Determination of the Phototoxicity Potential of
Commercially Available Tattoo Inks Using the 3T3-
neutral Red Uptake Phototoxicity Test

ALPERTUNGA, Buket; TÜRK, Elif Gözde UTKU; JANNUZZI, Ayse Tarbin

doi:10.4274/tjps.galenos.2021.86344

Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test

Elif Gözde UTKU TÜRK, (İstanbul Üniversitesi Eczacılık Fakültesi Farmasötik Toksikoloji Anabilim Dalı, İstanbul, Türkiye)

Ayse Tarbin JANNUZZI, (İstanbul Üniversitesi Eczacılık Fakültesi Farmasötik Toksikoloji Anabilim Dalı, İstanbul, Türkiye)

Buket ALPERTUNGA (İstanbul Üniversitesi Eczacılık Fakültesi Farmasötik Toksikoloji Anabilim Dalı, İstanbul, Türkiye)

Turkish Journal of Pharmaceutical Sciences

2 2

Yıl: 2022 Cilt: 19 Sayı: 1 Sayfa Aralığı: 70 - 75 Metin Dili: İngilizce DOI: 10.4274/tjps.galenos.2021.86344 İndeks Tarihi: 26-06-2022

Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test

Öz:

Objectives: Tattooing is an ancient practice and its popularity has been increasing in the recent years. After tattooing, complications may occur related to compose tattoo inks. In this study, the phototoxicity potential of the blue, red and black colors of the most commonly used three different commercially-available tattoo ink brands have been examined by performing in vitro 3T3-neutral red uptake (NRU) phototoxicity test. Materials and Methods: In the study, the phototoxicity of serial diluted concentrations of tattoo inks were evaluated with in vitro 3T3-NRU phototoxicity test method according to OECD guide 432. The data obtained from the NRU test result were uploaded to Phototox software (version 2.0) and the phototoxicity potentials of tattoo inks were determined via the calculation of the mean photo effect (MPE) and photo irritation factor (PIF) values. Results: The red, black and blue colors of three different commercially available tattoo inks did not cause a cytotoxic activity on BALB/c 3T3 cells with 3T3-NRU test. The IC50 values could not be determined +ultraviolet (UV) and -UV conditions. PIF values could not be calculated and MPE values were <0.1, which predicts the absence of phototoxic effect for all of the tested tattoo inks. Conclusion: All tested inks were evaluated as non-phototoxic according to the results of MPE values calculated using Phototox software. However, test results should be verified by other phototoxicity test methods to obtain a comprehensive evaluation of phototoxic complications of different tattoo inks.

Anahtar Kelime:

Piyasada Satılan Dövme Boyalarının 3T3-nötral Kırmızısı Alım Fototoksisite Testi ile Fototoksisite Potansiyelinin Belirlenmesi

Öz:

Amaç: Dövme yapmak eski zamanlardan beri süregelen bir uygulamadır ve popülaritesi son yıllarda artmaktadır. Dövme yapılmasından sonra dövme boyalarının bileşimine bağlı olarak çeşitli komplikasyonlar oluşabilir. Bu çalışmada, piyasada en çok kullanılan üç farklı dövme boyası markasının mavi, kırmızı ve siyah renklerinin fototoksisite potansiyeli in vitro 3T3-nötral kırmızı alım (NRU) fototoksisite testi yapılarak incelenmiştir. Gereç ve Yöntemler: Çalışmada, dövme boyalarının seri olarak seyreltilmiş konsantrasyonlarının fototoksisitesi, OECD kılavuz 432’ye göre in vitro 3T3- NRU fototoksisite test yöntemi ile değerlendirilmiştir. NRU test sonucundan elde edilen veriler Phototox yazılımıyla (sürüm 2.0) değerlendirilmiştir. Dövme boyalarının fototoksisite potansiyelleri medyan foto etki (MPE) ve foto irritan faktör (PIF) değerlerinin hesaplanmasıyla belirlenmiştir. Bulgular: Ticari olarak temin edilebilen üç farklı dövme boyasının kırmızı, siyah ve mavi renkleri, 3T3-NRU testi ile BALB/c 3T3 hücrelerinde sitotoksik aktiviteye neden olmamıştır. +Ultraviyole (UV) ve -UV koşullarında IC50 değerleri belirlenememiştir. PIF değerleri hesaplanamamıştır ve MPE değerleri <0,1 olarak belirlenmiştir. Bu sonuçlar test edilen tüm dövme boyalarının fototoksik etkilerinin olmadığını göstermektedir. Sonuç: Phototox yazılımı ile hesaplanan MPE değerlerinin sonuçlarına göre test edilen tüm boyaların fototoksik olmadığı sonucuna ulaşılmıştır. Bununla birlikte, farklı dövme boyalarının fototoksik komplikasyonlarının kapsamlı bir değerlendirmesini yapabilmek için test sonuçlarının diğer fototoksisite test yöntemleriyle doğrulanması gerekmektedir.

Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

1. Grant CA, Twigg PC, Baker R, Tobin DJ. Tattoo ink nanoparticles in skin tissue and fibroblasts. Beilstein J Nanotechnol. 2015;6:1183-1191.
2. Dirks M. Making innovative tattoo ink products with improved safety: possible and impossible ingredients in practical usage. Curr Probl Dermatol. 2015;48:118-127.
3. Laux P, Tralau T, Tentschert J, Blume A, Dahouk SA, Bäumler W, Bernstein E, Bocca B, Alimonti A, Colebrook H, de Cuyper C, Dähne L, Hauri U, Howard PC, Janssen P, Katz L, Klitzman B, Kluger N, Krutak L, Platzek T, Scott-Lang V, Serup J, Teubner W, Schreiver I, Wilkniß E, Luch A. A medical-toxicological view of tattooing. Lancet. 2016;387:395-402.
4. Lerche CM, Heerfordt IM, Serup J, Poulsen T, Wulf HC. Red tattoos, ultraviolet radiation and skin cancer in mice. Exp Dermatol. 2017;26:1091-1096.
5. Mang R, Stege H, Krutmann J. Mechanisms of phototoxic and photoallergic reactions in Contact Dermatitis (5th ed). Berlin; Springer. 2011;6:97-104.
6. Serup J, Niels H, Linnet JT, Møhl B, Olsen O, Westh H. Tattoos—health, risks and culture. With introduction of the” seamless prevention” strategy. Copenhagen: The Council on Health and Disease Prevention. 2015.
7. Union P. Regulation (EC) no 1223/2009 of the European Parliament and of the Council. The OJEU. 2009;342:59.
8. EC (European Commission). Full EU ban on animal testing for cosmetics enters into force. Press release. 2013.
9. Sreedhar D, Manjula N, Pise SA, Ligade V. Ban of cosmetic testing on animals: a brief overview. Int J Cur Res Rev. 2020;12:113-116.
10. Vinken M. 3Rs toxicity testing and disease modeling projects in the European Horizon 2020 research and innovation program. EXCLI J. 2020;19:775-784.
11. Liebsch M, Spielmann H, Pape W, Krul C, Deguercy A, Eskes C. UV- induced effects. Altern Lab Anim. 2005;(Suppl 1):131-146.
12. Spielmann H, Balls M, Dupuis J, Pape WJ, Pechovitch G, de Silva O, Holzhütter HG, Clothier R, Desolle P, Gerberick F, Liebsch M, Lovell WW, Maurer T, Pfannenbecker U, Potthast JM, Csato M, Sladowski D, Steiling W, Brantom P. The International EU/COLIPA In Vitro Phototoxicity Validation Study: Results of Phase II (Blind Trial). Part 1: the 3T3 NRU phototoxicity test. Toxicol In Vitro. 1998;12:305-327.
13. OECD. Test Guideline 432: In Vitro 3T3 NRU Phototoxicity Test. OECD Guidel Test Chem. 2019.
14. Juhas E, English JC 3rd. Tattoo-associated complications. J Pediatr Adolesc Gynecol. 2013;26:125-129.
15. Savitha AS. Composition of Tattoo. TATTOO-The Invaluable Compendium for Dermatologists. New Delhi; The Health Sciences; 2017;24.
16. Forte G, Petrucci F, Cristaudo A, Bocca B. Market survey on toxic metals contained in tattoo inks. Sci Total Environ. 2009;407:5997-6002.
17. Serup J. Seamless prevention of adverse events from tattooing: integrated strategy emphasising the customer-tattooist interaction. Curr Probl Dermatol. 2015;48:236-247.
18. Shinohara MM. Complications of decorative tattoo. Clin Dermatol. 2016;34:287-292.
19. Hutton Carlsen K, Serup J. Photosensitivity and photodynamic events in black, red and blue tattoos are common: A ‘Beach Study’. J Eur Acad Dermatol Venereol. 2014;28:231-237.
20. Kim K, Park H, Lim KM. Phototoxicity: Its Mechanism and Animal Alternative Test Methods. Toxicol Res. 2015;31:97-104. Erratum in: Toxicol Res. 2015;31:321.
21. Repetto G, del Peso A, Zurita JL. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nat Protoc. 2008;3:1125-1131.
22. Clothier R. The FRAME modified neutral red uptake cytotoxicity test. Invittox Protocol (3a). 1991.
23. Cudazzo G, Smart DJ, McHugh D, Vanscheeuwijck P. Lysosomotropic- related limitations of the BALB/c 3T3 cell-based neutral red uptake assay and an alternative testing approach for assessing e-liquid cytotoxicity. Toxicol In Vitro. 2019;61:104647.
24. Høgsberg T, Loeschner K, Löf D, Serup J. Tattoo inks in general usage contain nanoparticles. Br J Dermatol. 2011;165:1210-1218.
25. Kluger N, Phan A, Debarbieux S, Balme B, Thomas L. Skin cancers arising in tattoos: coincidental or not? Dermatology. 2008;217:219-221.
26. Kim SY, Seo S, Choi KH, Yun J. Evaluation of phototoxicity of tattoo pigments using the 3 T3 neutral red uptake phototoxicity test and a 3D human reconstructed skin model. Toxicol In Vitro. 2020;65:104813.
27. Regensburger J, Lehner K, Maisch T, Vasold R, Santarelli F, Engel E, Gollmer A, König B, Landthaler M, Bäumler W. Tattoo inks contain polycyclic aromatic hydrocarbons that additionally generate deleterious singlet oxygen. Exp Dermatol. 2010;19:e275-e281.
28. Bao L, Xu A, Tong L, Chen S, Zhu L, Zhao Y, Zhao G, Jiang E, Wang J, Wu L. Activated toxicity of diesel particulate extract by ultraviolet a radiation in mammalian cells: role of singlet oxygen. Environ Health Perspect. 2009;117:436-441.
29. Nisbet ICT, LaGoy PK. Toxic equivalency factors (TEFs) for polycyclic aromatic hydrocarbons (PAHs). Regul Toxicol Pharmacol. 1992;16:290- 300.
30. Arl M, Nogueira DJ, Schveitzer Köerich J, Mottim Justino N, Schulz Vicentini D, Gerson Matias W. Tattoo inks: Characterization and in vivo and in vitro toxicological evaluation. J Hazard Mater. 2019;364:548-561.
31. Wamer WG, Yin JJ. Photocytotoxicity in human dermal fibroblasts elicited by permanent makeup inks containing titanium dioxide. J Cosmet Sci. 2011;62:535-547.
32. Ceridono M, Tellner P, Bauer D, Barroso J, Alépée N, Corvi R, De Smedt A, Fellows MD, Gibbs NK, Heisler E, Jacobs A, Jirova D, Jones D, Kandárová H, Kasper P, Akunda JK, Krul C, Learn D, Liebsch M, Lynch AM, Muster W, Nakamura K, Nash JF, Pfannenbecker U, Phillips G, Robles C, Rogiers V, Van De Water F, Liminga UW, Vohr HW, Wattrelos O, Woods J, Zuang V, Kreysa J, Wilcox P. The 3T3 neutral red uptake phototoxicity test: practical experience and implications for phototoxicity testing--the report of an ECVAM-EFPIA workshop. Regul Toxicol Pharmacol. 2012;63:480-488

APA	TÜRK E, JANNUZZI A, ALPERTUNGA B (2022). Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test. , 70 - 75. 10.4274/tjps.galenos.2021.86344
Chicago	TÜRK Elif Gözde UTKU,JANNUZZI Ayse Tarbin,ALPERTUNGA Buket Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test. (2022): 70 - 75. 10.4274/tjps.galenos.2021.86344
MLA	TÜRK Elif Gözde UTKU,JANNUZZI Ayse Tarbin,ALPERTUNGA Buket Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test. , 2022, ss.70 - 75. 10.4274/tjps.galenos.2021.86344
AMA	TÜRK E,JANNUZZI A,ALPERTUNGA B Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test. . 2022; 70 - 75. 10.4274/tjps.galenos.2021.86344
Vancouver	TÜRK E,JANNUZZI A,ALPERTUNGA B Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test. . 2022; 70 - 75. 10.4274/tjps.galenos.2021.86344
IEEE	TÜRK E,JANNUZZI A,ALPERTUNGA B "Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test." , ss.70 - 75, 2022. 10.4274/tjps.galenos.2021.86344
ISNAD	TÜRK, Elif Gözde UTKU vd. "Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test". (2022), 70-75. https://doi.org/10.4274/tjps.galenos.2021.86344

APA	TÜRK E, JANNUZZI A, ALPERTUNGA B (2022). Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test. Turkish Journal of Pharmaceutical Sciences, 19(1), 70 - 75. 10.4274/tjps.galenos.2021.86344
Chicago	TÜRK Elif Gözde UTKU,JANNUZZI Ayse Tarbin,ALPERTUNGA Buket Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test. Turkish Journal of Pharmaceutical Sciences 19, no.1 (2022): 70 - 75. 10.4274/tjps.galenos.2021.86344
MLA	TÜRK Elif Gözde UTKU,JANNUZZI Ayse Tarbin,ALPERTUNGA Buket Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test. Turkish Journal of Pharmaceutical Sciences, vol.19, no.1, 2022, ss.70 - 75. 10.4274/tjps.galenos.2021.86344
AMA	TÜRK E,JANNUZZI A,ALPERTUNGA B Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test. Turkish Journal of Pharmaceutical Sciences. 2022; 19(1): 70 - 75. 10.4274/tjps.galenos.2021.86344
Vancouver	TÜRK E,JANNUZZI A,ALPERTUNGA B Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test. Turkish Journal of Pharmaceutical Sciences. 2022; 19(1): 70 - 75. 10.4274/tjps.galenos.2021.86344
IEEE	TÜRK E,JANNUZZI A,ALPERTUNGA B "Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test." Turkish Journal of Pharmaceutical Sciences, 19, ss.70 - 75, 2022. 10.4274/tjps.galenos.2021.86344
ISNAD	TÜRK, Elif Gözde UTKU vd. "Determination of the Phototoxicity Potential of Commercially Available Tattoo Inks Using the 3T3- neutral Red Uptake Phototoxicity Test". Turkish Journal of Pharmaceutical Sciences 19/1 (2022), 70-75. https://doi.org/10.4274/tjps.galenos.2021.86344