Keeping an Eye on Decellularized Corneas: A Review of Methods, Characterization and Applications

Wilson, Samantha; Sidney, Laura; Dunphy, Siobhán; Rose, James; Hopkinson, Andrew

doi:10.3390/jfb4030114

Cited by 100Tissue Engineering and Regenerative MedicineCorneal Surgery and Treatments3D Printing in Biomedical Research

Keeping an Eye on Decellularized Corneas: A Review of Methods, Characterization and Applications

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Wilson, Samantha, Sidney, Laura, Dunphy, Siobhán, Rose, James, Hopkinson, Andrew·2013·Journal of Functional Biomaterials

DOI 10.3390/jfb4030114PMC PMC4030906

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Abstract

The worldwide limited availability of suitable corneal donor tissue has led to the development of alternatives, including keratoprostheses (Kpros) and tissue engineered (TE) constructs. Despite advances in bioscaffold design, there is yet to be a corneal equivalent that effectively mimics both the native tissue ultrastructure and biomechanical properties. Human decellularized corneas (DCs) could offer a safe, sustainable source of corneal tissue, increasing the donor pool and potentially reducing the risk of immune rejection after corneal graft surgery. Appropriate, human-specific, decellularization techniques and high-resolution, non-destructive analysis systems are required to ensure reproducible outputs can be achieved. If robust treatment and characterization processes can be developed, DCs could offer a supplement to the donor corneal pool, alongside superior cell culture systems for pharmacology, toxicology and drug discovery studies.

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TEM elucidates ultrastructural details of tissues, including fibril diameter, spacing, and collagen fibril organization, making it a critical technique for understanding native tissue architecture.
“TEM elucidates ultrastructural details of tissues, including fibril diameter, spacing, and collagen fibril organization, establishing it as a critical technique for an in‐depth understanding of native tissue architecture.[ 94 ]”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [94]
Section 4.2.2, “Electron Microscopy,” says TEM reveals fibril diameter, interfibrillar spacing, and collagen fibril order, and calls it central to understanding corneal architecture. The source makes this point specifically about corneal tissue; the citing sentence extends it to native tissue architecture generally. (LLM, conf=0.84)
Experimental — accuracy not yet measured
OCT eliminates the need for fluorescent labeling, is relatively inexpensive compared to confocal and multiphoton techniques, and is well-suited for scanning decellularized tissues, facilitating non-destructive, rapid, sterile, in situ, real-time studies.
“OCT, eliminating the need for fluorescent labeling of tissues and being relatively inexpensive compared to confocal and multiphoton techniques, is well‐suited for scanning decellularized tissues and facilitates non‐destructive, rapid, sterile, in situ, real‐time studies.[ 94 ]”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [94]
Section 4.2.5 says OCT needs no fluorescent labeling, is relatively inexpensive, and allows fast, sterile, non-destructive, in situ, real-time investigation. It calls OCT suitable for monitoring decellularized corneas, but does not extend that suitability to decellularized tissues generally. (LLM, conf=0.82)
Experimental — accuracy not yet measured
Standard optical microscopy permits direct scaffold observation; however, its lower resolution restricts observation of microstructural details.
“Standard optical microscopy, which is frequently employed for structural and morphological analysis, permits direct scaffold observation; however, its lower resolution restricts observation of microstructural details.”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [94]
Histological and immunohistochemical staining are commonly used image-based techniques, but fixation, labeling, and processing steps may be destructive, potentially distorting cellular expression and tissue architecture, and tend to provide information for a limited tissue area.
“Histological and immunohistochemical staining are commonly used image‐based techniques. However, the fixation, labeling, and processing steps may be destructive, potentially distorting cellular expression and tissue architecture, and tend to provide information for a limited tissue area.”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [94]
Maurice and colleagues initially employed X-rays to investigate collagen fiber arrangement in corneal tissues, enabling quantification of orientation and content across the cornea and limbus, with changes detected via X-ray scattering patterns.
“Maurice and colleagues initially employed X‐rays to investigate structural features pertaining to the arrangement of collagen fibers in corneal tissues, enabling the quantification of the orientation and content of collagen fibers across the cornea and limbus, with changes in fiber arrangement being readily detected via X‐ray scattering patterns.[ 94 , 105 , 111 ]”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [94]

Critical point drying for SEM sample preparation is linked to varying degrees of sample shrinkage and may necessitate harsh fixatives.
“However, the critical point drying technique for SEM sample preparation is linked to varying degrees of sample shrinkage and may necessitate harsh fixatives.[ 94 ]”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [94]
Second harmonic generation (SHG) imaging delivers images with exceptional spatial resolution and contrast, which are crucial for visualizing tissue architecture.
“Second harmonic generation (SHG) imaging delivers images with exceptional spatial resolution and contrast, which are crucial for visualizing tissue architecture.[ 94 , 109 ]”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [94]
Polarized microscopy facilitates the quantitative analysis of collagen organization.
“Polarized microscopy facilitates the quantitative analysis of collagen organization.[ 94 ]”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [94]
SEM imaging demonstrates that collagen fiber alignment in decellularized bovine tendon is significantly distorted compared to the uniform and directionally stacked fibers in native tissues.
“For instance, SEM imaging demonstrates that collagen fiber alignment in decellularized bovine tendon is significantly distorted compared to the uniform and directionally stacked fibers in native tissues.[ 94 ]”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [94]
Section 4.2.2 discusses SEM imaging of corneal tissue, not a comparison of native and decellularized bovine tendon. Searches for bovine tendon and distorted collagen alignment found no passage reporting the claimed result; the source’s discussion of SEM does not establish it. (LLM, conf=0.75)
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Decellularization and recellularization of cornea: Progress towards a donor alternative
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Corneal Decellularization: A Method of Recycling Unsuitable Donor Tissue for Clinical Translation?
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Self-citation
Decellularized corneal-based 3D scaffolds: methods decellularization, characterization, mechanical properties, and species source
Akbaripour, V., Rezaei, L., Rostaminasab, G., Daneshgar, F., Bahiraee, O., & Rezakhani, L. (2025) · Regenerative Therapy · doi:10.1016/j.reth.2025.101044 · Cited by 2 · see citing works
Corneal decellularisation: recycling tissue for transplantation
WILSON, S. (2014) · Acta Ophthalmologica · doi:10.1111/j.1755-3768.2014.1723.x
Self-citation
Chemical, Physical, and Biological Corneal Decellularization Methods: A Review of Literature
Procházková, A., Poláchová, M., Dítě, J., Netuková, M., & Studený, P. (2024) · Journal of Ophthalmology · doi:10.1155/2024/1191462 · Cited by 15 · see citing works

Authors: Wilson, S., Sidney, L., Dunphy, S., Rose, J., & Hopkinson, A.
Affiliations: University of Nottingham (GB)
Corresponding author: Hopkinson, Andrew
ORCID: Wilson, Samantha
Sidney, Laura
Dunphy, Siobhán
Hopkinson, Andrew
Venue: Journal of Functional Biomaterials
Publisher: MDPI AG
Published: 2013-07-10
Volume: 4
Issue: 3
Pages: 114-161
ISSN: 2079-4983 (ISSN-L)
DOI: 10.3390/jfb4030114
PMC: PMC4030906
OpenAlex: W2149410257
Cited by: 100
References: 263
Citations by year: 2026: 7 · 2025: 6 · 2024: 11 · 2023: 8 · 2022: 9 · 2021: 6 · 2020: 11 · 2019: 11 · 2018: 4 · 2017: 10 · 2016: 8 · 2015: 7 · 2014: 1 · 2013: 1
FWCI: 5.59
Type: journal article
Language: en
Open access: gold
doi.org
Licence: cc-by
creativecommons.org/licenses/by/3.0/ (vor)
Locations: Journal of Functional Biomaterials (publishedVersion, cc-by)
Europe PMC (PubMed Central) (publishedVersion, cc-by)
PubMed Central (publishedVersion, cc-by)
PubMed (publishedVersion)
Repository@Nottingham (University of Nottingham) (publishedVersion, cc-by)
DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Loughborough University Institutional Repository (Loughborough University) (submittedVersion, cc-by)
Loughborough University Research Repository (Loughborough University) (submittedVersion, cc-by)
Repository@Nottingham (University of Nottingham) (submittedVersion, cc-by)
Funders: 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 tissue engineering, bioscaffolds, decellularization methods, immune rejection, corneal graft, corneal biomechanics, corneal ultrastructure, drug discovery

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Keeping an Eye on Decellularized Corneas: A Review of Methods, Characterization and Applications

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