Vacuum-assisted decellularization: an accelerated protocol to generate tissue-engineered human tracheal scaffolds
Open accessAbstract
Patients with large tracheal lesions unsuitable for conventional endoscopic or open operations may require a tracheal replacement but there is no present consensus of how this may be achieved. Tissue engineering using decellularized or synthetic tracheal scaffolds offers a new avenue for airway reconstruction. Decellularized human donor tracheal scaffolds have been applied in compassionate-use clinical cases but naturally derived extracellular matrix (ECM) scaffolds demand lengthy preparation times. Here, we compare a clinically applied detergent-enzymatic method (DEM) with an accelerated vacuum-assisted decellularization (VAD) protocol. We examined the histological appearance, DNA content and extracellular matrix composition of human donor tracheae decellularized using these techniques. Further, we performed scanning electron microscopy (SEM) and biomechanical testing to analyze decellularization performance. To assess the biocompatibility of scaffolds generated using VAD, we seeded scaffolds with primary human airway epithelial cells in vitro and performed in vivo chick chorioallantoic membrane (CAM) and subcutaneous implantation assays. Both DEM and VAD protocols produced well-decellularized tracheal scaffolds with no adverse mechanical effects and scaffolds retained the capacity for in vitro and in vivo cellular integration. We conclude that the substantial reduction in time required to produce scaffolds using VAD compared to DEM (approximately 9 days vs. 3-8 weeks) does not compromise the quality of human tracheal scaffold generated. These findings might inform clinical decellularization techniques as VAD offers accelerated scaffold production and reduces the associated costs.
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Vacuum-assisted decellularization, employing negative pressure to expedite the process, can be utilized in conjunction with other techniques to enhance outcomes.
“Vacuum‐assisted decellularization, employing negative pressure to expedite the process, can be utilized in conjunction with other techniques to enhance outcomes.”
From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [67]
The Materials and methods section describes applying a vacuum of 1 Torr alongside detergent and enzyme treatments. The Discussion reports that this protocol shortened preparation of human tracheal scaffolds to nine days, versus 3–8 weeks for the comparator, while producing comparable scaffold quality. It does not establish that combining vacuum assistance with other techniques improves scaffold outcomes beyond that acceleration. (LLM, conf=0.85)
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- Authors
- Butler, C. R., Hynds, R. E., Crowley, C., Gowers, K. H. C., Partington, L., Hamilton, N. J., Carvalho, C., Platé, M., Samuel, E. R., Burns, A. J., Urbani, L., Birchall, M. A., Lowdell, M. W., De Coppi, P., & Janes, S. M.
- Affiliations
- Great Ormond Street Hospital (GB)
- University College London (GB)
- The Royal Free Hospital (GB)
- Royal National Throat Nose and Ear Hospital (GB)
- Corresponding author
- Janes, Sam M.
- ORCID
- Venue
- Biomaterials
- Publisher
- Elsevier BV
- Published
- 2017-04
- Volume
- 124
- Pages
- 95-105
- ISSN
- 0142-9612 (ISSN-L)
- PMC
- PMC5332556
- OpenAlex
- W2587160918
- Cited by
- 99
- References
- 46
- Citations by year
- 2026: 4 · 2025: 10 · 2024: 10 · 2023: 14 · 2022: 7 · 2021: 20 · 2020: 12 · 2019: 9 · 2018: 5 · 2017: 8
- FWCI
- 9.53
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Biomaterials (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- EUR Research Repository (Erasmus University Rotterdam) (publishedVersion)
- EUR Research Repository (Erasmus University Rotterdam) (submittedVersion)
- RePub (Erasmus University Rotterdam) (submittedVersion, other-oa)
- UCL Discovery (University College London) (submittedVersion)
- RePub (Erasmus University Rotterdam) (submittedVersion)
- Funders
- MRC Regenvox (G1001539)
- MRC-funded RegenVOX II clinical trial (MR/K026453/1)
- Wellcome Senior Fellow in Clinical Science (WT107963AIA)
- The Roy Castle Lung Cancer Foundation
- Charitable Foundation
- Topics
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Tracheal and airway disorders
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Keywords
- decellularized extracellular matrix, tracheal replacement, airway reconstruction, biocompatibility, chick chorioallantoic membrane assay, subcutaneous implantation, tissue mechanics, DNA content
- MeSH terms
- Cell Fractionation, Cell-Free System, Cells, Cultured, Equipment Design, Extracellular Matrix, Humans, Trachea, Vacuum, Equipment Failure Analysis, Tissue Engineering, Tissue Scaffolds
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