Fully automatic flow-based device for monitoring of drug permeation across a cell monolayer

Abstract

A novel flow-programming setup based on the sequential injection principle is herein proposed for on-line monitoring of temporal events in cell permeation studies. The permeation unit consists of a Franz cell with its basolateral compartment mixed under mechanical agitation and thermostated at 37 °C. The apical compartment is replaced by commercially available Transwell inserts with a precultivated cell monolayer. The transport of drug substances across epithelial cells genetically modified with the P-glycoprotein membrane transporter (MDCKII-MDR1) is monitored on-line using rhodamine 123 as a fluorescent marker. The permeation kinetics of the marker is obtained in a fully automated mode by sampling minute volumes of solution from the basolateral compartment in short intervals (10 min) up to 4 h. The effect of a P-glycoprotein transporter inhibitor, verapamil as a model drug, on the efficiency of the marker transport across the cell monolayer is thoroughly investigated. The analytical features of the proposed flow method for cell permeation studies in real time are critically compared against conventional batch-wise procedures and microfluidic devices.

3D Printing: The Second Dawn of Lab-On-Valve Fluidic Platforms for Automatic (Bio)Chemical Assays

“…After detection, the acceptor volume was returned to the chamber in order to maintain the total volume unaltered and avoid changes in permeation equilibria. This is a crucial difference against previous online Franz-cell permeation tests − for which recalculation of the concentration of target analyte in the acceptor phase throughout time was called for. The above analytical protocol was repeated until no significant differences were encountered for the absorbance readouts of five consecutive readings.…”

“…In the present study, the kinetics of permeation of a dye model using an artificial membrane was explored in a fully automatic LOV mode as a proof of concept of dermal fibroblast or intestinal epithelial cell permeation-based in vitro bioavailability testing. − Transwell inserts with translucent polycarbonate membranes (diameter: 12 mm, pore size: 3.0 μm, growth surface area: 1.12 cm 2 ) from Corning (Baria s.r.o., Czech Republic) allowed for online passive diffusion monitoring. To the best of our knowledge, Transwell units are for the first time in this work allied to LOV approaches without the need of liberation units, the so-called Franz diffusion cell, because the luer lock port of the LOV print allowed to accommodate a 5 mL syringe body with appropriate dimensions for accommodation of the inset and with sufficient recipient volume for monitoring purposes.…”

Sequential Injection Analysis for Automation and Evaluation of Drug Liberation Profiles: Clotrimazole Liberation Monitoring

“…As Franz diffusion cells were used for liberation testing, the same units with different dimensions [12] can be applied not only for liberation testing, but also for permeation testing or other interaction studies where different artificial or model membranes and even inserts with cultivated cell lines [13] are placed in the Franz cell design. In the case of liberation, different technological parameters in pharmaceutical formulation development can be observed, e.g., enrofloxacin incorporation into liposomes [14], cannabidiol release through rabbit ear skin [15], and intercalation of diclofenac and naproxen to slow down their release [16].…”