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Optimizing the Decellularization of the Rodent Epigastric Free Flap: A Comparison of Automated SDS-Based Protocols
Benjamin K. Schilling, MS; Lei Chen, MD; Chiaki Komatsu, MD; Fuat Baris Bengur, MD; Kacey G. Marra, PhD; Lauren E. Kokai, PhD; Mario G. Solari, MD
University of Pittsburgh
2021-02-10
Presenter: Fuat Baris Bengur
Affidavit:
I certify that the material proposed for presentation in this abstract has not been published in any scientific journal or previously presented at a major meeting.
Director Name: Mario G Solari
Author Category: Fellow Plastic Surgery
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction
Purpose:
Though flaps can provide reliable reconstruction of complex wounds, they come at the cost of donor site morbidity. One tissue engineering strategy to avoid a donor site is flap decellularization. Sodium dodecyl sulfate (SDS)-based decellularization protocols are being used to remove whole cells to create a scaffold with an intact vascular network. This study aims to optimize the protocol for automated decellularization and scaffold preservation.
Methods:
A 3D-printed closed-system bioreactor capable of continuously perfusing fluid throughout the vasculature was used for decellularization. Three rat epigastric free flaps were evaluated in each group. The artery and vein were cannulated and a 1% SDS solution was perfused for different durations followed by 1 day of 1% Triton X-100 and 1 day of 1xPhosphate-buffered saline.
Results:
Histology was performed to assess architecture and locale of residual nuclei. Residual DNA was quantified by the fluorescent marker PicoGreen. 5 days of 1%SDS solution had the least residual DNA content followed by 10 days and 3 days (p<0.001). The DNA content ratio of skin over subcutaneous tissues was consistent across all protocols with skin having twice as much residual DNA after each protocol. The vascular network was visualized for qualitative assessment with the perfusion of hardening cast.
Conclusions:
The bioreactor is capable of running automated and repeatable protocols using several solutions and continuously perfusing fluid throughout the vasculature of a free flap. This compact and integrated system can decrease hands-on time and be used in the future for further recellularization of scaffolds.
