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Title: Rat epidermal keratinocyte organotypic culture (ROC) as a model for chemically induced skin irritation testing

Abstract

The potential of rat epidermal keratinocyte (REK) organotypic culture (ROC) with proper stratum corneum barrier as a model for screening skin irritants was evaluated. The test chemicals were selected from ECETOC database (1995) and the observed in vitro irritation potential was compared to ECETOC in vivo primary irritation index (PII), to EU risk phrases, and to the harmonized OECD criteria. Chemicals were applied onto the stratum corneum surface of ROC for 30 min and samples were taken from the underlying medium at 4 and 8 h after exposure. Cell membrane integrity (determined by LDH assay) and pro-inflammatory effect (determined by IL-1{alpha} release) were verified at both time points and correlated to PII values. The best correlation (R {sup 2} = 0.831) was seen with LDH leakage test. Based on obtained data, chemicals were classified according to criteria defined by EU and OECD. From 12 chemicals, only two were incorrectly classified according to OECD criteria when using LDH leakage and IL-1{alpha} release as irritation markers. At the end of experiment, chemical-treated ROC cultures were fixed and histological changes were assessed. Typical signs for irritation were lightly stained cytoplasm, condensed nuclei, cellular vacuolization, eosinophilic cytoplasms, and blebbing. These irritation effects of chemicalsmore » were graded visually into four classes (A-D). The extent of morphological perturbations of the cultures mostly correlated with PII. The present results indicate the validity of the ROC model in predicting skin irritation potential of chemicals and show that the use of set of irritation markers with different mechanistic responses gives more information on irritation than if only one marker was used.« less

Authors:
 [1];  [2];  [3];  [4];  [2];  [3]
  1. Department of Pharmaceutics, University of Kuopio, PO Box 1627, 70211 Kuopio (Finland). E-mail: sari.pappinen@uku.fi
  2. Department of Anatomy, University of Kuopio, PO Box 1627, 70211 Kuopio (Finland)
  3. Department of Pharmaceutics, University of Kuopio, PO Box 1627, 70211 Kuopio (Finland)
  4. (Finland)
Publication Date:
OSTI Identifier:
20722024
Resource Type:
Journal Article
Resource Relation:
Journal Name: Toxicology and Applied Pharmacology; Journal Volume: 208; Journal Issue: 3; Other Information: DOI: 10.1016/j.taap.2005.02.014; PII: S0041-008X(05)00110-9; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CELL CULTURES; CELL MEMBRANES; CHEMICAL INDUSTRY; CYTOPLASM; ECOLOGY; HEALTH HAZARDS; IN VITRO; IN VIVO; INFLAMMATION; LACTATE DEHYDROGENASE; MORPHOLOGY; RATS; SKIN; SODIUM; SULFATES

Citation Formats

Pappinen, Sari, Pasonen-Seppaenen, Sanna, Suhonen, Marjukka, Department of Pharmaceutics, Biopharmaceutical In Vitro Drug Evaluation Laboratory, University of Kuopio, PO Box 1627, 70211 Kuopio, Tammi, Raija, and Urtti, Arto. Rat epidermal keratinocyte organotypic culture (ROC) as a model for chemically induced skin irritation testing. United States: N. p., 2005. Web. doi:10.1016/j.taap.2005.02.014.
Pappinen, Sari, Pasonen-Seppaenen, Sanna, Suhonen, Marjukka, Department of Pharmaceutics, Biopharmaceutical In Vitro Drug Evaluation Laboratory, University of Kuopio, PO Box 1627, 70211 Kuopio, Tammi, Raija, & Urtti, Arto. Rat epidermal keratinocyte organotypic culture (ROC) as a model for chemically induced skin irritation testing. United States. doi:10.1016/j.taap.2005.02.014.
Pappinen, Sari, Pasonen-Seppaenen, Sanna, Suhonen, Marjukka, Department of Pharmaceutics, Biopharmaceutical In Vitro Drug Evaluation Laboratory, University of Kuopio, PO Box 1627, 70211 Kuopio, Tammi, Raija, and Urtti, Arto. Tue . "Rat epidermal keratinocyte organotypic culture (ROC) as a model for chemically induced skin irritation testing". United States. doi:10.1016/j.taap.2005.02.014.
@article{osti_20722024,
title = {Rat epidermal keratinocyte organotypic culture (ROC) as a model for chemically induced skin irritation testing},
author = {Pappinen, Sari and Pasonen-Seppaenen, Sanna and Suhonen, Marjukka and Department of Pharmaceutics, Biopharmaceutical In Vitro Drug Evaluation Laboratory, University of Kuopio, PO Box 1627, 70211 Kuopio and Tammi, Raija and Urtti, Arto},
abstractNote = {The potential of rat epidermal keratinocyte (REK) organotypic culture (ROC) with proper stratum corneum barrier as a model for screening skin irritants was evaluated. The test chemicals were selected from ECETOC database (1995) and the observed in vitro irritation potential was compared to ECETOC in vivo primary irritation index (PII), to EU risk phrases, and to the harmonized OECD criteria. Chemicals were applied onto the stratum corneum surface of ROC for 30 min and samples were taken from the underlying medium at 4 and 8 h after exposure. Cell membrane integrity (determined by LDH assay) and pro-inflammatory effect (determined by IL-1{alpha} release) were verified at both time points and correlated to PII values. The best correlation (R {sup 2} = 0.831) was seen with LDH leakage test. Based on obtained data, chemicals were classified according to criteria defined by EU and OECD. From 12 chemicals, only two were incorrectly classified according to OECD criteria when using LDH leakage and IL-1{alpha} release as irritation markers. At the end of experiment, chemical-treated ROC cultures were fixed and histological changes were assessed. Typical signs for irritation were lightly stained cytoplasm, condensed nuclei, cellular vacuolization, eosinophilic cytoplasms, and blebbing. These irritation effects of chemicals were graded visually into four classes (A-D). The extent of morphological perturbations of the cultures mostly correlated with PII. The present results indicate the validity of the ROC model in predicting skin irritation potential of chemicals and show that the use of set of irritation markers with different mechanistic responses gives more information on irritation than if only one marker was used.},
doi = {10.1016/j.taap.2005.02.014},
journal = {Toxicology and Applied Pharmacology},
number = 3,
volume = 208,
place = {United States},
year = {Tue Nov 01 00:00:00 EST 2005},
month = {Tue Nov 01 00:00:00 EST 2005}
}