Dual flow bioreactor with ultrathin microporous TEER sensing membrane for evaluation of nanoparticle toxicity

Abstract

Abstract:Permeability studies across biological barriers are of primary importance in drug delivery as well as in toxicology when investigating the absorption and translocation of a substance. The study of nanomaterial interaction with epithelial barriers is of particular interest given their growing use in nanomedicine as well as concerns about their potential hazard. Here we describe the design and fabrication of a new bioreactor with an ultrathin microporous sensing support for the study of nanoparticle toxicity in intestinal epithelial cells in conditions which better recapitulate the physiological environment. Thanks to the integration of 4 electrodes in the microporous membrane, the system allows real-time and continuous sensing of TEER (trans epithelial electrical resistance) during flow without interruption or perturbation of experiments. The TEER bioreactor was tested using Caco-2 cells as an in vitro model of intestinal epithelia. When exposed to silver nanoparticles, which are known to be toxic, the embedded electrodes enabled non-invasive evaluation of barrier impairment over time. This device can be used to study barrier integrity and the kinetics of nanomaterial induced damage to epithelial barriers in physiologically relevant conditions.

Real-time cellular impedance monitoring and imaging of biological barriers in a dual-flow membrane bioreactor

“…Similarly, a two-layer membrane-based device made of PDMS was proposed by Griep et al In their system a polycarbonate membrane separated the top channel from the bottom one and platinum wire electrodes with a diameter of 200 μm were placed in contact with the two channels to perform impedance measurements using a spectrum analyser (Griep et al, 2013). Recently, a milli-fluidic bioreactor was integrated with four platinum electrodes were recorded via a NI USB-6008 (National Instrument) acquisition card (Sbrana et al, 2016). Costello et.…”

“…Therefore, barrier-forming cells are often cultured in fluidic systems, or bioreactors, which are able to apply dynamic flow conditions (Giusti et al, 2014;Martin et al, 2004;Sbrana et al, 2016).…”

“…With the exception of the milli-fluidic device described in (Sbrana et al, 2016), all the bioreactors with integrated TEER/TEEI measurement systems are microfluidic devices and their electrodes are either interfaced with the EVOM or a laboratory impedance analyser, which are not particularly suitable for cellular measurements. Moreover, given their small volumes, micro-fluidic bioreactors have limited space to incorporate multiple electrodes.…”

3D‐Printed Bioreactor with Integrated Impedance Spectroscopy for Cell Barrier Monitoring

“…Often, integrated electrodes are only used to calculate resistance and not impedance. [26][27][28] In principle, integrated electrodes for resistance measurement can be used for EIS. The input signal needs to be changed from direct current or single frequency, as used in voltohmmeter (EVOM or Millicell), to a frequency spectrum.…”

Polymeric Microporous Nanofilms as Smart Platforms for <italic>in Vitro</italic> Assessment of Nanoparticle Translocation and Caco-2 Cell Culture

“…Such a system is unsuitable for nanoparticle translocation tests in an intestinelike barrier, since nanoparticles can cover a broad range of dimensions. Other groups reported the use of microporous membranes for lab-on-chip applications [18][19][20][21]. However, these systems are rather rigid and excessively costly, or too thick to prevent nanoparticles clogging in the pores.…”