Evaluation of a modified HET-CAM assay as a screening test for eye irritancy

Gilleron, L.; Coecke, S.; Sysmans, M.; Hansen, Erik W.; Oproy, S. Van; Marzin, Daniel; Cauteren, H. Van; Vanparys, Ph.

doi:10.1016/0887-2333(96)00021-5

Cited by 56 publications

(27 citation statements)

References 17 publications

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“…This indicates that there will also be a need for in vitro methods that can identify conjunctiva effects, and especially conjunctiva redness, to fully replace the in vivo Draize eye test if the animal data continue to be interpreted as currently expressed in UN GHS and EU CLP and the classifications obtained thereafter continue being used as reference in the evaluation/validation of alternative methods. The Hen’s Egg Test–Chorioallantoic Membrane (HET-CAM) (Luepke 1985; de Silva et al 1992; Gilleron et al 1996; Spielmann et al 1996) and similar methods like the Chorioallantoic Membrane Vascular Assay (CAMVA) (Bagley et al 1994) have been proposed to provide information on conjunctiva effects in vivo due to the similarity of the CAM to the conjunctiva. Both HET-CAM and CAMVA underwent multiple international validation studies (Bagley et al 1992b, 1999b; Spielmann et al 1993, 1996, 1997; Brantom et al 1997; Ohno et al 1999; Hagino et al 1999).…”

Section: Discussionmentioning

confidence: 99%

Retrospective analysis of the Draize test for serious eye damage/eye irritation: importance of understanding the in vivo endpoints under UN GHS/EU CLP for the development and evaluation of in vitro test methods

Adriaens¹,

Barroso

Eskes

et al. 2013

Arch Toxicol

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For more than two decades, scientists have been trying to replace the regulatory in vivo Draize eye test by in vitro methods, but so far only partial replacement has been achieved. In order to better understand the reasons for this, historical in vivo rabbit data were analysed in detail and resampled with the purpose of (1) revealing which of the in vivo endpoints are most important in driving United Nations Globally Harmonized System/European Union Regulation on Classification, Labelling and Packaging (UN GHS/EU CLP) classification for serious eye damage/eye irritation and (2) evaluating the method’s within-test variability for proposing acceptable and justifiable target values of sensitivity and specificity for alternative methods and their combinations in testing strategies. Among the Cat 1 chemicals evaluated, 36–65 % (depending on the database) were classified based only on persistence of effects, with the remaining being classified mostly based on severe corneal effects. Iritis was found to rarely drive the classification (<4 % of both Cat 1 and Cat 2 chemicals). The two most important endpoints driving Cat 2 classification are conjunctiva redness (75–81 %) and corneal opacity (54–75 %). The resampling analyses demonstrated an overall probability of at least 11 % that chemicals classified as Cat 1 by the Draize eye test could be equally identified as Cat 2 and of about 12 % for Cat 2 chemicals to be equally identified as No Cat. On the other hand, the over-classification error for No Cat and Cat 2 was negligible (<1 %), which strongly suggests a high over-predictive power of the Draize eye test. Moreover, our analyses of the classification drivers suggest a critical revision of the UN GHS/EU CLP decision criteria for the classification of chemicals based on Draize eye test data, in particular Cat 1 based only on persistence of conjunctiva effects or corneal opacity scores of 4. In order to successfully replace the regulatory in vivo Draize eye test, it will be important to recognise these uncertainties and to have in vitro tools to address the most important in vivo endpoints identified in this paper.

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Section: Discussionmentioning

confidence: 99%

Retrospective analysis of the Draize test for serious eye damage/eye irritation: importance of understanding the in vivo endpoints under UN GHS/EU CLP for the development and evaluation of in vitro test methods

Adriaens¹,

Barroso

Eskes

et al. 2013

Arch Toxicol

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“…Ex vivo toxicity evaluation of optimized NEG loaded with KP was carried out using chorioallantoic membrane (CAM), a highly vascularized membrane that surrounds the developing chick embryo (Gilleron et al, 1996). Hen's egg test-chorioallantoic membrane (HET-CAM) assay has been used to evaluate the blood vessels reaction and damage to the CAM of a fertilized hen's egg to determine the toxicity potential of chemicals and formulations in vitro.…”

Section: Irritation Testing Using Het-cam Methodsmentioning

confidence: 99%

Formulation development of novel in situ nanoemulgel (NEG) of ketoprofen for the treatment of periodontitis

2014

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The aim of the present study was to formulate and evaluate in situ gelling syringeable nanoemulgels (NEGs) of ketoprofen for periodontal delivery. Application of 3-factor 3-level design was employed using the Box-Behnken experimental design for the optimization of nanoemulsion using three independent variables such as percent concentration (v/v) of oil (X 1 ), S mix (mixture of surfactant and cosurfactant) (X 2 ) and water (X 3 ); while the particle size (nm) (Y 1 ), polydispersity index (Y 2 ) and zeta potential (mV) (Y 3 ) were used as dependent variables. The NEG was evaluated based on their drug content, pH measurement, mucoadhesion on the goat buccal mucosa, syringeability and inverted sol-gel transition temperature. The drug release data were analyzed for curve fitting based on the Korsmeyer-Peppas law, and the n-values of optimized A5 and A8 formulations were found 0.3721 and 0.3932, respectively, confirmed that both the formulations followed pseudo Fickian diffusion (n50.43). The formulation A8 with the optimal drug release was identified as the best NEG formulation. Results of rheological, mucoadhesion and syringeability studies showed the suitability of desired sol-gel property for periodontal drug delivery. The Herschel-Bulkley model was the best fit model to explain the flow behavior of optimized formulation. Using the HET-CAM method, significantly lower in vitro toxicity was indicated the suitability of developed NEG for intra-pocket delivery.

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“…Hen's egg test in chorioallantoic membrane (HET-CAM) was performed as described by Ref. 21 and adapted by Ref. 22.…”

Section: Evaluation Of Pnpp-19 Toxicity Through Hen's Egg Test In Chomentioning

confidence: 99%

PnPP-19 Peptide as a Novel Drug Candidate for Topical Glaucoma Therapy Through Nitric Oxide Release

Silva

Dourado

Lima

et al. 2020

Trans. Vis. Sci. Tech.

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Evaluation of a modified HET-CAM assay as a screening test for eye irritancy

Cited by 56 publications

References 17 publications

Retrospective analysis of the Draize test for serious eye damage/eye irritation: importance of understanding the in vivo endpoints under UN GHS/EU CLP for the development and evaluation of in vitro test methods

Retrospective analysis of the Draize test for serious eye damage/eye irritation: importance of understanding the in vivo endpoints under UN GHS/EU CLP for the development and evaluation of in vitro test methods

Formulation development of novel in situ nanoemulgel (NEG) of ketoprofen for the treatment of periodontitis

PnPP-19 Peptide as a Novel Drug Candidate for Topical Glaucoma Therapy Through Nitric Oxide Release

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