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BMJ Open Ophthalmol
BMJ Open Ophthalmol
bmjophth
bmjophth
BMJ Open Ophthalmology
2397-3269
BMJ Group BMA House, Tavistock Square, London, WC1H 9JR

37282691
bmjophth-2022-EEBA.18
10.1136/bmjophth-2022-EEBA.18
Oral abstracts
Theme 3 – Corneal storage and microbiological safety measures before/after transplant
1506
18 A porcine cornea and lamellar tissue model to investigate effects of storage conditions on corneal preservation
Rodella Umberto 1
Bosio Lorenzo 2
Giurgola Laura 1
Gatto Claudio 1
Rossi Orietta 1
Ferrari Stefano 2
Tothova Jana D’amato 1
1 Al.chi.mi.a. Srl, Ponte San Nicolò, Italy
2 Fondazione Banca degli Occhi del Veneto Onlus, Venezia, Italy
2022
16 11 2022
7 Suppl 2 Abstracts of the European Eye Bank Association Virtual Meeting, 3–5 March 2022 A8A8
© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.
2022
https://creativecommons.org/licenses/by-nc/4.0/ This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/ .

Background

Globally, more than 12 million people are awaiting corneal transplantation and cornea donor reduction has been observed since the outbreak of the COVID-19 pandemic, negatively influencing the availability of human corneas for research purposes as well. Therefore, the exploitation of ex vivo animal models in this field is of great value.

The present study aimed at the development of a novel experimental model of porcine cornea ex vivo and lamellar tissue preparation to investigate the effects of storage conditions on corneal preservation.

Methods

Twelve fresh porcine eye bulbs were disinfected by immersion in 10 mL of 5% povidone-iodine under orbital mixing for 5 minutes at room temperature. The corneoscleral rims were dissected, and stored in Tissue-C (Alchimia S.r.l., n=6) at 31°C and in Eusol-C (Alchimia S.r.l., n=6) at 4°C up to 14 days.

The evaluation of Endothelial Cell Density (ECD) and endothelial mortality was performed using vital dye Trypan Blue staining (TB-S, Alchimia S.r.l.). Digital 1X pictures of TB-stained corneal endothelium were acquired and percentage of stained area was quantified using FIJI ImageJ software. ECD and endothelial mortality were determined at 0, 3, 7 and 14 days.

Medium turbidity detected by naked eye was considered as proof of tissue contamination.

Additionally, non-vital staining of the endothelium with Alizarin Red (AR) was performed and the endothelial morphology was investigated at Day 14 in both whole corneas and dissected endothelial lamellae.

Results

The contamination rate of porcine corneas corresponded to <10% and 0% in Tissue-C and Eusol-C after 14 days, respectively.

Porcine corneas stored in Tissue-C and Eusol-C showed <10% and <20% mortality in Tissue-C and Eusol-C respectively at the end of storage.

Preliminary ECD determination (range 3700-4100 cells/mm2) at Day 0 aligned with data present in the literature (Meltendorf et al., Graefe’s Arch Clin Exp Ophthalmol, 2007).

Whole cornea and dissected lamellae stained with TB and AR showed comparable endothelial morphology after incubation in Tissue-C and Eusol-C for 14 days. The lamellar tissue allowed endothelium morphology analysis at higher magnification compared to whole cornea.

Conclusion

The presented ex vivo porcine model allows evaluation of the performance and safety of storage conditions. Future perspectives of this method will be the extension of the porcine corneas storage up to 28 days.

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