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Cited by 87Tissue Engineering and Regenerative MedicineCorneal Surgery and Treatments3D Printing in Biomedical Research

Corneal Decellularization: A Method of Recycling Unsuitable Donor Tissue for Clinical Translation?

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Wilson, Samantha L., Sidney, Laura E., Dunphy, Siobhán E., Dua, Harminder S., Hopkinson, Andrew·2016·Current Eye Research
DOI 10.3109/02713683.2015.1062114PMC PMC4926783
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Abstract

BACKGROUND: There is a clinical need for biomimetic corneas that are as effective, preferably superior, to cadaveric donor tissue. Decellularized tissues are advantageous compared to synthetic or semi-synthetic engineered tissues in that the native matrix ultrastructure and intrinsic biological cues including growth factors, cytokines and glycosaminoglycans may be retained. However, there is currently no reliable, standardized human corneal decellularization protocol. METHODS: Corneal eye-bank tissue unsuitable for transplantation was utilized to systematically compare commonly used decellularization protocols. Hypertonic sodium chloride; an ionic reagent, sodium dodecyl sulphate; a non-ionic detergent, tert-octylphenol polyoxyethylene (Triton-X); enzymatic disaggregation using Dispase; mechanical agitation; and the use of nucleases were investigated. Decellularization efficacy, specifically for human corneal tissue, was extensively evaluated. Removal of detectable cellular material was evidenced by histological, immunofluorescence and biochemical assays. Preservation of macroscopic tissue transparency and light transmittance was evaluated. Retention of corneal architecture, collagen and glycosaminoglycans was assessed via histological, immunofluorescence and quantitative analysis. Biocompatibility of the resulting scaffolds was assessed using cell proliferation assays. RESULTS: None of the decellularization protocols investigated successfully removed 100% of cellular components. The techniques with the least residual cellular material were most structurally compromised. Biochemical analysis of glycosaminoglycans demonstrated the stripping effects of the decellularization procedures. CONCLUSION: The ability to utilize, reprocess and regenerate tissues deemed "unsuitable" for transplantation allows us to salvage valuable tissue. Reprocessing the tissue has the potential to have a considerable impact on addressing the problems associated with cadaveric donor shortage. Patients would directly benefit by accessing greater numbers of corneal grafts and health authorities would fulfill their responsibility for the delivery of effective corneal reconstruction to alleviate corneal blindness. However, in order to progress, we may need to take a step back to establish a "decellularization" criterion; which should balance effective removal of immune reactive material with maintenance of tissue functionality.

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  • DNase is typically applied after chemical detergents to solubilize cell membranes and dissociate their inner structure, enhancing DNase infiltration and efficiency within tissues.

    “Deoxyribonuclease (DNase), a common nuclease, is typically applied after chemical detergents to solubilize cell membranes and dissociate their inner structure, enhancing DNase infiltration and efficiency within tissues [[152], [153], [154]].”

    From Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities (2024) · cites this paper as [152]

    The Methods (Human Cornea Decellularization) apply DNase and RNase after initial detergent treatment. The Discussion (Residual DNA) says nuclease treatment reduced DNA more successfully after decellularizing pretreatment because ECM disruption allowed nuclease access; another Discussion passage says SDS solubilizes cell membranes. The study supports the sequence and mechanism in corneas, but does not establish that this sequence is typical across tissues. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

  • Prolonged exposure to nucleases used in research settings, which may contain impurities, may negatively impact ECM structure and constituents, affecting the mechanical stability.

    “Prolonged exposure to nucleases used in research settings, which may contain impurities, may negatively impact ECM structure and constituents, affecting the mechanical stability [152,153].”

    From Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities (2024) · cites this paper as [152]

    The Discussion section on corneal transparency and structural integrity reports additional structural disruption after nuclease treatment, supporting the claim that nuclease treatment may harm ECM structure. The Methods describe a 48-hour nuclease treatment, but the paper does not attribute the disruption to prolonged exposure or impurities. Its discussion of possible loss of mechanical integrity concerns SDS treatment, not nuclease treatment. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

  • Thermolysin is less damaging to the ECM than other basement membrane protein-targeting enzymes.

    “Other enzymes, such as lipases, hydrolyze lipids, whereas dispersases and thermolysin target basement membrane proteins, with thermolysin proving less damaging to the ECM.”

    From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [50]

    The Discussion section on corneal transparency and structural integrity describes possible basement-membrane damage after Dispase treatment, but does not discuss thermolysin. Searches for thermolysin and for a comparison with other basement-membrane-targeting enzymes found no passage supporting the claimed lower ECM damage; page numbers were not available in our copy. (LLM, conf=0.76)

    Experimental — accuracy not yet measured

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Authors
Wilson, S. L., Sidney, L. E., Dunphy, S. E., Dua, H. S., & Hopkinson, A.
Affiliations
University of Nottingham (GB)
Corresponding author
Wilson, Samantha L.
ORCID
Venue
Current Eye Research
Publisher
Informa UK Limited
Published
2015-09-23
Volume
41
Issue
6
Pages
769-782
ISSN
  • 0271-3683 (ISSN-L)
  • 1460-2202
DOI
10.3109/02713683.2015.1062114
PMC
PMC4926783
OpenAlex
W1789148173
Cited by
87
References
68
Citations by year
2026: 3 · 2025: 9 · 2024: 12 · 2023: 12 · 2022: 9 · 2021: 8 · 2020: 11 · 2019: 8 · 2018: 5 · 2017: 5 · 2016: 5
FWCI
5.04
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Current Eye Research (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Repository@Nottingham (University of Nottingham) (publishedVersion, other-oa)
  • Repository@Nottingham (University of Nottingham) (submittedVersion, cc-by)
  • Loughborough University Institutional Repository (Loughborough University) (submittedVersion, cc-by)
  • Loughborough University Research Repository (Loughborough University) (submittedVersion, cc-by)
Funders
  • Medical Research Council (MC_PC_14102, MC_PC_13072)
  • Engineering and Physical Sciences Research Council (EP/I017801/1)
Topics
  • Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Corneal Surgery and Treatments
  • Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
Keywords
corneal graft, decellularization protocols, sodium dodecyl sulfate, Triton X-100, dispase, nucleases, glycosaminoglycans, collagen, corneal transparency, biocompatibility, corneal blindness
MeSH terms
Cornea, Eye Banks, Humans, Tissue Donors, Corneal Transplantation, Tissue Engineering, Cell Proliferation, Tissue Scaffolds

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