The PhysioHeart™ is a mature acute platform, based isolated slaughterhouse hearts and able to validate cardiac devices and techniques in working mode. Despite perfusion, myocardial edema and time-dependent function degradation are reported. Therefore, monitoring several variables is necessary to identify which of these should be controlled to preserve the heart function. This study presents biochemical, electrophysiological and hemodynamic changes in the PhysioHeart™ to understand the pitfalls of ex vivo slaughterhouse heart hemoperfusion.

Seven porcine hearts were harvested, arrested and revived using the PhysioHeart™. Cardiac output, SaO2, glucose and pH were maintained at physiological levels. Blood analyses were performed hourly and unipolar epicardial electrograms (UEG), pressures and flows were recorded to assess the physiological performance.

Normal cardiac performance was attained in terms of mean cardiac output (5.1 ± 1.7 l/min) and pressures but deteriorated over time. Across the experiments, homeostasis was maintained for 171.4 ± 54 min, osmolarity and blood electrolytes increased significantly between 10 and 80%, heart weight increased by 144 ± 41 g, free fatty acids (− 60%), glucose and lactate diminished, ammonia increased by 273 ± 76% and myocardial necrosis and UEG alterations appeared and aggravated. Progressively deteriorating electrophysiological and hemodynamic functions can be explained by reperfusion injury, waste product intoxication (i.e. hyperammonemia), lack of essential nutrients, ion imbalances and cardiac necrosis as a consequence of hepatological and nephrological plasma clearance absence.

The PhysioHeart™ is an acute model, suitable for cardiac device and therapy assessment, which can precede conventional animal studies. However, observations indicate that ex vivo slaughterhouse hearts resemble cardiac physiology of deteriorating hearts in a multi-organ failure situation and signalize the need for plasma clearance during perfusion to attenuate time-dependent function degradation. The presented study therefore provides an in-dept understanding of the sources and reasons causing the cardiac function loss, as a first step for future effort to prolong cardiac perfusion in the PhysioHeart™. These findings could be also of potential interest for other cardiac platforms.

• Biomarkers
• Cardiac electrophysiology
• Cardiac physiology
• Ex vivo
• Normothermic perfusion

To compare the effect of University of Wisconsin (UW) solution with or without metformin, an AMP-activated protein kinase (AMPK) activator, for preserving standard and marginal liver grafts of young and aged rats ex vivo by hypothermic machine perfusion (HMP).

Eighteen young (4 mo old) and 18 aged (17 mo old) healthy male SD rats were selected and randomly divided into three groups: control group, UW solution perfusion group (UWP), and UW solution with metformin perfusion group (MUWP). Aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), interleukin-18 (IL-18), and tumor necrosis factor-alpha (TNF-α) in the perfused liquid were tested. The expression levels of AMPK and endothelial nitric oxide synthase (eNOS) in liver sinusoidal endothelial cells were also examined. Additionally, microscopic evaluation of the harvested perfused liver tissue samples was done.

AST, ALT, LDH, IL-18 and TNF-α levels in the young and aged liver-perfused liquid were, respectively, significantly lower in the MUWP group than in the UWP group (P < 0.05), but no significant differences were found between the young and aged MUWP groups. Metformin increased the expression of AMPK and eNOS protein levels, and promoted the extracellular release of nitric oxide through activation of the AMPK-eNOS mediated pathway. Histological examination revealed that in the MUWP group, the extent of liver cells and tissue damage was significantly reduced compared with the UWP group.

The addition of metformin to the UW preservative solution for ex vivo HMP can reduce rat liver injury during cold ischemia, with significant protective effects on livers, especially of aged rats.

• Metformin
• AMP-activated protein kinase
• Cold ischemia injury
• Hypothermic machine perfusion
• Liver Grafts

• AMP-Activated Protein Kinases/metabolism
• Adenosine/pharmacology
• Alanine Transaminase/analysis
• Allopurinol/pharmacology
• Animals
• Aspartate Aminotransferases/analysis
• Cold Ischemia/adverse effects
• Glutathione/pharmacology
• Hepatocytes/drug effects
• Hepatocytes/pathology
• Hepatocytes/ultrastructure
• Humans
• Infusion Pumps
• Insulin/pharmacology
• L-Lactate Dehydrogenase/analysis
• Liver/cytology
• Liver/drug effects
• Liver/enzymology
• Liver Transplantation/adverse effects
• Liver Transplantation/methods
• Male
• Metformin/pharmacology
• Microscopy, Electron, Transmission
• Models, Animal
• Nitric Oxide Synthase Type III/metabolism
• Organ Preservation/methods
• Organ Preservation Solutions/pharmacology
• Perfusion/instrumentation
• Perfusion/methods
• Raffinose/pharmacology
• Rats
• Rats, Sprague-Dawley
• Reperfusion Injury/etiology
• Reperfusion Injury/prevention & control
• Tissue and Organ Harvesting/adverse effects
• Tissue and Organ Harvesting/methods

• aging
• donation after circulatory death
• heart transplantation
• immunosuppression
• ischemia-reperfusion injury
• rejection

• Cardioprotection
• Cold storage
• Donor heart preservation
• Myocardial ischaemia–reperfusion injury
• Preservation solutions

Hearts preserved ex vivo at 4°C undergo time-dependent irreversible injury due to extreme hypothermia. Studies using novel organ preservative solution SOMAH, suggest that hearts are optimally `preserved' at subnormothermic temperature of 21°C. Present study evaluates relative efficacy of SOMAH `cardioplegia' at 4 and 21°C in preservation of optimum heart function after in vitro storage at subnormothermia.

Porcine hearts arrested with SOMAH cardioplegia at 4 or 21°C were stored in SOMAH for 5-hour at 21°C (n = 5). At the end of storage, the weight of hearts was recorded and biopsies taken for cardiac tissue high energy phosphate level measurements. The hearts were then attached to a reperfusion apparatus and biochemical parameters including cardiac enzyme release and myocardial oxygen consumption and lactate production were determined in perfusate samples at regular intervals during ex vivo perfusion experiment. Functional evaluation of the hearts intraoperatively and ex vivo was performed by 2D echocardiography using trans-esophageal echocardiography probe.

Post-storage heart weights were unaltered in both groups, while available high-energy phosphates (HEP) were greater in the 21°C group. Upon ex vivo reperfusion, coronary flow was significantly greater (p < 0.05) in 21°C group. 2D echo revealed a greater cardiac output, fractional area change and ejection fraction in 21°C group that was not significantly different than the 4°C group. However, unlike 4°C hearts, 21°C hearts did not require inotropic intervention. Upon reperfusion, rate of cardiac enzyme release temporally resolved in 21°C group, but not in the 4°C group. 21°C working hearts maintained their energy state during the experimental duration but not the 4°C group; albeit, both groups demonstrated robust metabolism and function during this period.

Rapid metabolic switch, increased synthesis of HEP, decreased injury and optimal function provides evidence that hearts arrested at 21°C remain viably and functionally superior to those arrested at 4°C when stored in SOMAH at ambient temperature pre-transplant.

Cardioplegic arrest and preservation of hearts in SOMAH at ambient temperature efficiently conserves metabolism and function in in vitro porcine model of heart transplant.

The online version of this article (doi:10.1186/s13019-014-0155-z) contains supplementary material, which is available to authorized users.