TatS: a novel in vitro tattooed human skin model for improved pigment toxicology research

Abstract

Reports of tattoo-associated risks boosted the interest in tattoo pigment toxicity over the last decades. Nonetheless, the influence of tattoo pigments on skin homeostasis remains largely unknown. In vitro systems are not available to investigate the interactions between pigments and skin. Here, we established TatS, a reconstructed human full-thickness skin model with tattoo pigments incorporated into the dermis. We mixed the most frequently used tattoo pigments carbon black (0.02 mg/ml) and titanium dioxide (TiO 2 , 0.4 mg/ml) as well as the organic diazo compound Pigment Orange 13 (0.2 mg/ml) into the dermis. Tissue viability, morphology as well as cytokine release were used to characterize TatS. Effects of tattoo pigments were compared to monolayer cultures of human fibroblasts. The tissue architecture of TatS was comparable to native human skin. The epidermal layer was fully differentiated and the keratinocytes expressed occludin, filaggrin and e-cadherin. Staining of collagen IV confirmed the formation of the basement membrane. Tenascin C was expressed in the dermal layer of fibroblasts. Although transmission electron microscopy revealed the uptake of the tattoo pigments into fibroblasts, neither viability nor cytokine secretion was altered in TatS. In contrast, TiO 2 significantly decreased cell viability and increased interleukin-8 release in fibroblast monolayers. In conclusion, TatS emulates healed tattooed human skin and underlines the advantages of 3D systems over traditional 2D cell culture in tattoo pigment research. TatS is the first skin model that enables to test the effects of pigments in the dermis upon tattooing. Electronic supplementary material The online version of this article (10.1007/s00204-020-02825-z) contains supplementary material, which is available to authorized users.

Cutaneous Malignancies in Tattoos, a Case Series of Six Patients

“…More details are presented in Table S1. Interestingly, it was revealed in the study by Henrik Hering et al that fibroblasts absorb tattoo pigments and that the tattoo pigment TiO2 significantly decreases cell viability and increases interleukin-8 release in fibroblast monolayers [92].…”

Commensal-Related Changes in the Epidermal Barrier Function Lead to Alterations in the Benzo[ a ]Pyrene Metabolite Profile and Its Distribution in 3D Skin

“…Subsequently, cryosections were fixed for 10 min with ice-cold methanol (−20°C). Antibody (Ab) staining was performed as described by Hering et al ( 52 ). The following antibodies were used: rabbit Ab against cytokeratin 10 (18343-1-AP) and involucrin (55328-1-AP; Proteintech Group, Rosemont, IL, USA) and mouse Ab against filaggrin (NBP2-53245-20; Novus Biologicals, Littleton, CO, USA) and E-cadherin (33-4000; Thermo Scientific, Waltham, MA, USA) at a concentration of 5 μg/ml.…”

Evaluation of in vitro human skin models for studying effects of external stressors and stimuli and developing treatment modalities

“…[125] In the last decade, the prevalence of tattoos in western society has risen, and along with it, concerns of tattoo pigment toxicity. [27] Given the solubility of tattoo ink, transferring it into cell culture medium will only elucidate the toxicity of soluble ingredients. Whereas tattooing a prefabricated skin model will lead to significant damage that is unlikely to heal prior to culture expiration.…”