Assessment of Toxic Potential of Industrial Chemicals Using a Cultured Human Bioartificial Skin Model: Production of Interleukin 1α and Hydroxyeicosatetraenoic Acids

Abstract

Cytotoxicity assays using artificial skin are proposed as alternative methods for in vitro tests to minimize animals used in ocular and dermal irritation testing. The responses of the artificial skins were studied to a well-characterized chemical irritant, such as toluene, glutaraldehyde and sodium lauryl sulfate (SLS), and a nonirritant, such as polyethylene glycol. The evaluation of irritating and nonirritating test chemicals was also compared with responses seen in human dermal fibroblasts and human epidermal keratinocytes grown in monolayer culture. The responses monitored included the MTT mitochondrial functionality assay. In order to better understand the local mechanisms involved in skin damage and repair, the productions of several mitogenic proinflammatory mediators such as interleukin-1α (IL-1α), 12-hydroxyeicosatetraenoic acid (12-HETE) and 15-HETE were investigated. Dose-dependent increases in the levels of IL-1α and HETEs were observed in the underlying medium of the skin systems exposed to two skin irritants, glutaraldehyde and SLS. The results of the present study show that both human artificial skins can be used as efficient testing models for the evaluation of skin toxicity in vitro and for screening the contact skin irritancy in vitro.

Hydroxyeicosatetraenoic acids released through the cytochrome P-450 pathway regulate 3T6 fibroblast growth

“…These authors also detected 15-HETE, whereas 5-HETE was not encountered. Other studies found that interleukin-1 increased the pro-duction of 12-and 15-HETE in human dermal fibroblasts (11,12). Additionally, it has been demonstrated that fibroblasts can respond to LOX products such as LTs or HETEs.…”

Three‐dimensional organization of dermal fibroblasts by macromass culture

“…The medium was DMEM+10% FBS. For exposure to a known irritant [16,17], monolayers and macromasses were incubated in the presence of different concentrations of SDS (Sigma) or vehicle (water) added to the medium. Separate 80 μl volumes of 0, 0.1, 0.2, 0.3, 0.4 and 0.5% (w/v) SDS were added/ml of medium.…”

“…The monolayers and macromasses were exposed to the SDS for 24 h, after which cell mitochondrial function was assessed by the MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2 H ‐tetrazolium bromide; Sigma–Aldrich] assay. For exposure to a known non‐irritant, monolayers and three‐dimensional constructs were incubated in the presence of different concentrations of PEG 400 [16] or vehicle (water) added to the medium. Separate 80 μl volumes of 0, 20, 40, 60, 80 and 100% (v/v) PEG 400 (Merck) were added/ml of medium.…”

Hand hygiene and skin health

“…Detergents destroy the barrier and stimulate the release of proinflammatory cytokines from keratinocytes. 42,43 Antiseptic alcohols have lipid-depleting detergents and irritating fragrances added to them. The picture is further obscured by large variations in the sensitivity to barrier problems from one person to another.…”