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    Rocking Media Over Ex Vivo Corneas Improves This Model and Allows the Study of the Effect of Proinflammatory Cytokines on Wound Healing

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    Publication date
    2015
    Author
    Deshpande, P.
    Ortega, Í.
    Sefat, Farshid
    Sangwan, V.S.
    Green, N.H.
    Claeyssens, F.
    MacNeil, S.
    Keyword
    Cornea organ culture; Inflammation; Wound healing; Proinflammatory cytokines
    Rights
    © 2015 The Authors. Published by ARVO. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. (http://creativecommons.org/licenses/by-nc-nd/4.0/).
    Peer-Reviewed
    yes
    
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    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.
    URI
    http://hdl.handle.net/10454/11157
    Version
    Published version
    Citation
    Deshpande P, Ortega I, Sefat F, Sangwan V S, Green N, Claeyssens F and MacNeil S (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): 1553-61.
    Link to publisher’s version
    http://dx.doi.org/10.1167/iovs.14-15308
    Type
    Article
    Collections
    Engineering and Informatics Publications

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