Validating and troubleshooting ocular in vitro toxicology tests

Journal of Pharmacological and Toxicological Methods

Volumn 61, Issue 2, 2010, Pages 136-145

  Barile, Frank A ^a  

^a ST JOHN'S UNIVERSITY   (United States)

Author keywords

Alternative toxicology testing; ECVAM; ICCVAM; In vitro cytotoxicity; Methods; Ocular models; Organotypic

Indexed keywords

ANALYTIC METHOD; ANIMAL TESTING ALTERNATIVE; ARTICLE; BIOLOGICAL MODEL; BOVINE CORNEAL OPACITY AND PERMEABILITY ASSAY; CLINICAL PROTOCOL; CORNEA EPITHELIUM; CORNEA OPACITY; CORNEA PERMEABILITY; CYTOTOXICITY TEST; EYE EXAMINATION; EYE INFLAMMATION; EYE IRRITATION; EYE TOXICITY; HEN EGG TEST CHORIOALLANTOIC MEMBRANE ASSAY; HUMAN; IN VIVO STUDY; INTERMETHOD COMPARISON; ISOLATED CHICKEN EYE TEST METHOD; ISOLATED ORGAN; ISOLATED RABBIT EYE ASSAY; NONHUMAN; OPTOMETRY; PREDICTION; QUANTITATIVE ANALYSIS; SPECTROPHOTOMETRY; TOXICITY TESTING; VALIDATION PROCESS; VALIDATION STUDY;

ANIMAL TESTING ALTERNATIVES; ANIMALS; CATTLE; CHICKENS; CHORIOALLANTOIC MEMBRANE; CORNEAL OPACITY; EPITHELIUM, CORNEAL; EYE; EYE DISEASES; HUMANS; IRRITANTS; RABBITS; REPRODUCIBILITY OF RESULTS; TOXICITY TESTS;

BOVINAE; MAMMALIA; ORYCTOLAGUS CUNICULUS;

EID: 77950525080     PISSN: 10568719     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.vascn.2010.01.001     Document Type: Article

References (72)

1. Balls M., Botham P.A., Bruner L.H., Spielmann H. The EC/HO international validation study on alternatives to the Draize eye irritation test. Toxicology In Vitro 1995, 9:871-929.
2. Balls M., Amcoff P., Bremer S., Casati S., Coecke S., Clothier R., et al. The principles of weight of evidence validation of test methods and testing strategies. The report and recommendations of ECVAM workshop. Alternatives to Laboratory Animals (ATLA) 2006, 34:603-620.
3. Barile F.A. Continuous cell lines as a model for drug toxicity assessment. In vitro methods in pharmaceutical research 1997, 33-54. Academic Press, London, UK. J.V. Castell, M.J. Gómez-Lechón (Eds.).
4. Barile F.A. Chapter 13. Cell culture methods for acute toxicology testing. Principles of toxicology testing 2007, 175-202. Informa HealthCare, New York. F.A. Barile (Ed.).
5. Barile F.A., Cardona M. Acute cytotoxicity testing with cultured human lung and dermal cells. In vitro Cellular & Developmental Biology 1998, 34:631-635.
6. Blazka M., Harbell J.W., Klausner M., Merrill J.C., Kubilus J., Kloss C., et al. Evaluating the ocular irritation potential of 54 test articles using the EpiOcular human tissue construct model (OCL-200). The Toxicologist 2003, 72:221.
7. Budai P., Várnagy L. In vitro ocular irritation toxicity study of some pesticides. Acta Veternaria Hungarica 2000, 48:221-228.
8. Burton A.B.G. A method for the objective assessment of eye irritation. Food and Cosmetic Toxicology 1972, 10:209-217.
9. Burton A.B.G., York M., Lawrence R.S. The in vitro assessment of severe eye irritants. Food and Cosmetic Toxicology 1981, 19:471-480.
10. Calabro A.R., Konsoula R., Barile F.A. Evaluation of in vitro cytotoxicity and paracellular permeability of intact monolayers with mouse embryonic stem cells. Toxicology in Vitro 2008, 22:1273-1284.
11. Casterton P.L., Potts L.F., Klein B.D. A novel approach to assessing eye irritation potential using the bovine corneal opacity and permeability assay. Journal of Toxicology: Cutaneous & Ocular Toxicology 1996, 15:147-163.
12. Christian M.S., Diener R.M. Soaps and detergents-alternatives to animal eye irritation tests. Journal of the American College of Toxicology 1996, 15:1-44.
13. Collaborative study on the evaluation of alternative methods to the eye irritation test 1991, Commission of the European Communities (CEC), Doc. XI/632/91/V/E/1/131/91 Part I and II.
14. Cooper K.J., Earl L.K., Harbell J., Raabe H. Prediction of the ocular irritancy of prototype shampoo formulations by the isolated rabbit eye (IRE) test and bovine corneal opacity and permeability assay (BCOP) assay. Toxicology In Vitro 2001, 15:95-103.
15. Curren R., Evans M., Raabe H., Ruppalt R., Harbell J. Correlation of histopathology, opacity, and permeability of bovine corneas exposed in vitro to known ocular irritants. Veterinary Pathology 2000, 37:557-564.
16. De Wever B., Tornier C., Rosdy C., Beuerman R. In vitro corneal recovery assay using a reconstituted human corneal epithelial model. Fourth world congress on alternatives and animal use in the life sciences: Program and abstracts 2002, http://www.worldcongress.net [accessed January 2010].
17. Demirci B., Dadandi M., Paper D., Franz G., Baser K. Chemical composition of the essential oil of Phlomis linearis Boiss. & Bal., and biological effects on the CAM-assay: A safety evaluation. Zeitschrift für Naturforschung, Journal of Biosciences 2003, 58:826-829.
18. Demirci F., Paper D., Franz G., Baser K. Investigation of the Origanum onites L. essential oil using the chorioallantoic membrane (CAM) assay. Journal of Agriculture & Food Chemistry 2004, 52:251-254.
19. Draize J., Woodard G., Calvery H. Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membranes. Journal of Pharmacology & Experimental Therapeutics 1944, 82:377-390.
20. Manual of decisions for implementation of the sixth and seventh amendments to Directive 67/548/EEC on Dangerous Substances (Directives 79/831/EEC and 92/32/EEC) 2004, E.U.
21. Commission Directive 2001/59/EC of 6 August 2001 adapting to technical progress for the 28th time Council Directive 67/548/EEC on the approximation of the laws, regulations and administrative provisions relating to the classification, packaging and labeling of dangerous substances. Official Journal of European Communities 2001, L255:1-333. E.U. (European Union).
22. ESAC statements-Chemicals, cosmetics and pesticides; Statement on the conclusions of the ICCVAM retrospective study on organotypic in vitro assays as screening tests to identify potential ocular corrosives and severe irritants, 27 April 2007 2007, ECVAM Scientific Advisory Committee (ESAC), http://ecvam.jrc.it [accessed January 2010].
23. Ekwall B., Barile F.A. Chapter 11, Standardization and validation. Introduction to in vitro cytotoxicology: Mechanisms and methods 1994, 189-208. CRC Press, Inc, Boca Raton, FL. USA. F.A. Barile (Ed.).
24. Eskes C., Bessou S., Bruner L., Curren R., Harbell J., Jones P., et al. Eye irritation. Alternatives to Laboratory Animals (ATLA) 2005, 33(1):47-81.
25. Eskes C., Cole T., Hoffmann S., Worth A., Cockshott A., Gerner I., et al. The ECVAM international validation study on in vitro tests for acute skin irritation: Selection of test chemicals. Alternatives to Laboratory Animals (ATLA) 2007, 35:603-619.
26. Gautheron P., Dukic M., Alix D., Sina J.F. Bovine corneal opacity and permeability test: An in vitro assay of ocular irritancy. Fundamentals of Applied Toxicology 1992, 18:442-449.
27. Gettings S.D., Lordo R.A., Hintze K.L., Bagley D.M., Casterton P.L., Chudkowski M., et al. The CTFA evaluation of alternatives program: An evaluation of in vitro alternatives to the Draize primary rabbit eye irritation test. (Phase III) Surfactant-based formulations. Food and Chemical Toxicology 1996, 34:79-117.
28. Harbell J.W., Curren R.D. The bovine corneal opacity and permeability assay: Observations on assay performance. In Vitro & Molecular Toxicology 1998, 11:337-341.
29. Harbell J.W., Curren R.D. In vitro methods for the prediction of ocular and dermal toxicity. Handbook of toxicology 2002, 835-866. Boca Raton, FL, USA, CRC Press. M.J. Derelanko, M.A. Hollinger (Eds.).
30. Hartung T., Bremer S., Casati S., Coecke S., Corvi R., Fortaner S., et al. A modular approach to the ECVAM principles on test validity. Alternatives to Laboratory Animals (ATLA) 2004, 32:467-472.
31. ICCVAM, Validation and regulatory acceptance of toxicological test methods: A report of the ad hoc Interagency Coordinating Committee on the Validation of Alternative Methods. (1997). NIH Publication No.: 97-3981. National Toxicology Program (NTP) Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM), National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health (NIH), U.S. Public Health Service, Department of Health and Human Services.
32. ICCVAM Guidelines for the nomination and submission of new, revised, and alternative test methods. (2003). NIH Publication No: 03-4508. National Toxicology Program, NICEATM, NIEHS, NIH, U.S. Public Health Service, Department of Health and Human Services.
33. ICCVAM Test method evaluation report: In vitro ocular toxicity test methods for identifying severe irritants and corrosives. (2006). NIH Publication No.: 06-4511, National Toxicology Program (NTP), NICEATM, NIEHS, NIH, U.S. Public Health Service, Department of Health and Human Services.
34. ICCVAM, Background review document: Current status of in vitro test methods for identifying ocular corrosives and severe irritants: bovine corneal opacity and permeability test method. (2006). NIH Publication No.: 06-4512, National Toxicology Program (NTP), NICEATM, NIEHS, NIH, U.S. Public Health Service, Department of Health and Human Services. Available: [accessed January, 2010]. http://iccvam.niehs.nih.gov/methods/ocudocs/ocu_brd.htm#bcop.
35. ICCVAM, Background review document: Current status of in vitro test methods for identifying ocular corrosives and severe irritants: isolated chicken eye test method. (2006). NIH Publication No.: 06-4513, National Toxicology Program (NTP), NICEATM, NIEHS, NIH, U.S. Public Health Service, Department of Health and Human Services. Available: [Accessed January, 2010]. http://iccvam.niehs.nih.gov/methods/ocudocs/ocu_brd.htm#ice.
36. ICCVAM, Background review document: Current status of in vitro test methods for identifying ocular corrosives and severe irritants: Isolated rabbit eye test method. (2006). NIH Publication No.: 06-4514, National Toxicology Program (NTP), NICEATM, NIEHS, NIH, U.S. Public Health Service, Department of Health and Human Services. Available: [Accessed January, 2010]. http://iccvam.niehs.nih.gov/methods/ocudocs/ocu_brd.htm#ire.
37. ICCVAM, Background review document: Current status of in vitro test methods for identifying ocular corrosives and severe irritants: Hen's egg test-chorioallantoic membrane test method. (2006). NIH Publication No.: 06-4515, National Toxicology Program (NTP), NICEATM, NIEHS, NIH, U.S. Public Health Service, Department of Health and Human Services. Available: [Accessed January, 2010]. http://iccvam.niehs.nih.gov/methods/ocudocs/ocu_brd.htm#hetcam.
38. ICCVAM. (2009). Draft proposed ICCVAM test method recommendations: Evaluation of the validation status of alternative ocular safety testing methods and approaches. National Toxicology Program (NTP), NICEATM, NIEHS, NIH, U.S. Public Health Service, Department of Health and Human Services.
39. Kalweit S., Gerner I., Spielmann H. Validation project of alternatives for the Draize eye test. Molecular Toxicology 1987, 1:597-603.
40. Konsoula R., Barile F.A. Correlation of in vitro cytotoxicity with paracellular permeability in mortal rat intestinal cells. Journal of Pharmacological & Toxicological Methods 2007, 55:176-183.
41. Lordo R.A., Feder P.I., Gettings S.D. Comparing and evaluating alternative (in vitro) tests on their ability to predict the Draize maximum average score. Toxicology in Vitro 1999, 13:45-72.
42. Luepke N. Hen's egg chorioallantoic membrane test for irritation potential. Food & Chemical Toxicology 1985, 23:287-291.
43. Luepke N., Kemper F. The HET-CAM test: An alternative to the Draize eye test. Food & Chemical Toxicology 1986, 24:495-496.
44. Macián M., Seguer J., Infante M., Selve C., Vinardell M. Preliminary studies of the toxic effects of non-ionic surfactants derived from lysine. Toxicology 1996, 106:1-9.
45. McNamme P., Hibatallah J., Costabel-Farkas M., Goebel C., Araki D., Dufour E., et al. A tiered approach to the use of alternatives to animal testing for the safety assessment of cosmetics: Eye irritation. Regulatory Toxicology and Pharmacology 2009, 54:197-209.
46. Millichamp N.J. Species specificity: Factors affecting the interpretation of species differences in toxic responses of ocular tissues. Target organ toxicology series 1999, 89-114. Taylor and Francis, Philadelphia, PA, USA. 2nd ed. G.C.Y. Chiou (Ed.).
47. Muir C.K. Opacity of the bovine cornea in vitro induced by surfactants and industrial chemicals compared with ocular irritancy in vivo. Toxicology Letters 1985, 24:157-162.
48. Nguyen D.H., Beuerman R.W., De Wever B., Rosdy M. Three-dimensional construct of the human corneal epithelium for in vitro toxicology. Alternative methods for the new millennium, chapter 14 2003, 145-157. CRC Press, Boca Raton, FL, USA. H. Salem, S.A. Katz (Eds.).
49. Test No. 430. In vitro skin corrosion: Transcutaneous electrical resistance test (TER). OECD guidelines for testing of chemicals. Section 4: Health effects 2004, OECD, OECD Publishing, http://www.oecd.org/dataoecd/14/2/43295353.pdf. [accessed January, 2010].
50. Test No. 431. In vitro skin model. OECD guidelines for the testing of chemicals. Section 4: Health effects 2004, OECD, OECD Publishing, http://www.oecd.org/dataoecd/63/6/43302385.pdf. [accessed January, 2010].
51. Test No. 435. In vitro membrane barrier test method. OECD guideline for the testing of chemicals. Section 4: Health effects 2006, OECD, OECD Publishing, http://www.oecd.org/dataoecd/63/23/32037794.pdf. [accessed January, 2010].
52. Test No. 438: Isolated chicken eye test method for identifying ocular corrosives and severe irritants. OECD guidelines for the testing of chemicals, Section 4: Health effects 2009, OECD, OECD Publishing, http://dx.doi.org/10.1787/9789264076310-en, [accessed January, 2010].
53. Ohno Y., Kaneko T., Inoue T., Morikawa Y., Yoshida T., Fuji A., et al. Inter-laboratory validation of the in vitro eye irritation tests for cosmetic ingredients. (1) Overview of the validation study and Draize scores for the evaluation of the tests. Toxicology in Vitro 1999, 13:73-98.
54. Test No. 437: Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants. OECD guidelines for the testing of chemicals, Section 4: Health effects 2009, Organisation for Economic Co-operation and Development, OECD, OECD Publishing, http://dx.doi.org/10.1787/9789264076303-en [accessed January, 2010].
55. Parish W. Ability of in vitro (corneal injury-eye organ- and chorioallantoic membrane) tests to represent histopathological features of acute eye inflammation. Food & Chemical Toxicology 1985, 23:215-227.
56. Prinsen M.K., Koëter B.W.M. Justification of the enucleated eye test with eyes of slaughterhouse animals as an alternative to the Draize eye irritation test with rabbits. Food & Chemical Toxicology 1993, 31:69-76.
57. Rachui S.R., Robertson W.D., Duke M.A., Paller S., Ziets G.A. Predicting the ocular irritation potential of cosmetics and personal care products using two in vitro models. In Vitro Toxicology 1994, 7:45-52.
58. Scott L., Eskes C., Hoffmann S., Adriaens E., Alepée N., Bufo M., et al. A proposed eye irritation testing strategy to reduce and replace in vivo studies using bottom-up and top-down approaches. Toxicology In Vitro 2010, 24:1-9.
59. Sina J.F., Galer D.M., Sussman R.G., Gautheron P.D., Sargent E.V., Leong B., et al. A collaborative evaluation of seven alternatives to the Draize eye irritation test using pharmaceutical intermediates. Fundamental & Applied Toxicology 1995, 26:20-31.
60. Sina J.F., Gautheron P.D. A multitest approach to evaluating ocular irritation in vitro. Toxicology Methods 1994, 4:41-49.
61. Sina J.F., Gautheron P.D. Report from the bovine corneal opacity and permeability technical workshop (November 3-4, 1997): An historical perspective. In Vitro & Molecular Toxicology 1998, 11:316-326.
62. Sinn-Harlon J. Chickscope. University of Illinois-Urbana-Champagne 1998, Available: http://chickscope.itg.uiuc.edu/explore/embryology/day04/4Focuson_1.html [accessed January, 2010].
63. Spielmann H. Ocular irritation. In vitro methods in pharmaceutical research 1997, 265-287. Academic Press, London, UK. J.V. Castell, M.J. Gómez-Lechón (Eds.).
64. Steiling W., Bracher M., Courtellemont P., deSilva O. The HET-CAM, a useful in vitro assay for assessing the eye irritation properties of cosmetic formulations and ingredients. Toxicology In Vitro 1999, 13:375-384.
65. Sterzel W., Bartnik F., Matthies W., Kästner W., Künstler W. Comparison of two in vitro and two in vivo methods for the measurement of irritancy. Toxicology In Vitro 1990, 4:698-701.
66. Tufan A.C., Satiroglu-Tufan N.L. A chick embryo chorioallatoic membrane as a model system for the study of tumor angiogenesis, invasion, and development of anti-angiogenic agents. Current Cancer Drug Targets 2005, 5:249-266.
67. Ubels J.L., Paauw J.D., Casterton P.L., Kool D.J. A redesigned corneal holder for the bovine cornea opacity and permeability assay that maintains normal corneal morphology. Toxicology In Vitro 2002, 16:621-628.
68. Test for eye irritants 1995, U.S. Consumer Products Safety Commission (CPSC), 16CFR1500.42. Available at: http://www.access.gpo.gov/nara/cfr/waisidx_00/16cfr1500_00.html [accessed January, 2010].
69. Label review manual 1996, U.S. Environmental Protection Agency (EPA), U.S. Environmental Protection Agency, Washington, D.C, EPA737-B-96-001. 2nd ed.
70. Health effects test guideline, OPPTS 870.2400 Acute Eye Irritation 1998, U.S. EPA, U.S. Environmental Protection Agency, Washington, D.C, EPA712-C-98-195.
71. Globally harmonized system (GHS) of classification and labeling of chemicals 2003, United Nations (U.N.), United Nations Publications, New York, Available: http://www.unece.org/trans/danger/publi/ghs/officialtext.html [accessed January, 2010].
72. Weil C.S., Scala R.A. Study of intra- and inter-laboratory variability in the results of rabbit eye and skin irritation tests. Toxicology and Applied Pharmacology 1971, 19:276-360.