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Francesco Menduni, Tugce Ipek, Leon N Davies, Antonio Fratini, James Stuart Wolffsohn; Development of an ex-vivo animal model to evaluate novel pharmaceutical approaches to Dry Eye Disease management. Invest. Ophthalmol. Vis. Sci. 2018;59(9):3303. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Previous research has established the viability of maintaining, independently, either the corneal or crystalline lens tissue physiologically stable for a period of ~10 days. This study aimed to design and evaluate a novel complete anterior eye model that combines these two structures, allowing anterior chamber evaluation.
Four freshly enucleated porcine eyes were obtained from the local abattoir and transported to the laboratory in a de-swelling solution. Whole eyes were dipped in a 0.75% povidone iodine solution for 3 minutes to safely decontaminate the tissue, sectioned at the equator using a vibrating blade, and mounted in sealed perfusion chambers specially designed in SOLIDWORKS 2015 and manufactured in PTFE. All artificial anterior chambers were perfused with sanitised supplemented cell culture media at a flow rate of 15 ml/min and at a temperature of 37.5°C. Ocular surface irrigation was achieved via a custom-made automated spray system misting supplemented cell culture media over the ocular tissue every 20s (Figure 1). Corneal and conjunctival damage were assessed with sodium fluorescein and lissamine green on a tissue level. Trypan blue exclusion technique and Live/Dead staining were used to evaluate the model on a cellular level. Data were quantified using ImageJ software.
The model maintained the porcine anterior segment in good condition for the 10-day period during which the system was evaluated. No significant (p>0.05) increase in corneal or conjunctival fluorescein and lissamine green staining was observed, and both conjunctival cell viability and the number of trypan blue-stained corneal cells were not found to be significantly different during the whole evaluation period (p>0.05).
This ex-vivo anterior eye model may bridge the gap between in-vitro and in vivo investigation. Due to the possibility of varying parameters such as osmolarity and irrigation rate, this model could represent a reliable tool for the evaluation of novel anterior eye targeted pharmaceutical approaches. Moreover, this model may also play a key role in medical device research as the crystalline lens is preserved in loco.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
Ex-vivo eye model
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