Carotenuto R et al. (2021), Comparative Toxicological Evaluation of Tattoo ...

XB-ART-58769

Biology (Basel) 2021 Dec 09;1012:. doi: 10.3390/biology10121308.

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Comparative Toxicological Evaluation of Tattoo Inks on Two Model Organisms.

Carotenuto R, Fogliano C, Rienzi M, Siciliano A, Salvatore MM, De Tommaso G, Benvenuto G, Galdiero E, Guida M.

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Tattooing is a technique that introduces colored substances under the skin in order to color it permanently. Decomposition products of tattoo pigments produce numerous damages for the skin and other organs. We studied the effects of a commercial red ink tattoo, PR170, on Xenopus laevis embryos and Daphnia magna nauplii using concentrations of 10, 20, and 40 mg/L. For Xenopus, we applied the FETAX protocol analyzing survival, malformations, growth, heart rate, and the expression of genes involved in the development. In D. magna, we evaluated the toxicity with an immobilization test. Moreover, we investigated the production of ROS, antioxidant enzymes, and the expression of the ATP-binding cassette in both models. Our results indicate that PR170 pigment has nanoparticle dimensions, modifies the survival and the ATP-binding cassette activity, and induces oxidative stress that probably produces the observed effects in both models. Deformed embryos were observed in Xenopus, probably due to the modification of expression of genes involved in development. The expression of pro-inflammatory cytokines was also modified in this amphibian. We think that these effects are due to the accumulation of PR170 and, in particular, to the presence of the azoic group in the chemical structure of this pigment. Further studies needed to better understand the effects of commercial tattoo inks.

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Species referenced: Xenopus laevis

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	Figure 1. PR170 characterization. (A) TEM micrography of PR170 highlighted their polygonal shape and the formation of agglomerates (arrow). (B) Distribution of diameters of PR170 contained in tattoo ink. Average diameter is 110 nm.
	Figure 2. Graphical representations of DLS measurements of PR170. (A): hydrodynamic radii distribution (9 ± 1, 35 ± 9 and 113 ± 9); (B): normalized hydrodynamic radii distribution (9 ± 1 and 30 ± 1).
	Figure 3. Mortality, growth retardation, and heartrate evaluations after PR170 pigment treatment in X. laevis embryos. (A) Mortality percentage distributions of untreated and treated embryos display a peak at 20 mg/L and values not so far from the control for 10 mg/L and 40 mg/L. (B) Analysis revealed unmodified length of treated embryos if compared to the wild type (p > 0.05). (C) Treated embryos showed a growing tachycardia with maximum rate at 40 mg/L. p < 0.01, ** p < 0.0001.
	Figure 4. Daphnia magna exposure. Immobility of Daphnia magna after 48 h of exposure (A); D. magna exposed to PR170 samples at different concentration (B): Control (Ctr), 10 mg/L and 20 mg/L. In the control (Ctr) is visible a fasting gut, in the 10 mg/L and 20 mg/L exposures PR170 accumulation is visible inside the digestive system.
	Figure 5. Effects of treatment with PR170 on X. laevis embryos development. (A) Control. (B) Presence of embryos with fold tail (white line). (C) Red pigment invasion of the intestine (arrow). (D) Presence of head edema (asterisk) accompanied by fold tail (white line), misshapen eyes (empty arrowhead), and increase in pigment (white arrowhead). (E) Excessive pigmentation (white arrowhead) and presence of bigger eyes (empty arrowhead). (F) Widespread edema of head and abdomen (asterisks). (G) Intestine protrusion from the abdomen (double arrows).
	Figure 6. Histological analysis of X. laevis embryos treated with PR170 pigment. (A) Control eye. (A’) Embryos treated with PR170 pigment in some cases showed eyes with elongated shape (empty arrowhead). (B) Control intestine. (B’) Intestine of treated embryos showed the presence of pigment in the intestinal loops (arrows) with tight adhesion to the walls (B’’, arrow). (C) Gills of treated embryos were not invaded by pigment (asterisk). Magnification: (B,B’,C): 5×; (A,A’): 20×; (B’’): 40×.
	Figure 7. Gene expression in X. laevis and D. magna. Expression of genes involved in early embryonic development (A) and inflammation (B) of Xenopus laevis. ATP binding cassette mechanism expression was showed in Daphnia magna (C) and Xenopus laevis (B). Data are presented as mean with SD. Statistical significance was determined using t-tests Hold Siddak correction for multiple comparison. * p < 0.05, p < 0.01, * p < 0.001, **** p < 0.0001.
	Figure 8. Oxidative stress in X. laevis and D. magna. Effects of PR170 on (A) ROS production expressed as fluorescence intensity, (B) CAT activity, and (C) SOD activity. The level of significance was set at p < 0.01, * p < 0.001.

References [+] :

Arl, Tattoo inks: Characterization and in vivo and in vitro toxicological evaluation. 2019, Pubmed