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. 2001 Apr;85(4):450–453. doi: 10.1136/bjo.85.4.450

A simple corneal perfusion chamber for drug penetration and toxicity studies

M Thiel 1, N Morlet 1, D Schulz 1, H Edelhauser 1, J Dart 1, D Coster 1, K Williams 1
PMCID: PMC1723919  PMID: 11264136

Abstract

AIMS—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.
CONCLUSION—The new chamber device forms a reliable tool for in vitro drug penetration and toxicity studies in isolated perfused corneoscleral tissue.



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Figure 1  .

Figure 1  

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Corneal perfusion chamber shown without a mounted cornea. The clamping sleeve sits on top of the main body. The fixation ring that screws onto the main body is shown on the left.

Figure 2  .

Figure 2  

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Chamber design: (A) main body, (B) clamping sleeve, and (C) fixation ring. All parts were made of polycarbonate and stainless steel. Corneas are clamped between the main body and the clamping sleeve.

Figure 3  .

Figure 3  

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Corneal thickness (mean (SD)) as measured by pachymetry plotted against perfusion time for isolated perfused pig corneas mounted in the modified perfusion chamber. Toxic destruction of the endothelium by a brief perfusion of ethanol resulted in irreversible tissue swelling.

Figure 4  .

Figure 4  

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Specular microscopy (A, B) and histology (C) of a perfused pig cornea. (A) Endothelium before tissue dissection, (B) endothelium after 4 hours of perfusion (magnification ×40), (C) epithelium after perfusion for 10 hours, topical BSS eye drops every 20 minutes (periodic acid Schiff stain, magnification ×100).

Figure 5  .

Figure 5  

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Fluorescein penetration (mean (SD) of replicate determinations) through pig corneas with or without epithelium. No correction was applied for the dilution of fluorescein that occurred when a sample was removed for testing and was replaced by fresh BSS-Plus. The epithelial barrier was well preserved over the perfusion period of 10 hours.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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