Development of an Alternative Test for Skin Sensitization Using a Three-dimensional Human Skin Model Consisting of Dendritic Cells, Keratinocytes and Fibroblasts

Tadashi Uchino1, Toshiaki Takezawa2, Yoshiaki Ikarashi1 and Tetsuji Nishimura1

1National Institute of Health Sciences, Tokyo, Japan
2National Institute of Agrobiological Sciences, Ibaraki, Japan

AATEX 16(1):1-8, 2011

In order to evaluate water-insoluble chemicals using the skin model which is more similar to real skin and detectable interaction among three kind cells, we established a test method which is a three-dimensional human skin model consisting of normal fibroblasts, normal keratinocytes and normal dendritic cells utilizing a collagen vitrigel membrane (VG-KDF-Skin). Nine sensitizers and five non-sensitizers were then examined. After 24 hr, the amount of IL-1α and IL-4 release was measured, and then positive/negative outcomes were evaluated (VG-KDF-Skin method).
The accuracy, sensitivity and specificity of positive/negative outcomes of the VG-KDF-Skin method, whose indicator is IL-4 release vs. local lymph node assay (LLNA), were 93%, 89% and 100%, respectively. The accuracy, sensitivity and specificity of the VG-KDF-Skin method, whose indicator is IL-1α release vs. LLNA, were 50%, 56% and 40%, respectively.
In order to study the possibility of applying an established test method to cosmetic products such as milky lotion and cream, two model cosmetic samples containing a typical skin sensitizer [2,4-dinitrochlorobenzene (DNCB)] were made and IL-4 release evaluated. Significant IL-4 release was induced.
These results suggest that it is possible the VG-KDF-Skin-method using IL-4 as an indicator for skin sensitization potential would be useful for evaluating the skin sensitization potential of chemicals and cosmetic products.

key words: skin sensitization test, 3D human skin model, collagen vitrigel membrane


(AATEX: Altern. Animal Test. EXperiment.: Alternatives to Animal Testing and EXperimentation)