De-Epithelialization Protocol with Tapered Sodium Dodecyl Sulfate Concentrations Enhances Short-Term Chondrocyte Survival in Porcine Chimeric Tracheal Allografts

Authors

  • Kevin Xiang Zhou BMSc (Hons.). Schulich School of Medicine and Dentistry, 1151 Richmond St, London, ON N6A 5C1, Canada https://orcid.org/0000-0003-0332-9947
  • Fabio Gava Aoki PhD, MASc. Latner Thoracic Surgery Research Laboratories, Division of Thoracic Surgery, University Health Network, 101 College St., Toronto, ON M5G 1L7, Canada https://orcid.org/0000-0003-3387-5121
  • Alba Marin MD. Latner Thoracic Surgery Research Laboratories, Division of Thoracic Surgery, University Health Network, 101 College St., Toronto, ON M5G 1L7, Canada
  • Golnaz Karoubi PhD. Latner Thoracic Surgery Research Laboratories, Division of Thoracic Surgery, University Health Network, 101 College St., Toronto, ON M5G 1L7, Canada https://orcid.org/0000-0003-3764-3379
  • Siba Haykal MD, PhD, FRCS(C), FACS. Latner Thoracic Surgery Research Laboratories, Division of Thoracic Surgery, University Health Network, 101 College St., Toronto, ON M5G 1L7, Canada https://orcid.org/0000-0002-4247-2810
  • Thomas K. Waddell MD, PhD, MSc, FRCSC. Latner Thoracic Surgery Research Laboratories, Division of Thoracic Surgery, University Health Network, 101 College St., Toronto, ON M5G 1L7, Canada https://orcid.org/0000-0002-3235-9208

DOI:

https://doi.org/10.5195/ijms.2023.1437

Keywords:

Tissue Engineering, Decellularization, Allograft, Trachea, Bioreactor, Regenerative Medicine, Chondrocyte, Stem Cell, Graft, Transplantation, Transplant, Surgery, Bioengineering, Stenosis, Cartilage, Viability, Cell Viability, Medicine

Abstract

Background: Tracheal transplantation is indicated in cases where injury exceeds 50% of the organ in adults and 30% in children. However, transplantation is not yet considered a viable treatment option partly due to high morbidity and mortality associated with graft rejection. Recently, decellularization (decell) has been explored as a technique for creating bioengineered tracheal grafts. However, risk of post-operative stenosis increases due to the death of chondrocytes, which are critical to maintain the biochemical and mechanical integrity of tracheal cartilage. In this project, we propose a new de-epithelialization protocol that adequately removes epithelial, mucosal, and submucosal cells while maintaining a greater proportion of viable chondrocytes.

Methods: The trachea of adult male outbred Yorkshire pigs were extracted, decontaminated, and decellularized according to the original and new protocols before incubation at 37 °C in DMEM for 10 days. Chondrocyte viability was quantified immediately following post-decellularization and on days 1, 4, 7, and 10. Histology was performed pre-decellularization, post-decellularization, and post-incubation.

Results: The new protocol showed a significant (p < 0.05) increase in chondrocyte viability up to four days after de-ep when compared to the original protocol. We also found that the new protocol preserves ECM composition to a similar degree as the original protocol. When scaffolds created using the new protocol were re-epithelialized, cell growth curves were near identical to published data from the original protocol.

Conclusion: While not without limitations, our new protocol may be used to engineer chimeric tracheal allografts without the need for cartilage regeneration.

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Published

2023-03-08 — Updated on 2023-03-31

How to Cite

Zhou, K. X., Aoki, F. G., Marin, A., Karoubi , G., Haykal, S., & Waddell , T. K. (2023). De-Epithelialization Protocol with Tapered Sodium Dodecyl Sulfate Concentrations Enhances Short-Term Chondrocyte Survival in Porcine Chimeric Tracheal Allografts. International Journal of Medical Students, 11(1), 13–21. https://doi.org/10.5195/ijms.2023.1437

Issue

Vol. 11 No. 1 (2023)

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Original Article

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Copyright (c) 2023 Kevin Xiang Zhou, Fabio Gava Aoki, Alba Marin, Golnaz Karoubi , Siba Haykal, Thomas K. Waddell

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