Regulatory agencies require testing of chemicals and products to protect workers and consumers from potential eye injury hazards. This testing, referred to as ocular toxicity testing, represents one of the four most commonly conducted product safety tests. Ocular toxicity tests are conducted to identify ocular corrosives (substances that cause permanent eye tissue damage) and ocular irritants (substances that cause temporary eye tissue damage). Evaluation of test methods that reliably identify ocular corrosives and irritants while replacing, reducing, and refining animal use is a high priority for NICEATM and ICCVAM.
ICCVAM has recommended the use of in vitro ocular toxicity test methods and testing strategies that replace animal use to
NICEATM and ICCVAM also evaluated approaches to reduce and refine animal use for eye safety testing, including:
Summaries of these evaluations appear below. Click hyperlinked headings for more information about specific projects.
OptiSafe is an in vitro test method in which a test substance is applied to a semipermeable membrane. Damage to macromolecules in the membrane is measured to assess the test substance’s potential to cause eye irritation.
In 2013, at the request of the test method developer, Lebrun Labs, NICEATM identified a panel of reference chemicals against which to validate the OptiSafe test method. These chemicals represented a range of physical and chemical properties and hazard classes. Lebrun Labs used the panel in coded form in a validation study. NICEATM’s initial review of the validation study data indicated that the OptiSafe method compared favorably to other in vitro ocular toxicity testing methods.
ICCVAM reactivated its Ocular Toxicity Working Group in 2014 to provide guidance to Lebrun Labs on additional validation activities necessary to thoroughly evaluate this test method and determine its potential regulatory utility.
The short time exposure (STE) test is an in vitro test proposed for identifying ocular irritants. NICEATM compared STE test data with data generated using the traditional rabbit eye test. The results of the comparison, including comments from external reviewers, was included in a proposal to the Organisation for Economic Co-operation and Development (OECD) for a test guideline for the STE test.
Eye safety testing procedures vary among U.S. agencies. The U.S. Code of Federal Regulations (16 CFR 1500.42) provides criteria and procedures for identifying eye injury hazards based on rabbit eye test results. However, these procedures do not provide criteria to classify results obtained from a three-animal test.
NICEATM, in collaboration with ICCVAM, conducted an analysis to determine appropriate three-animal test classification criteria that were equivalent to the current testing procedure criteria cited in 16 CFR 1500.42. In 2012, ICCVAM recommended a procedure that uses 50% to 83% fewer animals than accepted testing procedures and harmonizes the number of animals used for ocular toxicity testing across U.S. regulatory agencies and international test guidelines based on their analysis.
ICCVAM considered comments from both the Scientific Advisory Committee on Alternative Toxicological Methods (SACATM) and from the public as it finalized its recommendations. These recommendations have been forwarded to Federal agencies.
The endorsement of several alternative testing methods by Federal agencies has provided regulated industries with important new tools for assessing potential eye injury hazards. The new test methods were the topic of a 2011 workshop organized by NICEATM and ICCVAM. The workshop identified best practices for chemical and product ocular toxicity testing using methods and approaches that replace, reduce, and refine animal use.
Despite progress in alternative ocular toxicity test method development, no alternative test method has been adopted by regulatory authorities as a complete replacement for the in vivo rabbit eye test. Accordingly, when the rabbit eye test is necessary, ICCVAM recommends that pain management procedures always be used. A test method protocol describing specific procedures to prevent or minimize animal pain and distress is included in the ICCVAM report evaluating the use of anesthetics, analgesics, and humane endpoints during the in vivo eye test.
ICCVAM evaluated and developed recommendations for five in vitro test methods used to identify ocular irritants:
ICCVAM recommended that the CM test method be used as a screening tool to identify:
ICCVAM concluded that the performance of the other four test methods must be improved before they can be used in regulatory safety testing to identify substances that may cause nonsevere eye injuries or that do not require eye irritation hazard labeling. The 2010 report on the ICCVAM evaluation of these test methods includes recommendations for future studies that could potentially improve these methods.
ICCVAM previously recommended the use of the BCOP and ICE test methods to identify substances that may cause severe or permanent eye injuries (see “In Vitro Test Methods for Detecting Ocular Corrosives and Severe Irritants”, below).
In 2008, NICEATM received a U.S. Environmental Protection Agency (EPA)-sponsored submission for an in vitro testing strategy to assess the eye irritation potential of antimicrobial cleaning products. This proposed testing strategy used the BCOP, CM, and EpiOcular test methods to determine the EPA hazard category and labeling requirements for this class of products. ICCVAM evaluated the proposed strategy and recommended further studies to characterize its usefulness and limitations. As part of this evaluation, ICCVAM recommended that the low volume eye test, a test method used to generate data supporting the proposed testing strategy validity, should not be used for future regulatory testing.
In 2006, ICCVAM forwarded its first recommendations for the use of in vitro test methods for eye injury hazard classification testing to Federal agencies.
ICCVAM recommended including the BCOP and ICE test methods in a weight-of-evidence approach to identify products with the potential to cause permanent or severe eye injuries, noting specific limitations for certain chemical classes and/or physical properties. Substances that test positive in these assays can be classified as ocular corrosives or severe irritants without further testing in animals. Furthermore, ICCVAM recommended that these in vitro test methods be considered for eye injury hazard classification testing before using animals, and used whenever appropriate.
Subsequent to acceptance of the ICCVAM recommendations by U.S. Federal agencies, the OECD formally adopted guidelines describing the use of the BCOP and ICE test methods. These guidelines, designated Test Guidelines 437 and 438, respectively, were based on the ICCVAM recommendations. Acceptance of Test Guidelines 437 and 438 permits the international use of these test methods to identify substances that may cause permanent or severe eye injuries without the need for animal testing.
NICEATM and ICCVAM worked with international partners to organize two 2005 scientific symposia on eye safety testing.
Symposium I (Mechanisms of Chemically Induced Ocular Injury and Recovery) identified research needed to address current knowledge gaps and to advance the development and validation of test methods for ocular toxicity testing that protect human health and replace, reduce, or refine animal use.
The participants in Symposium I recommended that:
Symposium II (Minimizing Pain and Distress in Ocular Toxicity Testing) reviewed the current understanding of the sources and mechanisms of pain and distress in ocular toxicity testing. Workshop presentations identified current best practices for recognizing, preventing, and alleviating ocular pain and distress and identified additional research, development, and validation studies necessary to support scientifically valid ocular toxicity testing procedures that avoid pain and distress.
The participants in Symposium II recommended that:
Both symposia were sponsored by ICCVAM, NICEATM, the European Union Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM), and the European Cosmetic, Toiletry and Perfumery Association. The symposia were organized by ICCVAM, NICEATM, and EURL ECVAM.