Toxicity evaluation of antiglaucoma drugs using stratified human cultivated corneal epithelial sheets

Invest Ophthalmol Vis Sci. 2012 Aug 3;53(9):5154-60. doi: 10.1167/iovs.12-9685.

Abstract

Purpose: To investigate the toxicity profiles of seven antiglaucoma topical eye drops and benzalkonium chloride (BAC) using stratified cultivated human corneal epithelial cell sheets (HCES) in a serum-free culture system.

Methods: A range of prostaglandin analogies and preservatives, including BAC, sofZia (SZ), sodium benzoate (SB), and polyquaternium-1 (PQ) were tested. The barrier function and cell viability were examined by a carboxyfluorescein permeability assay and WST-1 assay. Histological evaluation of the HCES was also performed after application of each solution.

Results: The carboxyfluorescein permeability assay had a higher sensitivity for the detection of toxicity of test solutions than the WST-1 assay or histological examination. Latanoprost BAC, latanoprost/timolol BAC, and 0.02% or higher concentration of BAC were the most toxic, followed by latanoprost SB, latanoprost preservative-free, BAC 0.002%, and travoprost/ latanoprost PQ. Travoprost SZ and tafluprost BAC (preserved with 0.001% BAC) was the least toxic in our experimental conditions.

Conclusions: The carboxyfluorescein permeability assay using HCES in a serum-free system was the most useful for the quantification of toxicity of ophthalmic solutions. Among the regimens examined, a BAC concentration of 0.001% or lower or non-BAC preservative sofZia was suggested to be the least toxic to the ocular surface.

MeSH terms

  • Antihypertensive Agents / toxicity*
  • Benzalkonium Compounds / toxicity
  • Cell Membrane Permeability
  • Cell Survival
  • Cells, Cultured
  • Culture Media, Serum-Free
  • Drug Combinations
  • Epithelium, Corneal / drug effects*
  • Fluoresceins / metabolism
  • Fluorescent Dyes / metabolism
  • Glaucoma / drug therapy
  • Humans
  • Membrane Proteins / metabolism
  • Ophthalmic Solutions / toxicity
  • Phosphoproteins / metabolism
  • Preservatives, Pharmaceutical / toxicity
  • Prostaglandins F, Synthetic / toxicity
  • Timolol / toxicity
  • Zonula Occludens-1 Protein

Substances

  • Antihypertensive Agents
  • Benzalkonium Compounds
  • Culture Media, Serum-Free
  • Drug Combinations
  • Fluoresceins
  • Fluorescent Dyes
  • Membrane Proteins
  • Ophthalmic Solutions
  • Phosphoproteins
  • Preservatives, Pharmaceutical
  • Prostaglandins F, Synthetic
  • TJP1 protein, human
  • Zonula Occludens-1 Protein
  • 6-carboxyfluorescein
  • Timolol