Ocular Irritancy Assessment of Cosmetics Formulations and Ingredients: Fluorescein Leakage Test

Zanvit, A.; Meunier, Pierre-Alain; Clothier, Richard H.; Ward, R.K.; Buiatti-Tcheng, M.

doi:10.1016/s0887-2333(98)00084-8

Cited by 11 publications

(7 citation statements)

References 7 publications

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“…One method for the evaluation of the tightness of the epithelial barriers formed, is the sodium fluorescein leakage assay (8)(9)(10)36). The advantage of this assay is that it does not damage the cells, so can be combined with a cell activity or cell viability assay (12,21), such that it can be repeated and exposure effects and recovery profiles can be generated.…”

Section: Discussionmentioning

confidence: 99%

“…Social and scientific criticism of the Draize eye test (2) has stimulated research to develop in vitro alternatives that provide a quantitative and mechanistic evaluation of potential ocular damage. Several techniques have been proposed to replace the Draize eye test, as recently reviewed (3,4), including the use of corneal epithelial cells (5-7) associated with specific assays, for example the fluorescein leakage assay (8)(9)(10), the resazurin reduction assay (11)(12)(13), and assays for glutathione (14) and total cellular protein (such as, the kenacid blue assay [15,16]). These are designed to evaluate both function/activity and viability on exposure to chemicals, and subsequent recovery from insult (17,18).…”

Section: Introductionmentioning

confidence: 99%

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Comparison of an Animal Product Free Medium and Normal Growth Supplement on the Growth and Barrier Integrity of a Human Corneal Epithelial Cell Line

Wilkinson

Clothier

2005

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With the development of defined media for general and specific use with cell cultures, and concern over the use of human cells and over potential prion infections associated with growth factor extracts such as bovine pituitary extract, an animal product-free medium has become available. The basic keratinocyte defined medium can be used with a choice of animal product-containing or animal product-free supplements. Human corneal epithelia cell lines were cultured in the media with these two types of supplement, and compared in terms of their growth rates, their capacity to form tight barriers, and calcium regulation of the location of a junction-associated protein, zonula occludins-1 (ZO-1). The growth rates were not different in the two media, as long as the recommended coating was applied to the culture flask for the animal product-free medium. The barrier function was equally effective for confluent cultures seeded at the same densities. A calcium concentration of 100μM or above resulted in ZO-1 localisation at the cell membrane in either medium. Hence, cultures in the media are comparable, when the coating is employed. Further experiments are being conducted to establish the comparability of responses to chronic treatment with surfactants.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Comparison of an Animal Product Free Medium and Normal Growth Supplement on the Growth and Barrier Integrity of a Human Corneal Epithelial Cell Line

Wilkinson

Clothier

2005

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“…The MDCK epithelial cell line has been used to predict the potential ocular irritancy of many chemicals (7,9,10,21). MDCK cells develop zona occludens and macula adherens cell junctions in vitro, forming an impermeable barrier to large molecules (7).…”

Section: Discussionmentioning

confidence: 99%

The Prediction of Human Skin Responses by Using the Combined In Vitro Fluorescein Leakage/Alamar Blue (Resazurin) Assay

et al. 2002

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A range of cosmetics formulations with human patch-test data were supplied in a coded form, for the examination of the use of a combined in vitro permeability barrier assay and cell viability assay to generate, and then test, a prediction model for assessing potential human skin patch-test results. The target cells employed were of the Madin Darby canine kidney cell line, which establish tight junctions and adherens junctions able to restrict the permeability of sodium fluorescein across the barrier of the confluent cell layer. The prediction model for interpretation of the in vitro assay results included initial effects and the recovery profile over 72 hours. A set of the hand-wash, surfactant-based formulations were tested to generate the prediction model, and then six others were evaluated. The model system was then also evaluated with powder laundry detergents and hand moisturisers: their effects were predicted by the in vitro test system. The model was under-predictive for two of the ten hand-wash products. It was over-predictive for the moisturisers, (two out of six) and eight out of ten laundry powders. However, the in vivo human patch test data were variable, and 19 of the 26 predictions were correct or within 0.5 on the 0–4.0 scale used for the in vivo scores, i.e. within the same variable range reported for the repeat-test hand-wash in vivo data.

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“…In the absence of a suitable prediction model for converting the percentage permeability data to a prediction of in vivo irritation potential, only the benchmark approach was evaluated with the FL assay. Other prediction models for this assay have been published (1,22), but these are based on dose-response measurements with various incubation times, and therefore could not be evaluated in this study. Zanvit et al (22) reported a good correlation between classifications derived from Draize test results and predictions from a classification model based on the concentration causing 20% fluorescein leakage 4 hours after a 15-minute treatment.…”

Section: Discussionmentioning

confidence: 99%

“…More recently, data have been reported for several other in vitro tests for ocular irritation, for example, from studies with 3-dimensional reconstructed models (13)(14)(15), the bovine corneal opacity and permeability (BCOP) assay (16)(17)(18)(19), and the fluorescein leakage (FL) assay (20)(21)(22), which indicate that these methods can be used for comparative product safety assessments. A particular feature of the organotypic models, which appears advantageous, is the provision of a more complex barrier than that found with simple monolayer culture cytotoxicity assays.…”

Section: Introductionmentioning

confidence: 99%

Comparative Evaluation of Five In Vitro Tests for Assessing the Eye Irritation Potential of Hair-care Products

et al. 2001

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This study compared five methods, the isolated rabbit eye (IRE), bovine corneal opacity and permeability (BCOP), EpiOcular™, fluorescein leakage (FL) and neutral red release (NRR) assays, for predicting the eye irritation potential of hair-care formulations. Ten shampoo and seven conditioner formulations of known ocular irritation potential were tested. Each group included a market-acceptable formulation as a comparative benchmark. Predictions of ocular irritation were made by using classification models (IRE, BCOP and EpiOcular assays) or by direct comparison with benchmarks (IRE, EpiOcular, FL and NRR assays). The BCOP assay was less sensitive than the IRE test in discriminating between formulations of different irritation potentials, and did not perform as well as the other assays in identifying mild formulations. All of the assays appeared to be better at discriminating correctly between the shampoos than between the conditioners. The EpiOcular assay showed the closest concordance between the in vivo results and the in vitro data from cell-based assays (particularly for shampoos). The FL assay also showed a high concordance (particularly for conditioners). There was a tendency for these in vitro assays to over-predict eye irritation potential, but there was no under-prediction and they were particularly successful at identifying mild formulations. The NRR assay was less predictive with both shampoos and conditioners. The results from this comparative evaluation fully support the continued use of the IRE test as a suitable alternative to in vivo eye irritation testing in rabbits, although it also over-predicted the irritancies of several of the formulations. The value of using concurrent benchmarks (reference standards), appropriate to the materials being tested, in interpreting the data obtained from in vitro tests, was also demonstrated. Overall, the results indicate that further comparisons of the IRE, EpiOcular and FL assays are warranted using much larger numbers of test materials.

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Ocular Irritancy Assessment of Cosmetics Formulations and Ingredients: Fluorescein Leakage Test

Cited by 11 publications

References 7 publications

Comparison of an Animal Product Free Medium and Normal Growth Supplement on the Growth and Barrier Integrity of a Human Corneal Epithelial Cell Line

Comparison of an Animal Product Free Medium and Normal Growth Supplement on the Growth and Barrier Integrity of a Human Corneal Epithelial Cell Line

The Prediction of Human Skin Responses by Using the Combined In Vitro Fluorescein Leakage/Alamar Blue (Resazurin) Assay

Comparative Evaluation of Five In Vitro Tests for Assessing the Eye Irritation Potential of Hair-care Products

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