Ocular Irritation Assays

In Vitro & Alternative Test Methods

What is the Draize Rabbit Eye Test?

The Draize Rabbit Eye Test is an acute toxicity assay developed in 1944 by Food and Drug Administration (FDA) and has been the basis of ocular irritation screening used by US and other regulatory agencies around the world. This test involves topically dosing a test material (liquid/solids) in to the eye and assessing the effects of the cornea, iris and conjunctiva.

A current challenge facing regulators is to eliminate the use of live animals in eye irritation testing, which will allow categorization of irritation from non-irritating to corrosive. Efforts to meet this challenge have been met with very limited success because, in vitro and alternative tests must satisfy the regulatory community (i.e., FDA , EPA, OECD, HMIS, etc.) with a level of conservatism as the current Draize Rabbit Eye test.

MB Research has been using and developed in vitro and alternative ocular irritation testing since the late 1980s. We offer a wide array of non-animal test methods to aid in the classification of consumer products, chemicals, pharmaceuticals, and cosmetics.

For more information about ocular irritation testing, please feel free to contact us.

In Vitro and Alternative Test Methods

Ocular Irritation Assays


OptiSafe™ - Eye Irritation Test

  • OptiSafe™ In Vitro Eye Irritation Test
  • An in vitro test method in which a test substance is applied to a semi-permeable membrane. Damage to macromolecules in the membrane is measured to assess the test substance’s potential to cause eye irritation.
  • Can be used to determine the irritation potential of cosmetics, creams, and a wide variety of consumer products. Results are presented as GHS, EPA classifications, an ocular irritation score and class

Porcine Cornea Reversibility Assay (PorCORA)

  • Uses excised porcine cornea normally discarded
  • Corneas maintained in culture for up to 21 Days
  • Measures "Days to Recover" after exposure to possible Irritants
  • Allows discrimination between Severe Irritant and Ocular Corrosive

Chorioallantoic Membrane Vascular Assay (CAMVA)

  • Uses fertilized chicken eggs on embryonic day 10 or 14 of development
  • Measures hemodynamic effects, injury, and anti-angiogenic effects to the membrane microvasculature
  • Especially useful for alcohol-containing formulations

HET-CAM - Hen's Egg Test - Chorioallantoic Membrane (HET-CAM)

  • Uses fertilized chicken eggs
  • INVITTOX PROTOCOL Number 47

Bovine Corneal Opacity and Permeability Assay (BCOP)

  • Uses excised bovine corneas normally discarded
  • Measures two endpoints; 1) changes in opacity, reflecting protein denaturation and corneal injury, and 2) fluorescein permeability reflecting damage to corneal epithelium
  • Optional histological endpoint is available (H & E staining)
  • Measurement of pro-inflammatory mediators in liberated corneal cells (by flow cytometry or ELISA)

Replacement Ocular Battery (ROBatt)

  • Uses tiered testing strategy of alternative ocular irritation assays (CAMVA, BCOP, PorCORA, PorFocal) to determine regulatory classification of a test material without the use of live animals.
  • Currently Under Development

3D Human Ocular Tissue Equivalent Systems

MatTek EpiOcular™, SkinEthic corneocyte models

  • These models mimic characteristics of the epithelium of the eye
  • Unique properties of 3D models allow test articles to be applied topically, allowing testing of solids, organics, and insoluble materials

Keratinocyte/Fibroblast Viability Assay (Neutral Red or MTT)


MB Research uses normal human primary epidermal keratinocytes or fibroblasts (or mouse keratinocytes and fibroblasts)
  • Cells are representative of the eye and skin epithelium
  • Human cells are derived from neonatal, adolescent, or aged donors
  • Cytokine expression or release (LDH, PGE-2, interleukins) can be measured
  • Low serum or serum free protocol options are available

Keratinocyte Proliferation (turnover) Assay

  • Using primary human cell or mouse cell lines in serum-free or low serum media
  • Rate of proliferation or apoptosis is measured by vital dye uptake (MTT or neutral red) or by BrdU incorporation and flow cytometry