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 (TiO2, 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, TiO2 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.
Highlights
Tattooing is an ancient body modification in which sharp tools are used to insert non-soluble pigments into the dermal layer of the skin
An in vitro model suited to investigate the effects of the tattoo pigments carbon black, T iO2 and the organic azo pigment P.O.13 on human skin is reported
Neither tissue viability nor cytokine secretion of tattooed reconstructed human skin equivalents (TatS) was altered in the presence of tattoo pigments
Summary
Tattooing is an ancient body modification in which sharp tools are used to insert non-soluble pigments into the dermal layer of the skin. The tattoo pigments reside in the dermis where they can be taken up by fibroblasts and macrophages (Anderson et al 1996; Zelickson et al 1994). Pigments are transported to the lymph nodes and likely to other organs (Schreiver et al 2017). Besides phototoxicity, which represents 60% of side effects (Hutton Carlsen and Serup 2014), the contamination and release of potential carcinogens and other toxic substances are possible risks related to tattooing (Hauri and Hohl 2015; Hering et al 2018; Schreiver et al 2016). Infections, allergies and foreign body granulomas are common (Serup et al 2016)
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