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Cited by 276Tissue Engineering and Regenerative MedicineHydrogels: synthesis, properties, applications3D Printing in Biomedical Research

The impact of decellularization methods on extracellular matrix derived hydrogels

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Fernández-Pérez, Julia, Ahearne, Mark·2019·Scientific Reports
DOI 10.1038/s41598-019-49575-2PMC PMC6797749
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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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  • GAGs and growth factor depletion are major issues when using ionic detergents due to their impact on compromising future recellularization.

    “GAGs and growth factor depletion are major issues that should be considered when using ionic detergents due to their impact on compromising future recellularization [87].”

    From Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine (2023) · cites this paper as [87]

    The Results (Biochemical characterization of decellularized material; page unavailable) report significant sGAG loss after treatment with SDS, an ionic detergent. The Results (Cytocompatibility; page unavailable) also report that SDS-treated hydrogels were cytotoxic, but attribute this to residual detergent, not GAG or growth-factor depletion. The cited passages do not establish growth-factor depletion or that depletion compromises future recellularization. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • 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 [87]

    The Results > Cytocompatibility section (page unavailable) reports that SDS-treated corneal ECM hydrogels were cytotoxic and that SDS remnants were detected in them. It does not establish cytotoxicity thresholds of 50 mg/L in washing solution or 10 µg/mg dry weight in biomaterials; searches for both quantities found no such measurements. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

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Authors
Fernández-Pérez, J., & Ahearne, M.
Affiliations
Trinity College Dublin (IE)
Corresponding author
Ahearne, Mark
ORCID
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2019-10-17
Volume
9
Issue
1
Pages
14933-14933
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/s41598-019-49575-2
PMC
PMC6797749
OpenAlex
W2981167732
Cited by
276
References
75
Citations by year
2026: 21 · 2025: 36 · 2024: 53 · 2023: 49 · 2022: 47 · 2021: 42 · 2020: 27 · 2019: 1
FWCI
27.95
Type
journal article
Language
en
Update notices
correction (2019-12-19)
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Arrow@dit (Dublin Institute of Technology) (submittedVersion, cc-by-nc-sa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • Trinity's Access to Research Output (TARA) (Trinity College Dublin) (submittedVersion, cc-by)
Funders
  • H2020 European Research Council, European Union (637460)
  • Science Foundation Ireland (15/ERC/3269)
Topics
  • Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Hydrogels: synthesis, properties, applications
  • Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
Keywords
decellularization, extracellular matrix hydrogel, porcine cornea, Triton X-100, corneal stromal cells, cytocompatibility, cytotoxicity, glycosaminoglycans, gelation kinetics, cell encapsulation, tissue engineering scaffolds
MeSH terms
Animals, Biocompatible Materials, Cells, Cultured, Collagen, Cornea, Detergents, Extracellular Matrix, Freeze Drying, Humans, Materials Testing, Sodium Dodecyl Sulfate, Porosity, Stromal Cells, Octoxynol, Hydrogels, Cryoelectron Microscopy, Tissue Engineering, Sus scrofa, Primary Cell Culture

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