Evaluation of the Skin Irritation Using a DNA Microarray on a Reconstructed Human Epidermal Model

Abstract

To avoid the need to use animals to test the skin irritancy potential of chemicals and cosmetics, it is important to establish an in vitro method based on the reconstructed human epidermal model. To evaluate skin irritancy efficiently and sensitively, we determined the gene expression induced by a topically-applied mild irritant sodium dodecyl sulfate (SDS) in a reconstructed human epidermal model LabCyte TM EPI-MODEL (LabCyte) using a DNA microarray carrying genes that were related to inflammation, immunity, stress and housekeeping. The expression and secretion of IL-1a a in reconstructed human epidermal culture is known to be induced by irritation. We detected the induction of IL-1a a expression and its secretion into the cell culture medium by treatment with 0.075% SDS for 18 h in LabCyte culture using DNA microarray, quantitative reverse-transcription polymerase chain reaction (RT-PCR) and ELISA. DNA microarray analysis indicated that the expression of 10 of the 205 genes carried on the DNA microarray was significantly induced in a LabCyte culture by 0.05% or 0.075% SDS irritation for 18 h. RT-PCR analysis confirmed that SDS treatment significantly induced the expressions of interleukin-1 receptor antagonist (IL-1RN), FOS-like antigen 1 (FOSL1), heat shock 70 kDa protein 1A (HSPA1) and myeloid differentiation primary response gene (88) (MYD88), as well as the known marker genes for irritation IL-1b b and IL-8 in a LabCyte culture. Our results showed that a DNA microarray is a useful tool for efficiently evaluating mild skin irritation using a reconstructed human epidermal model.

β-Cryptoxanthin Alleviates Diet-Induced Nonalcoholic Steatohepatitis by Suppressing Inflammatory Gene Expression in Mice

“…In our previous study, the fibrous DNA microarray efficiently detected the specific gene expression associated with lipid metabolism and inflammation with a high sensitivity [18], [20], [23]. To understand the effect of β-cryptoxanthin on the alteration of specific gene expression associated with cholesterol and other lipid metabolism, inflammation, and fibrosis, we then synthesized DNA oligonucleotide probes and prepared the fibrous DNA microarray carrying 174 genes (Table S3).…”

A novel model of inflammatory pain in human skin involving topical application of sodium lauryl sulfate

“…The pathophysiology of SLS inflammation and possible mechanism of sensitization of peripheral nociceptors in the skin remain to be clarified. Studies with human skin cells in vitro, animal skin in vivo and human skin have shown that SLS releases a plethora of inflammatory mediators [6], including cytokines such as TNF-a, IL-1b, and IL-6 as well as a variety of chemokines [4][5][6][7][8][9][10][11]. The cellular source of mediators causing nociceptor sensitization in SLS reactions, as well as other human models of inflammatory pain, remains to be elucidated.…”

“…However, several lines of circumstantial evidence indicate that SLS may modulate pain responses in the skin. In vitro studies with human keratinocytes, as well as studies in animal skin, have shown that SLS induces the release or up-regulation of a variety of cytokines, including several considered pivotal in peripheral pain modulation [2,3], including TNF-a, IL-1a, IL-6, IL-8, IL-10, and several chemokines [4][5][6][7]. SLS also induces the release of pro-inflammatory cytokines and chemokines in human skin as shown by superficial lymph vessel drainage techniques [8], the modified skin window technique [9], and macroscopic and microscopic evaluations [10,11].…”

Selected Biomarkers Revealed Potential Skin Toxicity Caused by Certain Copper Compounds

“…The search for new endpoints is necessary, given the complexity of skin irritation mechanism. Various potential biomarkers have been studied and identified using proteomic and toxicogenomic technologies242526. These technologies potentially allow the setup of an in vitro test system, which resembles the in vivo situation as closely as possible27.…”