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RECONSTITUTED 3-DIMENSIONAL HUMAN SKIN OF VARIOUS ETHNIC ORIGINS AS AN IN VITRO MODEL FOR STUDIES OF PIGMENTATION.

Yoon1,2, T.J., Lei2, T.C., Yamaguchi2, Y., Batzer3, J., Wolber3, R., Hearing2, V.J. 1Gyeongsang National University Hospital, Gyeongnam, South Korea, 2National Cancer Institute, Bethesda, MD, 3R&D Cosmed, Skin Research, Beiersdorf AG, Hamburg, Germany.
Abstract

Reconstituted 3-dimensional human skin equivalents containing melanocytes and keratinocytes on an artifcial dermal substitute are gaining popularity for studies of skin metabolism because they exhibit morphological and growth characteristics similar to human epidermis. In this study, we show that such a pigmented epidermis model can be used to assess the regulation of pigmentation by known melanogenic compounds. In monolayers or in melanocyte-keratinocyte co-cultures, melanocyte-keratinocyte interactions are missing or are spatially limited. The commercial skin equivalents used in this study were derived from epidermal cells obtained from donors of three different ethnic origins (African-American, Asian, and Caucasian),and they reflect those distinct skin phenotypes. We used these pigmented human epidermis models to test compounds for potential effects on pigmentation in a more physiologically relevant context, which allows further characterization and validation of interesting melanogenic factors. We used known melanogenic stimulators (a melanocyte-stimulating hormone and 3,4-dihydroxyphenylalanine) and inhibitors (hydroquinone, arbutin, kojic acid, and niacinamide) and examined their effects on the production of melanin and its distribution in upper layers of the skin. Our studies indicate that commercial skin equivalents provide a convenient and cost-effective alternative to animal testing for evaluating the regulation of mammalian pigmentation by melanogenic factors and for elucidating their mechanisms of action.

Keywords

3,4-dihydroxyphenylalanine (DOPA), 3-Dimensional skin equivalent, African-american, Alexa 594-conjugated goat antibody against mouse IgG, Alpha-MSH (Alpha-melanocyte stimulating hormone), Alpha-melanocyte-stimulating hormone (MSH), Arbutin, Asian, Basal layer, BioSkin, Black/brown eumelanins, Caucasian, Co-culture, DC, Dendritic cells, EpiDerm, EpiSkin, Fibroblasts, Heterogenous biopolymers, Human HMB-45, Hydroquinone, Keratinocyte, Kojic acid, L-NMM medium, Langerhans cells, MSH, MTT, MTT ET-50 tissue viability assay, MTT assay, Mel-A, Mel-B, Mel-C, Melanin, MelanoDerm, Melanocyte, Melanocyte stimulating hormone, Metabolism, Niacinamide, Phorbol 12-myristate 13-acetate (PMA), Phosphate-buffered saline (PBS), Photoprotective efficacy, Pigmentation, Reconstituted 3-D human epidermis, Red/yellow pheomelanins, Standardized testing of pigmentary regulators (STOPR), Stratum corneum, Superbasal layer, Tyrosinase

Materials Tested

3,4-dihydroxyphenylalanine (DOPA), Alpha-melanocyte-stimulating hormone (MSH), Arbutin, Hydroquinone, Kojic ccid, L-NMM medium, MTT Kit, Niacinamide, Phosphate-buffered saline (PBS)

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