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&Estaining 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 language | English (US) |
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Pages (from-to) | 100-107 |
Number of pages | 8 |
Journal | International Journal of Artificial Organs |
Volume | 41 |
Issue number | 2 |
DOIs | |
State | Published - Sep 1 2018 |
Keywords
- Airway
- Decellularization
- Detergent
- Regenerative
- Scaffold
ASJC Scopus subject areas
- Medicine (miscellaneous)
- Bioengineering
- Biomaterials
- Biomedical Engineering