Development of a novel ex vivo equine corneal model.
Abstract: To develop an ex vivo equine corneal organ culture model. Specifically, to assess the equine cornea's extracellular matrix and cellularity after 7 days using two different culture techniques: either (i) immersion system or (ii) air/liquid interface system, to determine the best ex vivo equine corneal model. Methods: Fourteen healthy equine corneas of various breeds. Methods: Equine corneas with 2 mm of perilimbal sclera were freshly harvested from 7 horses undergoing humane euthanasia. One corneal-scleral ring (CSR) from each horse was randomly placed in the (i) immersion condition organ culture system (IC), with the contralateral CSR being placed in the (ii) air/liquid interface organ culture system (ALC) for 7 days. All corneas were evaluated using serial daily gross photography, histology, qPCR, and TUNEL assay. Results: corneal-scleral rings placed in the IC (i) had complete loss of corneal transparency on gross photography by 7 days, showed a significant level of corneal stromal disorganization, significantly increased α-SMA levels on qPCR, and apoptosis on TUNEL assay compared to controls. The ALC (ii) had weak stromal disorganization on histopathologic examination and was not significantly different from normal equine corneal controls on all other evaluated parameters. Conclusions: The air-liquid interface organ culture system maintains the equine cornea's extracellular matrix and preserves corneal transparency, while the immersion condition results in near complete degradation of normal equine corneal architecture after 7 days in culture. The air-liquid organ culture is a viable option to maintain a healthy equine cornea in an ex vivo setting for wound healing studies.
© 2016 American College of Veterinary Ophthalmologists.
Publication Date: 2016-07-29 PubMed ID: 27471196PubMed Central: PMC5276793DOI: 10.1111/vop.12415Google Scholar: Lookup
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- Journal Article
Summary
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This research focuses on developing an effective method for maintaining horse corneas outside the body for scientific study, specifically comparing two different culture techniques: immersion and air/liquid interface system. The results indicate that the air/liquid interface organ culture system is more effective in preserving the structure and health of the equine cornea than the immersion culture system.
Methods
- The corneas, with 2 mm of surrounding sclera, were harvested from the eyes of 7 healthy equines of varying breeds which were being humanely euthanized for reasons unrelated to this study.
- For each horse, one corneal-scleral ring (a combination of the cornea and some sclera) was placed in an immersion condition organ culture system (IC), while the counterpart from the other eye was placed in an air/liquid interface organ culture system (ALC).
- These were both maintained for a period of 7 days, during which they were routinely monitored using various methods such as daily photography, histology, qPCR, and a TUNEL assay.
Results
- The results showed that after the 7 day period, the corneal-scleral rings in the immersion system had completely lost transparency.
- Histologically, significant levels of stromal disorganization were observed, along with increased α-SMA levels on qPCR, and cell death (apoptosis) presented in the TUNEL assay.
- Contrastingly, the corneas in the air/liquid interface system showed only weak stromal disorganization as observed in the histopathological examination, and on all other fronts, they were not significantly different from the normal equine cornea.
Conclusion
- Based on these findings, the researchers concluded that the air-liquid interface organ culture system was the superior method for maintaining structural integrity and health of the equine cornea in an experimental setting.
- This method offers promise for future studies on corneal disorders and wound healing, as it maintains the cornea’s extracellular matrix and transparency while leaving the underlying corneal structure largely undisturbed.
- Compared to the immersion system, which causes substantial degradation of the normal equine corneal architecture after 7 days in culture, the air-liquid interface system is much more potential for ongoing studies on equine corneal health.
Cite This Article
APA
Marlo TL, Giuliano EA, Sharma A, Mohan RR.
(2016).
Development of a novel ex vivo equine corneal model.
Vet Ophthalmol, 20(4), 288-293.
https://doi.org/10.1111/vop.12415 Publication
Researcher Affiliations
- Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri, Columbia, MO, USA.
- Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri, Columbia, MO, USA.
- Harry S. Truman Veterans Memorial Hospital, Columbia, MO, USA.
- Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri, Columbia, MO, USA.
- Harry S. Truman Veterans Memorial Hospital, Columbia, MO, USA.
- Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri, Columbia, MO, USA.
- Harry S. Truman Veterans Memorial Hospital, Columbia, MO, USA.
- Mason Eye Institute, University of Missouri, 1 Hospital Drive, Columbia, MO, 65212, USA.
MeSH Terms
- Animals
- Cornea / anatomy & histology
- Cornea / cytology
- Corneal Stroma
- Extracellular Matrix
- Horses
- Organ Culture Techniques / methods
- Organ Culture Techniques / veterinary
- Random Allocation
- Time Factors
Grant Funding
- I01 BX000357 / BLRD VA
- R01 EY017294 / NEI NIH HHS
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Citations
This article has been cited 8 times.- Gorbatyuk M, Sinha NR, Kumar R, Zhylkibayev A, Athar M, McNutt P, Mohan RR. Current progress in research on ocular injury caused by exposure to vesicants. Prog Retin Eye Res 2025 Nov;109:101413.
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