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Veterinary ophthalmology2021; 24(5); 509-519; doi: 10.1111/vop.12935

Interleukin-6 and lactate dehydrogenase expression in a novel ex vivo rocking model of equine corneal epithelial wound healing.

Abstract: To establish a physiologically relevant ex vivo model of equine corneal epithelial wound healing. Methods: Fourteen equine corneas were randomly assigned to one of two groups: wounded (n = 8) or unwounded (n = 6) controls. In the wounded group, the axial corneal epithelium was removed by applying a 6 mm filter paper disk soaked in 1N-NaOH for 60 s. Corneas were subsequently cultured using an air-liquid interface model. Evaluation of corneal healing was performed daily, and culture medium was collected. Corneas were randomly assigned to undergo processing via histopathology and RNAscope in situ hybridization for interleukin-6 (IL-6) and alpha-smooth muscle actin (αSMA) expression at T24, T48, and T72 h after wounding. Media of the cultured corneas were evaluated for the presence of lactate dehydrogenase (LDH) by a colorimetric assay. Results: The ulcerated area of the wounded corneas decreased over time and all corneas healed within 72 h. Histologically, normal corneal architecture was observed including healthy epithelium (in areas other than the ulcerated ones), minimal stromal edema, intact endothelium, and Descemet's membrane. IL-6 expression was increased in wounded corneas compared with unwounded controls. LDH expression was elevated for both wounded and unwounded corneas at T24 but decreased substantially and was not detected at T48 in media from wounded and unwounded corneas, respectively. No αSMA expression was detected from either wounded or unwounded corneas. Conclusions: The equine air-liquid interface, ex vivo, corneal epithelial wound healing model is effective and physiologically relevant. This model can be used in future studies evaluating various corneal therapies.
© 2021 The Authors. Veterinary Ophthalmology published by Wiley Periodicals LLC on behalf of American College of Veterinary Ophthalmologists.
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Publication Date: 2021-09-23 PubMed ID: 34553825DOI: 10.1111/vop.12935Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research is about the establishment of a novel ex vivo model for studying the process of corneal epithelial wound healing in horses, based on the analysis of interleukin-6 (IL-6) and lactate dehydrogenase (LDH) expression.

Method

Fourteen horse corneas were randomly divided into two groups, the wounded group consisting of eight corneas and the unwounded group which had six. A wound was created on the corneas in the wounded group by using a filter paper soaked in NaOH. Then, the wound healing process was studied under controlled conditions, by cultivating the corneas in an air-liquid interface model. Throughout the process, the corneal healing was evaluated at 24, 48, and 72 hours after wounding, also the culture medium was checked. The tissues were tested for the expression of IL-6 and alpha-smooth muscle actin (αSMA), with the help of RNA scope in situ hybridization. The presence of LDH in the medium collected from wounded and unwounded corneas was assessed using a colorimetric assay.

  • The wounded corneas showed a decrease in the ulcerated area over time, with complete healing observed within 72 hours.
  • Wounded samples revealed a significant rise in expression of IL-6, a signal of inflammation, in comparison to the unwounded controls.
  • No αSMA expression, a marker for wound healing and inflammatory response, was detected in either group.
  • Though the initial 24 hours showed elevated LDH levels in both wounded and unwounded samples, the levels decreased notably at 48 hours, and were undetectable beyond this time frame.

Conclusion

The research concludes that the designed air-liquid interface model for studying corneal epithelial wound healing in horses is effective and relevant in a physiological context. The increased IL-6 expression indicates inflammation due to wounding, and the accompanied decrease in LDH levels suggests the healing process. The absence of αSMA implies that the wound healing does not involve fibrosis. This model is suggested for future studies for examining different corneal therapies.

Cite This Article

APA
Wehrman RF, Genschel U, Charli A, Kanthasamy AG, Allbaugh RA, Ben-Shlomo G. (2021). Interleukin-6 and lactate dehydrogenase expression in a novel ex vivo rocking model of equine corneal epithelial wound healing. Vet Ophthalmol, 24(5), 509-519. https://doi.org/10.1111/vop.12935

Publication

Veterinary ophthalmology
ISSN: 1463-5224
NlmUniqueID: 100887377
Country: England
Language: English
Volume: 24
Issue: 5
Pages: 509-519

Researcher Affiliations

Wehrman, Rita Fay
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.
Genschel, Ulrike
  • Department of Statistics, College of Liberal Arts and Sciences, Iowa State University, Ames, Iowa, USA.
Charli, Adhithiya
  • Department of Biomedical Sciences, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.
Kanthasamy, Anumantha G
  • Department of Biomedical Sciences, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.
Allbaugh, Rachel Anne
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.
Ben-Shlomo, Gil
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.
  • Department of Biomedical Sciences, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.
  • Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.

MeSH Terms

  • Animals
  • Colorimetry / veterinary
  • Corneal Injuries / metabolism
  • Corneal Injuries / pathology
  • Corneal Injuries / veterinary
  • Disease Models, Animal
  • Female
  • Horses / injuries
  • Interleukin-6 / metabolism
  • L-Lactate Dehydrogenase / metabolism
  • Male
  • Primary Cell Culture / veterinary
  • Wound Healing

Grant Funding

  • American Quarter Horse Foundation
  • Iowa State University

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Citations

This article has been cited 2 times.
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  2. Hamza KH, El-Shanshory AA, Agwa MM, Abo-Alkasem MI, El-Fakharany EM, Abdelsattar AS, El-Bardan AA, Kassem TS, Mo X, Soliman HMA. Topically Applied Biopolymer-Based Tri-Layered Hierarchically Structured Nanofibrous Scaffold with a Self-Pumping Effect for Accelerated Full-Thickness Wound Healing in a Rat Model. Pharmaceutics 2023 May 17;15(5).
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