Patzer J F, Mazariegos G V, Lopez R, Molmenti E, Gerber D, Riddervold F, Khanna A, Yin W Y, Chen Y, Scott V L, Aggarwal S, Kramer D J, Wagner R A, Zhu Y, Fulmer M L, Block G D, Amiot B P
Department of Surgery, University of Pittsburgh, Pennsylvania 15213-2582, USA. patzer+@pitt.edu
Ann N Y Acad Sci. 1999 Jun 18;875:340-52. doi: 10.1111/j.1749-6632.1999.tb08516.x.
Preclinical safety and efficacy evaluation of a novel bioartificial liver support system (BLSS) was conducted using a D-galactosamine canine liver failure model. The BLSS houses a suspension of porcine hepatocytes in a hollow fiber cartridge with the hepatocytes on one side of the membrane and whole blood flowing on the other. Porcine hepatocytes harvested by a collagenase digestion technique were infused into the hollow fiber cartridge and incubated for 16 to 24 hours prior to use. Fifteen purpose-bred male hounds, 1-3 years old, 25-30 kg, were administered a lethal dose, 1.5 g/kg, of D-galactosamine. The animals were divided into three treatment groups: (1b) no BLSS treatment (n = 6); (2b) BLSS treatment starting at 24-26 h post D-galactosamine (n = 5); and (2c) BLSS treatment starting at 16-18 h post D-galactosamine (n = 4). While maintained under isoflurane anesthesia, canine supportive care was guided by electrolyte and invasive physiologic monitoring consisting of arterial pressure, central venous pressure, extradural intracranial pressure (ICP), pulmonary artery pressure, urinary catheter, and end-tidal CO2. All animals were treated until death or death-equivalent (inability to sustain systolic blood pressure > 80 mmHg for 20 minutes despite massive fluid resuscitation and/or dopamine administration), or euthanized at 60 hours. All animals developed evidence of liver failure at 12-24 hours as evidenced by blood pressure lability, elevated ICP, marked hepatocellular enzyme elevation with microscopic massive hepatocyte necrosis and cerebral edema, elevated prothrombin time, and metabolic acidosis. Groups 2b and 2c marginally prolong survival compared with Group 1b (pairwise log rank censored survival time analysis, p = 0.096 and p = 0.064, respectively). Since survival times for Groups 2b and 2c are not significantly different (p = 0.694), the groups were combined for further statistical analysis. Survival times for the combined active treatment Groups 2b and 2c are significantly prolonged versus Group 1b (p = 0.047). These results suggest the novel BLSS reported here can have a significant impact on the course of liver failure in the D-galactosamine canine liver failure model. The BLSS is ready for Phase I safety evaluation in a clinical setting.
使用D - 半乳糖胺犬肝衰竭模型对一种新型生物人工肝支持系统（BLSS）进行了临床前安全性和有效性评估。该BLSS在中空纤维盒中容纳猪肝细胞悬液，肝细胞位于膜的一侧，全血在另一侧流动。通过胶原酶消化技术收获的猪肝细胞被注入中空纤维盒，并在使用前孵育16至24小时。15只1 - 3岁、体重25 - 30 kg的专用雄性猎犬被给予致死剂量1.5 g/kg的D - 半乳糖胺。这些动物被分为三个治疗组：（1b）不进行BLSS治疗（n = 6）；（2b）在给予D - 半乳糖胺后24 - 26小时开始BLSS治疗（n = 5）；以及（2c）在给予D - 半乳糖胺后16 - 18小时开始BLSS治疗（n = 4）。在异氟烷麻醉下维持期间，犬的支持性护理以电解质和侵入性生理监测为指导，监测内容包括动脉压、中心静脉压、硬膜外颅内压（ICP）、肺动脉压、导尿管和呼气末二氧化碳。所有动物均接受治疗直至死亡或出现等效死亡情况（尽管进行了大量液体复苏和/或给予多巴胺，但收缩压仍无法维持> 80 mmHg达20分钟），或在60小时时实施安乐死。所有动物在12 - 24小时出现肝衰竭迹象，表现为血压不稳定、ICP升高、肝细胞酶显著升高伴显微镜下大量肝细胞坏死和脑水肿、凝血酶原时间延长以及代谢性酸中毒。与1b组相比，2b组和2c组的存活时间略有延长（成对对数秩检验删失生存时间分析，p分别为0.096和0.064）。由于2b组和2c组的存活时间无显著差异（p = 0.694），因此将这两组合并进行进一步统计分析。合并后的活性治疗组2b和2c的存活时间与1b组相比显著延长（p = 0.047）。这些结果表明，此处报道的新型BLSS可能对D - 半乳糖胺犬肝衰竭模型中的肝衰竭进程产生重大影响。该BLSS已准备好在临床环境中进行I期安全性评估。
