Title
Methyl green-pyronine staining of porcine organotypic skin explant cultures: An alternative model for screening for skin irritants
Author
Jacobs, J.J.L.
Lehé, C.
Cammans, K.D.A.
Das, P.K.
Elliott, G.R.
Publication year
2000
Abstract
We describe a new alternative method for screening for skin irritants by using fresh intact porcine skin biopsies. Test chemicals were applied to the epidermis of the biopsies, which were then incubated for different times in tissue culture medium at 37°C and with 5% carbon dioxide. A decrease in epidermal keratinocyte RNA staining, visualised in frozen sections by using a modified methyl-green pyronine (MGP) staining procedure, was employed as a marker of irritancy. If a decrease in epidermal RNA was observed after incubation for 4 hours (strong irritant), the chemical had an MGP score of 3; if after incubation for 24 hours (moderate irritant), the MGP score was 2; and if after incubation for 48 hours (weak irritant), the MGP score was 1. If no keratinocyte cytotoxicity was observed after incubation for 48 hours, the chemical was classified as non-irritant (MGP score = 0). At least three ears were used per chemical. The average MGP score was used to classify the chemical. Based on the MGP score for 20% sodium dodecyl sulphate, chemicals classified as strong or moderate irritants by using the MGP test were grouped together as category R38 chemicals. Weak irritants or non-irritants were not classified (NC). The MGP staining correctly identified 23 of 25 skin irritants for which reference data were available.
Subject
Methyl green, 54327-10-5
Pyronine, 92-32-0
Alternative method
Irritant testing
Methyl-green pyronine
Pig skin
Irritant agent
Animal tissue
Nonhuman
Screening test
Skin culture
Skin irritation
Skin toxicity
Swine
Toxicity testing
To reference this document use:
http://resolver.tudelft.nl/uuid:a1307abf-8d89-4332-b8a2-3ac4b4697d1e
TNO identifier
235385
ISSN
0261-1929
Source
ATLA Alternatives to Laboratory Animals, 28 (28), 279-292
Document type
article