Hybrid artificial liver support system for treatment of severe liver failure

Abstract

Aim: To construct a novel hybrid artificial liver support system (HALSS) and to evaluate its efficacy in patients with severe liver failure.

Methods: Hepatocytes were isolated from suckling pig by the modified Seglen's method. Isolated hepatocytes were cultured in a spinner flask for 24 h to form spheroids before use and the functions of spheroids were detected. HALSS consisted of a plasma separator, a hemo-adsorba and a bioreactor with hepatocytes spheroids in its extra-fiber space. HALSS was applied to 10 patients with severe liver failure. The general condition and the biochemical indexes of the patients were studied just before and after the treatment.

Results: The number of cells per liver was about 2-4 x 10(10) (mean, 3.1+/-1.5 x 10(10)). The cell viabilities were more than 95%. After 24 h of spheroid culture, most hepatocytes formed spheroids. The levels of albumin and urea in the medium of spheroid culture were higher than those in supernatant of petri dish culture (P = 0.0015 and 0.0001, respectively). The capacity of albumin production and urea synthesis remained stable for more than one wk and declined rapidly after two weeks in vitro. In HALSS group, the duration of HALSS treatment was 6-10 h each time. All patients tolerated the treatment well without any fatal adverse reaction. After HALSS treatment, the general condition, psychic state, encephalopathy and hepatic function of the patients were improved. The survival rate of the HALSS group, Plasmapheresis group and control group was 30% (3/10), 20% (2/10) and 0% (0/10), respectively (P = 0.024). Two weeks after treatment, Tbil and ALT decreased and the PTA level elevated in HALSS group and pasmapheresis group (P value: 0.015 vs 0.020, 0.009 vs 0.012 and 0.032 vs 0.041, respectively). But there was no significant change of blood albumin concentration before and after treatment in HALSS group and Plasmapheresis group.

Conclusion: The HALSS established by us is effective in supporting liver function of patients with severe liver failure.

Figure 1

Schematic diagram of HALSS.

Figure 2

Morphology of hepatocytes. A: freshly isolated single hepatocytes stained with trypan blue (×200); B: spheroid formation after 24 h of spheroid culture (×100).

Figure 3

Spheroid cultivation of hepatocytes at a density of 5×10⁶/mL cells. Control hepatocytes were cultivated in a petri dish at the same density. A: albumin production; B: urea synthesis.

Figure 4

Changes of total bilirubin, ALT, albumin and PTA index in patients in HALSS group, Plasmapheresis group and control group. Pre-: index before treatment. Post-: index at two weeks after treatment.