A simple corneal perfusion chamber for drug penetration and toxicity studies.
OBJECTIVE
Corneal perfusion chambers are important tools in the development and assessment of ophthalmic drugs. The aim of this study was to design and test a modified perfusion chamber suitable for topical application of drugs to isolated corneoscleral preparations, and which allowed continuous monitoring of endothelial cell function.


METHODS
A polycarbonate and stainless steel perfusion chamber was designed to clamp corneas in a horizontal plane suitable for topical drug delivery. Endothelial cell function was assessed by ultrasonic pachymetry and specular microscopy during perfusion. Epithelial barrier function was assessed by penetration of fluorescein. Leakage was examined by measuring penetration of a large protein, IgG. Tissue architecture after perfusion was examined by conventional histology.


RESULTS
Corneas maintained a functionally and morphologically intact endothelial monolayer during perfusion periods of up to 14 hours. The epithelial barrier function was well preserved. The tissue clamp sealed the preparation effectively against leakage of macromolecules.


CONCLUSIONS
The new chamber device forms a reliable tool for in vitro drug penetration and toxicity studies in isolated perfused corneoscleral tissue.
Department of Ophthalmology, Flinders University of South Australia, Adelaide, Australia.
M A
MA
Thiel
N
N
Morlet
D
D
Schulz
H F
HF
Edelhauser
J K
JK
Dart
D J
DJ
Coster
K A
KA
Williams
D000818:Animals; D001639:Bicarbonates; D015194:Diffusion Chambers, Culture; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation*; D004728:Endothelium, Corneal / Q000187:drug effects / Q000502:physiology; D004867:Equipment Design; D019793:Fluorescein / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D009883:Ophthalmic Solutions / Q000493:pharmacokinetics*; D010477:Perfusion / Q000295:instrumentation; D013193:Stainless Steel; D013552:Swine
D001639:Bicarbonates; D007074:Immunoglobulin G; D009883:Ophthalmic Solutions; D013193:Stainless Steel; D019793:Fluorescein