Tracheal Decellularization using a Combination of Chemical, Physical and Bioreactor Methods

Yourka D. Tchoukalova, Justin M. Hintze, Richard E Hayden, David G Lott

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Introduction: The purpose of this study was to compare different decellularization protocols with the conventional detergent enzymatic method (DEM) using continuous agitation. Methods: The first experiment compared conventional DEM with sonication and lyophilization+freeze-thaw cycles. A second experiment was carried out to compare time-adjusted DEM (2-hour instead of 4-hour incubations with 4% deoxycholate) to decellularization in a bioreactor. Cellularity was determined by DNA-quantitation, H&E-staining and immunostaining for major histocompatibility complex-1 (MHC-1). Results: Compared to untreated trachea, DNA content significantly decreased after 2 cycles in all groups in the first experiment and dropped below the minimal criteria for efficient decellularization (<50 ng dsDNA/mg dry weight) after 4 cycles. However, nuclei were seen in the cartilage and MCH-1 staining was detected in some submucosal areas, indicating presence of chondrocytes and cellular residues that may render the scaffold immunogenic. In the second experiment DNA content significantly decreased after 1 cycle in both groups; however, even after 4 cycles, DNA content was above the minimal criteria for efficient decellularization. While collagen-levels remained stable, glycosaminoglycans diminished significantly after the initial cycles. Conclusions: Efficient decellularization can be achieved after only 4 cycles of DEM compared to the 17 cycles previously reported. The use of a bioreactor can preserve the integrity of the extracellular matrix.

Original languageEnglish (US)
JournalInternational Journal of Artificial Organs
DOIs
StateAccepted/In press - Sep 1 2017

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Detergents
Bioreactors
DNA
Experiments
Deoxycholic Acid
Sonication
Staining and Labeling
Cartilage
Glycosaminoglycans
Collagen
Scaffolds
Freeze Drying
Chondrocytes
Trachea
Major Histocompatibility Complex
Extracellular Matrix
Weights and Measures

Keywords

  • Airway
  • Decellularization
  • Detergent
  • Regenerative
  • Scaffold

ASJC Scopus subject areas

  • Bioengineering
  • Medicine (miscellaneous)
  • Biomaterials
  • Biomedical Engineering

Cite this

Tracheal Decellularization using a Combination of Chemical, Physical and Bioreactor Methods. / Tchoukalova, Yourka D.; Hintze, Justin M.; Hayden, Richard E; Lott, David G.

In: International Journal of Artificial Organs, 01.09.2017.

Research output: Contribution to journalArticle

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