Multiple approaches to predicting oxygen and glucose consumptions by HepG2 cells on porous scaffolds in an axial-flow bioreactor.

In this study, the distribution of oxygen and glucose was evaluated along with consumption by hepatocytes using three different approaches. The methods include (i) Computational Fluid Dynamics (CFD) simulation, (ii) residence time distribution (RTD) analysis using a step-input coupled with segregation model or dispersion model, and (iii) experimentally determined consumption by HepG2 cells in an open-loop. Chitosan-gelatin (CG) scaffolds prepared by freeze-drying and polycaprolactone (PCL) scaffolds prepared by salt leaching technique were utilized for RTD analyses. The scaffold characteristics were used in CFD simulations i.e. Brinkman's equation for flow through porous medium, structural mechanics for fluid induced scaffold deformation, and advection-diffusion equation coupled with Michaelis-Menten rate equations for nutrient consumption. With the assumption that each hepatocyte behaves like a micro-batch reactor within the scaffold, segregation model was combined with RTD to determine exit concentration. A flow rate of 1 mL/min was used in the bioreactor seeded with 0.6 × 10(6) HepG2 cells/cm(3) on CG scaffolds and oxygen consumption was measured using two flow-through electrodes located at the inlet and outlet. Glucose in the spent growth medium was also analyzed. RTD results showed distribution of nutrients to depend on the surface characteristics of scaffolds. Comparisons of outlet oxygen concentrations between the simulation results, and experimental results showed good agreement with the dispersion model. Outlet oxygen concentrations from segregation model predictions were lower. Doubling the cell density showed a need for increasing the flow rate in CFD simulations. This integrated approach provide a useful strategy in designing bioreactors and monitoring tissue regeneration.

Modeling pressure drop using generalized scaffold characteristics in an axial-flow bioreactor for soft tissue regeneration.

The goal of this study was to better understand how analytical permeability models based on scaffold architecture can facilitate a non-invasive technique to real time monitoring of pressure drop in bioreactors. In particular, we evaluated the permeability equations for electrospun and freeze dried scaffolds via pressure drop comparison in an axial-flow bioreactor using computational fluid dynamic (CFD) and experimentation. The polycaprolactone-cellulose acetate fibers obtained by co-axial electrospinning technique and Chitosan-Gelatin scaffolds prepared using freeze-drying techniques were utilized. Initially, the structural properties (fiber size, pore size and porosity) and mechanical properties (elastic modulus and Poisson's ratio) of scaffolds in phosphate buffered saline at 37 °C were evaluated. The CFD simulations were performed by coupling fluid flow, described by Brinkman equation, with structural mechanics using a moving mesh. The experimentally obtained pressure drop values for both 1 mm thick and 2 mm thick scaffolds agreed with simulation results. To evaluate the effect of permeability and elastic modulus on pressure drop, CFD predictions were extended to a broad range of permeabilities spanning synthetic scaffolds and tissues, elastic moduli, and Poisson's ratio. Results indicated an increase in pressure drop with increase in permeability. Scaffolds with higher elastic modulus performed better and the effect of Poisson's ratio was insignificant. Flow induced deformation was negligible in axial-flow bioreactor. In summary, scaffold permeabilities can be calculated using scaffold microarchitecture and can be used in non-invasive monitoring of tissue regeneration.