Reconstructed skin models are suitable test systems for toxicity testing and for basic investigations on (patho-)physiological aspects of human skin. Reconstructed human skin, however, has clear limitations such as the lack of immune cells and a significantly weaker skin barrier function compared to native human skin. Potential reasons for the latter might be the lack of mechanical forces during skin model cultivation which is performed classically in static well-plate setups. Mechanical forces and shear stress have a major impact on tissue formation and, hence, tissue engineering. In the present work, a perfusion platform was developed allowing dynamic cultivation of in vitro skin models. The platform was designed to cultivate reconstructed skin at the air-liquid interface with a laminar and continuous medium flow below the dermis equivalent. Histological investigations confirmed the formation of a significantly thicker stratum corneum compared to the control cultivated under static conditions. Moreover, the skin differentiation markers involucrin and filaggrin as well as the tight junction proteins claudin 1 and occludin showed increased expression in the dynamically cultured skin models. Unexpectedly, despite improved differentiation, the skin barrier function of the dynamically cultivated skin models was not enhanced compared with the skin models cultivated under static conditions.

Keywords:
Skin models, Dynamic perfusion platform, Skin barrier function, Shear stress
1.
European Commission: Seventh Report on the Statistics on the Number of Animals used for Experimental and other Scientific Purposes in the Member States of the European Union. Brussels, European Commission, 2013.
2.
Seok J, Warren HS, Cuenca AG, Mindrinos MN, Baker H V, Xu W, et al: Genomic responses in mouse models poorly mimic human inflammatory diseases. Proc Natl Acad Sci USA 2013;110:3507-3512.
3.
Perrin S: Preclinical research: make mouse studies work. Nature 2014;507:423-425.
4.
de Vries RBM, Leenaars M, Tra J, Huijbregtse R, Bongers E, Jansen JA, et al: The potential of tissue engineering for developing alternatives to animal experiments: a systematic review. J Tissue Eng Regen Med 2015;9:771-778.
5.
Reisinger K, Hoffmann S, Alépée N, Ashikaga T, Barroso J, Elcombe C, et al: Systematic evaluation of non-animal test methods for skin sensitisation safety assessment. Toxicol Vitr 2015;29:259-272.
6.
OECD: OECD Guideline No. 431 for the Testing of Chemicals: in vitro Skin Corrosion: Reconstructed Human Epidermis (RhE) Test Method. Paris, OECD, 2004.
7.
OECD: OECD Guideline No. 439 for the Testing of Chemicals: in vitro Skin Irritation: Reconstructed Human Epidermis Test Method. Paris, OECD, 2010.
8.
OECD: OECD Guideline No. 428 for the Testing of Chemicals: Skin Absorption: in vitro Method. Paris, OECD, 2004.
9.
Zöller NN, Kippenberger S, Thaçi D, Mewes K, Spiegel M, Sättler A, et al: Evaluation of beneficial and adverse effects of glucocorticoids on a newly developed full-thickness skin model. Toxicol In Vitro 2008;22:747-759.
10.
Hönzke S, Wallmeyer L, Ostrowski A, Radbruch M, Mundhenk L, Schäfer-Korting M, et al: Influence of Th2 cytokines on the cornified envelope, tight junction proteins, and β-defensins in filaggrin-deficient skin equivalents. J Invest Dermatol 2016;136:631-639.
11.
van Drongelen V, Haisma EM, Out-Luiting JJ, Nibbering PH, El Ghalbzouri A: Reduced filaggrin expression is accompanied by increased Staphylococcus aureus colonization of epidermal skin models. Clin Exp Allergy 2014;44:1515-1524.
12.
Berroth A, Kühnl J, Kurschat N, Schwarz A, Stäb F, Schwarz T, et al: Role of fibroblasts in the pathogenesis of atopic dermatitis. J Allergy Clin Immunol 2013;131:1547-1554.
13.
Ponec M, Boelsma E, Gibbs S, Mommaas M: Characterization of reconstructed skin models. Skin Pharmacol Appl Skin Physiol 2002;15(suppl 1):4-17.
14.
Vávrová K, Henkes D, Strüver K, Sochorová M, Skolová B, Witting MY, et al: Filaggrin deficiency leads to impaired lipid profile and altered acidification pathways in a 3D skin construct. J Invest Dermatol 2014;134:746-753.
15.
Schäfer-Korting M, Bock U, Diembeck W, Düsing H-J, Gamer A, Haltner-Ukomadu E, et al: The use of reconstructed human epidermis for skin absorption testing: results of the validation study. Altern Lab Anim 2008;36:161-187.
16.
Küchler S, Strüver K, Friess W: Reconstructed skin models as emerging tools for drug absorption studies. Expert Opin Drug Metab Toxicol 2013;9:1255-1263.
17.
Shiu Y-T: Mechanical forces on cells; in Bronzino JD (ed): Tissue Engineering and Artificial Organs. Boca Raton, CRC Press, Taylor & Francis Group, 2006.
18.
Hronik-Tupaj M, Kaplan DL: A review of the responses of two- and three-dimensional engineered tissues to electric fields. Tissue Eng B Rev 2012;18:167-180.
19.
Huh D, Matthews BD, Mammoto A, Montoya-Zavala M, Hsin HY, Ingber DE: Reconstituting organ-level lung functions on a chip. Science 2010;328:1662-1668.
20.
Ingber DE: Tensegrity II. How structural networks influence cellular information processing networks. J Cell Sci 2003;116:1397-1408.
21.
Davisson T, Kunig S, Chen A, Sah R, Ratcliffe A: Static and dynamic compression modulate matrix metabolism in tissue engineered cartilage. J Orthop Res 2002;20:842-848.
22.
Zhang X, Wang X, Keshav V, Wang X, Johanas JT, Leisk GG, et al: Dynamic culture conditions to generate silk-based tissue-engineered vascular grafts. Biomaterials 2009;30:3213-3223.
23.
Hronik-Tupaj M, Kaplan DL: A review of the responses of two- and three-dimensional engineered tissues to electric fields. Tissue Eng B Rev 2012;18:167-180.
24.
Ataç B, Wagner I, Horland R, Lauster R, Marx U, Tonevitsky AG, et al: Skin and hair on-a-chip: in vitro skin models versus ex vivo tissue maintenance with dynamic perfusion. Lab Chip 2013;13:3555-3561.
25.
Griffith LG, Swartz MA: Capturing complex 3D tissue physiology in vitro. Nat Rev Mol Cell Biol 2006;7:211-224.
26.
Gysler A, Lange K, Korting HC, Schäfer-Korting M: Prednicarbate biotransformation in human foreskin keratinocytes and fibroblasts. Pharm Res 1997;14:793-797.
27.
Eckl K-M, Alef T, Torres S, Hennies HC: Full-thickness human skin models for congenital ichthyosis and related keratinization disorders. J Invest Dermatol 2011;131:1938-1942.
28.
Küchler S, Henkes D, Eckl K-M, Ackermann K, Plendl J, Korting H-C, et al: Hallmarks of atopic skin mimicked in vitro by means of a skin disease model based on FLG knock-down. Altern Lab Anim 2011;39:471-480.
29.
Leist M, Hartung T: Inflammatory findings on species extrapolations: humans are definitely no 70-kg mice. Arch Toxicol 2013;87:563-567.
30.
Ponec M: Skin constructs for replacement of skin tissues for in vitro testing. Adv Drug Deliv Rev 2002;54(suppl 1):S19-S30.
31.
van Smeden J, Boiten WA, Hankemeier T, Rissmann R, Bouwstra JA, Vreeken RJ: Combined LC/MS-platform for analysis of all major stratum corneum lipids, and the profiling of skin substitutes. Biochim Biophys Acta 2014;1841:70-79.
32.
Mammoto T, Ingber DE: Mechanical control of tissue and organ development. Development 2010;137:1407-1420.
33.
Mammoto A, Mammoto T, Ingber DE: Mechanosensitive mechanisms in transcriptional regulation. J Cell Sci 2012;125:3061-3073.
34.
Proksch E, Brandner JM, Jensen JM: The skin: an indispensable barrier. Exp Dermatol 2008;17:1063-1072.
35.
McAleer MA, Irvine AD: The multifunctional role of filaggrin in allergic skin disease. J Allergy Clin Immunol 2013;131:280-291.
36.
Cabanillas B, Novak N: Atopic dermatitis and filaggrin. Curr Opin Immunol 2016;42:1-8.
37.
Candi E, Schmidt R, Melino G: The cornified envelope: a model of cell death in the skin. Nat Rev Mol Cell Biol 2005;6:328-340.
38.
Nagy E, Hadik P: Analysis of mass transfer in hollow-fiber membranes. Desalination 2002;145:147-152.
39.
Wickramasinghe SR, Garcia JD, Han B: Mass and momentum transfer in hollow fibre blood oxygenators. J Memb Sci 2002;208:247-256.
40.
Bridge MJ, Broadhead KW, Hitchcock RW, Webb K, Tresco PA: A simple instrument to characterize convective and diffusive transport of single hollow fibers of short length. J Memb Sci 2001;183:223-233.
© 2017 S. Karger AG, Basel
2017
Copyright / Drug Dosage / Disclaimer
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.
You do not currently have access to this content.