A Device and Methodology for Continuous Hypothermic Perfusion of Explanted Large Mammalian Hearts, Followed by In Vitro Langendorff Reanimation: Pilot Studies

The current methodologies of clinical heart transplantation limit the ischemic window to 4–6 h. Periods longer than this can induce dysfunction in the organ and can lead to increased patient morbidity and mortality. An alternative to the current methods of static cold storage (CS) is continuous hypothermic perfusion (CHP), where a hypothermic oxygenated crystalloid solution is mechanically perfused through the coronary arteries. This has been shown to preserve the function for up to 72 h, but the techniques have yet to be optimized. We have developed an apparatus and methodology for performing CHP on large mammalian hearts, followed by reanimation in our in vitro Langendorff apparatus (The Visible HeartTM). We are also investigating the utility of the cardioprotective agents docosahexaenoic acid and [D-Ala2, D-Leu5] enkephalin, both of which have shown cardioprotective effects in our laboratory, and we believe that their addition to the preservation solution can further extend the transplant window. A series of pilot studies has been performed to date, with modestly successful results. Hearts preserved with CHP seem to show better functionality than CS hearts but far worse functionality than hearts reanimated immediately after explant. We hope to use this system to optimize CHP methodology and eventually develop a system for prolonging the window for heart transplantation.

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Cold Storage of Hearts from Brain-dead Donors using a Preservation Solution, Supplemented by a Mesenchymal Stem Cell Conditioned Medium Improves Contractile Function of Graft after Heart Transplantation in Rats

The Thoracic and Cardiovascular Surgeon ◽

10.1055/s-0038-1627835 ◽

2018 ◽

Vol 66 (S 01) ◽

pp. S1-S110

Author(s):

S. Korkmaz-Icöz ◽

K. Li ◽

M. Ruppert ◽

S. Loganathan ◽

T. Radovits ◽

...

Keyword(s):

Stem Cell ◽

Mesenchymal Stem Cell ◽

Heart Transplantation ◽

Conditioned Medium ◽

Cold Storage ◽

Contractile Function ◽

Preservation Solution ◽

Brain Dead ◽

Cell Conditioned Medium

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The investigation of infarct-limiting effects of preservation solutions in the rat model of long-time static cold storage of isolated rat heart

Regional blood circulation and microcirculation ◽

10.24884/1682-6655-2015-14-3-78-81 ◽

2015 ◽

Vol 14 (3) ◽

pp. 78-81 ◽

Cited By ~ 1

Author(s):

S. M. Minasian ◽

Yu. V. Dmitriev ◽

M. M. Galagudza ◽

A. A. Karpov ◽

E. A. Bobrova ◽

...

Keyword(s):

Cold Storage ◽

Rat Model ◽

Isolated Rat Heart ◽

Limiting Factor ◽

Preservation Solution ◽

Heart Preservation ◽

Long Time ◽

Preservation Solutions ◽

Cardioprotective Effects ◽

Krebs Henseleit Buffer

One of the most important aims of modern transplantation is the preservation of the morphofunctional state of donor organs for as long as possible time after removing them from the donor prior to implantation into a patient. To solve this problem, heart preservation solutions are used. However, despite the large number of prescriptions of preservation solutions, the time, limited to six hours, after removing donor heart is still a limiting factor in determining the possibility of implanting heart into a patient. In this study, cardioprotective effects of widely used in clinical practice preservation solution HTK and new Krebs-Henseleit buffer-based preservation solution were studied in the rat model of prolonged static cold storage. It was shown that new Krebs-Henseleit buffer-based preservation solution protected the heart significantly even after an eight-hour global ischemia and has the advantage over Custodiol in the infarct-limiting action.

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Effectiveness Assessment of a Modified Preservation Solution Containing Thyrotropin or Follitropin Based on Biochemical Analysis in Perfundates and Homogenates of Isolated Porcine Kidneys after Static Cold Storage

International Journal of Molecular Sciences ◽

10.3390/ijms22168360 ◽

2021 ◽

Vol 22 (16) ◽

pp. 8360

Author(s):

Aneta Ostróżka-Cieślik ◽

Barbara Dolińska ◽

Florian Ryszka

Keyword(s):

Protective Effect ◽

Cold Storage ◽

Biochemical Parameters ◽

Biochemical Analysis ◽

Structural Integrity ◽

Biochemical Tests ◽

Preservation Solution ◽

Kidney Perfusion ◽

Effectiveness Assessment

In this paper, we assess the nephroprotective effects of thyrotropin and follitropin during ischaemia. The studies were performed in vitro in a model of isolated porcine kidneys stored in Biolasol (FZNP, Biochefa, Sosnowiec, Poland) and modified Biolasol (TSH: 1 µg/L; FSH 1 µg/L). We used the static cold storage method. The study was carried out based on 30 kidneys. The kidneys were placed in 500 mL of preservation solution chilled to 4 °C. The samples for biochemical tests were collected during the first kidney perfusion (after 2 h of storage) and during the second perfusion (after 48 h of storage). The results of ALT, AST, and LDH activities confirm the effectiveness of Biolasol + p-TSH in maintaining the structural integrity of renal cell membranes. Significantly reduced biochemical parameters of kidney function, i.e., creatinine and protein concentrations were also observed after 48 h storage. The protective effect of Biasol + p-TSH is most pronounced after 2 h of storage, suggesting a mild course of damage thereafter. A mild deterioration of renal function was observed after 48 h. The results of our analyses did not show any protective effect of Biolasol + p-FSH on the kidneys during ischaemia.

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Structural integrity after cold storage of human epidermal model in hypothermosol: A correlative ultrastructural and fluorescent assay

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100169080 ◽

1994 ◽

Vol 52 ◽

pp. 270-271

Author(s):

Henry H. Eichelberger ◽

John G. Baust ◽

Robert G. Van Buskirk

Keyword(s):

Cold Storage ◽

Structural Integrity ◽

Culture Media ◽

Cell Structure ◽

Natural State ◽

Sodium Cacodylate ◽

Ruthenium Tetroxide ◽

Cacodylate Buffer ◽

Before And After

For research in cell differentiation and in vitro toxicology it is essential to provide a natural state of cell structure as a benchmark for interpreting results. Hypothermosol (Cryomedical Sciences, Rockville, MD) has proven useful in insuring the viability of synthetic human epidermis during cold-storage and in maintaining the epidermis’ ability to continue to differentiate following warming.Human epidermal equivalent, EpiDerm (MatTek Corporation, Ashland, MA) consisting of fully differentiated stratified human epidermal cells were grown on a microporous membrane. EpiDerm samples were fixed before and after cold-storage (4°C) for 5 days in Hypothermosol or skin culture media (MatTek Corporation) and allowed to recover for 7 days at 37°C. EpiDerm samples were fixed 1 hour in 2.5% glutaraldehyde in sodium cacodylate buffer (pH 7.2). A secondary fixation with 0.2% ruthenium tetroxide (Polysciences, Inc., Warrington, PA) in sodium cacodylate was carried out for 3 hours at 4°C. Other samples were similarly fixed, but with 1% Osmium tetroxide in place of ruthenium tetroxide. Samples were dehydrated through a graded acetone series, infiltrated with Spurrs resin (Polysciences Inc.) and polymerized at 70°C.

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Custodiol-N, a novel organ preservation solution reduces ischemia/reperfusion injury after heart transplantation

The Thoracic and Cardiovascular Surgeon ◽

10.1055/s-0029-1247060 ◽

2010 ◽

Vol 58 (S 01) ◽

Author(s):

S Loganathan ◽

T Radovits ◽

K Hirschberg ◽

S Korkmaz ◽

A Koch ◽

...

Keyword(s):

Heart Transplantation ◽

Reperfusion Injury ◽

Organ Preservation ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Preservation Solution ◽

Organ Preservation Solution

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Impact of Histidine-Tryptophan-Ketoglutarate Preservation Solution in Heart Transplantation with Extended Distance

The Journal of Heart and Lung Transplantation ◽

10.1016/j.healun.2021.01.650 ◽

2021 ◽

Vol 40 (4) ◽

pp. S226

Author(s):

F.A. Rodrigues Gonçalves ◽

E.M. Trindade ◽

S. Mangini ◽

F.A. Gaiotto ◽

F. Bacal

Keyword(s):

Heart Transplantation ◽

Preservation Solution

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Melatonin membrane receptor 2 activation is a key determinant for melatonin-mediated cardioprotection in cardiac ischaemia-reperfusion injury

European Heart Journal ◽

10.1093/ehjci/ehaa946.2573 ◽

2020 ◽

Vol 41 (Supplement_2) ◽

Author(s):

K Singhanat ◽

N Apaijai ◽

T Jaiwongkam ◽

S Kerdphoo ◽

S.C Chattipakorn ◽

...

Keyword(s):

Infarct Size ◽

Mitochondrial Dynamics ◽

Melatonin Receptors ◽

Lv Function ◽

H9c2 Cell ◽

Cardiac Ischaemia ◽

Beneficial Effects ◽

Ischaemia Reperfusion ◽

Cardioprotective Effects

Abstract Background Cardiac ischaemia/reperfusion (I/R) injury has been an economic and health burden worldwide. Previous studies have reported the beneficial effects of melatonin when given prior to cardiac ischaemia in animals with cardiac I/R injury. However, the effects of melatonin on the hearts when it is given after ischaemia or at the onset of reperfusion, which is more relevant to the clinical setting, is not known. Moreover, the mechanisms responsible for the potential benefits of melatonin and the roles of melatonin receptors on the heart during cardiac I/R injury have not been fully investigated. Purpose We tested the hypothesis that in rats with cardiac I/R injury, melatonin exerts cardioprotective effects even when it is given after ischaemia via an activation of both melatonin receptors 1 (MT1) and 2 (MT2), leading to decreased mitochondrial dysfunction, mitochondrial dynamics imbalance, excessive mitophagy, cardiomyocyte death and finally resulting in decreased infarct size and improved left ventricular (LV) function. Methods Male Wistar rats were subjected to cardiac I/R (30 min of LAD ligation and 120 min of reperfusion). These rats were divided into 4 interventions (n=12/group) including vehicle, pretreatment with melatonin, melatonin treatment during ischaemia, or at the onset of reperfusion. Melatonin was given to the rats at the dose of 10 mg/kg via intravenous injection. In addition, either a non-specific melatonin receptor blocker (Luzindole) or specific MT2 blocker (4-PPDOT) at 1 mg/kg was given intravenously to 2 additional sets of rats (n=12/set) prior to melatonin and cardiac I/R induction. At the end of cardiac I/R, infarct size, LV function, and molecular mechanisms were determined. Furthermore, in vitro experiment was conducted in MT1 or MT2 silenced H9C2 cell with hypoxia/reoxygenation (H/R) to investigate the mechanism underlying cardioprotective effects of melatonin during cardiac I/R. Results Rats in all melatonin-treated groups had similarly reduced cardiac I/R injury as indicated by reduced infarct size (Fig. 1A), arrhythmia score. Melatonin-treated rats also had decreased mitochondrial ROS production, mitochondrial depolarization and swelling, decreased p-Drp1/Drp1 ratio (Fig. 1B) and increased Mfn1, Mfn2, and OPA1, and decreased apoptosis, leading to increased %LVEF. Luzindole and 4-PPDOT abolished these protective effects of melatonin (Fig. 1A). In in vitro study, melatonin increased %cell viability (Fig. 1C), reduced mitochondrial dynamics imbalance and cardiomyocyte apoptosis in H9C2 cells with H/R. However, these beneficial effects of melatonin were abrogated only in MT2 silenced H9C2 cell with H/R. Conclusion Melatonin exerted both preventive and treatment effects in reducing cardiac I/R injury. Its cardioprotective effects were dependent upon the activation of MT2 receptor. Figure 1 Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): National Science and Technology Development Agency of Thailand

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Hypertrophy-Reduced Autophagy Causes Cardiac Dysfunction by Directly Impacting Cardiomyocyte Contractility

Cells ◽

10.3390/cells10040805 ◽

2021 ◽

Vol 10 (4) ◽

pp. 805

Author(s):

Christiane Ott ◽

Tobias Jung ◽

Sarah Brix ◽

Cathleen John ◽

Iris R. Betz ◽

...

Keyword(s):

Cardiomyocyte Hypertrophy ◽

Aortic Constriction ◽

Cell Contractility ◽

Neonatal Cardiomyocytes ◽

Cardiomyocyte Contractility ◽

Cardioprotective Effects ◽

Modified Proteins ◽

Cardiomyocyte Autophagy

Cardiac remodeling and contractile dysfunction are leading causes in hypertrophy-associated heart failure (HF), increasing with a population’s rising age. A hallmark of aged and diseased hearts is the accumulation of modified proteins caused by an impaired autophagy-lysosomal-pathway. Although, autophagy inducer rapamycin has been described to exert cardioprotective effects, it remains to be shown whether these effects can be attributed to improved cardiomyocyte autophagy and contractility. In vivo hypertrophy was induced by transverse aortic constriction (TAC), with mice receiving daily rapamycin injections beginning six weeks after surgery for four weeks. Echocardiographic analysis demonstrated TAC-induced HF and protein analyses showed abundance of modified proteins in TAC-hearts after 10 weeks, both reduced by rapamycin. In vitro, cardiomyocyte hypertrophy was mimicked by endothelin 1 (ET-1) and autophagy manipulated by silencing Atg5 in neonatal cardiomyocytes. ET-1 and siAtg5 decreased Atg5–Atg12 and LC3-II, increased natriuretic peptides, and decreased amplitude and early phase of contraction in cardiomyocytes, the latter two evaluated using ImageJ macro Myocyter recently developed by us. ET-1 further decreased cell contractility in control but not in siAtg5 cells. In conclusion, ET-1 decreased autophagy and cardiomyocyte contractility, in line with siAtg5-treated cells and the results of TAC-mice demonstrating a crucial role for autophagy in cardiomyocyte contractility and cardiac performance.

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Efficacy of coupling gamma irradiation with calcium chloride and lemongrass oil in maintaining guava fruit quality and inhibiting fungal growth during cold storage

Folia Horticulturae ◽

10.2478/fhort-2018-0007 ◽

2018 ◽

Vol 30 (1) ◽

pp. 67-78 ◽

Cited By ~ 2

Author(s):

Ramadan A. Hassanein ◽

Ehab A. Salem ◽

Ahmed A. Zahran

Keyword(s):

Vitamin C ◽

Fruit Quality ◽

Cold Storage ◽

Fungal Growth ◽

Alternaria Solani ◽

Titratable Acidity ◽

Soluble Solids ◽

Γ Irradiation ◽

Lemongrass Oil

AbstractThis study was performed to explore the efficacy of combining more than one postharvest treatment in maintaining some quality attributes and reducing fungal pathogenicity in cold-stored guava fruits. The investigated postharvest treatments included the control, CaCl2(4%), lemongrass oil (2 dm3kg−1), gamma (γ) irradiation (0.2, 0.4 and 0.6 kGy), 0.4 kGy γ irradiation + CaCl2(4%), and 0.4 kGy γ irradiation + lemongrass oil (2 dm3kg−1). The studied physiochemical attributes included weight loss, decay percentage, fruit firmness, total soluble solids (TSS), titratable acidity (TA), and vitamin C content. Different fungal species were also isolated from decayed fruits and were identified asAlternaria alternata,Alternaria solani,Aspergillus niger,Botrytis cinerea,Fusarium solaniandRhizopus stolonifer. The severity of infection for the different fungi was determined, and anin vitroantifungal assay was conducted for lemongrass oil. All the investigated treatments generally reduced decay and water loss percentages, and controlled TSS, TA and vitamin C decrements that occurred during cold storage. On the other hand, higher irradiation doses generally increased fruit softness, and the 0.4 kGy γ dose did not contribute to the overall fruit quality when coupled with CaCl2and lemongrass oil, compared to CaCl2and lemongrass oil treatments alone.

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