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Ex-vivo Normothermic Limb Perfusion Preserves Limb Viability and Function

Eliana F R Duraes, Maria Madajka, Basem Soliman, Cagri Cakmakoglu, Stephanie Kortyka, Addison Barnett, Russell Frautschi, Kashyap Tadisina, Qiang Liu, Toshihiro Okamoto, Cristiano Quintini, Francis Papay, Antonio Rampazzo, Bahar Bassiri Gharb
Cleveland Clinic Foundation
2016-02-01

Presenter: Eliana F R Duraes

Affidavit:
Dr. Eliana Duraes was involved in all the steps of this study.

Director Name: Steven Bernard

Author Category: Fellow Plastic Surgery
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction

Purpose – The aim of this study was to establish an ex-vivo limb perfusion protocol, as an alternative to cold storage, decreasing the ischemia time following amputation and preserving viability and function for replantation and transplantation.
Method – Porcine forelimbs were perfused using a circuit driven by a roller pump. Circuit and solution parameters were progressively adjusted to achieve 12 hours of perfusion maintaining muscle contractility(MsC), normal compartment pressure(CP), minimal percentage of weight gain, physiologic arterial pressure(BP), pH, glucose, and electrolyte levels. The final protocol included: perfusion at 39oC using an oxygenated colloid solution mixed with washed red blood cells. Partial perfusate exchange was introduced to counteract electrolyte derangement and hemolysis. Perfusion quality was assessed by tissue histology, indocyanine green angiography, thermography, tissue oximetry using near infrared spectroscopy, MsC, glucose and oxygen consumption, and levels of myoglobin, lactate, and creatine kinase(CK).
Results - 18 limbs were perfused, 5 with the final established protocol. Of these 5, all reached the 12 hour target with preserved MsC; four had no weight gain. During the perfusion, muscle temperature, 33.52±1.88oC; K+, 5.66±1.61 mmol/ml; Na+, 162.5±9.25 mmol/L; lactate, 9.15±4.07 mmol/L; hemoglobin 4.37±0.89 g/dL; arterial oxygen partial pressure, 133.93±46.6 mmHg; pulse oximetry, 91.9±7.38%; and tissue oximetry readings, 58.67±9.56%. ICG angiography showed uniform perfusion in three limbs, and areas of hypo perfusion in the other two limbs. Average values of final CP, myoglobin, and CK were 16.25±7.10 mmHg, 875±291.4 ng/mL, and 53344±14850.34 U/L, respectively.
Conclusions – Ex-vivo normothermic limb perfusion preserved limb physiology and function for at least 12 hours.

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