Vacuum-assisted decellularization: an accelerated protocol to generate tissue-engineered human tracheal scaffolds

Colin R. Butler, Robert E. Hynds, Claire Crowley, Kate H C Gowers, Leanne Partington, Nicholas J. Hamilton, Carla Carvalho, Manuela Platé, Edward R. Samuel, Alan J. Burns, Luca Urbani, Martin A. Birchall, Mark W. Lowdell, Paolo De Coppi, Sam M. Janes

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)95-105
Number of pages11
JournalBiomaterials
Volume124
DOIs
StatePublished - Apr 1 2017
Externally publishedYes

Keywords

  • Epithelial cells
  • Tissue engineering
  • Tissue scaffolds
  • Trachea
  • Transplantation

ASJC Scopus subject areas

  • Bioengineering
  • Ceramics and Composites
  • Biophysics
  • Biomaterials
  • Mechanics of Materials

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  • Cite this

    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. (2017). Vacuum-assisted decellularization: an accelerated protocol to generate tissue-engineered human tracheal scaffolds. Biomaterials, 124, 95-105. https://doi.org/10.1016/j.biomaterials.2017.02.001