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      Comparative analysis of in situ versus ex situ perfusion on flow and microcirculation in kidney procurement: research on a porcine model

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          Abstract

          Background

          The first crucial step in transplantation appears to be the effective rinsing of the graft during organ procurement. Even though there is strong suspicion that ex situ perfusion results in better rinsing of the graft, there is no proof for this hypothesis. The aim of this study was to analyse the differences of in situ and ex situ kidney perfusion in a porcine model.

          Methods

          Standardised multiorgan procurement was performed in 15 German landrace pigs. Perfusion was carried out using histidine–tryptophan–ketoglutarate solution (HTK) under the application of pressure. In one kidney, in situ perfusion via the aorta was carried out while the second kidney received ex situ perfusion via the renal artery (RA). Perfusate flow inside the aorta and the RA was recorded at different pressure steps. In order to visualise the effect on the microcirculation, different coloured microparticles (MPs; 10 μm) were administered via the aorta or RA. Subsequently, frozen sections of the explanted kidneys were analysed histologically and MPs were evaluated quantitatively.

          Results

          Ex situ kidney perfusion resulted in significantly improved flow rates ( P<0.0001) compared with in situ perfusion. By applying ex situ perfusion it was even possible to attain physiological flow levels on the RA under the application of external pressure of 150 to 200 mmHg. The amount of MPs was able to highlight the positive impact of ex situ perfusion on microcirculation of the kidney graft ( P<0.0001).

          Conclusions

          The use of MPs represents a valuable tool for quantitative investigation and illustration of kidney perfusion in experimental setups. Additional e x situ perfusion is able to improve the quality of kidney perfusion.

          Translated abstract

          Abstract in German language
          Hintergrund

          Der erste und essentielle Schritt für eine Organtransplantation ist die effektive Spülung des Organs im Rahmen der Organentnahme. Obwohl es einige Hinweise dafür gibt, dass eine ex situ Perfusion die Spülung des Organs verbessern würde, gibt es bisher keinen Beweis für diese Hypothese. Das Ziel dieses Versuches war es daher die Unterschiede zwischen ex situ und in situ Perfusion eines Nierentransplantates in einem Großtiermodell zu untersuchen.

          Methodik

          Eine standardisierte Multi-Organentnahme wurde an 15 Schweinen Deutscher Landrasse durchgeführt. Als Perfusionslösung wurde Histidin-Tryptophan-Ketoglutarat (HTK) verwendet. Auf den Perfusionsbeutel wurde schrittweise externer Druck appliziert. Eine Niere wurde in situ über die Aorta perfundiert, die andere Niere wurde ex situ über die Nierenarterie perfundiert. Die Flussraten der Perfusion wurden unter den verschiedenen Druckstufen gemessen. Um den Surrogatparameter Fluss zu visualisieren, wurden gefärbte Mikropartikel (MPs; 10 μm) appliziert. Die Gefrierschnitte der Nieren wurden histologisch untersucht und die MPs quantifiziert.

          Ergebnisse

          Die ex situ Perfusion resultierte in einer signifikant erhöhten Flussrate ( P<0.0001). Ferner war es unter der ex situ Perfusion durch eine zusätzliche Druckapplikation von 150 bis 200 mmHg möglich physiologische Flusswerte auf der Nierenarterie zu erreichen. Die gemessene Anzahl der MPs konnte den positiven Effekt der ex situ Perfusion auf die Mikrozirkulation im Nierenparenchym verdeutlichen ( P<0.0001).

          Schlussfolgerungen

          Der Einsatz von MPs ermöglicht die Visualisierung der Nierenperfusion in einem experimentellen Versuchsaufbau. Die ex situ Perfusion der Niere bietet eine höhere Perfusionsqualität als die in situ Perfusion.

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          Most cited references12

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          Principles of solid-organ preservation by cold storage.

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            Evaluation of nonradioactive, colored microspheres for measurement of regional myocardial blood flow in dogs.

            Measurement of regional myocardial blood flow (RMBF) is crucial in experimental studies of myocardial ischemia and reperfusion in dogs. The standard measurement technique uses radioactive microspheres; however, not all institutions are able to dispose of radioactive waste and therefore cannot make use of this method. We tested a new, nonradioactive microsphere, labeled with colors instead of nuclides. Simultaneous blood flow measurements with two nuclide-labeled and two colored microspheres were performed after coronary occlusion in dogs. Both techniques show a within-method correlation of r greater than 0.98. Duplicate variability for paired RMBF values in 80 samples was 8.7 +/- 0.1% when computed with radioactive microspheres and 13.2 +/- 1.8% when computed with colored microspheres. There was a good correlation in the measurement of RMBF between the radioactive- and colored-microsphere methods (r = 0.98). The best-fitting linear regression line was expressed by the formula: Colored-microsphere RMBF = 1.11 (radioactive-microsphere RMBF)-0.02. When measured by colored microspheres, RMBF was approximately 8% higher than when computed with radioactive microspheres for blood flow values of 0-2 ml/min/g. When blood flow was increased pharmacologically to levels of 2-7.5 ml/min/g, colored microspheres yielded blood flow values 39% higher than the values computed by radioactive microspheres. We conclude that the nonradioactive, colored-microsphere method correlates with the radioactive technique, but at high flows, it yields values greater than those obtained with radioactive microspheres.
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              Holmium-166 poly(L-lactic acid) microsphere radioembolisation of the liver: technical aspects studied in a large animal model

              Objective To assess the accuracy of a scout dose of holmium-166 poly(L-lactic acid) microspheres (166Ho-PLLA-MS) in predicting the distribution of a treatment dose of 166Ho-PLLA-MS, using single photon emission tomography (SPECT). Methods A scout dose (60 mg) was injected into the hepatic artery of five pigs and SPECT acquired. Subsequently, a ‘treatment dose’ was administered (540 mg) and SPECT, computed tomography (CT) and magnetic resonance imaging (MRI) of the total dose performed. The two SPECT images of each animal were compared. To validate quantitative SPECT an ex vivo liver was instilled with 166Ho-PLLA-MS and SPECT acquired. The liver was cut into slices and planar images were acquired, which were registered to the SPECT image. Results Qualitatively, the scout dose and total dose images were similar, except in one animal because of catheter displacement. Quantitative analysis, feasible in two animals, tended to confirm this similarity (r 2 = 0.34); in the other animal the relation was significantly better (r 2 = 0.66). The relation between the SPECT and planar images acquired from the ex vivo liver was strong (r 2 = 0.90). Conclusion In the porcine model a scout dose of 166Ho-PLLA-MS can accurately predict the biodistribution of a treatment dose. Quantitative 166Ho SPECT was validated for clinical application.
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                Author and article information

                Contributors
                Journal
                Transplant Res
                Transplant Res
                Transplantation Research
                BioMed Central
                2047-1440
                2013
                9 July 2013
                : 2
                : 13
                Affiliations
                [1 ]Department of Transplantation and Hepatobiliopancreatic Surgery, University Medical Centre, Johannes Gutenberg University, Langenbeckstraße 1, 55131, Mainz, Germany
                [2 ]Institute of Neurosurgical Pathophysiology, University Medical Centre, Johannes Gutenberg University, Langenbeckstraße 1, 55131, Mainz, Germany
                [3 ]Institute of Medical Biostatistics, Epidemiology and Informatics, University Medical Centre, Johannes Gutenberg University, Langenbeckstraße 1, 55131, Mainz, Germany
                [4 ]Institute of Pathology, University Medical Centre, Johannes Gutenberg University, Langenbeckstraße 1, 55131, Mainz, Germany
                Article
                2047-1440-2-13
                10.1186/2047-1440-2-13
                3748828
                23837545
                2c1037de-aaca-43a2-adf1-cbb80d0c0047
                Copyright © 2013 Foltys et al.; licensee BioMed Central Ltd.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Categories
                Research

                Transplantation

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