Year: 2023, EEBA

Authors: Rodella U.; Bosio L.; Ferrari S.; Gatto C.; Giurgola L.; Rossi O.; Ciciliot S.; Ragazzi E.; Ponzin D.; D’Amato Tóthová J.

Considering the growing shortage of corneal tissues for research, the present study aimed to develop and optimize a porcine cornea model with qualitative features comparable to those of human tissues.

A new decontamination procedure of porcine eye bulbs was set up and its efficacy as well as endothelial mortality were evaluated. Human corneas unsuitable for transplant and porcine corneas were then compared after storage under hypothermic (4–8°C, Eusol-C, AL.CHI.MI.A. S.R.L) or organ-culture (31–35°C, Tissue-C, AL.CHI.MI.A. S.R.L) storage conditions for 14 days. A new method, based on the semi-automatic analysis of Trypan-blue stained endothelial areas by Fiji software, was developed to quantify the whole endothelium viability.
Corneas were assessed for central corneal thickness (CCT), corneal transparency, endothelial morphology, and endothelial cell density (ECD) at days 0, 7, and 14 of storage. Portions of lamellar tissues consisting of Descemet’s membrane and endothelial cells were prepared for histological investigations.

The new decontamination procedure of porcine eye bulbs resulted in 18% versus 89% (“no decontamination” control) of corneas still contaminated after 28 days of storage at 31°C. The decontamination protocol did not affect endothelium viability, as assessed by the new Fijibased method. ECD (porcine: 3156 ± 144 cells/mm2; human: 2287 ± 152 cells/mm2), CCT (porcine: 1073 ± 151 μm; human: 581 ± 39 μm), transparency (porcine: 88.6 ± 11.0 %; human: 76.3 ± 5.4 %), and morphology score (porcine: 4.0 ± 0.0; human: 3.2 ± 0.4) measured in the porcine cornea at day 0 were significantly higher than in human corneas. Nonetheless, the qualitative parameters of porcine and human corneas showed comparable trends during the storage under hypothermic (4–8°C) and organ-culture (31–35°C) conditions for 14 days.

The presented porcine cornea model represents a reliable and alternative model to human donor tissues for preliminary investigations and can be used for testing new media, substances, drugs, or preservation conditions and their impact on corneal tissue quality and safety. Furthermore, the quantitative method to assess whole endothelium mortality can be implemented at eye banks for the evaluation of corneas intended for transplantation.