A superfusion apparatus for ex vivo human eye irritation investigations

Toxicol In Vitro. 2015 Oct;29(7):1619-27. doi: 10.1016/j.tiv.2015.06.022. Epub 2015 Jun 20.

Abstract

A superfusion apparatus (SA) was developed to maintain isolated human corneas ex vivo under conditions which mimic the natural eye environment in vivo, including controlled temperature, tear flow and intraocular pressure. The SA was designed, developed and tested for use in ophthalmic pre-clinical research and to test new pharmaceutical formulations. Corneas undergo an equilibration process in the new physiological environment for one day. The test was then initiated by the application of the test substance, incubation, and temporal assessment of corneal damage using various parameters. The effects of mild and severe irritant concentrations of NaOH (2% and 8%, respectively) on corneal opacity, swelling and epithelial integrity were studied, and the inflammatory status assessed using F4/80 and MPO as macrophages and neutrophils markers, respectively. The SA was then used to test new artificial tear formulations supplemented with silver ions as an active constituent, showing different degrees of inflammatory responses as indicated by the migration of MPO and F4/80 positive cells towards the epithelium. The human cornea superfusion apparatus was proposed as a model for acute eye irritation research.

Keywords: AgNO(3); Alternative method; Cornea; Ex vivo; NaOH.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animal Testing Alternatives*
  • Antigens, Differentiation / metabolism
  • Ascorbic Acid / toxicity
  • Caustics / toxicity*
  • Cornea / drug effects*
  • Cornea / pathology
  • Corneal Opacity
  • Humans
  • In Vitro Techniques
  • Irritants / toxicity*
  • Lubricant Eye Drops / toxicity
  • Ophthalmic Solutions
  • Peroxidase / metabolism
  • Silver Nitrate / toxicity
  • Sodium Hydroxide / toxicity*

Substances

  • Antigens, Differentiation
  • Caustics
  • Irritants
  • Lubricant Eye Drops
  • Ophthalmic Solutions
  • monocyte-macrophage differentiation antigen
  • Sodium Hydroxide
  • Silver Nitrate
  • Peroxidase
  • Ascorbic Acid