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Evaluation of Normothermic Flush and Perfusion Techniques, Using a Novel Normothermic Preservation Solution, in an Isolated Porcine Haemoperfusion Model

By Mark Kay


The basis of the work in this thesis is an assessment of renal preservation techniques. The first chapter comprises a review of the pathophysiology of ischaemic injury and the role of hypothermic preservation. In the second chapter a review of the various preservation solutions in use is presented as well as the constituents of preservation solutions. The role of the organ flush is then presented, comparing warm and cold flush. The effect of viscosity and pressure is then examined followed by a review of hypothermic and normothermic preservation methods.\ud The five main experimental chapters are then presented as a series of papers.\ud The first of these aimed to assess the rate of organ cooling during back-table flush using different preservation solutions. In this paper, porcine kidneys were flushed with UW or Soltran solution, and the perfusion rate and cooling of the kidney was assessed. The second paper follows on comparing organ cooling using either a cold flush or a warm flush followed by a cold flush to compare flush rates and cooling times. The third paper assesses different flush techniques using a novel normothermic preservation solution, AQIX® RS-I, to assess the optimal flush conditions and functional outcomes. The fourth paper compares functional results of AQIX with the more commonly used cold flush solutions, Soltran and University of Wisconsin solution. The final experiment assesses the effectivness of complete normothermic preservation using AQIX® RS-I. An isolated organ perfusion system using cardiopulmonary bypass equipment was used for normothermic preservation and as a surrogate for transplantation. Porcine kidneys were used throughout all experiments as large animal models best reflect the human condition

Publisher: University of Leicester
Year: 2010
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  1. (1967). 24-hour and 72-hour preservation of canine kidneys. Lancet
  2. (1976). 72 hour canine kidney preservation without continuous perfusion. Transplantation
  3. A comparison of cadaver kidney storage methods: pump perfusion and cold storage solutions. Clin Transplant 1993;7:
  4. (2004). A comparison of renal preservation by cold storage and machine perfusion using a porcine autotransplant model. Transplantation
  5. A comparison of the results of renal transplantation from non-heart-beating, conventional cadaveric and living donors.
  6. (1974). A fibrinogen –free plasma perfusate for preservation of kidneys for one hundred and twenty hours. Surg Gynae Obstet
  7. (2003). A Guide for the preparation and use of buffers in biological systems. Calbriochem R, EMD Biosciences INC.
  8. A lipid-phase separation model of low-temperature damage to biological membranes.
  9. A multicenter, randomized, controlled trial of Celsior for flush and hypothermic storage of cardiac allografts.
  10. (2002). A paired study comparing the efficacy of renal preservation by normothermic autologous blood perfusion and hypothermic pulsatile perfusion. Transplant Proc.
  11. (2004). A pathophysiologic study of the kidney tubule to optimise organ preservation solutions. Kidney International
  12. (1992). A preliminary report of the HTK randomised multicentre study comparing kidney graft preservation with HTK and EuroCollins solutions. Transpl Intl 5:S429,
  13. A prospective controlled trial of cold-storage versus machine-perfusion preservation in cadaveric renal transplantation.
  14. A randomised prospective trial of cold storage versus pulstaile machine perfusion for cadaver kidney preservation. Transplantation 1987;43:
  15. An automated and portable low-flow perfusion system for organ preservation.
  16. Analysis of the optimal perfusion pressure and flow rate of the vascular resistance and oxygen consumption in the hypothermic perfused kidney.
  17. Are we frozen in time? Analysis of the utilization and efficacy of pulsatile perfusion in renal transplantation.
  18. (2002). ATP supplies in the cold preserved liver: A long neglected factor of organ viability. Hepatology
  19. Beneficial effects of adenosine and phosphate in kidney preservation.
  20. (1992). Beneficial effects of prewarming the donor liver for rat liver transplantation.
  21. (1973). Benefits of methylprednisolone in the isolated perfused organ. Ann Surg
  22. (1999). Biphasic role for nitric oxide in experimental renal warm ischemia-reperfusion injury.
  23. (1991). Carolina rinse solution--a new strategy to increase survival time after orthotopic liver transplantation in the rat. Transplantation
  24. (2004). Choosing the right preservation solution. Global Surgery
  25. (2000). Churchill Conservation of phosphorylation state of cardiac phosphofructokinase during in vitro hypothermic hypoxia.
  26. (2000). Cold preservation induced sensitivity of rat hepatocyte function to rewarming injury and its prevention by short-term reperfusion. Hepatology
  27. (1989). Combined cold storage-perfusion preservation of the kidney with a new synthetic perfusate. Transplant Proceedings
  28. (2006). Comparative efficacy of renal preservation solutions to limit functional impairment after warm ischemic injury. Kidney International
  29. (2006). Comparison of histidine-tryptophan-ketoglutarate and University of Wisconsin in living-donor liver transplantation. Transplant Proceedings
  30. Comparison of simple hypothermic storage, pulsatile perfusion with Belzers gluconate-albumin solution, and pulsatile perfusion with UW solution for renal allograft preservation. Transplant Proc 1991;23:
  31. (2006). Comparison of two gene transfer models for the attenuation of myocardial ischemiareperfusion injury following preservation for cardiac transplantation. Eur J Cardiothorac Surg
  32. Compartmentalization of neutrophils in the kidney and lung following acute ischemic kidney injury.
  33. Consideration of the inorganic and organic composition of mammalian perfusion solutions.
  34. Custodiol HTK solution for Kidney/Liver preservation. Prescribing information
  35. Defence strategies against hypoxia and hypothermia.
  36. Degree and time sequence of hypothermic protection against experimental ischaemic acute renal failure.
  37. (1997). Deletion of hydroxyethylstarch from University of Wisconsin solution induces cell shrinkage and proteolysis during and after cold storage of rat liver. Hepatology
  38. (2008). Donor preconditioning with taurine protects kidney grafts from injury after experimental transplantation.
  39. (1997). Effect of flushperfusion on vascular endothelial and smooth muscle function. Ann Thoarc Surg
  40. Effect of oxidized and reduced glutathione in liver preservation. Transplantation 1990;50: 948
  41. (1992). Effect of preservation solution on results of cadaveric kidney transplantation. The European Multicentre Study Group. Lancet
  42. Effect of temperature on hepatic and renal uptake of components from university of Wisconsin solution.
  43. (2006). Effects of arterial pressure in an experimental isolated haemoperfused porcine kidney preservation system. Br J Surg.
  44. (1991). Effects of calcium depletion and calcium paradox on the ultrastructure of th frog heart.
  45. Effects of Carolina rinse and adenosine on micovascular perfusion and intrahepatic leukocyteendothelium interaction after liver transplantation in the rat.
  46. (1996). Effects of different cold preservation solutions on restoration of hepatic energy metabolism during cold reperfusion. Cryobiology
  47. Effects of hypothermia on canine kidney mitochondria.
  48. (1988). Effects of perfusion pressure during flushing on the viability of the procured liver using noninvasive fluorometry. Transplantation
  49. Effects of warm Carolina rinse on microvascular reperfusion injury in rat liver transplantation.
  50. Endotheilial cell injury induced by preservation solutions: a confocal microscopy study.
  51. Endothelin up-regulation and localisation following renal ischemia and reperfusion.
  52. Englesbe MJ. Comparison of histidine-tryptophan-ketoglutarate and University of Wisconsin preservation in renal transplantation.
  53. Enhanced resistance to the effects of normothermic ischaemia in kidneys using pulsatile machine perfusion.
  54. (2004). Ergothioneine pretreatment protects the liver from ischemia-reperfusion injury caused by increasing hepatic heat shock protein 70. J Surg Res,
  55. (2003). et al Hepatic preconditioning after prolonged warm ischemia by means of S-adenosyl-L-methionine administration in pig liver transplantation from non-heart-beating donors. Transplantation.
  56. (2000). et al Porcine liver transplantation from non-heart-beating cadaver donor: effect of initial flushing with cold versus warm UW solution. Transplant Proc.
  57. Euro-Collins, low-potassium dextran, and Krebs-Henseleit solutions for hypothermic lung preservation.
  58. Evaluation of eight preservation solutions for Endothelial in situ preservation.
  59. (1994). Evaluation of preservation conditions and various solutions for small bowel preservation. Transplantation
  60. (2007). Evaluation of preservation solutions by ESR-spectroscopy: superior effects of University of Wisconsin over Histidine-Tryptophan-Ketoglutarate in reducing renal reactive oxygen species. Kidney International
  61. Evidence that late perfusion does not influence delayed allograft function.
  62. (1973). Experimental and clinical results of continuous albumin perfusion of kidneys.
  63. Experimental evaluation of Celsior, a new heart preservation solution.
  64. (2008). Experimental renal preservation by normothermic resuscitation perfusion with autologous blood.
  65. (2002). Extended preservation of non-heart-beating donor livers with normothermic machine perfusion.Br J Surg.
  66. (2002). Extended preservation of rat liver graft by induction of heme oxygenase-1. Hepatology,
  67. Fibrinolytic preflush upon liver retrieval from non-heart-beating donors to enhance post-preservation viability and energetic recovery upon reperfusion.
  68. Forty-eight to 96-hour preservation of canine kidneys by initial perfusion and hypothermic storage using the Euro-Collins solution.
  69. (2002). Friend PJ Advantages of normothermic perfusion over cold storage in liver preservation. Transplantation.
  70. (2007). Glutamine donor pretreatment in rat kidney transplants with severe preservation reperfusion injury.
  71. (1999). Gravity perfusion versus high-pressure perfusion in kidney transplantation: results from a prospective randomized study. Transplant Proceedings
  72. (2003). Heme oxygenase-1 attenuates ischemia/reperfusion-induced apoptosis and improves survival in rat renal allografts. Kidney Int,
  73. (1998). Hepatic blood flow and oxygen extraction ratio during normothermic recirculation and total body cooling as viability predictors in non-heart-beating donor pigs. Transplantation
  74. (2007). Histidine-tryptophanketoglutarate solution vs. University of Wisconsin solution for liver transplantation: a systematic review. Liver Transpl.
  75. (2003). Hyperaggregating effect of hydroxyethyl starch components and University of Wisconsin solution on human red blood cells: a risk of impaired graft perfusion in organ procurement? Transplantation
  76. (2001). Hypothermia--a limiting factor in using warm ischemically damaged kidneys.Am J Transplant.
  77. (1989). Hypothermic perfusion of rabbit livers: effect of perfusate composition (Ca and Lactobionate) on enzyme release and tissue swelling. Cryobiology
  78. Hypothermic perfusion preservation of liver: the role of phosphate in stimulating ATP synthesis studied by 31P NMR Transplant International1995;
  79. Hypothermic perfusion preservation: The future of organ preservation revisited?
  80. (1989). Hypothermic preservation of hepatocytes. I. Role of cell swelling. Cryobiology
  81. Immediate function and cost comparison between static and pulsatile preservation in kidney recipients.
  82. Impact of organ preservation using HTK for graft flush and subsequent storage in UW in rat kidney transplantation. European Surg Res.
  83. Importance of glutathione and adenosine in cold storage of the kidney.
  84. (1990). Importance of glutathione and adenosine in cold storage of the kidney. Transplant
  85. Improved porcine renal preservation with a simple extracellular solution- PBS140.
  86. (2005). Improved rat liver preservation by hypothermic continous machine perfusion using polysol, a new, enriched preservation solution. Liver Transpl.
  87. Improvement of microvascular graft equilibration and preservation in non-heartbeating donors by warm preflush with streptokinase.
  88. (2004). In: Anaesthesia and Intensive care, principles and practice;
  89. Increase in survival of liver grafts after rinsing with warm ringers solution due to improvement of hepatic microcirculation.
  90. (1998). Inferior outcome of cadaveric kidneys preserved for more than 24 hours in HTK. Transplantation
  91. (1996). Influence of pulmonary arterial pressure during flushing on lung preservation. Transplantation
  92. (2007). Influence of pulsatile perfusion preservation on outcomes in kidney transplantation from expanded criteria donors.
  93. Initial blood washout during organ procurement determines liver injury and function after preservation and reperfusion.
  94. (2006). Intermediate-term outcomes with expanded criteria deceased donors in kidney transplantation: a spectrum or specter of quality? Ann Surg
  95. (1998). Intracellular volumes and membrane permeability in rat hearts during prolonged hypothermic preservation with St Thomas and University of Wisconsin solutions.
  96. (1996). Intravital studies on beneficial effects of warm Ringer's lactate rinse in liver transplantation.
  97. Ischaemia-reperfusion injury.
  98. (2000). Ischemia-reperfusion associated myocardial contractile dysfunction may depend on Ca (2+)-activated cytoskeleton protein degradation. Cardiovasc Res
  99. (1969). Kidney preservation for transportation. Initial perfusion and 30 hours ice storage. Lancet
  100. (1999). Kidney preservation in the next millenium. Transpl Int
  101. (2001). Kidney preservation with university of Wisconsin and Celsior solution: a prospective multicenter randomized study. Transplantation
  102. Kidney transplantation from elderly donors: a prospective randomized study comparing celsior and UW solutions.
  103. (2003). Koostra G Induction of heme oxygenase-1 in kidneys during ex vivo warm perfusion.Transplantation.
  104. (2005). Koostra G.Repair of damaged organs in vitro.Am J Transplant.
  105. (1994). Kootstra G: University of Wisconsin solution is superior to histidine-tryptophan-ketoglutarate for preservation of ischaemically damaged kidneys. Transplantation 58:979-84,
  106. (1990). Lactobionate as the most important component in UW solution for liver preservation. Transplant Proc
  107. (2000). lactobionate, and histidine: cryptic inhibitors of matrix metalloproteinases contained in University of Wisconsin and histidine/tryptophan/ketoglutarate liver preservation solutions. Hepatology
  108. Lactobionic acid as an iron chelator: a rationale for its effectiveness as an organ preservant.
  109. Late perfusion; A simple remedy for renal allograft primary nonfunction.
  110. Late refush in clinical renal transplantation. Protection against delayed graft function not observed.
  111. (2006). Leucocyte depletion improves renal function during reperfusion using an experimental isolated haemoperfused organ preservation system.
  112. (1938). Lindbergh CA. The culture of organs.
  113. (1998). Lipid-mediated gene transfer of viral IL-10 prolongs vascularized cardiac allograft survival by inhibiting donor-specific cellular and humoral immune responses. Gene Therapy
  114. (2001). Liver transplantation after organ preservation with normothermic extracorporeal perfusion. Ann Surg.
  115. (1997). Liver Transplantation From Non–Heart Beating Donors in Rats: Influence of Viscosity and Temperature of Initial Flushing Solutions on Graft Function. Liver Transpl Surg
  116. Low viscosity histidine-tryptophanketoglutarate graft flush improves subsequent extended cold storage in University of Wisconsin solution in an extracorporeal rat liver perfusion and rat liver transplantation model.
  117. (2009). Machine perfusion or cold storage in deceased-donor kidney transplantation.
  118. (2006). Machine perfusion preservation of the pig liver using a new preservation solution, polysol. Transplant Proc.
  119. (1996). Membrane damage and the Ca2+- paradox in the perfused rat kidney.
  120. (2001). Methods of Renal Preservation. Transplantation reviews
  121. Microcirculatory disturbances and leucocyte adherence in transplanted livers after cold storage in EuroCollins, UW and HTK solutions.
  122. Microvascular dysfunction induced by reperfusion injury and protective effect of ischaemic preconditioning. Free Radic. Biol Med.
  123. Mosca F Pancreas preservation with University of Wisconsin and Celsior solutions.
  124. (2007). Multicenter analysis of kidney preservation. Transplantation
  125. (2003). Multigene adenoviral therapy for the attenuation of ischemia-reperfusion injury after preservation for cardiac transplantation. J Thorac Cardiovasc Surg
  126. (2002). Myocardial preservation using Celsior solution in cardiac transplantation: early results and 5-year follow-up of a multicenter prospective study of 70 cardiac transplantations. Ann Thorac Surg.
  127. Neutrophil and Kupffer cell-induced oxidant stress and ischaemia-reperfusion injury in the rat liver.
  128. New concepts in organ preservation. Clinical Transplantation1997;7:
  129. (2002). New Solutions in Organ Preservation Transplantation Reviews
  130. (2008). Nicholson ML The relative effects of warm and cold ischemic injury in an experimental model of non-heart-beating donor kidneys. Transplantation
  131. (2007). Nicholson ML.Static normothermic preservation of renal allografts using a novel non- phosphate buffered preservation solution. Transplant International
  132. (1995). Non-heart beating kidney donation in uncontrolled donor procedures. Transplant Proceedings
  133. Normothermic recirculation reduces primary graft dysfunction of kidneys obtained from non-heartbeating donors.
  134. (2003). NOS: the underlying mechanism preserving vascular integrity and during ex vivo warm kidney perfusion.
  135. (2002). Optimisation of bile production during normothermic preservation of porcine livers.
  136. organ-preservation solution on Haemorheology. Transplant Int.
  137. (2002). Overcoming severe renal ischemia: the role of ex vivo warm perfusion.Transplantation.
  138. Oxygen consumption and blood flow in the hypothermic perfused kidney.
  139. (2004). Pancreas preservation with University of Wisconsin and Celsior solutions: a single-center, prospective, randomized pilot study. Transplantation
  140. Parada B Efficacy of renal preservation: comparative study of Celsior and University of Wisconsin solutions.Transplant
  141. (1999). Persijin GG: Eurotransplant randomized multicenter kidney graft preservation study comparing HTK with UW and Euro-Collins. Transpl Int 12(6):447-53,
  142. (2004). Phosphate increases mitochondrial reactive oxygen species release Free Radic.Res
  143. (1996). Physiological and histological characterization of a pig kidney in vitro perfusion model for xenotransplantation studies.
  144. (2004). Pig kidney: anatomical relationships between the intrarenal arteries and kidney collecting system. Applied study for urological research and surgical training.
  145. (2003). Preconditioning with tin-protoporphyrin IX attenuates ischemia/reperfusion injury in the rat kidney. Kidney Int.
  146. (1992). Preservation and reperfusion injuries in liver allografts. An overview and synthesis of current studies. Transplantation
  147. (1970). Preservation and transplantation of human cadaver kidneys: A two year experience. Ann Surg
  148. (1997). Preservation by simple hypothermia.
  149. Preservation damage in liver transplantation. Influence of rapid cooling.
  150. Preservation methods do not affect renal allograft outcome. The SEOPF prospective study 1977-1982. Transplant Proc 1984;14:
  151. Preservation of pig liver allografts after warm ischaemia: normothermic perfusion versus cold storage.
  152. (2004). Preservation of porcine nonheart-beating donor livers by sequential cold storage and warm perfusion.Transplantation.
  153. Preservation of the canine liver for 24-48 hours using simple cold storage with UW solution. Transplantation 1988;46:517-
  154. Preservation of the liver graft with Celsior solution.
  155. (2001). Pretransplantation prognostic testing on damaged kidneys during ex vivo warm perfusion.Transplantation.
  156. (1999). Prolongation of cardiac allograft survival with intracoronary viral interleukin-10 gene transfer.Transplant Proc.
  157. (2004). Prolonged hypothermic machine perfusion preserves hepatocellular function but potentiates endothelial cell dysfunction in rat livers. Transplantation
  158. (1993). Prospective randomized comparison of University of Wisconsin and UW-modified, lacking hydroxyethyl-starch, cold-storage solutions in kidney transplantation. Transplantation
  159. (2008). Protective effects of apocynin and allopurinol on ischemia/reperfusion-induced liver injury in mice. World
  160. (1994). Protective effects of various preservation solutions on cultured endothelial cells. Annals of Thoracic Surgery
  161. Pulsatile machine perfusion versus cold storage of kidneys for transplantation: a rapid and systematic review.
  162. (2006). Pulsatile perfusion reduces the incidence of delayed graft function in expanded criteria donor kidney transplantation.
  163. Pulsatile preservation in renal transplantation.
  164. (2006). Randomized clinical assay for hepatic grafts preservation with University of Wisconsin or histidine-tryptophan-ketoglutarate solutions in liver transplantation.
  165. (1997). Randomized study on in situ liver perfusion techniques: gravity perfusion vs high-pressure perfusion. Transplant Proceedings
  166. Reactive oxygen metabolites and reperfusion injury: aberrant triggering of reticuloendothelial function.
  167. (1991). Reactive oxygen species and rat renal epithelial cells during hypoxia and reoxygenation. Kidney Int
  168. Reduced renal vascular injury following warm ischaemia and preservation by hypothermic machine perfusion. Transplant Proc 1997;29:
  169. (1997). Reduction of proteolysis by venous-systemic oxygen persufflation during rat liver preservation and improved functional outcome after transplantation. Transplantation
  170. Regeneration of ATP in kidney slices after warm ischaemia and hypothermic preservation.
  171. (1998). Relationship between flushing pressure and nitric oxide production in preserved lungs. Transplantation
  172. Renal ischaemia-reperfusion injury.
  173. Renal preservation by ice cooling. An experimental study relating to kidney transplantation from cadavers.
  174. (2001). Renal preservation.
  175. (1996). Retrovirus-mediated transfer of viral IL-10 gene prolongs murine cardiac allograft survival.
  176. (1996). Shumakov V: Histidine-tryptophan-ketoglutarate versus EuroCollins for preservation of kidneys from non-heart-beating donors. Transplant Proc 28:202,
  177. Single-donor cold storage versus machine perfusion in cadaver kidney transplantation.
  178. (1984). Six-day canine kidney preservation. Hypothermic perfusion combined with isolated blood perfusion. Transplantation.
  179. Successful 72 hour storage of dog kidneys with UW solution.
  180. (2002). Successful extracorporeal porcine liver perfusion for 72 hr. Transplantation.
  181. (1989). Successful five-day perfusion preservation of the canine kidney. Transplantation
  182. (1993). Successful seven-day perfusion of the canine kidney.
  183. (2005). The effect of normothermic recirculation is mediated by ischemic preconditioning in NHBD liver transplantation.
  184. The effects of allopurinol in hepatic ischemia and reperfusion: experimental study in rats. Eur Surg Res.
  185. (1988). The effects of cooling rates and storage temperature on the function of 24-hour cold-preserved rabbit kidneys. Transplantation
  186. The endothelial damage of pulsatile renal preservation and its relationship to perfusion pressure and colloid osmotic pressure.
  187. (1991). The function of a colloid in liver cold-storage preservation. Transplantation
  188. (1991). The functional effects of suppression of hypothermia-induced cell swelling in liver preservation by cold storage. Cryobiology
  189. (1993). The influence of the temperature of storage on survival and ultrastructure of transplanted UW-preserved rat liver grafts. Transplantation
  190. (1971). The no reflow phenomenon in renal ishaemia. Lab Invest
  191. (2002). The optimal pressure for initial flush with UW solution in heart procurement.
  192. (1994). The pig as a potential xenograft donor. Vet Immuno Immunopathology
  193. The problem of renal preservation.
  194. The reflush effect- A prospective analysis of late perfusion.
  195. (1983). The role of oxygen free radicals in human disease processes. Surgery
  196. The use of non-heart-beating cadaver donors in experimental liver transplantation.
  197. (2003). Trends in Organ Preservation. Transplant International
  198. (2002). UW) cold storage solution. Prescribing information
  199. (1998). Warm flush at 37 degrees C following cold storage attenuates reperfusion injury in preserved rat livers.
  200. Warm preflush with streptokinase improves microvascular procurement and tissue integrity in liver graft retrieval from non-heart-beating donors.
  201. (2005). Wash-out of the non-heart-beating donor liver: A comparison between Ringer Lactate, HTK and Polysol. Transplant Proceedings
  202. (2004). Weight Increase during machine perfusion may be an indicator of organ and in particular, Vascular damage. Ann Transplant

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