A redesigned corneal holder for the bovine cornea opacity and permeability assay that maintains normal corneal morphology

• Published in: Toxicology in vitro 2002-10, Vol.16 (5), p.621-628
• Main Authors: Ubels, J.L., Paauw, J.D., Casterton, P.L., Kool, D.J.
• Format: Article
• Language: English
• Subjects: Alternative methods; Animal Testing Alternatives - instrumentation; Animals; Biological and medical sciences; Bovine cornea opacity and permeability assay; Cattle; Cell Membrane Permeability; Chemical and industrial products toxicology. Toxic occupational diseases; Cornea; Cornea - cytology; Cornea - pathology; Corneal endothelium; Corneal epithelium; Corneal Opacity - etiology; Corneal Opacity - pathology; Epithelium, Corneal - cytology; Epithelium, Corneal - drug effects; Epithelium, Corneal - pathology; Equipment and Supplies; Irritants - toxicity; Medical sciences; Reproducibility of Results; Time Factors; Toxicology; Various organic compounds
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/S0887-2333(02)00046-2

The bovine cornea opacity and permeability assay (BCOP) has been in use for nearly 10 years but has not been submitted for regulatory approval. In previous reports we have presented corneal hydration and endothelial damage as additional endpoints in this assay and have suggested that the design of the BCOP's corneal holder should be modified. The standard holder used in the BCOP assay induces physical damage to the cornea because it contacts clear cornea causing edge damage to the epithelial, stromal and endothelial layers. Second, by forcing a curved, oval-shaped bovine cornea into a flat, circular opening, corneal wrinkling occurs which can alter the cornea's optical characteristics and, most importantly, induces endothelial damage. We now report on a redesigned BCOP corneal holder that clamps onto the sclera, maintains normal corneal shape and does not cause damage to the endothelium. This ensures that irritancy tests are conducted using healthy, anatomically normal tissue. Tests of this holder using acetone, trichloroacetic acid, isopropanol and benzalkonium chloride show that it is now possible to evaluate effects of chemical substances on the endothelium. The effects of these compounds on corneal opacity and hydration in the new holder are similar to their effects on the cornea in the standard holder.