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Khoa D. Tran, Zhexun Jason Sun, David Putnam, Christopher Sales; Ex-vivo Testing Apparatus and Method to Examine the Effects of Materials on Human Corneal Endothelial Cell Loss.. Invest. Ophthalmol. Vis. Sci. 2020;61(7):1463.
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© ARVO (1962-2015); The Authors (2016-present)
To establish and test a novel assay that can be used to examine the effects of materials on the corneal endothelium.
Donor corneas with healthy endothelium were placed endothelium side up on a punch block. Punches (3 mm -9 mm) of materials such as rubber from a nitrile glove, viscoelastic, and polysaccharide hydrogels were placed directly onto the corneal endothelium. Subsequently, the cornea and attached materials were mounted on an artificial anterior chamber (AAC) and filled with air to bring the pressure to approximately 30 mmHg. Contact between the test articles and endothelium were examined by optical coherence tomography (OCT). After 15 minutes, the cornea was carefully dismounted from the AAC and the materials were removed. Each experiment was reproduced three times. The cornea was stained for 1 minute with Trypan blue, imaged, and returned to cold storage in Optisol-GS for 24 hours. The cornea was subsequently stained with Calcein-AM and imaged using a fluorescence microscope.
The nitrile glove material caused endothelial cell loss when in contact with the endothelium. Viscoelastic gels can be used to prevent this damage and acts as a negative control. Preliminary testing using different materials at different sizes revealed endothelial cell loss where there is contact between the test articles and the endothelium.
This simple and inexpensive assay can be used to examine the effect of materials on the corneal endothelium. It is sensitive enough to provide preliminary results for human endothelial cell loss due to contact with foreign materials.
This is a 2020 ARVO Annual Meeting abstract.
Figure 1: Description of test apparatus and method to analyse endothelial cell loss caused by different materials.
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