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      Mechanical Stretch on Human Skin Equivalents Increases the Epidermal Thickness and Develops the Basement Membrane

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          Abstract

          All previous reports concerning the effect of stretch on cultured skin cells dealt with experiments on epidermal keratinocytes or dermal fibroblasts alone. The aim of the present study was to develop a system that allows application of stretch stimuli to human skin equivalents (HSEs), prepared by coculturing of these two types of cells. In addition, this study aimed to analyze the effect of a stretch on keratinization of the epidermis and on the basement membrane. HSEs were prepared in a gutter-like structure created with a porous silicone sheet in a silicone chamber. After 5-day stimulation with stretching, HSEs were analyzed histologically and immunohistologically. Stretch-stimulated HSEs had a thicker epidermal layer and expressed significantly greater levels of laminin 5 and collagen IV/VII in the basal layer compared with HSEs not subjected to stretch stimulation. Transmission electron microscopy revealed that the structure of the basement membrane was more developed in HSEs subjected to stretching. Our model may be relevant for extrapolating the effect of a stretch on the skin in a state similar to an in vivo system. This experimental system may be useful for analysis of the effects of stretch stimuli on skin properties and wound healing and is also expected to be applicable to an in vitro model of a hypertrophic scar in the future.

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          Living tissue formed in vitro and accepted as skin-equivalent tissue of full thickness.

          Yigal Ehrlich, David J Buttle, T Nakatsuji … (1981)
          Living skin-equivalent grafts consisting of fibroblasts cast in collagen lattices and seeded with epidermal cells were successfully grafted onto the donors of the cells. The grafts were vascularized, did not evoke a homograft reaction, inhibited wound contraction, filled the wound space, and persisted.
          Article 0 comments Cited 142 times – based on 0 reviews     Review now
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            c-Jun and JunB antagonistically control cytokine-regulated mesenchymal-epidermal interaction in skin.

            S Andrecht, A Szabowski, N Fusenig … (2000)
            Interactions between mesenchymal and epithelial cells are responsible for organogenesis and tissue homeostasis. This mutual cross-talk involves cell surface proteins and soluble factors, which are mostly the result of regulated transcription. To elucidate dimer-specific functions of the AP-1 family of transcription factors, we reconstituted skin by combining primary human keratinocytes and mouse wild-type, c-jun(-/-), and junB(-/-) fibroblasts. We have discovered an antagonistic function of these AP-1 subunits in the fibroblast-mediated paracrine control of keratinocyte proliferation and differentiation, and traced this effect to the IL-1-dependent regulation of KGF and GM-CSF. These data suggest that the relative activation state of these AP-1 subunits in a non-cell-autonomous, transregulatory fashion directs regeneration of the epidermis and maintenance of tissue homeostasis in skin.
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              Proliferation and collagen production of human patellar tendon fibroblasts in response to cyclic uniaxial stretching in serum-free conditions.

              C. Wang, Guoguang Yang, Dana Crawford (2004)
              We studied the effect of cyclic mechanical stretching on the proliferation and collagen mRNA expression and protein production of human patellar tendon fibroblasts under serum-free conditions. The role of transforming growth factor-beta1 (TGF-beta1) in collagen production by cyclically stretched tendon fibroblasts was also investigated. The tendon fibroblasts were grown in microgrooved silicone dishes, where the cells were highly elongated and aligned with the microgrooves. Cyclic uniaxial stretching with constant frequency and duration (0.5 Hz, 4 h) but varying magnitude of stretch (no stretch, 4%, and 8%) was applied to the silicone dishes. Following the period of stretching, the cells were rested for 20 h in stretching-conditioned medium to allow for cell proliferation. In separate experiments, the cells were stretched for 4h and then rested for another 4 h. Samples of the medium, total cellular RNA and protein were used for analysis of collagen and TGF-beta1 gene expression and production. It was found that there was a slight increase in fibroblast proliferation at 4% and 8% stretch, compared to that of non-stretched fibroblasts, where at 8% stretch the increase was significant. It was also found that the gene expression and protein production of collagen type I and TGF-beta1 increased in a stretching-magnitude-dependent manner. And, levels of collagen type III were not changed, despite gene expression levels of the protein being slightly increased. Furthermore, the exogenous addition of anti-TGF-beta1 antibody eliminated the increase in collagen type I production under cyclic uniaxial stretching conditions. The results suggest that mechanical stretching can modulate proliferation of human tendon fibroblasts in the absence of serum and increase the cellular production of collagen type I, which is at least in part mediated by TGF-beta1.
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                Author and article information

                Contributors
                Christophe Egles: Role: Editor
                Journal
                Journal ID (nlm-ta): PLoS One
                Journal ID (iso-abbrev): PLoS ONE
                Journal ID (publisher-id): plos
                Journal ID (pmc): plosone
                Title: PLoS ONE
                Publisher: Public Library of Science (San Francisco, CA USA )
                ISSN (Electronic): 1932-6203
                Publication date (Electronic): 3 November 2015
                Publication date Collection: 2015
                Volume: 10
                Issue: 11
                Electronic Location Identifier: e0141989
                Affiliations
                [1 ]The Department of Plastic and Reconstructive Surgery, Okayama University Graduate School of Medicine, Okayama, Japan
                [2 ]Menicon Co., Ltd., Aichi, Japan
                [3 ]The Department of Cardiovascular Physiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
                Université de Technologie de Compiègne, FRANCE
                Author notes

                Competing Interests: The authors have read the journal's policy and the authors of this manuscript have the following competing interests: Yusuke Nagai is an employee of Menicon Co., Ltd. and Keiji Naruse is a research/adviser of Menicon Co., Ltd. There are no patents, products in development, or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials.

                Conceived and designed the experiments: ET YN KT YK KN. Performed the experiments: ET YN KT YK KN. Analyzed the data: ET YN KT YK KN. Contributed reagents/materials/analysis tools: ET YN KT YK KN. Wrote the paper: ET YK KN.

                Article
                Publisher ID: PONE-D-15-16883
                DOI: 10.1371/journal.pone.0141989
                PMC ID: 4631345
                PubMed ID: 26528823
                SO-VID: 33344442-cf57-47d8-8779-b16c2289ac0e
                Copyright statement: Copyright @ 2015
                License:

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                Date received : 18 April 2015
                Date accepted : 15 October 2015
                Page count
                Figures: 6, Tables: 0, Pages: 12
                Funding
                This study was supported by a Grant-in-Aid for Scientific Research (C), 24592713 provided by the Japan Society for the Promotion of Science (JSPS) and Menicon Co., Ltd. provided support in the form of salaries for authors YN and KN (who is a research/adviser for Menicon Co., Ltd). These funders did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.
                Categories
                Subject: Research Article
                Custom metadata
                Data Availability All relevant data are within the paper.

                ScienceOpen disciplines: Uncategorized
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                ScienceOpen disciplines: Uncategorized

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