Methods of tissue decellularization used for preparation of biologic scaffolds and in vivo relevance
Open accessAbstract
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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Decellularization protocols vary and are typically tailored based on tissue-specific factors like density and geometry.
“Decellularization protocols vary and are typically tailored based on tissue-specific factors like density and geometry [4, 18, 21, 22].”
From Effects of Various Decellularization Methods for the Development of Decellularized Extracellular Matrix from Tilapia (<i>Oreochromis niloticus</i>) Viscera (2024) · cites this paper as [22]
Collagen, the predominant structural protein in the ECM, provides an appropriate 3D microenvironment that promotes cell adhesion, proliferation, and differentiation, thereby facilitating tissue regeneration and repair.
“Collagen, the predominant structural protein in the ECM, provides an appropriate 3D microenvironment that promotes cell adhesion, proliferation, and differentiation, thereby facilitating tissue regeneration and repair.[ 119 ]”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [119]
Quantification of residual DNA as a potent immunogen and indirect index of remaining cell materials is a practical tool to estimate remaining immunogen amount throughout dECM.
“The quantification of residual DNA as a potent immunogen and indirect index of remaining cell materials is a practical tool to estimate the remaining immunogen amount throughout the dECM [54–56].”
From Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine (2023) · cites this paper as [56]
TX100 decellularization efficacy is highly dependent on target tissue configuration; complete decellularization has been reported in thin tissues such as nerves and pericardium, while TX100 application alone does not seem successful in removing immunogen cell debris from denser tissues such as arteries.
“It is of interest to further note that TX100 decellularization efficacy is highly dependent on the target tissue configuration. Complete decellularization has been reported in thin tissues such as nerves and pericardium, while TX100 application alone does not seem successful in removing immunogen cell debris from denser tissues such as arteries [56, 75, 83].”
From Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine (2023) · cites this paper as [56]
SDS concentrations over 50 mg/L in washing solution or remaining SDS in ECM-based biomaterials beyond 10 μg/mg dry weight have shown cytotoxicity.
“SDS concentrations over 50 mg/L in washing solution or remaining SDS in ECM-based biomaterials beyond 10 µg/mg dry weight have shown cytotoxicity [56, 87].”
From Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine (2023) · cites this paper as [56]
Ionizing radiations, including γ-irradiation and electron beam irradiation, are prominent physical sterilizers that induce nucleic acid denaturation to terminate pathogen activities.
“Ionizing radiations, including γ-irradiation and electron beam irradiation, are prominent physical sterilizers, inducing nucleic acid denaturation to terminate pathogen activities.”
From Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine (2023) · cites this paper as [56]
Mild irradiation induces crosslinking of dECM scaffolds, while heavy irradiation denatures and degrades ECM, resulting in inferior mechanical strength and cell attachment capability.
“Mild irradiation induces crosslinking of dECM scaffolds, while heavy irradiation denatures and degrades ECM, resulting in inferior mechanical strength and cell attachment capability.”
From Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine (2023) · cites this paper as [56]
25 kGy is the optimal Gamma dose for sterilization of dECM scaffolds with favorable ECM preservation and induction of minimal immune response.
“In this regard, 25 kGy is the optimal Gama dose for sterilization of dECM scaffolds with favorable ECM preservation and induction of minimal immune response.[56, 222].”
From Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine (2023) · cites this paper as [56]
Ionizing radiations may damage ECM ultrastructure and produce lipid-derived cytotoxic particles, limiting their application.
“They may damage ECM ultrastructure and produce lipid-derived cytotoxic particles that have limited their application.”
From Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine (2023) · cites this paper as [56]
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- Authors
- Keane, T. J., Swinehart, I. T., & Badylak, S. F.
- Affiliations
- University of Pittsburgh (US)
- McGowan Institute for Regenerative Medicine (US)
- Corresponding author
- Badylak, Stephen F.
- Venue
- Methods
- Publisher
- Elsevier BV
- Published
- 2015-08
- Volume
- 84
- Pages
- 25-34
- ISSN
- 1046-2023 (ISSN-L)
- OpenAlex
- W2078246403
- Cited by
- 670
- References
- 12
- Citations by year
- 2026: 39 · 2025: 62 · 2024: 81 · 2023: 78 · 2022: 65 · 2021: 75 · 2020: 61 · 2019: 60 · 2018: 55 · 2017: 48 · 2016: 32 · 2015: 13 · 2014: 1
- FWCI
- 9.44
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Methods (acceptedVersion)
- PubMed (publishedVersion)
- Funders
- National Science Foundation (DGE-1247842)
- Topics
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Hydrogels: synthesis, properties, applications
- Keywords
- extracellular matrix scaffolds, biological scaffolds, tissue remodeling, inflammation, decellularization methods
- MeSH terms
- Animals, Biocompatible Materials, Cell Separation, Extracellular Matrix, Humans, Materials Testing, Sterilization, Tissue Engineering, Tissue Scaffolds
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