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      Dermal Peptide Delivery Using Enhancer Molecules and Colloidal Carrier Systems – Part IV: Search for an Alternative Model Membrane for Future ATR Permeation Studies Using PKEK as the Model Substance

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          Abstract

          The main barrier of the human skin is the stratum corneum (SC). Its properties (also depending on the health and age of the individual) and its influence on improved penetration of active ingredients into the skin are the subject of many research projects. Since the availability of human skin, as the ideal model membrane, is limited, the aim of this study was to find a suitable alternative model membrane from the animal kingdom. The alternative model membrane should be used in subsequent permeation experiments with the Teflon diffusion cell instead of human SC. Previous studies have already investigated the permeation properties of pig, snake, and human skin, but not in a Teflon diffusion cell using ATR. Therefore, it first had to be proven that comparable results can be achieved with animal membranes even under these measurement conditions. This is the precondition for meaningful future permeation experiments with potential enhancers. For this purpose, permeation experiments on various model membranes (human isolated SC, sunburned SC, pig isolated SC, and shed snake skin) by means of FTIR-ATR in a Teflon diffusion cell containing the acceptor and the donor compartment as well as the model membrane were conducted and concentration-time courses of the model peptide PKEK determined. These concentration-time courses were used to calculate and compare the pharmacokinetic parameters (permeation coefficients, lag time, and flux). The starting point was a 10% PKEK solution in D<sub>2</sub>O. It turned out that snake skin is the appropriate alternative model membrane for this type of permeation test.

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          Most cited references31

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          Porcine ear skin: an in vitro model for human skin.

          Porcine ear skin is used in studies of percutaneous penetration as a substitute for human skin. The structure of this tissue, including hair follicles, was studied qualitatively and quantitatively in comparison with human skin. Sections of shock-frozen biopsies, biopsies embedded in paraffin and cyanoacrylate skin surface biopsies were investigated using microscopy. The thickness of the different skin layers and the follicular characteristics were determined. The thickness of the stratum corneum was 17-28 microm, whereas the viable epidermis was 60-85 microm thick. On 1 cm(2), 11-25 hairs were detected, showing a diameter of 58-97 microm and a maximal extension depth of 0.96-1.38 mm into the skin. The orifices of the porcine infundibula showed a diameter of approximately 200 microm. The results obtained are similar to those of human skin, indicating the suitability of this porcine tissue as a model for human skin.
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            Comparison of human skin or epidermis models with human and animal skin in in-vitro percutaneous absorption.

            For the study of in-vitro skin penetration of candidate drugs, excised animal skin is frequently used as a replacement for human skin. Reconstructed human skin or epidermis equivalents have been proposed as alternatives. We compared the penetration properties of human, pig and rat skin with the Graftskin LSE (living skin equivalent) and the Skinethic HRE (human reconstructed epidermis) models using four topical dermatological drugs (salicylic acid, hydrocortisone, clotrimazole and terbinafine) with widely varying polarity. In agreement with published data, pig skin appeared as the most suitable model for human skin: the fluxes through the skin and concentrations in the skin were of the same order of magnitude for both tissues, with differences of at most two- or fourfold, respectively. Graftskin LSE provided an adequate barrier to salicylic acid, but was very permeable for the more hydrophobic compounds (e.g. about 900-fold higher flux and 50-fold higher skin concentrations of clotrimazole as compared to human skin), even more than rat skin. In the case of the Skinethic HRE, we found similar concentrations of salicylic acid as in human skin and an approximately sevenfold higher flux. In contrast, the permeation of hydrophobic compounds through the epidermal layer was vastly higher than through split-thickness human skin (up to a factor of about 800). To conclude, currently available reconstituted skin models cannot be regarded as generally useful for in-vitro penetration studies.
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              PREPARATION OF ISOLATED SHEETS OF HUMAN STRATUM CORNEUM.

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                Author and article information

                Journal
                SPP
                Skin Pharmacol Physiol
                10.1159/issn.1660-5527
                Skin Pharmacology and Physiology
                S. Karger AG
                1660-5527
                1660-5535
                2019
                May 2019
                03 April 2019
                : 32
                : 3
                : 151-161
                Affiliations
                [_a] aHospital Pharmacy, Carl-von-Basedow-Klinikum Saalekreis gGmbH, Merseburg, Germany
                [_b] bInstitute of Pharmacy, Martin Luther University of Halle-Wittenberg, Halle (Saale), Germany
                [_c] cInstitute of Applied Dermatopharmacy, Martin Luther University of Halle-Wittenberg, Halle (Saale), Germany
                Author notes
                *Prof. Dr. Dr. h.c. Reinhard H.H. Neubert, Institute of Applied Dermatopharmacy, Martin Luther University of Halle-Wittenberg, Weinbergweg 23, DE–06120 Halle (Saale) (Germany), E-Mail reinhard.neubert@pharmazie.uni-halle.de
                Article
                495991 Skin Pharmacol Physiol 2019;32:151–161
                10.1159/000495991
                30943494
                943672a2-e2c9-4e98-9b72-53c74b3b9ab9
                © 2019 S. Karger AG, Basel

                Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

                History
                : 29 May 2018
                : 27 November 2018
                Page count
                Figures: 7, Tables: 2, Pages: 11
                Categories
                Research Article

                Oncology & Radiotherapy,Pathology,Surgery,Dermatology,Pharmacology & Pharmaceutical medicine
                Stratum corneum equivalents,Peptide,Skin permeation,Shed snake skin,PKEK,Pig isolated stratum corneum,FTIR-ATR diffusion cell,Stratum corneum

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