation of Organic Tattoo Pigments Releases Carcinogens with 3,3′- Dichlorobenzidine Inducing DNA Strand Breaks in Human Skin Cells Henrik Hering,

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Laser Irradiation of Organic Tattoo Pigments Releases Carcinogens with 3,3′- Dichlorobenzidine Inducing DNA Strand Breaks in Human Skin Cells Henrik Hering, Anja Yu Sung, Nadine Röder, Christoph Hutzler, Hans-Peter Berlien, Peter Laux, Andreas Luch, Ines Schreiver Journal of Investigative Dermatology Volume 138, Issue 12, Pages 2687-2690 (December 2018) DOI: 10.1016/j.jid.2018.05.031 Copyright © 2018 The Authors Terms and Conditions

Figure 1 Laser decomposition products of quinophthalone pigment yellow (P.Y.) 138 and the diazo pigment orange (P.O.) 13 in pig skin and their cyto- and genotoxic effects on human HaCaT and BJ skin cell lines. (a) Microscopic pictures of postmortem tattooed pig skin sections displayed as bright field (BF) or DAPI staining of the cell nuclei in overlay with autofluorescence (AF) of corresponding pigments (scale bar = 100 μm). (b) Quantity of decomposition products of both organic pigments after irradiation with three laser wavelengths in pig skin. Data are displayed as mean ± SD of three independent replicates at two separate days (N = 6). Values were rounded to two significant figures. (c) The level of cytotoxicity of the carcinogens hexachlorobenzene (HCB), aniline, and 3,3′-dichlorobenzidine (DCBD) is calculated from the activity of lactate dehydrogenase (LDH) in the supernatant divided by total LDH activity in the supernatant and lysed cells. (d) Induction of DNA double-strand breaks measured by flow cytometry using an antibody against the phosphorylated histone H2AX (γH2AX). Gating strategy is exemplarily displayed for HaCaT cells treated with DCBD for 72 hours. (e) γH2AX positive cells after DCBD, HCB, and aniline treatment. Data are displayed as mean + SD. Three independent experiments were carried out (N = 3). Asterisks indicate significance as identified by a two-way ANOVA with Tukey’s multiple comparison test between substance treatments and the respective control (*P < 0.05). Treatment of 100 μM DCBD was only significant against the smallest treatment concentration after 24 hours in BJ cells (+P < 0.05). ANOVA, analysis of variance; HCN, hydrogen cyanide; LOQ, limit of quantification; SD, standard deviation. Journal of Investigative Dermatology 2018 138, 2687-2690DOI: (10.1016/j.jid.2018.05.031) Copyright © 2018 The Authors Terms and Conditions