Epidermal-skin-test 1,000 (EST-1,000)--a new reconstructed epidermis for in vitro skin corrosivity testing.
The determination of a possible corrosive or irritative potential of certain products and ingredients is necessary for their classification and labeling requirements. Reconstructed skin as a model system provides fundamental advantages to single cell culture testing and leads to promising results as shown by different validation studies (for review: Fentem, J.H., Botham, P.A., 2002. ECVAM's activities in validating alternative tests for skin corrosion and irritation. ATLA 30(Suppl. 2), 61-67). In this study we introduce our new reconstructed epidermis "Epidermal-Skin-Test" (EST-1,000). This fully grown epidermis consists of proliferating as well as differentiating keratinocytes. EST-1,000 shows a high comparability to normal human skin as shown by histological and immunohistochemical data. Characteristic markers (KI-67, CK 1/10/5/14, transglutaminase, collagen IV, involucrin, beta 1 integrin) can be identified easily. The main focus of this work was to characterize EST-1,000 especially with respect to its barrier function by testing several substances of known corrosive potential. Skin corrosion was detected by the cytotoxic effect of the substances on a reconstructed epidermis after short-term application to the stratum corneum. The effect was determined by standard MTT assay and accompanying histological analysis. Hence EST-1,000 shows a very high predictive potential and closes the gap between animal testing and the established full-thickness model Advanced-Skin-Test 2,000 (AST-2,000) (Noll, M., Merkle, M.-L., Kandsberger, M., Matthes, T., Fuchs, H., Graeve, T., 1999. Reconstructed human skin (AST-2,000) as a tool for pharmaco-toxicology. ATLA 27, 302).
Hoffmann
J
J
Advanced CellSystems GmbH, Mülheimerstr. 26, D-53840 Troisdorf, Germany. jhoffmann@advancedcellsystems.com
Heisler
E
E
Karpinski
S
S
Losse
J
J
Thomas
D
D
Siefken
W
W
Ahr
H-J
HJ
Vohr
H-W
HW
Fuchs
H W
HW
D000179:Acrylates / Q000633:toxicity; D000826:Animal Testing Alternatives; D002210:Caprylates / Q000633:toxicity; D002424:Caustics / Q000145:classification / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D023381:Endpoint Determination; D004817:Epidermis / Q000187:drug effects / Q000473:pathology; D006801:Humans; D006878:Hydroxides / Q000633:toxicity; D007509:Irritants / Q000145:classification / Q000633:toxicity*; D009924:Organ Culture Techniques; D017680:Potassium Compounds / Q000633:toxicity; D015203:Reproducibility of Results; D012867:Skin / Q000187:drug effects* / Q000473:pathology; D023422:Skin Irritancy Tests / Q000379:methods*
D000179:Acrylates; D002210:Caprylates; D002424:Caustics; D006878:Hydroxides; D007509:Irritants; D017680:Potassium Compounds; C036658:acrylic acid; C031492:octanoic acid; C029943:potassium hydroxide