Deshpande, P., Ortega, Í., Sefat, F. et al. (4 more authors) (2015) Rocking media over ex vivo corneas improves this model and allows the study of the effect of proinflammatory cytokines on wound healing. Investigative Ophthalmology & Visual Science, 56 (3). 1553 - 1561.
Abstract
PURPOSE: The aim of this work was to develop an in vitro cornea model to study the effect of proinflammatory cytokines on wound healing. METHODS: Initial studies investigated how to maintain the ex vivo models for up to 4 weeks without loss of epithelium. To study the effect of cytokines, corneas were cultured with the interleukins IL-17A, IL-22, or a combination of IL-17A and IL-22, or lipopolysaccharide (LPS). The effect of IL-17A on wound healing was then examined. RESULTS: With static culture conditions, organ cultures deteriorated within 2 weeks. With gentle rocking of media over the corneas and carbon dioxide perfusion, the ex vivo models survived for up to 4 weeks without loss of epithelium. The cytokine that caused the most damage to the cornea was IL-17A. Under static conditions, wound healing of the central corneal epithelium occurred within 9 days, but only a single-layered epithelium formed whether the cornea was exposed to IL-17A or not. With rocking of media gently over the corneas, a multilayered epithelium was achieved 9 days after wounding. In the presence of IL-17A, however, there was no wound healing evident. Characterization of the cells showed that wherever epithelium was present, both differentiated cells and highly proliferative cells were present. CONCLUSIONS: We propose that introducing rocking to extend the effective working life of this model and the introduction of IL-17A to this model to induce aspects of inflammation extend its usefulness to study the effects of agents that influence corneal regeneration under normal and inflamed conditions.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2015 The Association for Research in Vision and Ophthalmology, Inc. This is an author produced version of a paper subsequently published in Investigative Ophthalmology & Visual Science. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | cornea organ culture; inflammation; wound healing |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Engineering (Sheffield) > Department of Materials Science and Engineering (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 09 Jun 2015 08:14 |
Last Modified: | 16 Nov 2016 09:22 |
Published Version: | http://dx.doi.org/10.1167/iovs.14-15308 |
Status: | Published |
Publisher: | Association for Research in Vision and Opthalmology |
Refereed: | Yes |
Identification Number: | 10.1167/iovs.14-15308 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:85908 |