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Ex vivo Normothermic Limb Perfusion with an HBOC-201 Based Perfusate.
Sayf Said, Carlos Ordeñana, Majid Razaie, Brian Figueroa, Vahe Fahradyan, Michael Annunziata, Lynn Orfahli, Antonio Rampazzo, Bahar Bassiri.
Research center training program.
2019-02-15
Presenter: Sayf Said, MD
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: Bahar Bassiri, MD PhD
Author Category: Fellow Plastic Surgery
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction
Background:
Ex-Vivo Normothermic Limb Perfusion (EVNLP) provide better viability and function preservation than cold storage. Perfusates containing Red Blood Cells (pRBC) as an oxygen carrier improve outcomes when compared to acellular perfusates. However, limitations of pRBC-based perfusion are scarcity, need for cross-matching and hemolysis. Hemopure is a polymerized bovine Hemoglobin-Based Oxygen Carrier (HBOC) of low immunogenicity, no risk of hemolytic reactions, lower viscosity, and a higher oxygen-carrying capacity. Here we investigate the feasibility of using HBOC as an oxygen carrier in EVNLP.
Methods:
Eight porcine limbs were procured, four of which were perfused using our established EVNLP model and a Hemopure-based perfusate, the contralateral limbs were kept in cold storage as the control. A custom-made circuit system composed of a roller pump, oxygenator, heat exchanger, a reservoir, and a dome cover. Electrolytes, limb temperature, weight, compartment pressure, nerve conduction, and perfusion, indicated by indocyanine green (ICG) angiography and Infra-Red (IR) thermography, were monitored. Histologic evaluation of muscle tissue was performed with Hematoxylin and Eosin and electron microscopy.
Results:
The perfusion duration was 21.75±1.9 hours. Muscle contractility was preserved for 17±6.1 hours. There was no muscle contraction immediately after amputation in the control limbs. IR thermography and ICG angiography confirmed the preservation of peripheral perfusion throughout the experiments. Additionally, preservation of the mitochondrial ultrastructure was evident at 12 hours in contrast to crystallization and destruction features in the control samples.
Conclusion:
Hemopure based perfusion has similar outcomes to RBC based perfusion with preservation of muscle contractility and mitochondrial structure.
