Sodium Hydrosulfide Treatment During Porcine Kidney Ex Vivo Perfusion and Transplantation.

Background: In rodents, hydrogen sulfide (H2S) reduces ischemia-reperfusion injury and improves renal graft function after transplantation. Here, we hypothesized that the benefits of H2S are conserved in pigs, a more clinically relevant model. Methods: Adult porcine kidneys retrieved immediately or after 60 min of warm ischemia (WI) were exposed to 100 µM sodium hydrosulfide (NaHS) (1) during the hypothermic ex vivo perfusion only, (2) during WI only, and (3) during both WI and ex vivo perfusion. Kidney perfusion was evaluated with dynamic contrast-enhanced MRI. MRI spectroscopy was further employed to assess energy metabolites including ATP. Renal biopsies were collected at various time points for histopathological analysis. Results: Perfusion for 4 h pig kidneys with Belzer MPS UW + NaHS resulted in similar renal perfusion and ATP levels than perfusion with UW alone. Similarly, no difference was observed when NaHS was administered in the renal artery before ischemia. After autotransplantation, no improvement in histologic lesions or cortical/medullary kidney perfusion was observed upon H2S administration. In addition, AMP and ATP levels were identical in both groups. Conclusions: In conclusion, treatment of porcine kidney grafts using NaHS did not result in a significant reduction of ischemia-reperfusion injury or improvement of kidney metabolism. Future studies will need to define the benefits of H2S in human, possibly using other molecules as H2S donors.

Subnormothermic Ex Vivo Porcine Kidney Perfusion Improves Energy Metabolism: Analysis Using 31P Magnetic Resonance Spectroscopic Imaging.

The ideal preservation temperature for donation after circulatory death kidney grafts is unknown. We investigated whether subnormothermic (22 °C) ex vivo kidney machine perfusion could improve kidney metabolism and reduce ischemia-reperfusion injury. Methods: To mimic donation after circulatory death procurement, kidneys from 45-kg pigs underwent 60 min of warm ischemia. Kidneys were then perfused ex vivo for 4 h with Belzer machine perfusion solution UW at 22 °C or at 4 °C before transplantation. Magnetic resonance spectroscopic imaging coupled with LCModel fitting was used to assess energy metabolites. Kidney perfusion was evaluated with dynamic-contrast enhanced MRI. Renal biopsies were collected at various time points for histopathologic analysis. Results: Total adenosine triphosphate content was 4 times higher during ex vivo perfusion at 22 °C than at 4 °C perfusion. At 22 °C, adenosine triphosphate levels increased during the first hours of perfusion but declined afterward. Similarly, phosphomonoesters, containing adenosine monophosphate, were increased at 22 °C and then slowly consumed over time. Compared with 4 °C, ex vivo perfusion at 22 °C improved cortical and medullary perfusion. Finally, kidney perfusion at 22 °C reduced histological lesions after transplantation (injury score: 22 °C: 10.5 ± 3.5; 4 °C: 18 ± 2.25 over 30). Conclusions: Ex vivo kidney perfusion at 22°C improved graft metabolism and protected from ischemia-reperfusion injuries upon transplantation. Future clinical studies will need to define the benefits of subnormothermic perfusion in improving kidney graft function and patient's survival.

DCD pigs' kidneys analyzed by MRI to assess ex vivo their viability.

BACKGROUND: Magnetic resonance imaging (MRI) gadolinium-perfusion was applied in simulated Donation after Cardiac Death (DCD) in porcine kidneys to measure intrarenal perfusion. Adenosine triphosphate (ATP) resynthesis during oxygenated hypothermic perfusion was compared to evaluate the "ex vivo organ viability". Adenine nucleotide (AN) was measured by P nuclear magnetic resonance (NMR) spectroscopy. Whereas this latter technique requires sophisticated hardware, gadolinium-perfusion can be realized using any standard proton-MRI scanner. The aim of this work was to establish a correlation between the two methods. METHODS: Twenty-two porcine kidneys presenting up to 90 min warm ischemia were perfused with oxygenation at 4 °C using our magnetic resonance-compatible machine. During the perfusion, P NMR spectroscopy and gadolinium-perfusion sequences were performed. Measures obtained from the gadolinium-perfusion were the speed of elimination of the cortical gadolinium and the presence or absence of a corticomedullar shunt. For ATP resynthesis analysis, P chemical shift imaging was acquired and analyzed. All the kidneys have been submitted to histologic examination. RESULTS: ATP resynthesis was observed in all organs presenting a cortical gadolinium elimination slope of (-) 23° or greater. In organs with lower gadolinium elimination, no AN or only precursors were detected. This study reveals a link between the two methods and demonstrates ex vivo viability in 93% of the analyzed kidneys. Benefits and side effects of both methods are discussed. CONCLUSION: Oxygenated hypothermic perfusion enables the evaluation of kidneys in DCD simulated situation; gadolinium-perfusion can be introduced into any center equipped with a proton-MRI scanner allowing results superposable with ATP measurement.

Detection of ATP by "in line" 31P magnetic resonance spectroscopy during oxygenated hypothermic pulsatile perfusion of pigs' kidneys.

OBJECT: To demonstrate that adenosine triphosphate (ATP), which provides a valuable biomarker for kidney viability in the context of donation after cardiac death (DCD) transplantation, can be detected by means of (31)P magnetic resonance spectroscopy (MRS) if kidneys are perfused with oxygenated hypothermic pulsatile perfusion (O(2)+HPP). MATERIALS AND METHODS: Porcine kidney perfusion was carried out using a home made, MR-compatible HPP-machine. Consequently, kidney perfusion could be performed continuously during magnetic resonance imaging and magnetic resonance spectroscopy recording. (31)P MR spectroscopy consisted of 3-dimensional chemical shift imaging (CSI), which allowed for the detection of ATP level in line. (31)P CSI was performed at 3 tesla in 44 min with a nominal voxel size of 6.1 cc. RESULTS: (31)P CSI enabled the detection of renal ATP when pO(2) was equal to 100 kPa. With pO(2) of 20 kPa, only phosphomonoester, inorganic phosphate and nicotinamide adenine dinucleotide could be found. Semi-quantitative analysis showed that ATP level was 1.3 mM in normal kidney perfused with pO(2) of 100 kPa. CONCLUSIONS: This combined technology may constitute a new advance in DCD organ diagnostics prior to transplantation, as it allows direct assessment of ATP concentration, which provides a reliable indicator for organ bioenergetics and viability. In this study, kidneys presenting no warm ischemia were tested in order to establish values in normal organs. The test could be easily integrated into the clinical environment and would not generate any additional delay into the transplantation clinical workflow.

Oxygenated hypothermic pulsatile perfusion versus cold static storage for kidneys from non heart-beating donors tested by in-line ATP resynthesis to establish a strategy of preservation.

AIM: The scarcity of kidneys for transplantation impels an expansion of the donor source to include the use of organs from Maastricht II and Maastricht I non heart-beating donors. The aim of this study was to establish the best method to preserve kidneys from non heart-beating donors (NHBD): cold static storage (CSS) or perfusion. ATP production during kidney preservation has been retained as a measure of their energetic levels and, consequently, their viability. The presence of warm ischemia with both types of preservation was studied. METHODS: Porcine kidneys presenting no warm ischemia or 30 minutes of warm ischemia were submitted to immediate oxygenated hypothermic pulsatile perfusion or immediate cold static storage. The study was divided into four groups. ATP resynthesis was measured after 8 h. of perfusion. ATP was assessed by in-line (31)P nuclear magnetic resonance spectroscopy (NMRS) during the preservations. RESULTS: Only oxygenated perfusion could restore ATP in organs with warm ischemia. Initial cold static storage seems deleterious on organs having suffered from warm ischemia. DISCUSSION: Only oxygenated perfusion could restore ATP in organs with warm ischemia. Initial cold static storage seems deleterious to organs having suffered from warm ischemia. Oxygenated perfusion must be introduced immediately after kidney removal from non heart-beating donors. In organs without warm ischemia, any kind of preservation is equivalent.