Imitation of drug metabolism in human liver and cytotoxicity assay using a microfluidic device coupled to mass spectrometric detection

Abstract

In this work, we developed a microfluidic device for the imitation of drug metabolism in human liver and its cytotoxicity on cells. The integrated microfluidic device consists of three sections: (1) bioreactors containing poly(ethylene) glycol (PEG) hydrogel encapsulated human liver microsomes (HLMs); (2) cell culture chambers for cytotoxicity assay; and (3) integrated micro solid-phase extraction (SPE) columns to desalt and concentrate the products of enzymatic reaction. To verify the feasibility of the integrated microchip, we studied uridine 5'-diphosphate-glucuronosyltransferase (UGT) metabolism of acetaminophen (AP) and the cytotoxicity of products on HepG2 cells. The products of the reaction in one region of the device were injected into the cell culture chamber for cytotoxicity assay, while those in another region were directly detected online with an electrospray ionization quadrupole time-of-flight mass spectrometer (ESI-Q-TOF MS) after micro-SPE pre-treatment. Semiquantitative analysis achieved in the experiments could be related to the drug-induced HepG2 cell cytotoxicity. Total analysis time for one product was about 30 min and only less than 4 μg HLM protein was required for one reaction region. The results demonstrated that the established platform could be used to imitate drug metabolism occurring in the human liver, thereby replacing animal experiments in the near future. In addition, the integrated microchip will be a useful tool for drug metabolism studies and cytotoxicity assays, which are pivotal in drug development.

High-throughput screening approaches and combinatorial development of biomaterials using microfluidics

“…Furthermore, it is difficult to retrieve a sample or to integrate a sensor, and even when this is possible, valuable spatial information is lost. Attempts have been made to adapt the conventional analytical techniques to the requirements of microfluidics, for example by developing customized interfaces, or by the use of single-cell or low cell number genomic or proteomic analyzes [5,209,210]. However, microfluidic platforms, as with many other high-throughput screening platforms, will continue to largely rely on biomolecular engineering techniques coupled with microscopy-based imaging.…”

“…Nevertheless, the validity of such assays, or the evidence that they are at least as reliable as conventional assays is needed for microfluidic platforms to be explored to the maximum extent. Alternatively to development of new assays, conventional analytical tools can be rendered applicable to microfluidic systems by means of customized interfacing [5].…”

A membrane-based microfluidic device for mechano-chemical cell manipulation

“…Despite the fact that conventional macroscopic assays for drug development are still widely used, microfluidic technology has attracted increasing interest in the past years (Nguyen et al 2013;Abkarian et al 2006;Dai et al 2010;Adamo et al 2012;Mao et al 2012;Ma et al 2009). …”

Pressure-Driven Microfluidic Perfusion Culture Device for Integrated Dose-Response Assays

“…Miniaturized high-throughput and high-content cell-based assays have been reported [14][15][16][17] and applied to analyses of liver drug metabolism and toxicity. 18,19 Assay miniaturization provides the opportunity for creating a variety of microenvironments, such as soluble factors and extracellular matrixes, within the microfluidic devices. 20,21 The fabrication of microfluidic devices with biomimetic tissues is another approach for cell-based assays.…”