38th Scientific
Meeting (GV - SOLAS)
Society for Laboratory Animal Science
(Essen, 2000)
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38th Scientific
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(Essen, 2000) |
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Kidney Abstract 1 Effects of Different Perfusates on Functional Parameters of Isolated Perfused Dog Kidneys J. Höchel (1), V. Unger (2), S. Dittrich (3), D. Lehmann (1),C. Große-Siestrup (2), C. Fehrenberg (2), H. Hartmann (1) (1)Freie Universität Berlin, Institut für Veterinär-Physiologie; (2) Humboldt-Universität zu Berlin, Charité, Campus Virchow-Klinikum, Tierexperimentelle Einrichtung; (3)Deutsches Herzzentrum Berlin, Abt. Angeborene Herzfehler Introduction Ischemia-reperfusion injury (IRI) is a major cause of renal dysfunction in both native kidneys and renal allografts. The isolated perfused kidney (IPK) is a useful model to study the cascade of events associated with IRI and its possible diminution or even prevention. The purpose of this study was to compare parameters of renal function in IPK from pigs and dogs in order to detect potential species differences, and to investigate the effect of the dog's age on the extent of the IRI. Material and Methods Over a period of 2.5 years 155 porcine slaughterhouse kidneys were normothermic haemoperfused with autologous blood after cold storage with Baeyer II solution. Duration of warm ischaemia was 18 ± 7 min, kidneys were stored over 5 ± 2 h. In the reperfusion system the diluted (Hct = 0.2) autologous blood recirculated continuously through the kidney and was dialyzed against a ten-fold volume of Tyrode solution. For the dog study 34 kidneys were harvested from animals, which had to be euthanized in veterinary practices for miscellanous reasons unrelated to our study. The age of the dogs ranged from 1.5 to 17 years (<3 years n=4; 3-8 years n=6; >8 years n=7). In the dogs the duration of warm ischaemia was 12 ± 7 min and of cold storage 4.5 ± 2 h. Reperfusion was performed in analogy to the pig study. Results In order to assess renal function during reperfusion parameters concerning (1) haemodynamics, (2) glomerular filtration and (3) tubular secretion and reabsorption have to be considered. In both studies 120 parameters were measured or calculated. After reaching a haemodynamically steady state perfusate flow rate was 1.2 ± 0.7 ml/min/g kidney weight (kw) in pigs compared to 1.1 ± 0.5 ml/min/g kw in dogs. Interquartile ranges of GFR measured as creatinine clearance were 0.01 to 0.25 and 0.002 to 0.032 ml/min/g kw, respectively, and of resulting filtration fraction 4 to 16 and 0.002 to 0.027 %. FENa interquartile ranges were 0.06 to 0.4 and 0.3 to 0.6, values for FEK were 0.6 to 1.4 and 1.3 to 2.9 in pigs and dogs, respectively. There was no statistically significant correlation between the age of the dogs and the above parameters.
Conclusions In respect to the parameters listed here canine kidneys seem to be more susceptible to IRI than porcine kidneys under comparable ischaemia and reperfusion conditions. Beside species differences the generally higher age of the dogs compared to the slaughterhouse pigs has to be considered. In vivo data concerning the kidney function from the same dogs as used for the ischaemia-reperfusion experiments supports this aspect. top Abstract 2 Isolated Pig Kidney Perfusion: 4-hour-cold-Preservation Superior to Fresh Harvested Unpreserved Unger V., von Baeyer H., Große-Siestrup C. Institute of Transfusion Medicine and Immunhaematology and Animal Experimental Facilities of Charite´, Campus Virchow Klinikum, Humboldt University, Berlin, Germany Introduction Isolated pig organ hemoperfusion can be used as a standardized method for pharmaka and medical device testing. The use of slaughterhouse organs helps to reduce or to replace animal experiments. However the influence of organ harvesting (surgical trauma, warm ischemic time) on organ function may be decisive for the results. Another important influence on organ function is the conservation method (conservation fluid, cold ischemic time). In this study the function of organs originating from slaughterhouse (SLA) is compared with organs explanted surgically in the operation-room (OP) under controlled conditions. Methods This retrospective study on 120 kidney perfusions was performed with the following groups: A (n=18) OP, immediately perfused, no preservation; B (n=12) OP, 4 hrs cold preservation; C (n=84) SLA, 4 hrs cold preservation; D (n=6), SLA, 1 day cold preservation. Standard perfusion technique [H. von Baeyer et al.: Biomed.Technik, Bd.42, p.61, 1997] with an optimized conservation medium [C. Fehrenberg et al.: Proc. 36th GV-SOLAS Scient.Meeting 1998,Print: Lab.Animals, ISBN 0-901334-12-x] was used with a steady state perfusion time over 90 min. Results The best organ function was in group B and the poorest function values in group D.
Both 4-h-cold preserved groups (B, C) show superior values (related to organ weight). Discussion Not the organs of group A (freshly perfused, unpreserved) but those of group B and C (4 h-cold-preserved organs) are more comparable to normal incorporated organs. Diuresis and mineral handling is even more physiologic in the slaughterhouse organs (group C). Thus slaughterhouse kidneys, harvested and perfused with a standardized technique and preserved with a special composed conservation fluid offer a realistic, economic, reliable and physiologic replacement of animal experiments. (Abstract für GV-Solas, 38th Sc. Meeting, 11.-14.9.00, Essen: Workshop Isolated Organs) top Abstract 3 Renal Ischaemia-Reperfusion Injuries - Influence of Species and Age on Functional Parameters in the Isolated Perfused Kidney from Dogs and Pigs Höchel J, Unger V, Dittrich S. Lehmann D. Große-Siestrup C. Fehrenberg, C. Harmann H. Institute of Transfusion Medicine and Immunhaematology and Animal Experimental Facilities of Charite´, Campus Virchow Klinikum, Humboldt University, Berlin, Germany Introduction Ischemia-reperfusion injury (IRI) is a major cause of renal dysfunction in both native kidneys and renal allografts. The isolated perfused kidney (IPK) is a useful model to study the cascade of events associated with IRI and its possible diminution or even prevention. The purpose of this study was to compare parameters of renal function in IPK from pigs and dogs in order to detect potential species differences, and to investigate the effect of the dog's age on the extent of the IRI. Material and Methods Over a period of 2.5 years 155 porcine slaughterhouse kidneys were normothermic haemoperfused with autologous blood after cold storage with Baeyer II solution. Duration of warm ischaemia was 18 ± 7 min, kidneys were stored over 5 ± 2 h. In the reperfusion system the diluted (Hct = 0.2) autologous blood recirculated continuously through the kidney and was dialyzed against a ten-fold volume of Tyrode solution. For the dog study 34 kidneys were harvested from animals, which had to be euthanized in veterinary practices for miscellanous reasons unrelated to our study. The age of the dogs ranged from 1.5 to 17 years (<3 years n=4; 3-8 years n=6; >8 years n=7). In the dogs the duration of warm ischaemia was 12 ± 7 min and of cold storage 4.5 ± 2 h. Reperfusion was performed in analogy to the pig study. Results In order to assess renal function during reperfusion parameters concerning (1) haemodynamics, (2) glomerular filtration and (3) tubular secretion and reabsorption have to be considered. In both studies 120 parameters were measured or calculated. After reaching a haemodynamically steady state perfusate flow rate was 1.2 ± 0.7 ml/min/g kidney weight (kw) in pigs compared to 1.1 ± 0.5 ml/min/g kw in dogs. Interquartile ranges of GFR measured as creatinine clearance were 0.01 to 0.25 and 0.002 to 0.032 ml/min/g kw, respectively, and of resulting filtration fraction 4 to 16 and 0.002 to 0.027 %. FENa interquartile ranges were 0.06 to 0.4 and 0.3 to 0.6, values for FEK were 0.6 to 1.4 and 1.3 to 2.9 in pigs and dogs, respectively. There was no statistically significant correlation between the age of the dogs and the above parameters. Conclusions In respect to the parameters listed here canine kidneys seem to be more susceptible to IRI than porcine kidneys under comparable ischaemia and reperfusion conditions. Beside species differences the generally higher age of the dogs compared to the slaughterhouse pigs has to be considered. In vivo data concerning the kidney function from the same dogs as used for the ischaemia-reperfusion experiments supports this aspect. top Abstract 4 New experiences with xenoperfusion by the use of a pig- to- human setting Aslan T, Grosse- Siestrup C, Unger V, von Baeyer H., Salama A. Institute of Transfusion Medicine and Immunhaematology and Animal Experimental Facilities of Charite´, Campus Virchow Klinikum, Humboldt University, Berlin, Germany Introduction In vitro organ perfusions present a useful tool to study hyperacute xenograft rejection (HXR). Recently, we have described an in vitro autologous pig kidney perfusion model (von Baeyer, Biomed. Tefchnik, Bd. 42, p. 61, 1997). In this model the organ is connected to an artificial circulatory circuit including a standard dialysis module, which carries out gas exchange and dialysis simultaneously to allow a perfusion at physiologic pressure, temperature and oxygenation conditions. Here, we describe our new experiences with this perfusion system. Method After autologous haemoperfusion all studied kidneys were perfused with human whole blood (n= 15). Autologous perfused kidneys served as control (n= 5). To exclude interferences of the perfusate with the tubing system of the perfusion circuit we performed two experiments without a kidney. These control-perfusions showed no signs of haemolysis or coagulation activation. Results Immediately after entrance of human blood into the pig kidney, the organ function declined with increasing vascular resistance and makrohaematuria. The maximum renal blood flow (RBF) and the fractioned sodium reabsorption declined similary. Histological analysis revealed glomerular stasis with formation of fibrin clots and bleedings into the bowmann capsula. Immunohistochemical staining demonstrated a homogen deposition of IgM along the glomerular and peritubular endothelium. These findings show humoral and cellular reactions corresponding to HXR. Surprisingly, we found interindividual factors influencing the course of HXR- manifestation. Impressing differences were observed between human individuals, which mostly seem to be dependent on the broad variation of xeno-antibody titers. Additionaly pig kidneys with a weight of more than 200 ± 30 g showed a delayed hyperacute rejection. We believe that our in vitro xenoperfusion model will be useful for further investigations to elucidate these new experiences. (Key words: isolated organ haemoperfusion, xenoperfusion, pig- to- human, HXR, pig kidney) top |
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| August 6, 2000 | Copyright
© 2000 (UNI - Klinikum |
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