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Experimental bio-artificial liver:Importance of the architectural design on ammonia detoxification performance

World Journal of Hepatology 2018年第10期10卷 719-730页

作者： María Dolores Pizarro María Eugenia Mamprin Lucas Damián Daurelio joaquín valentín rodriguez María Gabriela Mediavilla Consejo nacional de Investigaciones Científicas y Técnicas(COnICET) Rosario S2002 LRKArgentina Laboratorio de Investigaciones en Fisiología y Biología Molecular Vegetal(LIFiBVe) Facultad de Ciencias AgrariasUniversidad Nacional del LitoralEsperanza 3080Argentina Facultad de Ciencias Bioquímicas y Farmacéuticas Universidad Nacional de RosarioRosario S2002 LRKArgentina Centro Binacional de Criobiología Clínica y Aplicada(CAIC) Universidad Nacional de RosarioRosario S2011 BXNArgentina Instituto de Biología Molecular y Celular de Rosario(IBR CONICET-UNR)Consejo Nacional de Investigaciones Científicas y Tecnológicasy Universidad Nacional de RosarioRosario S2002 LRKArgentina

AIM To determine the influence of the construction design over the biological component's performance in an experimental bio-artificial liver(BAL) *** Two BAL models for liver microorgans(LMOs) were constructed. First, we constructed a cylindrical BAL and tested it without the biological component to establish its correct functioning. Samples of blood and biological compartment(BC) fluid were taken after 0, 60, and 120 min of perfusion. Osmolality, hematocrit, ammonia and glucose concentrations, lactate dehydrogenase(LDH) release(as a LMO viability parameter), and oxygen consumption and ammonia metabolizing capacity(as LMO functionality parameters) were determined. CPSI and OTC gene expression and function were measured. The second BAL, a "flat bottom" model, was constructed using a 25 cm2 culture flask while maintaining all other components between the models. The BC of both BALs had the same capacity(approximately 50 cm3) and both were manipulated with the same perfusion system. The performances of the two BALs were compared to show the influence of *** The cylindrical BAL showed a good exchange of fluids and metabolites between blood and the BC, reflected by the matching of osmolalities, and glucose and ammonia concentration ratios after 120 min of perfusion. no hemoconcentration was detected, the hematocrit levels remained stable during the whole study, and the minimal percentage of hemolysis(0.65% ± 0.10%) observed was due to the action of the peristaltic pump. When LMOs were used as biological component of this BAL they showed similar values to the ones obtained in a normothermic Reoxygenation System(nRS) for almost all the parameters assayed. After 120 min, the results obtained were: LDH release(%): 14.7 ± 3.1 in the BAL and 15.5 ± 3.2 in the nRS(n = 6); oxygen consumption(μmol/min?g wet tissue): 1.16 ± 0.21 in the BAL and 0.84 ± 0.15 in the nRS(n = 6); relative expression of Cps1 and Otc: 0.63 ± 0.12 and 0.67 ± 0.20, respectively,

关键词： Bio-artificial liver Ammonia detoxification Device design Ornithine Transcarbamylase Rat liver microorgans Carbamyl Phosphate SynthetaseⅠ

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Performance of cold-preserved rat liver Microorgans as the biological component of a simplified prototype model of bioartificial liver

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World Journal of Hepatology 2016年第33期8卷 1442-1451页

作者： María Dolores Pizarro María Gabriela Mediavilla Alejandra Beatriz Quintana ángel Luis Scandizzi joaquín valentín rodriguez María Eugenia Mamprin Centro Binacional de Criobiología Clínica y Aplicada Instituto de Biología Molecular y Celular de Rosario Consejo nacional de Investigaciones Científicas y Técnicas Morfología Facultad de Ciencias Bioquímicas y Farmacéuticas área Farma-cología Facultad de Ciencias Bioquímicas y FarmacéuticasUniversidad Nacional de Rosario

AIMTo develop a simplified bioartificial liver (BAL) device prototype, suitable to use freshly and preserved liver Microorgans (LMOs) as biological component. METHODSThe system consists of 140 capillary fibers through which goat blood is pumped. The evolution of hematocrit, plasma and extra-fiber fluid osmolality was evaluated without any biological component, to characterize the prototype. LMOs were cut and cold stored 48 h in BG35 and ViaSpan® solutions. Fresh LMOs were used as controls. After preservation, LMOs were loaded into the BAL and an ammonia overload was added. To assess LMOs viability and functionality, samples were taken to determine lactate dehydrogenase (LDH) release and ammonia detoxification capacity. RESULTSThe concentrations of ammonia and glucose, and the fluids osmolalities were matched after the first hour of perfusion, showing a proper exchange between blood and the biological compartment in the minibioreactor. After 120 min of perfusion, LMOs cold preserved in BG35 and ViaSpan® were able to detoxify 52.9% ± 6.5% and 53.6% ± 6.0%, respectively, of the initial ammonia overload. no significant differences were found with Controls (49.3% ± 8.8%, P ® cold preserved LMOs, respectively (n = 6, P COnCLUSIOnThis prototype relied on a simple design and excellent performance. It’s a practical tool to evaluate the detoxification ability of LMOs subjected to different preservation protocols.

关键词： Rat liver Microorgans Cold preservation BG35 preservation solution Bioartificial liver device Acute liver failure

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