The aim of this investigation was to determine the optimal storage medium for testicular hypothermic transportation and identify the ideal concentration for the application of the protective agent 5-aminolevulinic acid (5-ALA). Furthermore, this study aimed to explore the underlying mechanism of the protective effects of 5-ALA. First, we collected and stored mouse testicular fragments in different media, including Hank's balanced salt solution (HBSS; n = 5), Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12; n = 5), and alpha-minimum essential medium (αMEM; n = 5). Storage of testicular tissue in HBSS preserved the integrity of testicular morphology better than that in the DMEM/F12 group ( P < 0.05) and the αMEM group ( P < 0.01). Testicular fragments were subsequently placed in HBSS with various concentrations of 5-ALA (0 [control], 1 mmol l -1 , 2 mmol l -1 , and 5 mmol l -1 ) to determine the most effective concentration of 5-ALA. The 2 mmol l -1 5-ALA group ( n = 3) presented the highest positive rate of spermatogonial stem cells compared with those in the control, 1 mmol l -1 , and 5 mmol l -1 5-ALA groups. Finally, the tissue fragments were preserved in HBSS with control ( n = 3) and 2 mmol l -1 5-ALA ( n = 3) under low-temperature conditions. A comparative analysis was performed against fresh testes ( n = 3) to elucidate the underlying mechanism of 5-ALA. Gene set enrichment analysis (GSEA) for WikiPathways revealed that the p38 mitogen-activated protein kinase (MAPK) signaling pathway was downregulated in the 2 mmol l -1 5-ALA group compared with that in the control group (normalized enrichment score [NES] = -1.57, false discovery rate [FDR] = 0.229, and P = 0.019). In conclusion, these data suggest that using 2 mmol l -1 5-ALA in HBSS effectively protected the viability of spermatogonial stem cells upon hypothermic transportation.
Male ; Animals ; Testis/cytology* ; Aminolevulinic Acid/pharmacology* ; Mice ; Organ Preservation/methods* ; Organ Preservation Solutions/pharmacology* ; Cryopreservation/methods*

BACKGROUND: Histidine-tryptophan-ketoglutarate (HTK) is a solution commonly used for organ transplantation. However, there is no certified fixed regimen for on-pump heart surgery in neonates. We aimed to retrospectively evaluate the outcomes related to different HTK dosages and to analyze the safety of high-dosage perfusion. METHODS: A total of 146 neonates who underwent on-pump heart surgery with single-shot HTK perfusion were divided into two groups according to HTK dosages: a standard-dose (SD) group (n = 63, 40 mL/kg < HTK ≤ 60 mL/kg) and a high-dose (HD) group (n = 83, HTK >60 mL/kg). Propensity score matching (PSM) was performed to control confounding bias. RESULTS: The SD group had a higher weight (3.7 ± 0.4 vs. 3.4 ± 0.4 kg, P < 0.0001), a lower proportion of complete transposition of the great artery (69.8% vs. 85.5%, P = 0.022), a lower cardiopulmonary bypass (CPB) time (123.5 [108.0, 136.0] vs. 132.5 [114.8, 152.5] min, P = 0.034), and a lower aortic x-clamp time (82.9 ± 27.1 vs. 95.5 ± 26.0 min, P = 0.005). After PSM, 44 patients were assigned to each group; baseline characteristics and CPB parameters between the two groups were comparable. There were no significant differences in peri-CPB blood product consumption after PSM (P > 0.05). The incidences of post-operative complications were not significantly different between the two groups. There were no significant differences in ventilation time, intensive care unit stay, and post-operative hospital stay (P > 0.05). Follow-up echocardiography outcomes at 1 month, 3 to 6 months, and 1 year showed that left ventricular ejection fraction and end-diastolic dimension were comparable between the two groups. CONCLUSIONS In neonatal on-pump cardiac surgery patients, single-shot HD (>60 mL/kg) HTK perfusion had a comparable heart protection effect and short-term post-operative prognosis as standard dosage perfusion of 40 to 60 mL/kg. Thus, this study provides supporting evidence of the safety of HD HTK perfusion.
Glucose/therapeutic use* ; Histidine ; Humans ; Infant, Newborn ; Mannitol ; Organ Preservation Solutions ; Potassium Chloride/therapeutic use* ; Prognosis ; Retrospective Studies ; Stroke Volume ; Tryptophan ; Ventricular Function, Left

OBJECTIVETo study the feasibility of sustaining the viable status of a liver graft in at least 96 h by extracorporeal perfusion using autologous blood.

METHODSEight extracorporeal porcine liver perfusions using autologous blood were performed, each for 96 h with hepatectomy, cold preservation, cannulation of vessels, and initiation of perfusion with normothermic oxygenated porcine blood. The graft viability was assessed by metabolic, synthetic, hemodynamic, and histologic parameters.

RESULTSAfter 96 h of normothermic, extracorporeal perfusion using autologous blood, the isolated livers maintained normal physiological levels of pH and electrolytes with sustained hepatic protein synthesis (complement and factor V) throughout the perfusion. Hemodynamic parameters maintained normal physiological ranges. Histological inspection demonstrated good preservation of the liver with a good architectural integrity.

CONCLUSIONIt is possible to sustain the viable status of a liver graft within 96 h by extracorporeal perfusion using autologous blood.

Animals ; Extracorporeal Circulation ; Liver ; Liver Transplantation ; Organ Preservation ; Organ Preservation Solutions ; Swine

BACKGROUNDOrgan preservation keeps the quality of the organs under prolonged ischemia. Continuous machine perfusions are gaining an important position in clinical research and practice. The aim of this study was to evaluate the protective effect of continuous hypothermic machine perfusion transport system (AirdriveTM) on cold ischemic injury of canine kidney.

METHODSTen kidneys of five healthy preserving canines were taken out after general anesthesia. Five kidneys were stored using common cold preservation (CCP group) by immersing it in the organ preservation solution, mixed with water and ice, and kept in a cold room at 4°C. The other five kidneys were stored using continuous machine perfusion preservation (CMP group) and were placed into the Airdrive(TM) continuous machine perfusion device at room temperature. The renal tissues were examined by histopathology, electron microscopy, and mitochondrial activity check at different time points.

RESULTSHistologic sections showed that the structures of the ten renal tissues were similar during the first 24 hours. After 48 hours, the CCP group showed more pronounced changes, as the renal tubular epithelial cells were more obvious than those in the glomeruli. Oxygen consumption rate of state III and IV respiration in the CCP group decreased after 12-48 hours and increased at 48 hours, respectively, when compared to the CMP group (P < 0.05). Cortex respiratory control ratio and phosphorus oxygen ratio were significantly higher in the CMP group at 48 hours.

CONCLUSIONWith prolonged storage time, the effect of continuous hypothermic machine perfusion transport system is better than that of common cold preservation on canine kidney.

Animals ; Dogs ; Kidney ; Kidney Transplantation ; Male ; Organ Preservation ; methods ; Organ Preservation Solutions

Adolescent ; Age Factors ; Child ; Dental Pulp ; diagnostic imaging ; Dentition, Permanent ; Humans ; Organ Preservation Solutions ; Periodontal Ligament ; diagnostic imaging ; Radiography ; Time Factors ; Tissue Preservation ; Tooth Avulsion ; surgery ; Tooth Replantation ; methods

OBJECTIVETo investigate the protective effect of luteolin on isolated rat heart in hypothermic preservation.

METHODSForty male SD rats were randomly divided into 4 groups (n = 10): control group, luteolin low-dose group (7.5 micromol/L), middle-dose group (15 micromol/L) and high dose group (30 micromol/L). Langendorff model of isolated rat heart was used. After 30 min basal perfusion, the hearts were stored in University of Wisconsin solution (UW solution) at 4 degrees C with luteolin (7.5, 15 and 30 micromol/L) or without luteolin for 12 h and followed by 60 min reperfusion. The recovery of cardiac contractile and diastolic function, coronary flow (CF), creatine kinase (CK) leakage in the coronary effluent, myocardial water content were determined. The myocardial ultrastructure was also observed.

RESULTSThe results revealed that luteolin improved the recovery of left ventricular peak systolic pressure and +/- dp/dtmax dose-dependently and increased coronary flow. The leakage of creatine kinase in the coronary effluent was significantly reduced in luteolin-added hearts. Impairment of myocardial ultrastructure after 12 h hypothermic preservation was obviously alleviated in hearts luteolin-added group compared with that in control group. There were no differences between the groups in myocardial water contents.

CONCLUSIONLuteolin as a supplementation in cardiac preservation solution can significantly improve the hypothermic preservation effects on rat heart and have myocardial protection effect, especially in luteolin-added with 30 micromol/L.

Animals ; Cryopreservation ; In Vitro Techniques ; Luteolin ; pharmacology ; Male ; Myocardium ; Organ Preservation ; methods ; Organ Preservation Solutions ; Rats

BACKGROUNDThe composition of the lung preservation solution used in lung graft procurement has been considered the key to minimize lung injury during the period of ischemia. Low-potassium dextran glucose (LPDG), an extracellular-type solution, has been adopted by most lung transplantation centers, due to the experimental and clinical evidences that LPDG is superior to intracellular-type solutions. Ulinastatin has been shown to attenuate ischemia-reperfusion (I/R) injury in various organs in animals. We supposed that the addition of ulinastatin to LPDG as a flushing solution, would further ameliorate I/R lung injury than LPDG solution alone.

METHODSTwelve male New Zealand white rabbits were randomly divided into 2 groups. Using an alternative in situ lung I/R model, the left lung in the control group was supplied and preserved with LPDG solution for 120 minutes. In the study group 50,000 U/kg of ulinastatin was added to the LPDG solution for lung preservation. Then re-ventilation and reperfusion of the left lung were performed for 90 minutes. Blood gas analysis (PaO₂, PaCO₂), mean pulmonary artery pressure (MPAP) and serum TNF-α level were measured intermittently. The pulmonary water index (D/W), tissue myeloperoxidase (MPO) activity, tissue malondialdehyde (MDA) content and morphologic changes were analyzed.

RESULTSThe study group showed significantly higher PaO₂ and lower MPAP at the end of reperfusion. Serum TNF-α level, left lung tissue MPO and MDA in the study group were significantly lower than those in the control group. D/W and pathologic evaluation were also remarkably different between the two groups.

CONCLUSIONSThis study indicated that better lung preservation could be achieved with the use of an ulinastatin modified LPDG solution. Ulinastatin further attenuated lung I/R injury, at least partly by reducing oxidative reactions, inhibiting the release of inflammatory factors and neutrophils immigration.

Animals ; Glycoproteins ; pharmacology ; Lung ; drug effects ; metabolism ; Lung Transplantation ; Male ; Organ Preservation Solutions ; chemistry ; pharmacology ; Rabbits ; Random Allocation ; Reperfusion Injury ; prevention & control

OBJECTIVETo investigate the effect of diazoxide (DE) on the myocardial ultrastructure and opening of maitochondrial permeability transition pore (MPTP) in donor rat heart suffered from long-term hypothermic preservation.

METHODSThe Langendorff model of isolated rat heart was used. The hearts were stored in 4 degrees C Celsior solution containing different concentration of DE (15, 30, or 45 micromol/L) for 9 h followed by 60 min of reperfusion. The recovery of rate-pressure product (RPP) was observed. The opening of MPTP and myocardial mitochondria ultrastructure were also evaluated.

RESULTS(1) As compared with the celsior solution preserved group, DE (30 micromol/L) increased recovery of RPP during reperfusion and inhibited the opening of MPTP. DE also alleviated the myocardial mitochondrial ultrastucture damage induced by long-term hypothermic preservation. (2) The above effects of DE were attenuated by a mitoK(ATP) channel inhibitor 5-hydroxydecanoate and a MPTP opener atractyloside.

CONCLUSIONIn the donor rat heart, DE protects myocardial mitochondria ultrastructure against long-term hypothermic preservation injury via inhibiting the opening of MPIP.

Animals ; Cryopreservation ; Diazoxide ; pharmacology ; Heart ; In Vitro Techniques ; Male ; Mitochondria, Heart ; physiology ; ultrastructure ; Mitochondrial Membrane Transport Proteins ; drug effects ; metabolism ; Organ Preservation Solutions ; pharmacology ; Potassium Channels ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

BACKGROUNDPulmonary artery perfusion during cardiopulmonary bypass (CPB) is a novel adjunctive method, which can minimize the lung ischemic-reperfusion injury and inflammatory response. This study evaluated the protective effect of pulmonary perfusion with hypothermic HTK solution in corrections of congenital heart defects with pulmonary hypertension.

METHODSBetween June 2009 and December 2009, 24 consecutive infants with congenital heart defects and pulmonary hypertension were randomly divided into perfused group (n = 12) and control group (n = 12). Oxygen index, alveolar-arterial O2 gradient, serum levels of malondialchehyche (MDA), interleukin (IL)-6, -8, -10, soluble intercellular adhesion molecule-1 (sICAM-1), and P-selectin were measured before commencement and serially for 48 hours after termination of bypass.

RESULTSOxygenation values were better preserved in the perfused group than in the control group. The serum levels of IL-6 increased immediately after CPB in both groups and returned to baseline at 48 hours after CPB,but it was restored faster and earlier in the perfused group. The serum levels of IL-8, sICAM-1, and MDA remained at baseline at each point after CPB in the perfused group and elevated significantly immediately after CPB in the control group, except for sICAM-1. The serum level of IL-10 increased immediately after CPB and decreased to baseline at 48 hours after CPB in both groups, but the IL-10 level in the perfused group was significantly higher than in the control group at 12 hours after CPB. The serum P-selectin levels in the control group immediately after CPB were significantly higher than prebypass levels. Moreover, there were no significant differences in postoperative clinical characters, except for the intubated time.

CONCLUSIONIn infants with congenital heart defects, pulmonary perfusion with hypothermic HTK solution during cardiopulmonary bypass could ameliorate lung function and reduce the inflammatory response.

Cardiopulmonary Bypass ; adverse effects ; Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; blood ; surgery ; Humans ; Hypertension, Pulmonary ; blood ; Intercellular Adhesion Molecule-1 ; blood ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Lung Injury ; blood ; etiology ; prevention & control ; Male ; Organ Preservation Solutions ; therapeutic use ; P-Selectin ; blood ; Perfusion ; methods ; Pulmonary Artery

OBJECTIVETo evaluate the effects of tannic acid (TA) treatment on the physico-chemical properties of glutaraldehyde (Glut)-fixed bovine jugular vein (BJV).

METHODSFresh BJVs were treated with Glut or Glut/TA, respectively. The shrinkage temperature, resistance to collagenase or elastase digestion, bio-mechanical properties, and molecular structure of these prepared BJVs were evaluated by Fourier transform infrared spectroscopy.

RESULTSTA treatment resulted in higher shrinkage temperature (P < 0.01), higher resistance to collagenase or elastase digestion (P < 0.01), slightly increased tensile strength (P < 0.01), and elongation at break (P < 0.05) in Glut/TA BJV walls when compared with those of Glut group. Chemical bonds existed between TA and JBV tissue.

CONCLUSIONTA treatment can effectively improve the physicochemical properties of Glut-fixed BJVs.

Animals ; Cattle ; Chemical Phenomena ; drug effects ; Elasticity ; drug effects ; Glutaral ; pharmacology ; Jugular Veins ; pathology ; Organ Preservation Solutions ; pharmacology ; Tannins ; pharmacology ; Tensile Strength ; drug effects ; Tissue Fixation ; methods ; Tissue Preservation ; methods