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Study of a rat skin in vivo micronucleus test: data generated by mitomycin C and methyl methanesulfonate
T Nishikawa1, M Haresaku, K Adachi
1Human Safety Evaluation Center, Lion, 100 Tajima, Odawara-shi, Kanagawa 256-0811, Japan. ntaka@lion.co.jp
Abstract:
We have developed an in vivo micronucleus (MN) test that uses rat skin as the target organ. Sample preparation involves cold-treating the epidermis with trypsin, peeling it off with a fine forceps, treating it in hypotonic solution, and staining it with acridine orange (A.O.). We evaluated the assay using mitomycin C (MMC) and methyl methanesulfonate (MMS) as model clastogens, applying them as single and repeat treatments. Both chemicals induced a significant, dose-dependent increase in MN frequency in basal cells. One treatment per day for 3 days was optimal for MN induction.
Insights
This study introduces a new in vivo micronucleus (MN) test using rat skin. The assay effectively detects genotoxic effects of chemicals like mitomycin C (MMC) and methyl methanesulfonate (MMS) in skin cells.
Area of Science:
- Toxicology
- Genetics
- Dermatology
Background:
- The micronucleus (MN) test is a standard assay for detecting genotoxicity.
- Existing MN tests often target internal organs, necessitating invasive procedures.
- A non-invasive or less invasive skin-based MN test could offer advantages in toxicological assessments.
Purpose of the Study:
- To develop and validate a novel in vivo micronucleus (MN) test utilizing rat skin as the target organ.
- To assess the sensitivity of the rat skin MN test to known clastogenic agents.
Main Methods:
- Epidermal tissue was isolated from rats after cold trypsin treatment and forceps peeling.
- Isolated epidermis was subjected to hypotonic treatment and stained with acridine orange (A.O.).
- The assay was validated using mitomycin C (MMC) and methyl methanesulfonate (MMS) as model clastogens, with single and repeat dosing regimens.
Main Results:
- Both MMC and MMS induced a significant, dose-dependent increase in the frequency of micronuclei in basal epidermal cells.
- A repeat treatment regimen of one dose per day for three consecutive days was found to be optimal for MN induction.
Conclusions:
- The developed rat skin in vivo micronucleus test is a sensitive and viable method for genotoxicity assessment.
- This assay provides a valuable tool for evaluating the genotoxic potential of chemicals targeting the skin.