Kinetic modelling of in vitro cell-based assays to characterize non-specific bindings and ADME processes in a static and a perfused fluidic system

Abstract

Recently, physiologically based perfusion in vitro systems have been developed to provide cell culture environment close to in vivo cell environment (e.g., fluidic conditions, organ interactions). In this work, we model and compare the fate of a chemical, benzo[ a]pyrene (B[ a]P), in a perfusion and a standard (static well-plate) system. These in vitro systems are composed of Caco-2 and HepG2 cells so as to mimic absorption across the small intestine and intestinal and hepatic metabolism. Compartmental models were developed and calibrated with B[ a]P kinetics data in the culture medium to estimate the apparent permeability of Caco-2 cells, the in vitro biotransformation of B[ a]P, as well as the different routes of loss by non-specific adsorption. Our results show that non-specific binding is the main process responsible for the depletion of B[ a]P in the culture media: at steady state, only 40% and 24% of the total concentration of B[ a]P are bioavailable in the static and perfused systems, respectively. We also showed that Caco-2 permeability in the perfused culture system is closer to in vivo conditions than the one obtained in the static system and that higher cellular metabolic activities are observed in static conditions. Perfused in vitro systems combined with kinetic modelling are promising tools for studying in vitro the different processes involved in the toxicokinetics of xenobiotics.