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      Ex Vivo Dermis Mechanical Behavior in Relation to Decellularization Treatment Length

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          Abstract

          Background:

          The dermis is a commonly used source tissue for biologic scaffolds; all cellular and nuclear materials need to be removed to limit the inflammatory immune response by the host organism. The decellularization is critical because it must preserve the structural integrity of the extracellular matrix. This work has analyzed a decellularization procedure commonly followed for the dermal tissue that is a chemical treatment with sodium hydroxide. The goal of this work is to identify the optimal treatment length on the basis of structural properties.

          Methods:

          Tensile tests have been performed on the native tissue and on tissues decellularized for 1-7 weeks in sodium hydroxide. The collected data have been analyzed through Tukey-Kramer test to assess if the mechanical properties (ultimate tensile stress and elastic modulus) of decellularized tissues were significantly different from the properties of the native tissue. These tests have been performed on specimens cut along two orthogonal directions (parallel and perpendicular to Langer’s lines).

          Results:

          The decellularization treatment performed with sodium hydroxide in general weakens the tissue: both the ultimate stress and the elastic modulus get lower. The structural properties along Langer lines orientation are more strongly impacted, while the structural properties orthogonal to Langer lines can be preserved with an optimal duration of the decellularization treatment that is 5-6 weeks.

          Conclusion:

          The duration of the decellularization treatment is critical not only to reach a complete decellularization, but also to preserve the mechanical properties of the tissue; 5-6 week treatment performed with sodium hydroxide allows preserving the mechanical properties of the native tissue perpendicularly to Langer lines orientation, and minimizing the impact of the decellularization process on the mechanical properties along the Langer lines orientation.

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          Most cited references16

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          Decellularization of tissues and organs.

          Decellularized tissues and organs have been successfully used in a variety of tissue engineering/regenerative medicine applications, and the decellularization methods used vary as widely as the tissues and organs of interest. The efficiency of cell removal from a tissue is dependent on the origin of the tissue and the specific physical, chemical, and enzymatic methods that are used. Each of these treatments affect the biochemical composition, tissue ultrastructure, and mechanical behavior of the remaining extracellular matrix (ECM) scaffold, which in turn, affect the host response to the material. Herein, the most commonly used decellularization methods are described, and consideration give to the effects of these methods upon the biologic scaffold material.
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            Methods of tissue decellularization used for preparation of biologic scaffolds and in vivo relevance.

            Biologic scaffolds composed of extracellular matrix (ECM) are widely used in both preclinical animal studies and in many clinical applications to repair and reconstruct tissues. Recently, 3-dimensional ECM constructs have been investigated for use in whole organ engineering applications. ECM scaffolds are prepared by decellularization of mammalian tissues and the ECM provides natural biologic cues that facilitate the restoration of site appropriate and functional tissue. Preservation of the native ECM constituents (i.e., three-dimensional ultrastructure and biochemical composition) during the decellularization process would theoretically result in the ideal scaffold for tissue remodeling. However, all methods of decellularization invariably disrupt the ECM to some degree. Decellularization of tissues and organs for the production of ECM bioscaffolds requires a balance between maintaining native ECM structure and the removal of cellular materials such as DNA, mitochondria, membrane lipids, and cytosolic proteins. These remnant cellular components can elicit an adverse inflammatory response and inhibit constructive remodeling if not adequately removed. Many variables including cell density, matrix density, thickness, and morphology can affect the extent of tissue and organ decellularization and thus the integrity and physical properties of the resulting ECM scaffold. This review describes currently used decellularization techniques, and the effects of these techniques upon the host response to the material.
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              Matrix composition and mechanics of decellularized lung scaffolds.

              The utility of decellularized native tissues for tissue engineering has been widely demonstrated. Here, we examine the production of decellularized lung scaffolds from native rodent lung using two different techniques, principally defined by use of either the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) or sodium dodecyl sulfate (SDS). All viable cellular material is removed, including at least 99% of DNA. Histochemical staining and mechanical testing indicate that collagen and elastin are retained in the decellularized matrices with CHAPS-based decellularization, while SDS-based decellularization leads to loss of collagen and decline in mechanical strength. Quantitative assays confirm that most collagen is retained with CHAPS treatment but that about 80% of collagen is lost with SDS treatment. In contrast, for both detergent methods, at least 60% of elastin content is lost along with about 95% of native proteoglycan content. Mechanical testing of the decellularized scaffolds indicates that they are mechanically similar to native lung using CHAPS decellularization, including retained tensile strength and elastic behavior, demonstrating the importance of collagen and elastin in lung mechanics. With SDS decellularization, the mechanical integrity of scaffolds is significantly diminished with some loss of elastic function as well. Finally, a simple theoretical model of peripheral lung matrix mechanics is consonant with our experimental findings. This work demonstrates the feasibility of producing a decellularized lung scaffold that can be used to study lung matrix biology and mechanics, independent of the effects of cellular components. Copyright © 2011 S. Karger AG, Basel.
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                Author and article information

                Journal
                Open Biomed Eng J
                Open Biomed Eng J
                TOBEJ
                The Open Biomedical Engineering Journal
                Bentham Open
                1874-1207
                8 April 2016
                2016
                : 10
                : 34-42
                Affiliations
                [1 ]Department of Mechanical and Aerospace Engineering, Politecnico di Torino, Torino, Italy
                [2 ]Skin Bank, AOU Città della Salute e della Scienza, Torino, Italy
                [3 ]Department of Engineering, University of Perugia, Perugia, Italy
                Author notes
                [* ] Address correspondence to this author at the Department of Engineering, University of Perugia, Via Duranti 67, 06125 Perugia, Italy; Tel:+39 075 5853748; Fax: +39 075 5853606; E-mail: elisabetta.zanetti@ 123456unipg.it
                Article
                TOBEJ-10-34
                10.2174/1874120701610010034
                5395843
                28484575
                2fcd3291-c6b9-48d6-a081-79ed672cf4c2
                © Terzini et al.; Licensee Bentham Open.

                This is an open access article licensed under the terms of the Creative Commons Attribution-Non-Commercial 4.0 International Public License (CC BY-NC 4.0) ( https://creativecommons.org/licenses/by-nc/4.0/legalcode), which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

                History
                : 12 October 2015
                : 17 December 2015
                : 17 February 2016
                Categories
                Article

                Biomedical engineering
                acellularized dermis,allografts,decellularization treatment,elastic modulus,extracellular matrix,human dermis,tensile tests,ultimate stress

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