Hypothermic Ex Situ Perfusion Of Human Limbs With Acellular Solution For 24 Hours
Valentin F. M. Haug, M.D..
Brigham and Women's Hospital, Boston, MA, USA.
To evaluate the benefit of extracorporeal machine perfusion (MP) of human upper extremities with an oxygenated colloid-enriched acellular solution for 24 hours compared to the current standard of care, static cold storage (SCS).
Six human upper extremities were procured and assigned to either hypothermic MP (n=3), or SCS (n=3) for 24 hours. Perfusion solution samples were collected at 2-hourly intervals for analysis of muscle injury biomarkers. Muscle biopsy samples were analyzed for protein expression of hypoxia-inducible factor alpha (HIF-1α) and used for histological analysis. Flow rates, pressure, temperature, and oxygen partial pressure were continuously recorded.
Mean ischemia time before starting MP was 187±52 minutes. Myoglobin, creatine-kinase (CK) and lactate showed increased levels at the start of MP (myoglobin:
6064±3486 ng/ml, CK: 2086±1542 U/L, lactate: 6.6±1.2 mmol/L), peaking after six hours (myoglobin: 9268±1138 ng/ml, CK: 4802±3370 U/L) and plateauing for the rest of the experiment. Lactate levels decreased to 2.8±1.1 over time. Expression of HIF-1α peaked in the SCS-group after 8 hours, followed by a decrease, most likely to cell death. Increased HIF-1α expression in the MP-group was delayed until 20 hours. Perfusion pressure, temperature and circuit flow were maintained at mean of 30.88 mmHg, 9.77°Celsius, and 31.13 ml/min, respectively. Limb weight increased on average 1.4% in the SCS-group and 4.3% in the MP-group over 24 hours.
Hypothermic machine perfusion with an oxygenated acellular solution may be a promising approach to extend the allowable ex-vivo preservation of human limbs compared to SCS.
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