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Cited by 64Tissue Engineering and Regenerative Medicine3D Printing in Biomedical ResearchNerve injury and regeneration

Development of an apoptosis-assisted decellularization method for maximal preservation of nerve tissue structure

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Cornelison, R.C., Wellman, S.M., Park, J.H., Porvasnik, S.L., Song, Y.H., Wachs, R.A., Schmidt, C.E.·2018·Acta Biomaterialia
DOI 10.1016/j.actbio.2018.07.009PMC PMC6126930
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Abstract

Preservation of tissue structure is often a primary goal when optimizing tissue and organ decellularization methods. Many current protocols nonetheless rely on detergents that aid extraction of cellular components but also damage tissue architecture. It may be more beneficial to leverage an innate cellular process such as apoptosis and promote cell removal without the use of damaging reagents. During apoptosis, a cell detaches from the extracellular matrix, degrades its internal components, and fragments its contents for easier clearance. We have developed a method that leverages this process to achieve tissue decellularization using only mild wash buffers. We have demonstrated that treating peripheral nerve tissue with camptothecin induced both an early marker of apoptosis, cleaved caspase-3 expression, as well as a late stage marker, TUNEL+ DNA fragmentation. Clearance of the cellular components was then achieved in an apoptosis-dependent manner using a gentle wash in hypertonic phosphate buffered saline followed by DNase treatment. This wash paradigm did not significantly affect collagen or glycosaminoglycan content, but it was sufficient to remove any trace of the cytotoxic compound based on conditioned media experiments. The resulting acellular tissue graft was immunogenically tolerated in vivo and exhibited an intact basal lamina microarchitecture mimicking that of native, unprocessed nerve. Hence, ex vivo induction of apoptosis is a promising method to decellularize tissue without the use of harsh reagents while better preserving the benefits of native tissue such as tissue-specific composition and microarchitecture. Statement of significance Tissue decellularization has expanded the ability to generate non-immunogenic organ replacements for a broad range of health applications. Current technologies typically rely on the use of harsh agents for clearing cellular debris, altering the tissue structure and potentially diminishing the pro-regenerative effects. We have developed a method for effectively, yet gently, removing cellular components from peripheral nerve tissue while preserving the native tissue architecture. The novelty of this process is in the induction of programmed cell death - or apoptosis - via a general cytotoxin, thereby enabling antigen clearance using only hypertonic wash buffers. The resulting acellular nerve scaffolds are nearly identical to unprocessed tissue on a microscopic level and elicit low immune responses comparable to an isograft negative control in a model of subcutaneous implantation.

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  • Achieving complete cellular clearance often necessitates the inclusion of additional surfactants or enzymes, such as DNase, which can eliminate over 95% of DNA when used in conjunction with hypertonic washes.

    “Achieving complete cellular clearance often necessitates the inclusion of additional surfactants or enzymes, such as DNase, which can eliminate over 95% of DNA when used in conjunction with hypertonic washes.”

    From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [39]

    The Results report a 95% reduction in average DNA content when DNase followed a hypertonic wash in apoptosis-treated nerve tissue; the Discussion describes this as >95% DNA removal. The source does not establish that complete cellular clearance generally requires additional surfactants or enzymes, and its reported result depends on prior apoptosis induction. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • Combining DNase with detergents can achieve significant DNA clearance rates.

    “Combining DNase with detergents can achieve significant DNA clearance rates.”

    From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [39]

    The Results (Induction of apoptosis facilitates cellular antigen removal; page not available) report a significant reduction in nerve-tissue DNA after DNase treatment, including an average 95% reduction under specified conditions. But the tested combination was DNase with a hypertonic wash after apoptosis induction, not DNase with detergents; the Methods (Wash parameter optimization; page not available) explicitly say the protocol sought to avoid detergents. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • Apoptosis-assisted decellularization utilizes agents such as camptothecin to induce cell death and is especially promising for neural tissue and vascular structures.

    “Apoptosis‐assisted decellularization utilizes agents such as camptothecin to induce cell death and is especially promising for neural tissue and vascular structures.”

    From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [39]

    The Introduction and Discussion report that camptothecin induces apoptosis in peripheral nerve tissue and enables decellularization while preserving nerve architecture. The paper does not establish that the method is especially promising for vascular structures; its discussion of cardiovascular tissue concerns a different decellularization method. Page numbers are unavailable in our copy. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • Although hypertonic solutions are efficient in removing proteins and hypotonic solutions are efficient in removing nuclei and DNA, they may not completely eliminate cellular residues and may require additional treatment to facilitate removal of cellular debris.

    “Although hypertonic solutions are efficient in removing proteins and hypotonic solutions are efficient in removing nuclei and DNA, they may not completely eliminate cellular residues and may require additional treatment to facilitate the removal of cellular debris [117,119,120].”

    From Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities (2024) · cites this paper as [120]

    Split into 2 atoms — 1 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

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  • Immersion and agitation constitute a relatively straightforward decellularization method commonly used for thin tissues and smaller organs lacking vascular access.

    “Immersion and agitation constitute a relatively straightforward decellularization method commonly used for thin tissues and smaller organs lacking vascular access, such as the esophagus, small intestine submucosa, urinary bladder, blood vessels, trachea, skin, cornea, and nerve [117,119,120,126,152,[163], [164], [165], [166], [167]].”

    From Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities (2024) · cites this paper as [120]

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Authors
Cornelison, R. C., Wellman, S. M., Park, J. H., Porvasnik, S. L., Song, Y. H., Wachs, R. A., & Schmidt, C. E.
Affiliations
University of Florida (US)
Corresponding author
  • Wachs, R.A.
  • Schmidt, C.E.
ORCID
Venue
Acta Biomaterialia
Publisher
Elsevier BV
Published
2018-09
Volume
77
Pages
116-126
ISSN
1742-7061 (ISSN-L)
DOI
10.1016/j.actbio.2018.07.009
PMC
PMC6126930
OpenAlex
W2879455943
Cited by
64
References
1
Citations by year
2026: 3 · 2025: 6 · 2024: 18 · 2023: 10 · 2022: 8 · 2021: 9 · 2020: 7 · 2019: 3
FWCI
4.94
Type
journal article
Language
en
Open access
Licence
Locations
  • Acta Biomaterialia (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
National Institute of Biomedical Imaging and Bioengineering (R21EB013358)
Topics
  • Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
  • Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Nerve injury and regeneration
Keywords
decellularization, apoptosis, peripheral nerves, camptothecin, caspase-3, TUNEL, DNase treatment, tissue architecture preservation, basement membrane, collagen, glycosaminoglycans
MeSH terms
Animals, Basement Membrane, Camptothecin, Detergents, Extracellular Matrix, Glycosaminoglycans, Macrophages, Male, Nerve Tissue, Rats, Inbred Lew, Sciatic Nerve, Rats, Sprague-Dawley, Apoptosis, Peripheral Nervous System, Tissue Engineering, Rats, Caspase 3, DNA Fragmentation, Tissue Scaffolds

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