Search Results (16)

Microphysiological systems (MPSs), such as organ-on-a-chip platforms, are promising alternatives to animal testing for drug development and physiological research. The BioStellar™ Plate is a commercial MPS platform featuring an open-top culture chamber design with on-chip stirrer pumps that circulate culture medium through six independent, dual microchannel-connected chamber multiorgan units. Although this design enables a circular flow, the open-top culture chamber format prevents the application of fluidic shear stress, a force that cells experience in vivo, which affects their behavior and function. To address this, we developed two fluidic shear stress attachments for the BioStellar™ Plate. These attachment channel fluids provide controlled mechanical stimulation to cultured cells. The flow dynamics were simulated using COMSOL Multiphysics to estimate shear stress levels. The attachments were fabricated and validated through fluorescent bead tracking and biological assays. The FSSA-D is designed for flat-bottom standard cell cultures, while the FSSA-I is designed for epithelial monolayers, enabling the application of fluidic shear stress across the basal membrane. Experiments with intestinal epithelial cells (Caco-2) demonstrated that both attachments enhanced cell barrier function under a fluidic environment, as indicated by higher transepithelial electrical resistance (TEER). These findings demonstrate that the attachments are practical tools for mechanobiology research with MPS platforms. Full article

An obstacle for many microfluidic developments is the fabrication of its structures, which is often complex, time-consuming, and expensive. Additive manufacturing can help to reduce these barriers. This study investigated whether the results of a microfluidic assay for the detection of the promyelocytic leukemia (PML)-retinoic acid receptor α (RARα) fusion protein (PML::RARA), and thus for the differential diagnosis of acute promyelocytic leukemia (APL), could be transferred from borosilicate glass microfluidic structures to additively manufactured fluidics. Digital light processing (DLP) and stereolithography (SLA) printers as well as different photopolymerizable methacrylate-based resins were tested for fabrication of the fluidics. To assess suitability, both print resolution and various physical properties, serializability, biocompatibility, and functionalization with biological molecules were analyzed. The results show that additively manufactured microfluidics are suitable for application in leukemia diagnostics. This was demonstrated by transferring the microfluidic sandwich enzyme-linked immunosorbent assay (ELISA) for PML::RARA onto the surface of magnetic microparticles from a glass structure to three-dimensional (3D)-printed parts. A comparison with conventional glass microstructures suggests lower sensitivity but highlights the potential of additive manufacturing for prototyping microfluidics. This may contribute to the wider use of microfluidics in biotechnological or medical applications. Full article

In the preclinical phase of drug development, it is necessary to determine how the active compound can pass through the biological barriers surrounding the target tissue. In vitro barrier models provide a reliable, low-cost, high-throughput solution for screening substances early in the drug candidate development process, thus reducing more complex and costly animal studies. In this pilot study, the transport properties of TB501, an antimycobacterial drug candidate, were characterized using an in vitro barrier model of VERO E6 kidney cells. The compound was delivered into the apical chamber of the transwell insert, and its concentration passing through the barrier layer was measured through the automated sampling of the basolateral compartment, where media were replaced every 30 min for 6 h, and the collected samples were stored for further spectroscopic analysis. The kinetics of TB501 concentration obtained from VERO E6 transwell cultures and transwell membranes saturated with serum proteins reveal the extent to which the cell layer functions as a diffusion barrier. The large number of samples collected allows us to fit a detailed mathematical model of the passive diffusive currents to the measured concentration profiles. This approach enables the determination of the diffusive permeability, the diffusivity of the compound in the cell layer, the affinity of the compound binding to the cell membrane as well as the rate by which the cells metabolize the compound. The proposed approach goes beyond the determination of the permeability coefficient and offers a more detailed pharmacokinetic characterization of the transwell barrier model. We expect the presented method to be fruitful in evaluating other compounds with different chemical features on simple in vitro barrier models. The proposed mathematical model can also be extended to include various forms of active transport. Full article

Current available animal and in vitro cell-based models for studying brain-related pathologies and drug evaluation face several limitations since they are unable to reproduce the unique architecture and physiology of the human blood–brain barrier. Because of that, promising preclinical drug candidates often fail in clinical trials due to their inability to penetrate the blood–brain barrier (BBB). Therefore, novel models that allow us to successfully predict drug permeability through the BBB would accelerate the implementation of much-needed therapies for glioblastoma, Alzheimer’s disease, and further disorders. In line with this, organ-on-chip models of the BBB are an interesting alternative to traditional models. These microfluidic models provide the necessary support to recreate the architecture of the BBB and mimic the fluidic conditions of the cerebral microvasculature. Herein, the most recent advances in organ-on-chip models for the BBB are reviewed, focusing on their potential to provide robust and reliable data regarding drug candidate ability to reach the brain parenchyma. We point out recent achievements and challenges to overcome in order to advance in more biomimetic in vitro experimental models based on OOO technology. The minimum requirements that should be met to be considered biomimetic (cellular types, fluid flow, and tissular architecture), and consequently, a solid alternative to in vitro traditional models or animals. Full article

Electrical impedance spectroscopy (EIS) is widely recognized as a powerful tool in biomedical research. For example, it allows detection and monitoring of diseases, measuring of cell density in bioreactors, and characterizing the permeability of tight junctions in barrier-forming tissue models. However, with single-channel measurement systems, only integral information is obtained without spatial resolution. Here we present a low-cost multichannel impedance measurement set-up capable of mapping cell distributions in a fluidic environment by using a microelectrode array (MEA) realized in 4-level printed circuit board (PCB) technology including layers for shielding, interconnections, and microelectrodes. The array of 8 × 8 gold microelectrode pairs was connected to home-built electric circuitry consisting of commercial components such as programmable multiplexers and an analog front-end module which allows the acquisition and processing of electrical impedances. For a proof-of-concept, the MEA was wetted in a 3D printed reservoir into which yeast cells were locally injected. Impedance maps were recorded at 200 kHz which correlate well with the optical images showing the yeast cell distribution in the reservoir. Blurring from parasitic currents slightly disturbing the impedance maps could be eliminated by deconvolution using an experimentally determined point spread function. The MEA of the impedance camera can in future be further miniaturized and integrated into cell cultivation and perfusion systems such as organ on chip devices to augment or even replace light microscopic monitoring of cell monolayer confluence and integrity during the cultivation in incubation chambers. Full article

Microfluidic paper-based analytical devices have broadened the scope of microfluidics by offering low-cost, simple, faster fabrication, biodegradability, and environmentally benign devices capable of medical diagnostics, environmental sensing, and food quality control. These devices are incredibly versatile, which is one of their most notable features. Despite recent advancements in paper-fluidics, creating slow flow channels in paper still remains a challenge. Herein, we propose viscous barriers of various concentrations embedded along the paper channel to control its flow velocity by altering the pore size. We used sugar concentrations in the range of 0–40% dried in porous media and then recorded flow behaviors of water and castor oil. From experiments, it was observed that by increasing the sugar concentration, delay time also increased. Moreover, changing the type of fluid (w.r.t viscosity) also varies the flow delays as castor oil took a much longer time to cover the same channel length as compared to water. We believe that our proposed method will play an important part in improving flow delays and can be applied for food quality applications such as time–temperature indicators due to its simple fabrication and cost-effective technique. Full article

A helium (He) dielectric barrier discharge plasma jet (DBD jet) was used for the first time for treating graphite foil as the current collector of a paper-based fluidic aluminum-air battery. The main purpose was to improve the distribution of the catalyst layer through modification and functionalization of the graphite foil surface. The plasma functionalized the graphite foil surface to enhance the wettability where the more hydroxyl could be observed from XPS results. The 30 s-He DBD jet treatment on the graphite foil significantly improved the battery performance. The best current density of 85.6 mA/cm² and power density of 40.98 mW/cm² were achieved. The energy density was also improved to 720 Wh/kg. Full article

The fluidic barrier in centrifugal microfluidic platforms is a newly introduced concept for making multiple emulsions and microparticles. In this study, we focused on particle generation application to better characterize this method. Because the phenomenon is too fast to be captured experimentally, we employ theoretical models to show how liquid polymeric droplets pass a fluidic barrier before crosslinking. We explain how secondary flows evolve and mix the fluids within the droplets. From an experimental point of view, the effect of different parameters, such as the barrier length, source channel width, and rotational speed, on the particles’ size and aspect ratio are investigated. It is demonstrated that the barrier length does not affect the particle’s ultimate velocity. Unlike conventional air gaps, the barrier length does not significantly affect the aspect ratio of the produced microparticles. Eventually, we broaden this concept to two source fluids and study the importance of source channel geometry, barrier length, and rotational speed in generating two-fluid droplets. Full article

Static in vitro permeation experiments are commonly used to gain insights into the permeation properties of drug substances but exhibit limitations due to missing physiologic cell stimuli. Thus, fluidic systems integrating stimuli, such as physicochemical fluxes, have been developed. However, as fluidic in vitro studies display higher complexity compared to static systems, analysis of experimental readouts is challenging. Here, the integration of in silico tools holds the potential to evaluate fluidic experiments and to investigate specific simulation scenarios. This study aimed to develop in silico models that describe and predict the permeation and disposition of two model substances in a static and fluidic in vitro system. For this, in vitro permeation studies with a 16HBE cellular barrier under both static and fluidic conditions were performed over 72 h. In silico models were implemented and employed to describe and predict concentration–time profiles of caffeine and diclofenac in various experimental setups. For both substances, in silico modeling identified reduced apparent permeabilities in the fluidic compared to the static cellular setting. The developed in vitro–in silico modeling framework can be expanded further, integrating additional cell tissues in the fluidic system, and can be employed in future studies to model pharmacokinetic and pharmacodynamic drug behavior. Full article

The wide use of 3D-organotypic cell models is imperative for advancing our understanding of basic cell biological mechanisms. For this purpose, easy-to-use enabling technology is required, which should optimally link standardized assessment methods to those used for the formation, cultivation, and evaluation of cell aggregates or primordial tissue. We thus conceived, manufactured, and tested devices which provide the means for cell aggregation and online monitoring within a hanging drop. We then established a workflow for spheroid manipulation and immune phenotyping. This described workflow conserves media and reagent, facilitates the uninterrupted tracking of spheroid formation under various conditions, and enables 3D-marker analysis by means of 3D epifluorescence deconvolution microscopy. We provide a full description of the low-cost manufacturing process for the fluidic devices and microscopic assessment tools, and the detailed blueprints and building instructions are disclosed. Conclusively, the presented compilation of methods and techniques promotes a quick and barrier-free entry into 3D cell biology. Full article

Endothelial and epithelial cellular barriers play a vital role in the selective transport of solutes and other molecules. The properties and function of these barriers are often affected in case of inflammation and disease. Modelling cellular barriers in vitro can greatly facilitate studies of inflammation, disease mechanisms and progression, and in addition, can be exploited for drug screening and discovery. Here, we report on a parallelizable microfluidic platform in a multiwell plate format with ten independent cell culture chambers to support the modelling of cellular barriers co-cultured with 3D tumor spheroids. The microfluidic platform was fabricated by microinjection molding. Electrodes integrated into the chip in combination with a FT-impedance measurement system enabled transepithelial/transendothelial electrical resistance (TEER) measurements to rapidly assess real-time barrier tightness. The fluidic layout supports the tubeless and parallelized operation of up to ten distinct cultures under continuous unidirectional flow/perfusion. The capabilities of the system were demonstrated with a co-culture of 3D tumor spheroids and cellular barriers showing the growth and interaction of HT29 spheroids with a cellular barrier of MDCK cells. Full article

Neurology has always been one of the therapeutic areas with higher attrition rates. One of the main difficulties is the presence of the blood–brain barrier (BBB) that restricts access to the brain for major drugs. This low success rate has led to an increasing demand for in vitro tools. The shear stress, which positively affects endothelial cell differentiation by mimicking blood flow, is required for a more physiological in vitro BBB model. We created an innovative device specifically designed for cell culture under shear stress to investigate drug permeability. Our dynamic device encompasses two compartments communicating together via a semi-permeable membrane, on which human cerebral microvascular endothelial (hCMEC/D3) cells were seeded. The fluidic controlled environment ensures a laminar and homogenous flow to culture cells for at least seven days. Cell differentiation was characterized by immunodetection of inter-endothelial junctions directly in the device by confocal microscopy. Finally, we performed permeability assay with lucifer yellow in both static and dynamic conditions in parallel. Our dynamic device is suited to the evaluation of barrier function and the study of drug transport across the BBB, but it could also be used with other human cell types to reproduce intestinal or kidney barriers. Full article

Unstable liquid flow in syringe pump-driven systems due to the low-speed vibration of the step motor is commonly observed as an unfavorable phenomenon, especially when the flow rate is relatively small. Upon the design of a convenient and cost-efficient microfluidic standing air bubble system, this paper studies the physical principles behind the flow stabilization phenomenon of the bubble-based hydraulic capacitors. A bubble-based hydraulic capacitor consists of three parts: tunable microfluidic standing air bubbles in specially designed crevices on the fluidic channel wall, a proximal pneumatic channel, and porous barriers between them. Micro-bubbles formed in the crevices during liquid flow and the volume of the bubble can be actively controlled by the pneumatic pressure changing in the proximal channel. When there is a flowrate fluctuation from the upstream, the flexible air-liquid interface would deform under the pressure variation, which is analogous to the capacitive charging/discharging process. The theoretical model based on Euler law and the microfluidic equivalent circuit was developed to understand the multiphysical phenomenon. Experimental data characterize the liquid flow stabilization performance of the flow stabilizer with multiple key parameters, such as the number and the size of microbubbles. The developed bubble-based hydraulic capacitor could minimize the flow pulses from syringe pumping by 75.3%. Furthermore, a portable system is demonstrated and compared with a commercial pressure-driven flow system. This study can enhance the understanding of the bubble-based hydraulic capacitors that would be beneficial in microfluidic systems where the precise and stable liquid flow is required. Full article

The blood–brain barrier (BBB) plays critical role in the human physiological system such as protection of the central nervous system (CNS) from external materials in the blood vessel, including toxicants and drugs for several neurological disorders, a critical type of human disease. Therefore, suitable in vitro BBB models with fluidic flow to mimic the shear stress and supply of nutrients have been developed. Neurological disorder has also been investigated for developing realistic models that allow advance fundamental and translational research and effective therapeutic strategy design. Here, we discuss introduction of the blood–brain barrier in neurological disorder models by leveraging a recently developed microfluidic system and human organ-on-a-chip system. Such models could provide an effective drug screening platform and facilitate personalized therapy of several neurological diseases. Full article

Paper discusses a device belonging into an interesting and yet little-known family of no-moving-part active fluidic rectifiers. The generated steady component of flow and pressure are driven by input alternating flow from an external source. The absence of moving components results in the unique capability of unlimited life and reliability, especially useful for safety devices. In the experiment, the rectifier generated a pressure keeping dangerous liquid in the active zone. When the driving oscillation stops (like, e.g., due to coolant loss), the liquid leaves the zone under gravity, stopping the performed reaction. This safety facility is simple, inexpensive, and extremely reliable. Full article