Preservation Of Rat Hearts Research Articles

Hypothermic preservation of rat hearts using antifreeze glycoprotein.

Antifreeze proteins are an effective additive for low-temperature preservation of solid organs. Here, we compared static hypothermic preservation with and without antifreeze glycoprotein (AFGP), followed by nonfreezing cryopreservation of rat hearts. The heart was surgically extracted and immersed in one of the cardioplegia solutions after cardiac arrest. Control rat hearts (n=6) were immersed in University of Wisconsin (UW) solution whereas AFGP-treated hearts (AFGP group) (n=6) were immersed in UW solution containing 500 ?g/ml AFGP. After static hypothermic preservation, a Langendorff apparatus was used to reperfuse the coronary arteries with oxygenated Krebs-Henseleit solution. After 30, 60, 90, and 120 min, the heart rate (HR), coronary flow (CF), cardiac contractile force (max dP/dt), and cardiac diastolic force (min dP/dt) were measured. Tissue water content (TWC) and tissue adenosine triphosphate (ATP) levels in the reperfused preserved hearts were also assessed. All the parameters were compared between the control and AFGP groups. Compared with the control group, the AFGP group had significantly (p<0.05) higher values of the following parameters: HR at 60, 90, and 120 min; CF at all four time points; max dP/dt at 90 min; min dP/dt at 90 and 120 min; and tissue ATP levels at 120 min. TWC did not differ significantly between the groups. The higher HR, CF, max dP/dt, min dP/dt, and tissue ATP levels in the AFGP compared with those in control hearts suggested that AFGP conferred superior hemodynamic and metabolic functions. Thus, AFGP might be a useful additive for the static/nonfreezing hypothermic preservation of hearts.

Cardioprotection via Metabolism for Rat Heart Preservation Using the High-Pressure Gaseous Mixture of Carbon Monoxide and Oxygen.

The high-pressure gas (HPG) method with carbon monoxide (CO) and oxygen (O2) mixture maintains the preserved rat heart function. The metabolites of rat hearts preserved using the HPG method (HPG group) and cold storage (CS) method (CS group) by immersion in a stock solution for 24 h were assessed to confirm CO and O2 effects. Lactic acid was significantly lower and citric acid was significantly higher in the HPG group than in the CS group. Moreover, adenosine triphosphate (ATP) levels as well as some pentose phosphate pathway (PPP) metabolites and reduced nicotinamide adenine dinucleotide phosphate (NADPH) were significantly higher in the HPG group than in the CS group. Additionally, reduced glutathione (GSH), which protects cells from oxidative stress, was also significantly higher in the HPG group than in the CS group. These results indicated that each gas, CO and O2, induced the shift from anaerobic to aerobic metabolism, maintaining the energy of ischemic preserved organs, shifting the glucose utilization from glycolysis toward PPP, and reducing oxidative stress. Both CO and O2 in the HPG method have important effects on the ATP supply and decrease oxidative stress for preventing ischemic injury. The HPG method may be useful for clinical application.

Heavy Water (D2O) Containing Preservation Solution Reduces Hepatic Cold Preservation and Reperfusion Injury in an Isolated Perfused Rat Liver (IPRL) Model.

Background: Heavy water (D2O) has many biological effects due to the isotope effect of deuterium. We previously reported the efficacy of D2O containing solution (Dsol) in the cold preservation of rat hearts. Here, we evaluated whether Dsol reduced hepatic cold preservation and reperfusion injury. Methods: Rat livers were subjected to 48-hour cold storage in University of Wisconsin (UW) solution or Dsol, and subsequently reperfused on an isolated perfused rat liver. Graft function, injury, perfusion kinetics, oxidative stress, and cytoskeletal integrity were assessed. Results: In the UW group, severe ischemia and reperfusion injury (IRI) was shown by histopathology, higher liver enzymes leakage, portal resistance, and apoptotic index, oxygen consumption, less bile production, energy charge, and reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio (versus control). The Dsol group showed that these injuries were significantly ameliorated (versus the UW group). Furthermore, cytoskeletal derangement was progressed in the UW group, as shown by less degradation of α-Fodrin and by the inactivation of the actin depolymerization pathway, whereas these changes were significantly suppressed in the Dsol group. Conclusion: Dsol reduced hepatic IRI after extended cold preservation and subsequent reperfusion. The protection was primarily due to the maintenance of mitochondrial function, cytoskeletal integrity, leading to limiting oxidative stress, apoptosis, and necrosis pathways.

Different effects of partial pressure in a high-pressure gaseous mixture of carbon monoxide and oxygen for rat heart preservation

We maintained the function of an extracted rat heart after 24–48 h preservation in a high-pressure gaseous mixture of carbon monoxide (CO) and oxygen (O2). Here, we assessed the effects of different partial pressures of hyperbaric CO and O2 for 24–48 h at 4 °C on rat heart preservation and compared conditions including immersion in University of Wisconsin solution. Preserved hearts were transplanted into recipient rats via heterotopic cervical heart transplantation for in vivo evaluation and perfused using the Langendorff system for ex vivo evaluation. The survival rate of transplanted hearts was 100% at postoperative day 7 in the CO + O2 (PCO:PO2 = 1.5:2.0 atm) group but only 33% in the CO + O2 (PCO:PO2 = 2.0:1.5 atm) group. Langendorff system and histopathological analysis revealed that the left ventricular pressure of preserved hearts in the CO + O2 (PCO:PO2 = 1.5:2.0 atm) group was better than the CO + O2 (PCO:PO2 = 2.0:1.5 atm). We demonstrate that exposure of rat hearts to hyperbaric CO and O2 is superior to the immersion method and that partial pressure of hyperbaric CO and O2 is crucial to preservation.

The Snow Flea Antifreeze Protein and Cryopreservation

Hypogastrura harveyi (snow fleas) produce a glycine‐rich antifreeze protein which enables it to survive in environments that would otherwise freeze its body fluid. Snow flea antifreeze protein (sfAFP) is a 6.5 kDa monomeric protein made of six alpha helices linked by hydrogen bonds and disulfide bridges. The protein's amino acid sequence has a frequent glycine‐x‐y amino acid pattern, where the x amino acid is also glycine 50% of the time and the y amino acid is either hydrophobic or hydrophilic. The hydrophobic side of sfAFP binds to the surface of seed ice crystals, breaking up the ice surface into smaller areas and preventing the further binding of water molecules due to the Kelvin effect. Other AFP's have been shown to be efficacious in improving the preservation of rat hearts, leading to the yet‐untested hypothesis that sfAFP's are also able to prolong the cryopreservation of organs. The Ashbury MSOE Center for BioMolecular Modeling SMART Team used 3‐D modeling and printing technology to examine structure‐function relationships of sfAFP when bonding to an ice lattice.Support or Funding InformationMSOE ‐ Center for Biomolecular Modelling ‐ co‐sponsor Diane MunzenmaierThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Preservation of rat hearts in subfreezing temperature isochoric conditions to – 8 °C and 78 MPa

Isochoric (constant volume) preservation at subfreezing temperatures is being investigated as a novel method for preserving cells and organs. This study is a first initial effort to evaluate the efficacy of this method for heart preservation, and to provide a preliminary outline of appropriate preservation parameters. To establish a baseline for further studies, rat hearts were preserved in a University of Wisconsin (UW) intracellular solution for one hour under isochoric conditions at: 0 °C (atmospheric pressure - 0.1 MPa), - 4 °C (41 MPa), - 6 °C (60 MPa) and – 8 °C (78 MPa). The viability of the heart was evaluated using Langendorff perfusion and histological examination. The physiological performance of hearts preserved at – 4 °C (41 MPa) was comparable to that of a heart preserved on ice at atmospheric pressure, with no statistically significant difference in histological injury score. However, hearts preserved at −4 °C displayed substantially reduced interstitial edema compared to hearts preserved by conventional hypothermic preservation in UW on ice at atmospheric pressure, suggesting significant protection from increased vascular permeability following preservation. Hearts preserved at – 6 °C (60 MPa) suffered injury from cellular swelling and extensive edema, and at – 8 °C (78 MPa) hearts experienced significant morphological disruption. To the best of our knowledge, this is the first publication showing that a mammalian organ can survive low subfreezing temperatures without the use of a cryoprotective additive. Lowering the preservation temperature reduces metabolism and improves preservation quality, and these results suggest that improvements in preservation are possible at subzero temperatures with low to moderate pressures observed at −4 °C. Notably, tissue damage was observed at lower temperatures (−6 °C or below) accompanying further elevation of pressure associated with isochoric preservation that may prove detrimental. Therefore, subfreezing temperature isochoric preservation protocols should optimize, a combination of temperature and pressure that will minimize the negative effects of elevated pressure while retaining the beneficial effect of lower temperatures and reduced metabolism.

Subzero 24-hr Nonfreezing Rat Heart Preservation

A new supercooling system using a variable magnetic field has been recently developed. Subzero nonfreezing preservation has been believed to be a beneficial method because of the lower metabolic rate. The purpose of this study was to evaluate the hemodynamic and metabolic effects of rat heart preservation in a variable magnetic field without cryoprotectants. Rat hearts were perfused ex vivo for 120 min after 24-hr preservation in two groups (n=6 each): (1) conventional storage group, in which the hearts were stored at 4°C, and (2) the subzero group, in which the hearts were preserved at -3°C in a variable magnetic field. Reperfusion cardiac performance after preservation was significantly preserved in the subzero group compared with the conventional group with respect to heart rate, coronary flow, the peak positive dP/dt, and the peak negative dP/dt (P<0.05). Edema after reperfusion was significantly decreased (P<0.05), and the adenosine triphosphate level was higher in the subzero group (P<0.05). The rat hearts preserved in a variable magnetic field at -3°C showed better hemodynamic and metabolic performance than those preserved using conventional storage at 4°C.

Heat shock protein 90 mediates anti-apoptotic effect of diazoxide by preventing the cleavage of Bid in hypothermic preservation rat hearts

Successful organ preservation is the premise for clinical organ transplantation. The present study investigated whether heat shock protein 90 (Hsp90) is important in the anti-apoptotic effect of diazoxide in hypothermic preservation rat hearts. Isolated rat hearts were preserved in Celsior solution, with or without diazoxide, for 3 to 9 hours, followed by 60 minutes of reperfusion. Cell apoptosis was assessed by terminal deoxynucleotide transferase-mediated deoxy uridine triphosphate nick-end labeling. The left ventricular developed pressure (LVDP) was recorded. Expression of Hsp90 protein and cleavage of Bid were detected by Western blot and polymerase chain reaction. After hypothermic preservation for 3 to 9 hours, the LVDP recovery rate significantly decreased and cardiomyocyte apoptosis index increased in a time-dependent manner. When compared with the 9-hour preservation group, Celsior solution supplemented with diazoxide significantly enhanced the LVDP recovery rate and decreased the apoptosis index. The cleavage of Bid increased after 9 hours of hypothermic preservation, which was inhibited by Celsior solution supplemented with diazoxide. Hypothermic preservation of rat hearts for 9 hours decreased the expression of Hsp90, whereas diazoxide supplementation significantly increased the expression of Hsp90. The Hsp90 inhibitor 17-allylamino-17-demethoxy-geldanamycin inhibited the diazoxide-induced decrease in cleavage of Bid, improvement of cardiac function, and decrease of apoptosis. Hsp90 inhibitor had no effect on the diazoxide-induced increase of total Cx43 protein expression in hearts preserved 9 hours, but inhibited the diazoxide-induced increase of mitochondrial Cx43 protein level. Hsp90 might mediate diazoxide-induced cardioprotection against apoptosis in hypothermic preservation heart by preventing the cleavage of Bid.

Possible Role of Hydrogen Sulfide on the Preservation of Donor Rat Hearts

Objective The aim of this study was to observe the preservative effect of hydrogen sulfide (H 2S) on donor rat hearts. Materials and Methods The hearts of 24 Sprague-Dawley rats were perfused on a Langendorff perfusion column for 30 minutes. We calculated and recorded the left ventricular-developed pressure (LVDP), and positive and negative derivatives of left ventricular systolic pressure (LVSP; +dP/dt and −dP/dt). Hearts were then arrested and stored for 6 hours at 4°C: group 1, Krebs-Henseleit (KH) solution; group 2, KH solution with 1 μmol/L NaHS; group 3, KH solution with 1 μmol/L NaHS and 10 μmol/L glibenclamide; group 4, St. Thomas II solution. Hearts were transferred back to the Langendorff column. After stabilizing for 30 minutes, LV performance was assessed as before. The donor hearts were kept for pathological study including myocardial water ratio, ATP content, and myocyte apoptosis index. Results The recovery rates of +dp/dtmax, −dp/dtmax, and LVDP of groups 2 and 4 were much better than those of groups 1 and 3. The hearts contracted immediately after reperfusion in group 4. Ventricular fibrillation was seen before contraction in the other 3 groups, with the longest duration in group. No significant difference in myocardial water ratio was found. The ATP content was the highest in group 2. Apoptosis was observed in the 4 groups with the lowest apoptosis index in group 2. Conclusions H 2S has a protective effect on rat donor hearts at the concentration of 1 μmol/L. The protective effect is better than that of St. Thomas II solution. The protective effect of H 2S can be blocked by glibenclamide.

Evaluation of preservation solutions by ESR-spectroscopy: Superior effects of University of Wisconsin over Histidine–Tryptophan–Ketoglutarate in reducing renal reactive oxygen species

Despite the causative role of oxidative stress in renal ischemia-reperfusion (I-R) injury effects of preservation solutions on reactive oxygen species (ROS) release have not been sufficiently evaluated. We compared the effects of most common solutions in kidney transplantation, University of Wisconsin (UW) and Histidine-Tryptophan-Ketoglutarate (HTK). ROS formation in isolated perfused rat kidney was detected by electron spin resonance spectroscopy using spin label 1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethyl-pyrrolidine. Donor kidneys from Lewis rats were pretreated with saline (controls), in therapeutic groups, kidneys underwent 18 h of cold storage (CS) preserved by HTK or UW solution. Experimental protocol included a stabilization period followed by additional I-R. Kidneys preserved by HTK produced highest ROS values in the control period after CS, whereas levels in UW and control group did not vary significantly. A peak release induced by additional I-R was also significantly highest in HTK kidneys, and UW did not differ from controls. During reperfusion, levels in HTK exceeded control and UW values. Renal vascular resistance, caspase-3-activity, and tissue hydration were enhanced in HTK compared with UW group, whereas ATP concentration was less reduced in UW-preserved tissue. These data show the greater antioxidative potential of UW solution, which also attenuated organ impairment after CS in the early reperfusion period.

Taurine prevents myocardial ischemia/reperfusion-induced oxidative stress and apoptosis in prolonged hypothermic rat heart preservation

Taurine (2-aminoethanesulfonic acid) is a potent antioxidant and inhibits cell apoptosis in ischemic reperfusion injury. In this study we evaluated whether addition of taurine to St. Thomas' cardioplegic solution enhances its myocardial protective effects in prolonged hypothermic heart preservation in rats. Hearts isolated from male Sprague-Dawley rats were mounted on a Langendorff apparatus to estimate baseline cardiac function, then arrested and stored in St. Thomas' cardioplegic solution, with taurine (10 mM; taurine group, n = 8) or without taurine (control group, n = 8), for 6 h at 4 degrees C. After storage, the hearts were reperfused and heart rate (HR), coronary flow (CF), left ventricular developed pressure (LVP), and positive maximum left ventricular developing pressure (max LV dp/dt) were measured. The LV tissue was examined immunohistochemically for determining DNA oxidative stress and cell apoptosis. Compared with control groups, recovery of LVP (P < 0.001), max LV dp/dt (P < 0.001), and coronary flow (P < 0.001) were significantly enhanced, whereas glutamic oxaloacetic transaminase (P < 0.01), lactate dehydrogenase (P < 0.05), creatine phosphate kinase (P < 0.01), 8-hydroxy-2'-deoxyguanosine index (P < 0.01), caspase-3 mRNA expression (P < 0.05), and percentage of TUNEL-positive cardiomyocytes (P < 0.05) were reduced in the taurine group. Addition of taurine to St. Thomas' cardioplegic solution improved cardiac function recovery for prolonged hypothermic rat heart preservation by suppressing DNA oxidative stress and cell apoptosis.

Supplementation of Nucleoside-Nucleotide Mixture Enhances Functional Recovery and Energy Metabolism Following Long-Time Hypothermic Heart Preservation

OG-VI, a well-balanced mixture of nucleoside and nucleotides, has been demonstrated to have a favorable effect on energy metabolism. In this study, we tested the hypothesis that addition of OG-VI to the University of Wisconsin solution can improve the cardiac functional recovery following long-time hypothermic preservation. Forty-two male Wistar rats were randomized into four groups. After 30-min of isolated working heart perfusion, the rat hearts were arrested with St. Thomas cardioplegic solution and preserved at 4 degrees C in saline, OG-VI, UW, and UW+OG-VI, respectively. After 12-h of preservation, the hearts were reperfused for 60-min during which the recovery of cardiac functions were monitored continuously. Myocardial adenine nucleotides were analyzed using high-performance liquid chromomatograph. In the UW+OG-VI group, the recovery of cardiac output, coronary flow, aortic flow, rate-pressure product, left ventricle stroke volume, and stroke work were significantly higher than other groups (P < 0.05). Furthermore, all phosphate high-energy compounds were significantly higher in the UW+OG-VI group than in the other groups (P < 0.05). Coronary vascular resistance and myocardial wet/dry weight ratio were obviously lower in the UW+OG-VI group, compared to the other groups (P < 0.05). Heart function was better recovered when nucleoside-nucleotide mixture was added to UW solution during long-time hypothermic rat heart preservation. The mechanism is not totally clear, but enhancement of high-energy phosphate production is possible.

Activation of Mitochondrial ATP-Sensitive Potassium Channel Contributes to Protective Effect in Prolonged Myocardial Preservation.

To find a better strategy for effective donor organ preservation, here we used a model of long-term hypothermia preservation of rat hearts to investigate the cardioprotective effects of diazoxide, a selective opener of mitochondrial ATP-sensitive potassium channel (mitoKATP ). Cardiac function was impaired by 8-hour ischemic preservation following 30 minutes of reperfusion. Treatment with diazoxide significantly attenuated the decline of myocardial contractility, and decreased the lactate dehydrogenase leakage and myocardial edema. Diazoxide also prevented the loss of activity of mitochondrial superoxide dismutases and sarcolemmal Na+/K+ ATPase during ischemia or reperfusion period, respectively. These effects of diazoxide could be abolished by a selective mitoKATp blocker 5-hydroxydecanoate. The results suggest that diazoxide, as a supplementation in cardioplegic solution, could enhance myocardial protection by opening mitoKATp channel, and better maintenance of mitochondrial anti-oxidative enzyme.

Cardioprotective effects of FK409, a nitric oxide donor, after isolated rat heart preservation for 16 hours

Background. We examined the cardioprotective effects of FK409, a nitric oxide donor, after isolated rat heart preservation. Methods. FK409 was administered to the hearts in pretreatment (FK409-1 group), during ischemia (FK409-2), or during reperfusion (FK409-3). The combined nitrate and nitrite level, coronary flow, cardiac function, coronary vasodilatory response, creatine kinase (CK), and myocardial water content were evaluated after the hearts had been preserved in University of Wisconsin solution at 0°C for 16 hours. Results. The release of nitrate and nitrite increased in reperfusion between the 20-to-40-second measurement and the measurement at 40 minutes, and the recovery of cardiac function was significantly improved in the FK409 groups. The coronary vasodilatory response to acetylcholine chloride was enhanced in the FK409-1 and FK409-2 groups. CK release decreased in FK409 groups after 15 minutes in reperfusion. Conclusions. This study suggests that FK409 has the best protective effect on cardiac function and coronary endothelial function when it is administered in the ischemic period, a less protective effect when administered during pretreatment, and the least protective effect when FK409 is given during reperfusion after heart preservation for 16 hours.

Fructose-1,6-diphosphate and a glucose-free solution enhances functional recovery in hypothermic heart preservation

Background Fructose-1,6-diphosphate (FDP) has been shown to protect tissue during hypoxia under various ischemic conditions, including isolated heart perfusion. We tested the hypothesis that adding FDP to St. Thomas solution can extend hypothermic heart preservation time. Methods Sixteen adult Sprague-Dawley rats were used. Under general anesthesia, the hearts were removed and preserved at 4°C in St. Thomas solution (30 ml/kg) for 12 hours. FDP (5 mM) was added to the St. Thomas solution in the study group ( n = 8), whereas no FDP was used in the control group ( n = 10). The hearts were reperfused after 12 hours of preservation using a working heart model. Results In the study group, cardiac output ranged from 13.00 ± 2.34 to 17.66 ± 1.71 ml/min, maximum aortic flow was 3.40 ± 1.99 to 9.26 ± 1.72 ml/min, left ventricular stroke volume ranged from 0.074 ± 0.014 to 0.092 ± 0.009 ml, left ventricular stroke work ranged from 6.22 ± 0.39 to 7.95 ± 0.44 ml/mmHg, and maximum left ventricular generated power was 14.38 ± 2.94 to 20.16 ± 2.49 Joules/min. All of these parameters were higher than those in the control group ( p < 0.001). Coronary vascular resistance and myocardial tissue wet/dry weight ratio were lower in the study group than in the control group ( p < 0.05). Conclusions Heart function was better preserved when FDP was added to St. Thomas solution during hypothermic rat heart preservation. The mechanism is not totally clear, but enhancement of high-energy phosphate production during ischemia is possible. Key words: heart, procurement, hypothermia, fructose-1,6-diphosphate.

Beneficial effects of ginsenosides of stems and leaves on cardiac and coronary vascular functions after 12-hour rat heart preservation

Beneficial effects of ginsenosides of stems and leaves on cardiac and coronary vascular functions after 12-hour rat heart preservation

Enhancement of Hypothermic Heart Preservation with Fructose 1,6-Diphosphate

Background. We hypothesized that the addition of fructose 1,6-diphosphate (FDP) to a hypothermic heart preservation solution could improve metabolic recovery because it has several beneficial effects.Materials and Methods. Twenty adult Sprague-Dawley rats were used to study hypothermic heart preservation. The hearts were removed under general anesthesia and preserved at 4°C in Euro-Collins solution (30 ml/kg) for 8 h. In the study group (N = 10), FDP (5 mM) was added to the Euro-Collins solution. In the control group (N = 10), no FDP was added. Heart function was studied after preservation using a working heart model. The ability of various concentrations of fructose 1,6-phosphate to passively diffuse through an egg phosphatidylcholine multilamellar vesicle (MLV) membrane bilayer was examined.Results. Cardiac output ranged from 17.0 ± 1.9 to 24.9 ± 1.6 ml/min in the study group vs 2.0 ± 1.0–12.3 ± 1.7 ml/min for controls, average aortic flow was 10.8 ± 1.4 ml/min in the study group vs −1.3 ± 1.6 ml/min for controls, and maximum LV generated power was 22.8 ± 1.7 J/min vs 10.1 ± 1.6 J/min for controls. Coronary flow, left ventricular stroke volume and stroke work, and myocardial oxygen consumption were much higher in the study group than in the control group. Coronary vascular resistance was lower in the study group than in the control group. Electron microscopic study indicated that many myocytes displayed patches of swollen mitochondria in the control group, but was rarely observed in the study group. The addition of 50 mM FDP caused substantial changes in MLV permeability. No dose of sucrose buffers outside the vesicles resulted in a significant changes of MLV permeability.Conclusions. Our results indicate that the addition of FDP to Euro-Collins solution significantly improves hypothermic rat heart preservation, and FDP appeared to cross the membrane bilayer.

The effects of mannitol, albumin, and cardioplegia enhancers on 24-h rat heart preservation.

During 24 h in vitro heart preservation and reperfusion, tissue damage occurs that seriously reduces cardiac function. Prevention of free radical production during preservation and reperfusion of ischemic tissue using free radical scavengers is of primary importance in maintaining optimal heart function in long-term preservation protocols. We examined whether mannitol (68 mM) and albumin (1.4 microM) in combination with other cardioplegia enhancers decreased free radical formation and edema and increased cardiac function during 24-h cold (5 degrees C) heart preservation and warm (37 degrees C) reperfusion in the Langendorff-isolated rat heart. The performance of mannitol-treated hearts was significantly decreased compared with that of hearts without mannitol treatment after 24 h of preservation with regard to recovery of diastolic pressure, contractility (+dP/dt), relaxation (-dP/dt), myocardial creatine kinase release, coronary flow, and lipid peroxidation. Albumin-treated hearts demonstrated higher cardiac function (contractility and coronary flow especially) than hearts not treated with albumin or hearts treated with mannitol, and this appears to be due to the positive effects of increased cellular metabolism and the enhancement of membrane stability.

Continuous Perfusion Improves Preservation of Donor Rat Hearts: Importance of the Implantation Phase

Background. Continuous hypothermic perfusion of donor hearts may provide extra protection for long ischemic times and suboptimal donors. The aim of three separate studies was to assess the effect of continuous hypothermic perfusion during simulated donor heart storage and implantation. Methods. In study 1 twelve isolated rat hearts underwent 10 minutes of normothermic ischemia to simulate the effect of brain death on the heart and 5 hours of cardioplegic arrest, using University of Wisconsin solution. Six hearts were statically stored in University of Wisconsin solution at 2°C, and six were perfused with University of Wisconsin solution. To assess the effect of simulated implantation, in study 2 an additional 12 hearts were statically stored for 5.5 hours in University of Wisconsin solution, six of which were rewarmed to a mean of 16°C over the last 30 minutes of arrest. To assess the effect of simulated perfusion, in study 3 during implantation 12 hearts were rewarmed to a mean of 16°C over the last 30 minutes of arrest, during which time six were perfused with 2°C solution. Results. Hearts perfused during storage demonstrated greater recovery of prearrest power, 85.8% ± 1.8%, than hearts preserved by static storage, 72.7% ± 3.0% ( p < 0.01). The simulated warm implantation period reduced recovery of power from 68.3% ± 5.1% to 40.2% ± 2.0% ( p < 0.001). Perfusion during warm implantation improved recovery to 61.8% ± 3.9% ( p < 0.01). In all experiments improved function was accompanied by improved metabolic energy status. Conclusions. During the implantation period of heart transplantation the donor heart sustains injury that could amount to 50% of total ischemic injury. Continuous perfusion during the cold storage phase and during simulated implantation improves recovery of the donor heart.

Which injury is the most important for rat heart preservation: Flush, cold storage, and/or reperfusion injury? The effect of each injury on coronary endothelial function

Which injury is the most important for rat heart preservation: Flush, cold storage, and/or reperfusion injury? The effect of each injury on coronary endothelial function