Abstract
Background: The isolated perfused porcine kidney (IPPK) model has been the method of choice for the early preclinical evaluation of kidney graft preservation techniques. The preferred reperfusion conditions have not yet been determined. Here, we examined the effects of pressure- or flow-controlled perfusion and oxygenation by pure oxygen or carbogen (95% O<sub>2</sub>/5% CO<sub>2</sub>) on normothermic reperfusion in the IPPK model. Methods: Porcine kidneys were cold-stored for 24 h in histidine-tryptophan-ketoglutarate solution and reperfused for 1 h with normothermic whole blood/Krebs-Henseleit buffer medium (20/80%). Kidneys (n = 5/group) were flow-controlled reperfused with pure oxygen (1 ml/min/g; Flow-O<sub>2</sub>) or pressure-controlled reperfused (85 mm Hg mean arterial pressure) and oxygenated with either pure oxygen (Pressure-O<sub>2</sub>) or carbogen (Pressure-O<sub>2</sub>/CO<sub>2</sub>). Renal function and damage were assessed during reperfusion and NGAL and HIF-1α levels were analyzed using an ELISA. Results: Pressure-O<sub>2</sub> and Pressure-O<sub>2</sub>/CO<sub>2</sub> were associated with significantly better renal hemodynamics and acid-base homeostasis compared to Flow-O<sub>2</sub>. Urine protein concentrations and the fractional excretion of sodium were lower with both Pressure-O<sub>2</sub> and Pressure-O<sub>2</sub>/CO<sub>2</sub> than with Flow-O<sub>2</sub>. NGAL and HIF-1α levels were also lower with Pressure-O<sub>2</sub> and Pressure-O<sub>2</sub>/CO<sub>2</sub> than with Flow-O<sub>2</sub>. Only Pressure-O<sub>2</sub>/CO<sub>2</sub> could demonstrate a significantly increased urine production compared to Flow-O<sub>2</sub>. The structural integrity was well preserved in the Pressure-O<sub>2</sub> and Pressure-O<sub>2</sub>/CO<sub>2</sub> groups, whereas diffuse and global glomerular destruction was observed in the Flow-O<sub>2</sub> group. Conclusion: In the IPPK model, the application of pressure-controlled reperfusion with carbogen oxygenation, and to a lesser extent with pure oxygen, maintained physiological renal function for 1 h, thus providing a reliable and reproducible ex vivo evaluation of kidney preservation quality.
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