Changes in rabbit corneal epithelial membrane permeability caused by locally applied Pseudomonas aeruginosa cytotoxin: a microfluorometric examination in vivo.

The effects of a pore-forming protein from Pseudomonas aeruginosa on the rabbit cornea were tested in vivo by measuring intraepithelial carboxyfluorescein accumulation. Carboxyfluorescein diacetate and subsequently the P. aeruginosa cytotoxin were applied by means of contact lenses with a spherical cavity on the concave surface. This allowed the application of defined concentrations of carboxyfluorescein diacetate and cytotoxin on a defined area of the corneal epithelium. Starting at 0.5 microM, cytotoxin increased the epithelial cell membrane permeability for the intracellular carboxyfluorescein within 1 min. At higher concentrations cells were shed from the epithelium. Corresponding morphological changes of the cellular structure of the corneal epithelium were observed and documented by fluorescence photomicrography. The healing process of toxified corneal epithelium appeared to be complete within 3 days. The data presented here indicate the possible role of cytotoxin-induced changes in epithelial permeability in P. aeruginosa infections. In this context, the role of soft contact lenses as a possible cytotoxin reservoir is discussed.