| In Vivo Toxicology Assays |
Acute Eye Irritation
- EPA-OPPTS, OECD
- MB Protocol - 1200-02, 2200-02
- Purpose: Assessment and evaluation of the toxic characteristics of a substance, determination of the irritant and/or corrosive effects on the eyes. Information derived from this test serves to indicate the existence of possible hazards likely to arise from exposure of the eyes and associated mucous membranes to the test substance.
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Primary Eye Irritation
- OECD
- MB Protocol - 3200
- Purpose: Assessment and evaluation of the toxic characteristics of a substance, determination of the irritant and/or corrosive effects on the eyes. Information derived from this test serves to indicate the existence of possible hazards likely to arise from exposure of the eyes and associated mucous membranes to the test substance.
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In Vitro and Alternative Assays |
1) Three dimensional Human Ocular Tissue Equivalent Systems
- MatTek EpiOcularTM, SkinEthics 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
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2) Chorioallantoic Membrane Vascular Assay (CAMVA)
- Draize Rabbit Eye Test Alternative
- 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
- Can be used in combination with BCOP
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3) Hen's Egg Test-Chorioallantoic Membrane (HET-CAM)
- Draize Rabbit Eye Test Alternative
- 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
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4) Bovine Corneal Opacity and Permeability Assay (BCOP)
- Draize Rabbit Eye Test Alternative
- 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)
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5) 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
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6) 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
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