title: Functional testing of topical skin formulations using an optimised ex vivo skin organ culture model.
abstract: A number of equivalent-skin models are available for investigation of the ex vivo effect of topical application of drugs and cosmaceuticals onto skin, however many have their drawbacks. With the March 2013 ban on animal models for cosmetic testing of products or ingredients for sale in the EU, their utility for testing toxicity and effect on skin becomes more relevant. The aim of this study was to demonstrate proof of principle that altered expression of key gene and protein markers could be quantified in an optimised whole tissue biopsy culture model. Topical formulations containing green tea catechins (GTC) were investigated in a skin biopsy culture model (n = 11). Punch biopsies were harvested at 3, 7 and 10 days, and analysed using qRT-PCR, histology and HPLC to determine gene and protein expression, and transdermal delivery of compounds of interest. Reduced gene expression of α-SMA, fibronectin, mast cell tryptase, mast cell chymase, TGF-β1, CTGF and PAI-1 was observed after 7 and 10 days compared with treated controls (p < 0.05). Histological analysis indicated a reduction in mast cell tryptase and chymase positive cell numbers in treated biopsies compared with untreated controls at day 7 and day 10 (p < 0.05). Determination of transdermal uptake indicated that GTCs were detected in the biopsies. This model could be adapted to study a range of different topical formulations in both normal and diseased skin, negating the requirement for animal models in this context, prior to study in a clinical trial environment.
authors.affiliation: Plastic and Reconstructive Surgery Research, Faculty of Medicine and Human Sciences, Manchester Academic Health Science Centre, Institute of Inflammation and Repair, University of Manchester, Stopford Building, Manchester, M13 9PT, UK.|Centre for Dermatology Research, Institute of Inflammation and Repair, University of Manchester, Manchester, UK.|Healthcare Science Research Centre, Manchester Metropolitan University, Manchester, UK.
authors.forename: G P
authors.initials: GP
authors.lastname: Sidgwick
authors.affiliation: Grosvenor Nuffield Hospital, Wrexham Road, Chester, UK.
authors.forename: D
authors.initials: D
authors.lastname: McGeorge
authors.affiliation: Plastic and Reconstructive Surgery Research, Faculty of Medicine and Human Sciences, Manchester Academic Health Science Centre, Institute of Inflammation and Repair, University of Manchester, Stopford Building, Manchester, M13 9PT, UK. ardeshir.bayat@manchester.ac.uk.|Centre for Dermatology Research, Institute of Inflammation and Repair, University of Manchester, Manchester, UK. ardeshir.bayat@manchester.ac.uk.
authors.forename: A
authors.initials: A
authors.lastname: Bayat
mesh_terms: D000199:Actins / Q000378:metabolism; D000279:Administration, Cutaneous; D023401:Animal Use Alternatives / Q000379:methods*; D001706:Biopsy; D028241:Camellia sinensis / Q000737:chemistry*; D002392:Catechin / Q000008:administration & dosage*; D053818:Chymases / Q000378:metabolism; D055513:Connective Tissue Growth Factor / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008407:Mast Cells / Q000378:metabolism; D009924:Organ Culture Techniques; D010936:Plant Extracts / Q000008:administration & dosage*; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D053773:Transforming Growth Factor beta1 / Q000378:metabolism; D053802:Tryptases / Q000378:metabolism
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; C526317:CCN2 protein, human; D005353:Fibronectins; D010936:Plant Extracts; D017395:Plasminogen Activator Inhibitor 1; C051321:SERPINE1 protein, human; D053773:Transforming Growth Factor beta1; D055513:Connective Tissue Growth Factor; D002392:Catechin; D053818:Chymases; D053802:Tryptases
keywords: Dermal delivery; Dermatology; Extracellular matrix; Inflammation; Mast cells; Skin scarring