An ex vivo skin model to probe modulation of local cutaneous arachidonic acid inflammation pathway

Heard, Charles M. ORCID: https://orcid.org/0000-0001-9703-9777 2020. An ex vivo skin model to probe modulation of local cutaneous arachidonic acid inflammation pathway. Journal of Biological Methods 7 (4) , e138. 10.14440/jbm.2020.319

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Abstract

There is a need for inexpensive and reliable means to determine the modulation of cutaneous inflammation. The method outlined in this article draws together a number of scientific techniques and makes use of generally unwanted porcine ear skin as a means of determining skin inflammation ex vivo, and focuses on aspects of the arachidonic acid inflammation pathway. Freshly excised skin contains elevated levels of short-lived, inducible COX-2 and under viable conditions, COX-2 and its eicosanoid products will continue to be produced until tissue necrosis, providing a window of time in which relative levels can be probed to determine exacerbation due to some upregulating factor or reduction due the presence of an anti-inflammatory agent. Ex vivo porcine skin, mounted in Franz diffusion cells, is dosed topically with the xenobiotic challenge and at then techniques such as Western blotting and immunohistochemistry can then be used to probe relative COX-2 levels on a semi-quantitative or qualitative level. ELISA can be used to determine relative PGE-2 levels. Thus far the technique has been used to examine the effects of topically applied anti-inflammatories (betamethasone, ibuprofen, ketoprofen and methotrexate), natural products (fish oil, Devil’s Claw extract and pomegranate rind extract) and drug delivery vehicle (polyNIPAM nanogels). Topically applied xenobiotics that modulate factors such as COX-2 and PGE-2 must penetrate the intact skin, and thus provides indirect evidence of overcoming the barrier function of the stratum corneum in order to target the viable epidermis in sufficient levels to be able to elicit an effect. This system has particular potential as a pre-clinical screening tool for those working on the development of topical delivery systems, and has the additional benefit of being in line with 3 Rs philosophy.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Pharmacy
Additional Information:	This work is licensed under a Creative Commons Attribution-Non-Commercial-ShareAlike 4.0 International License: http://cre-ativecommons.org/licenses/by-nc-sa/4.0
ISSN:	2326-9901
Date of First Compliant Deposit:	11 November 2020
Date of Acceptance:	17 May 2020
Last Modified:	04 May 2023 20:27
URI:	https://orca.cardiff.ac.uk/id/eprint/136265

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