The impact of decellularization methods on extracellular matrix derived hydrogels

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Abstract

Tissue-derived decellularized biomaterials are ideal for tissue engineering applications as they mimic the biochemical composition of the native tissue. These materials can be used as hydrogels for cell encapsulation and delivery. The decellularization process can alter the composition of the extracellular matrix (ECM) and thus influence the hydrogels characteristics. The aim of this study was to examine the impact of decellularization protocols in ECM-derived hydrogels obtained from porcine corneas. Porcine corneas were isolated and decellularized with SDS, Triton X-100 or by freeze-thaw cycles. All decellularization methods decreased DNA significantly when measured by PicoGreen and visually assessed by the absence of cell nuclei. Collagen and other ECM components were highly retained, as quantified by hydroxyproline content and sGAG, by histological analysis and by SDS-PAGE. Hydrogels obtained by freeze-thaw decellularization were the most transparent. The method of decellularization impacted gelation kinetics assessed by turbidimetric analysis. All hydrogels showed a fibrillary and porous structure determined by cryoSEM. Human corneal stromal cells were embedded in the hydrogels to assess cytotoxicity. SDS decellularization rendered cytotoxic hydrogels, while the other decellularization methods produced highly cytocompatible hydrogels. Freeze-thaw decellularization produced hydrogels with the overall best properties.

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An overview of tissue and whole organ decellularization processes.

Peter M Crapo, Thomas W Gilbert, Stephen F Badylak (2011)

Biologic scaffold materials composed of extracellular matrix (ECM) are typically derived by processes that involve decellularization of tissues or organs. Preservation of the complex composition and three-dimensional ultrastructure of the ECM is highly desirable but it is recognized that all methods of decellularization result in disruption of the architecture and potential loss of surface structure and composition. Physical methods and chemical and biologic agents are used in combination to lyse cells, followed by rinsing to remove cell remnants. Effective decellularization methodology is dictated by factors such as tissue density and organization, geometric and biologic properties desired for the end product, and the targeted clinical application. Tissue decellularization with preservation of ECM integrity and bioactivity can be optimized by making educated decisions regarding the agents and techniques utilized during processing. An overview of decellularization methods, their effect upon resulting ECM structure and composition, and recently described perfusion techniques for whole organ decellularization techniques are presented herein. Copyright © 2011 Elsevier Ltd. All rights reserved.

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3D printed complex tissue construct using stem cell-laden decellularized extracellular matrix bioinks for cardiac repair.

Jinah Jang, Hun-Jun Park, Seok-Won Kim … (2017)

Stem cell therapy is a promising therapeutic method for the treatment of ischemic heart diseases; however, some challenges prohibit the efficacy after cell delivery due to hostile microenvironment of the injured myocardium. 3D printed pre-vascularized stem cell patch can enhance the therapeutic efficacy for cardiac repair through promotion of rapid vascularization after patch transplantation. In this study, stem cell-laden decellularized extracellular matrix bioinks are used in 3D printing of pre-vascularized and functional multi-material structures. The printed structure composed of spatial patterning of dual stem cells improves cell-to-cell interactions and differentiation capability and promotes functionality for tissue regeneration. The developed stem cell patch promoted strong vascularization and tissue matrix formation in vivo. The patterned patch exhibited enhanced cardiac functions, reduced cardiac hypertrophy and fibrosis, increased migration from patch to the infarct area, neo-muscle and capillary formation along with improvements in cardiac functions. Therefore, pre-vascularized stem cell patch provides cardiac niche-like microenvironment, resulting in beneficial effects on cardiac repair.

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A hydrogel derived from decellularized dermal extracellular matrix.

Matthew Wolf, Kerry Daly, Ellen P Brennan-Pierce … (2012)

The ECM of mammalian tissues has been used as a scaffold to facilitate the repair and reconstruction of numerous tissues. Such scaffolds are prepared in many forms including sheets, powders, and hydrogels. ECM hydrogels provide advantages such as injectability, the ability to fill an irregularly shaped space, and the inherent bioactivity of native matrix. However, material properties of ECM hydrogels and the effect of these properties upon cell behavior are neither well understood nor controlled. The objective of this study was to prepare and determine the structure, mechanics, and the cell response in vitro and in vivo of ECM hydrogels prepared from decellularized porcine dermis and urinary bladder tissues. Dermal ECM hydrogels were characterized by a more dense fiber architecture and greater mechanical integrity than urinary bladder ECM hydrogels, and showed a dose dependent increase in mechanical properties with ECM concentration. In vitro, dermal ECM hydrogels supported greater C2C12 myoblast fusion, and less fibroblast infiltration and less fibroblast mediated hydrogel contraction than urinary bladder ECM hydrogels. Both hydrogels were rapidly infiltrated by host cells, primarily macrophages, when implanted in a rat abdominal wall defect. Both ECM hydrogels degraded by 35 days in vivo, but UBM hydrogels degraded more quickly, and with greater amounts of myogenesis than dermal ECM. These results show that ECM hydrogel properties can be varied and partially controlled by the scaffold tissue source, and that these properties can markedly affect cell behavior. Copyright © 2012 Elsevier Ltd. All rights reserved.

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Author and article information

Contributors

Mark Ahearne: ahearnm@tcd.ie

Journal

Journal ID (nlm-ta): Sci Rep

Journal ID (iso-abbrev): Sci Rep

Title: Scientific Reports

Publisher: Nature Publishing Group UK (London )

ISSN (Electronic): 2045-2322

Publication date (Electronic): 17 October 2019

Publication date PMC-release: 17 October 2019

Publication date Collection: 2019

Volume: 9

Electronic Location Identifier: 14933

Affiliations

[1 ]ISNI 0000 0004 1936 9705, GRID grid.8217.c, Department of Mechanical and Manufacturing Engineering, School of Engineering, Trinity College Dublin, , the University of Dublin, ; Dublin, Ireland

[2 ]ISNI 0000 0004 1936 9705, GRID grid.8217.c, Trinity Centre for Biomedical Engineering, Trinity Biomedical Science Institute, Trinity College Dublin, , the University of Dublin, ; Dublin, Ireland

Author information

Julia Fernández-Pérez https://orcid.org/0000-0002-8375-2907

Article

Publisher ID: 49575

DOI: 10.1038/s41598-019-49575-2

PMC ID: 6797749

PubMed ID: 31624357

SO-VID: 66d56ab6-4e04-44c8-83c5-47e55b0f800f

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Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

History

Date received : 20 June 2019

Date accepted : 28 August 2019

Funding

Funded by: H2020 European Research Council, European Union

Award ID: 637460

Award Recipient : Julia Fernández-Pérez Mark Ahearne

Funded by: FundRef http://dx.doi.org/10.13039/501100001602, Science Foundation Ireland;

Award ID: 15/ERC/3269

Award Recipient : Mark Ahearne

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ScienceOpen disciplines: Uncategorized

Keywords: biomedical engineering,biomaterials,bioinspired materials

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ScienceOpen disciplines: Uncategorized

Keywords: biomedical engineering, biomaterials, bioinspired materials

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The impact of decellularization methods on extracellular matrix derived hydrogels

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Most cited references 52

An overview of tissue and whole organ decellularization processes.

3D printed complex tissue construct using stem cell-laden decellularized extracellular matrix bioinks for cardiac repair.

A hydrogel derived from decellularized dermal extracellular matrix.

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Cited by 72

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