Marco Caronna

A computational model for the optimization of transport phenomena in a rotating hollow fiber bioreactor for artificial liver

A comprehensive computational study modelling the operation of a rotating hollow-fiber bioreactor for artificial liver (BAL) was performed to explore the interactions between the oxygenated culture medium and the cultured hepatocytes. Computational fluid dynamics investigations were carried out using two-dimensional (2D) and 3D time-dependent numerical simulations, integrating calculations of diffusion, convection, and multiphase fluid dynamics. The analysis was aimed at determining the rotational speed value of the chamber to ensure homogenous distribution of the floating microcarrier-attached aggregated cells (microCAACs) and avoid their sedimentation and excessive packing, analyzing oxygen (O(2)) delivery and cellular O(2) consumption as an index of cellular metabolic activity, and analyzing the fluid-induced mechanical stress experienced by cells. According to our results, homogeneous distribution of cells is reached at a rotational speed of 30 rpm; spreading of cellular concentration at around the initial value of 12% was limited (median = 11.97%, 5th percentile = 10.94%, 95th percentile = 13.2%), resulting in uniform suspension of microCAACs, which did not appear to be excessively packed. Mixing within the rotating fluid caused a maximum fluid-induced stress value of 0.05 Pa, which was neither endangering for liver-specific functions of cultured cells, nor causing disruption of the floating aggregates. Moreover, an inlet medium flow rate of 200 mL/m with a partial pressure of oxygen (pO(2)) value of 160 mmHg was found to guarantee an adequate O(2) supply for the hepatocytes (2.7 x 10(8) hepatocytes are simulated); under such conditions, the minimum pO(2) value (23 mmHg) is above the critical threshold value, causing the onset of cellular hypoxia (10 mmHg). We proved that numerical simulation of transport phenomena is a valuable tool for the computer-aided design of BALs, helping overcome the unsolved issues in optimizing the cell-environment conditioning procedure in rotating BALs.

Introduction of a Novel Prototype Bioartificial Liver Support System Utilizing Small Human Hepatocytes in Rotary Culture

The aim of this study was to establish a stand-alone, perfused, rotary cell culture system using small human hepatocytes (SH) for bioartificial liver (BAL) support. SH were grown on cytodex 3 microcarriers (beads) to a maximum density of 1.2 +/- 0.3 x 10(7) cells per mL within 12 days. Size of aggregates formed by up to 15 beads was regulated by rotation speed. Cell function was proven by treatment with ammonia and galactose, and metabolism was analyzed. Treatment strategy was comprised of two phases, namely growth phase and treatment phase. Cells were grown for 6 days and subsequently incubated with ammonia or galactose for 2 days, followed by a 2-day regeneration period and another 2-day treatment phase. Consumption of glucose, release of lactate dehydrogenase, formation of lactate, and production of urea and albumin were determined regularly. Mean galactose consumption was 50 microg per 106 cells per hour, ammonia-induced urea formation was 3.6 microg per 106 cells per hour, and albumin production was 110 ng per 106 cells per hour. All metabolic parameters followed a logarithmic trend and were found to be very stable in the second half of the culture period when cells were treated with ammonia or galactose. Dissolved oxygen (%DO), pH, and temperature were monitored in-stream at intervals of 7 min, and the values were logged. Viability and morphology of cells were monitored via confocal microscopy. Viability was around 95% in controls and 90% during treatment. Promising results were obtained in support of our ongoing efforts to establish a fully autonomous BAL support device utilizing SH as a bridge to transplantation.

A CFD Computational Model for the Optimization of Transport Phenomena in a Rotating Hollow Fiber Bioreactor for Artificial Liver

Over the last decades, many attempts have been made aimed at developing bioartificial systems able for providing temporary hepatic support to patients with liver failure [1].Copyright