Integration of single-layer skin hollow fibers and scaffolds develops a three-dimensional hybrid bioreactor for bioartificial livers

摘要

Bioartificial liver support systems are expected to be an effective therapy as a "bridge" for liver transplantation or reversible acute liver disease. A major roadblock in the application of bioartificial livers is the need for a bioreactor that fully meets the requirements of hepatocyte culture, mass transfer and immunobarriers. In this study, we developed a three-dimensional hybrid bioreactor (3DHB) on a base of single-layer skin polyethersulfone hollow fibers by integrating with polyurethane scaffolds. The mass transfer of bilirubin and albumin from the in-tracapillary space to the extracapillary space of the hollow fibers was not significantly different between 3DHBs and hollow fiber bioreactors (HFBs). Cell viability staining showed that high-density hepatocytes were uniformly found in different regions of the 3DHB after 7 days of culture. Liver-specific functions of human mature hepatocytes cultured in the 3DHB, such as albumin secretion, urea production, ammonia removal rate and cytochrome P450 activity, were maintained stably and were significantly higher compared with the HFB. These results indicated that the 3DHB has good mass transfer and improves cell distribution and liver-specific functions. Meanwhile, the ammonia and unconjugated bilirubin concentrations in plasma from patients with liver failure were significantly decreased during 6 h of circulation by hepatocytes cultured in the 3DHB. Most hepatocytes in the 3DHB were viable after 6 h exposure to the patient plasma. We further demonstrated that bioartificial liver systems with 3DHB can remove toxins from and endure the deleterious effects of the patient plasma. Therefore, the 3DHB has the potential to accomplish different actions for the clinical application of bioartificial livers.