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Use of CAM-TBS assay as the alternative methods to Draize eye irritation test
( Chae Wook Kim ),( Kyoung Mi Jung ),( Byung Fhy Shu ),( Bae Hwan Kim ) 한국동물실험대체법학회 2007 한국동물실험대체법학회 학술대회집 Vol.2007 No.1
The hen`s egg test-chorioallantoic membrane (HET-CAM) assay, which employs the vascular membrane of a fertile hen`s egg, has been actively studied by several countries as an alternative method to the Draize rabbit eye irritation test. However, the scoring system of the original HET-CAM test is not satisfactory because of its lack of objectivity and quantitativeness. Moreover, the test required skill when the effects of test substance are to be regarded. To overcome these disadvantages, Hagino et al. developed and objective evaluation technique using the amount of trypan blue absorbed at the site of treatment as an indicator of injury to the CAM (Chorioallantoic membrane – trypan blue staining, CAM-TBS). In this study, we evaluate the CAM-TBS methods and used three materials (1%, 2%, 5% SLS, tween 20 and PBS) to evaluate the correlation of CAM-TBS with HET-CAM test. The amounts of trypan blue adsorbed with the CAM showed a good correlation with the HET-CAM test score(r = 0.958). According to these results, we suppose that the trypan blue staining assay using CAM-TBS may be a good method to replace the HET-CAM test as an alternative to Draize rabbit eye irritation test. Despite our results, additional studies with more materials are necessary to validate this method.
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