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

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Wehrman, Rita Fay
Genschel, Ulrike
Charli, Adhithiya
Kanthasamy, Anumantha G.
Ben-Shlomo, Gil
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Purpose 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.
This is the published version of the following article: Wehrman, Rita Fay, Ulrike Genschel, Adhithiya Charli, Anumantha G. Kanthasamy, Rachel Anne Allbaugh, and Gil Ben‐Shlomo. "Interleukin‐6 and lactate dehydrogenase expression in a novel ex vivo rocking model of equine corneal epithelial wound healing." Veterinary Ophthalmology 24, no. 5 (2021): 509-519. DOI: 10.1111/vop.12935. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivsLicense, which permits use and distribution in anymedium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.© 2021 The Authors. Posted with permission.