Efficacy of double filtration plasmapheretic cross-circulation using a high permeability membrane between totally hepatectomized dogs and donor pigs.

BACKGROUND: Major obstacles to develop a bioartificial liver are xenogeneic immune reactions and viral infection from donor pigs. To solve these problems, we studied the effect of xenogeneic double filtration plasmapheretic cross-circulation (DFPCC) using a high performance semipermeable membrane on totally hepatectomized dogs. METHODS: Mongrel dogs, weighing 12-15 kg, underwent total hepatectomy in one stage (n=18). One hr after total hepatectomy, the femoral vein and the jugular vein were cannulated in both dogs and pigs by using the blood access catheter tubes that were connected to the DFPCC system. In the DFPCC circuit, filtrated dog plasma and pig plasma counterflowed in a hollow fiber cartilage at a rate of 25 ml/min for 6 hr and met through a semipermeable membrane with 100 kd nominal molecular weight cut-off (n=5). In control dogs, the circuit was not connected to the pig (n=13). RESULTS: In vitro mass transfer study suggested that very little immunoglobulins crossed the semipermeable membrane. During and after 6 hr of DFPCC, anhepatic dogs had significantly lower blood ammonia and aromatic amino acid levels than did controls. DFPCC-treated dogs demonstrated decreased intracranial pressure and survived significantly longer than control dogs (20.75+/-3.80 hr vs. 14.75+/-1.30 hr, P<0.05). Histology showed no xenogeneic rejection in both dogs and pigs. CONCLUSIONS: Our DFPCC systems with a high permeability membrane demonstrated detoxification-function and contributed to intracranial decompression and longer animal survivals without adverse immune reaction and the possibility of zoonosis.

Efficacy of double-filtration plasmapheretic cross-circulation with a high-permeability membrane using canine harvested liver in porcine fulminant hepatic failure model.

INTRODUCTION: The use of bioartificial liver devices requires. A sufficient liver cell mass to provide adequate metabolic support, reduction of xenogeneic immune reactions, and avoidance of viral transmission. We have developed a plasmapheresis system using a semipermeable membrane combined with canine whole liver perfusion (PMCWLP). In this study, we investigated the efficacy of our system in a porcine fulminant hepatic failure (FHF) model. METHODS: The porcine FHF model was established by intraportal administration of alpha-amanitin (0.1 mg/kg) and lipopolysaccharide (1 microg/kg). Nine hours after drug injection, xenogenic perfusion treatment was performed twice within 6 hours (n = 5). As the plasmapheresis device, we used a hollow-fiber module with cellulose diacetate porous fibers (pore size, 0.05 microm, surface area, 2 m2). The canine whole liver was perfused with modified Krebs solution, which is commonly used in many laboratories, containing albumin (2 g/dL) and glucose (300 mg/dL). Control pigs (n = 10), had the circuit not connected to the whole canine liver. RESULTS: The survival of FHF pigs was significantly increased by the treatment (58.9 +/- 21.8 hour) compared with the controls (22.3 +/- 8.1 hour). Mean blood ammonia levels and intracranial pressure during treatment were significantly lower compared with control groups. CONCLUSION: Treatment of FHF pigs with the system significantly increased survival time, suggesting that this method may have applications as a clinical liver assist device.

In vitro evaluation of cross-circulation system using semipermeable membrane combined with whole liver perfusion.

INTRODUCTION: Many types of isolated hepatocytes-based bioartificial liver have been developed. However, to maintain hepatocyte-specific functions for a long period is still a significant challenge. The possibilities of rejection or viral transmission still remain as untackled obstacles. We developed a cross-circulation system, using a semipermeable membrane combined with whole liver perfusion. Detoxifying functions of the extracorporeal porcine liver and molecular movements across the membrane were evaluated in vitro. METHODS: The hollow-fiber module has a molecular cutoff of 100 kD. A spiked solution containing 500 mL low molecular dextran solution spiked with 12 mg ammonium chloride, 500 mg D-galactose, and 300 mg lidocaine, which mimicked a patient, was recirculated through the inner fiber space. The extracorporeal liver perfusion circuit consisted of an extra-fiber spaces. A reservoir containing 1000 mL healthy pig plasma, a membrane oxygenator, and a porcine whole liver. Both circuits circulated in the opposite direction for 6 hours. RESULT: In 6 hours, 47.3% +/- 10.2% of ammonia, 89.5% +/- 1.7% of D-galactose, and 95.5% +/- 1.0% of lidocaine were eliminated from the circuits; 66.5 +/- 11.1 mg of urea were produced at the same time. Oxygen consumption was maintained between 0.248 and 0.259 mL/100 g liver/min for 6 hours. Movement of IgM was completely blocked by the 100-kD membrane, whereas albumin was freely transferred from the reservoir to the intrafiber space. CONCLUSION: The perfusion experiments showed the possibility of using a whole liver with oxygenated plasma perfusion in a bioartificial liver system in vitro.