Okurowska, K., Roy, S., Thokala, P. et al. (5 more authors) (2020) Establishing a porcine ex vivo cornea model for studying drug treatments against bacterial keratitis. Journal of Visualized Experiments.
Abstract
When developing novel antimicrobials, the success of animal trials is dependent on accurate extrapolation of antimicrobial efficacy from in vitro tests to animal infections in vivo. The existing in vitro tests typically overestimate antimicrobial efficacy as the presence of host tissue as a diffusion barrier is not accounted for. To overcome this bottleneck, we have developed an ex vivo porcine corneal model of bacterial keratitis using Pseudomonas aeruginosa as a prototypic organism. This article describes the preparation of the porcine cornea and protocol for establishment of the infection. Bespoke glass molds enable straightforward setup of the cornea for infection studies. The model mimics in vivo infection as bacterial proliferation is dependent on the ability of the bacterium to damage corneal tissue. Establishment of infection is verified as an increase in the number of colony forming units assessed via viable plate counts. The results demonstrate that infection can be established in a highly reproducible fashion in the ex vivo corneas using the method described here. The model can be extended in the future to mimic keratitis caused by microorganisms other than P. aeruginosa. The ultimate aim of the model is to investigate the effect of antimicrobial chemotherapy on the progress of bacterial infection in a scenario more representative of in vivo infections. In so doing, the model described here will reduce the use of animals for testing, improve success rates in clinical trials and ultimately enable rapid translation of novel antimicrobials to the clinic.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2020 Journal of Visualized Experiments (JoVE). This is an author-produced version of a paper subsequently published in Journal of Visualized Experiments. Available under the terms of the Creative Commons Attribution-NonCommercial Licence (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. You may not use the material for commercial purposes. |
Keywords: | Pseudomonas aeruginosa; keratitis; cornea; ex vivo; eye; porcine |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Engineering (Sheffield) > Department of Chemical and Biological Engineering (Sheffield) |
Funding Information: | Funder Grant number INNOVATE UK (TSB) TS/P011616/1 Medical Research Council MR/S004688/1 |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 15 May 2020 09:51 |
Last Modified: | 12 Nov 2020 01:39 |
Published Version: | https://www.jove.com/video/61156/establishing-porc... |
Status: | Published |
Publisher: | Journal of Visualized Experiments |
Refereed: | Yes |
Identification Number: | 10.3791/61156 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:160753 |