Search Results (17)

The possibility of tightly controlling the cellular microenvironment within microfluidic devices represents an important step toward precision analysis of cellular phenotypes in vitro. Microfluidic platforms that allow both long-term mammalian cell culture and dynamic modulation of the culture environment can support quantitative studies of cells’ responses to drugs. Here, we report the design and testing of a novel microfluidic device of simple production (single Polydimethylsiloxane layer), which integrates a micromixer with vacuum-assisted cell loading for long-term mammalian cell culture and dynamic mixing of four different culture media. Finite element modeling was used to predict flow rates and device dimensions to achieve diffusion-based fluid mixing. The device showed efficient mixing and dynamic exchange of media in the cell-trapping chambers, and viability of mammalian cells cultured for long-term in the device. This work represents the first attempt to integrate single-layer microfluidic mixing devices with vacuum-assisted cell-loading systems for mammalian cell culture and dynamic stimulation. Full article

Gas embolism is a rare but life-threatening process characterized by the presence of gas bubbles in the venous or arterial systems. These bubbles, if sufficiently large or numerous, can block the delivery of oxygen to critical organs, in particular the brain, and subsequently they can trigger a cascade of adverse biochemical reactions with severe medical outcomes. Despite its critical nature, gas embolism remains poorly understood, necessitating extensive investigation, particularly regarding its manifestations in the human body and its modulation by various biological conditions. However, given its elusive nature, as well as potential lethality, gas embolism is extremely difficult to study in vivo, and nearly impossible to be the subject of clinical trials. To this end, we developed a microfluidic device designed to study in vitro the impact of blood properties and vascular geometries on the formation and evolution of gas bubbles. The system features a biomimetic vascular channel surrounded by two pressure chambers, which induce the genesis of bubbles under varying circumstances. The bubble parameters were correlated with different input parameters, i.e., channel widths, wall thicknesses, viscosities of the artificial blood, and pressure levels. Smaller channel widths and higher equivalent hematocrit concentrations in synthetic blood solutions increased the nucleation density and bubble generation frequencies. Small channel widths were also more prone to bubble formation, with implications for the vulnerability of vascular walls, leading to increased risks of damage or compromise to the integrity of the blood vessels. Larger channel widths, along with higher equivalent hematocrit concentrations, translated into larger bubble volumes and decreased bubble velocities, leading to an increased risk of bubble immobilization within the blood vessels. This biomimetic approach provides insights into the impact of patient history and biological factors on the incidence and progression of gas embolism. Medical conditions, such as anemia, along with anatomical features related to age and sex—such as smaller blood vessels in women and children or larger vascular widths in adult men—affect the susceptibility to the initiation and progression of gas embolism, explored here in vitro through the development of a controlled, physiological-like environment. The analysis of the videos that recorded gas embolism events in vitro for systems where pressure is applied laterally on the microvasculature with thin walls, i.e., 50 μm or less, suggests that the mechanism of gas transfer for the pressure area to the blood is based on percolation, rather than diffusion. These findings highlight the importance of personalized approaches in the management and prevention of gas embolism. Full article

The currently available point-of-care hemostasis tests are burdened by criticisms concerning the use of different activators and inhibitors and the lack of dynamic flow. These operating conditions may constitute an impediment to the determination of the patient’s hemostatic condition. Hence, the diffusion of these tests in clinical practice is still limited to specific scenarios. In this work, we present a new method for analyzing the patient’s global hemostasis based on the visualization of the main components of the coagulation process and its computerized quantitative image analysis. The automated “Smart Clot” point-of-care system presents a micro-fluidic chamber in which whole blood flows, without the addition of any activator or inhibitor. In this micro-channel, platelet adhesion, activation and aggregation to the type I collagen-coated surface take place (primary hemostasis), leading to the production of endogenous thrombin on the surface of platelet aggregates and the consequent fibrin mesh and thrombus formation (secondary hemostasis). These observations are verified by inhibiting primary hemostasis with the antiplatelet drugs Indomethacin (−70% on platelet aggregation, −60% on fibrin(ogen) formation) and Tirofiban (complete inhibition of platelet aggregation and fibrin(ogen) formation) and secondary hemostasis with the antithrombin drugs Heparin (−70% on platelet aggregation, −80% on fibrin(ogen) formation) and Lepirudin (−80% on platelet aggregation, −90% on fibrin(ogen) formation). Smart Clot, through a single test, provides quantitative results concerning platelet aggregation and fibrin formation and is suitable for undergoing comparative studies with other coagulation point-of-care devices. Full article

Organ-on-a-chip technologies show exponential growth driven by the need to reduce the number of experimental animals and develop physiologically relevant human models for testing drugs. In vitro, microfluidic devices should be carefully designed and fabricated to provide reliable tools for modeling physiological or pathological conditions and assessing, for example, drug delivery through biological barriers. The aim of the current study was to optimize the utilization of three existing skin-on-a-chip microfluidic diffusion chambers with various designs. For this, different perfusion flow rates were compared using cellulose acetate membrane, polyester membrane, excised rat skin, and acellular alginate scaffold in the chips. These diffusion platforms were integrated into a single-channel microfluidic diffusion chamber, a multi-channel chamber, and the LiveBox2 system. The experimental results revealed that the 40 µL/min flow rate resulted in the highest diffusion of the hydrophilic model formulation (2% caffeine cream) in each system. The single-channel setup was used for further analysis by computational fluid dynamics simulation. The visualization of shear stress and fluid velocity within the microchannel and the presentation of caffeine progression with the perfusion fluid were consistent with the measured data. These findings contribute to the development and effective application of microfluidic systems for penetration testing. Full article

This study proposes a redesign of asymmetric single-chamber microbial fuel cells (a-SCMFCs) with the goal of optimizing energy production. In the present work, the new approach is based on the introduction of a novel intermediate microfluidic septum (IMS) inside the electrolyte chamber. This IMS was designed as a relatively simple and inexpensive method to optimize both electrolyte flow and species transfer inside the devices. a-SCMFCs, featuring the IMS, are compared to control cells (IMS-less), when operated with sodium acetate as the carbon energy source. Performances of cells are evaluated in terms both of maximum output potential achieved, and energy recovery (E_rec) as the ratio between the energy yield and the inner electrolyte volume. The a-SCMFCs with the novel IMS are demonstrated to enhance the energy recovery compared to control cells exhibiting E_rec values of (37 ± 1) J/m³, which is one order of magnitude higher than that achieved by control cells (3.0 ± 0.3) J/m³. Concerning the maximum output potential, IMS cells achieve (2.8 ± 0.2) mV, compared to control cells at (0.68 ± 0.07) mV. Furthermore, by varying the sodium acetate concentration, the E_rec and maximum potential output values change accordingly. By monitoring the activity of a-SCMFCs for over one year, the beneficial impact of the IMS on both the initial inoculation phase and the long-term stability of electrical performance are observed. These improvements support the effectiveness of IMS to allow the development of efficient biofilms, likely due to the reduction in oxygen cross-over towards the anode. Electrochemical characterizations confirm that the presence of the IMS impacts the diffusion processes inside the electrolytic chamber, supporting the hypothesis of a beneficial effect on oxygen cross-over. Full article

New microfluidic lab-on-a-chip capabilities are enabled by broadening the toolkit of devices that can be created using microfabrication processes. For example, complex geometries made possible by 3D printing can be used to approach microfluidic design and application in new or enhanced ways. In this paper, we demonstrate three distinct designs for microfluidic one-way (check) valves that can be fabricated using digital light processing stereolithography (DLP-SLA) with a poly(ethylene glycol) diacrylate (PEGDA) resin, each with an internal volume of 5–10 nL. By mapping flow rate to pressure in both the forward and reverse directions, we compare the different designs and their operating characteristics. We also demonstrate pumps for each one-way valve design comprised of two one-way valves with a membrane valve displacement chamber between them. An advantage of such pumps is that they require a single pneumatic input instead of three as for conventional 3D-printed pumps. We also characterize the achievable flow rate as a function of the pneumatic control signal period. We show that such pumps can be used to create a single-stage diffusion mixer with significantly reduced pneumatic drive complexity. Full article

Cellular chemotaxis has been the subject of a variety of studies due to its relevance in physiological processes, disease pathogenesis, and systems biology, among others. The migration of cells towards a chemical source remains a closely studied topic, with the Boyden chamber being one of the earlier techniques that has successfully studied cell chemotaxis. Despite its success, diffusion chambers such as these presented a number of problems, such as the quantification of many aspects of cell behaviour, the reproducibility of procedures, and measurement accuracy. The advent of microfluidic technology prompted more advanced studies of cell chemotaxis, usually involving the social amoeba Dictyostelium discoideum (D. discoideum) as a model organism because of its tendency to aggregate towards chemotactic agents and its similarities to higher eukaryotes. Microfluidic technology has made it possible for studies to look at chemotactic properties that would have been difficult to observe using classic diffusion chambers. Its flexibility and its ability to generate consistent concentration gradients remain some of its defining aspects, which will surely lead to an even better understanding of cell migratory behaviour and therefore many of its related biological processes. This paper first dives into a brief introduction of D. discoideum as a social organism and classical chemotaxis studies. It then moves to discuss early microfluidic devices, before diving into more recent and advanced microfluidic devices and their use with D. discoideum. The paper then closes with brief opinions about research progress in the field and where it will possibly lead in the future. Full article

Several ex vivo and in vitro skin models are available in the toolbox of dermatological and cosmetic research. Some of them are widely used in drug penetration testing. The excised skins show higher variability, while the in vitro skins provide more reproducible data. The aim of the current study was to compare the chemical composition of different skin models (excised rat skin, excised human skin and human-reconstructed epidermis) by measurement of ceramides, cholesterol, lactate, urea, protein and water at different depths of the tissues. The second goal was to compile a testing system, which includes a skin-on-a-chip diffusion setup and a confocal Raman spectroscopy for testing drug diffusion across the skin barrier and accumulation in the tissue models. A hydrophilic drug caffeine and the P-glycoprotein substrate quinidine were used in the study as topical cream formulations. The results indicate that although the transdermal diffusion of quinidine is lower, the skin accumulation was comparable for the two drugs. The various skin models showed different chemical compositions. The human skin was abundant in ceramides and cholesterol, while the reconstructed skin contained less water and more urea and protein. Based on these results, it can be concluded that skin-on-a-chip and confocal Raman microspectroscopy are suitable for testing drug penetration and distribution at different skin layers within an exposition window. Furthermore, obese human skin should be treated with caution for skin absorption testing due to its unbalanced composition. Full article

Skin equivalents and skin explants are widely used for dermal penetration studies in the pharmacological development of drugs. Environmental parameters, such as the incubation and culture conditions affect cellular responses and thus the relevance of the experimental outcome. However, available systems such as the Franz diffusion chamber, only measure in the receiving culture medium, rather than assessing the actual conditions for cells in the tissue. We developed a sampling design that combines open flow microperfusion (OFM) sampling technology for continuous concentration measurements directly in the tissue with microfluidic biosensors for online monitoring of culture parameters. We tested our design with real-time measurements of oxygen, glucose, lactate, and pH in full-thickness skin equivalent and skin explants. Furthermore, we compared dermal penetration for acyclovir, lidocaine, and diclofenac in skin equivalents and skin explants. We observed differences in oxygen, glucose, and drug concentrations in skin equivalents compared to the respective culture medium and to skin explants. Full article

There is an increasing demand for transdermal transport measurements to optimize topical drug formulations and to achieve proper penetration profile of cosmetic ingredients. Reflecting ethical concerns the use of both human and animal tissues is becoming more restricted. Therefore, the focus of dermal research is shifting towards in vitro assays. In the current proof-of-concept study a three-layer skin equivalent using human HaCaT keratinocytes, an electrospun polycaprolactone mesh and a collagen-I gel was compared to human excised skin samples. We measured the permeability of the samples for 2% caffeine cream using a miniaturized dynamic diffusion cell (“skin-on-a-chip” microfluidic device). Caffeine delivery exhibits similar transport kinetics through the artificial skin and the human tissue: after a rapid rise, a long-lasting high concentration steady state develops. This is markedly distinct from the kinetics measured when using cell-free constructs, where a shorter release was observable. These results imply that both the established skin equivalent and the microfluidic diffusion chamber can serve as a suitable base for further development of more complex tissue substitutes. Full article

Hypoxia switches the metabolism of tumor cells and induces drug resistance. Currently, no therapeutic exists that effectively and specifically targets hypoxic cells in tumors. Development of such therapeutics critically depends on the availability of in vitro models that accurately recapitulate hypoxia as found in the tumor microenvironment. Here, we report on the design and validation of an easy-to-fabricate tumor-on-a-chip microfluidic platform that robustly emulates the hypoxic tumor microenvironment. The tumor-on-a-chip model consists of a central chamber for 3D tumor cell culture and two side channels for medium perfusion. The microfluidic device is fabricated from polydimethylsiloxane (PDMS), and oxygen diffusion in the device is blocked by an embedded sheet of polymethyl methacrylate (PMMA). Hypoxia was confirmed using oxygen-sensitive probes and the effect on the 3D tumor cell culture investigated by a pH-sensitive dual-labeled fluorescent dextran and a fluorescently labeled glucose analogue. In contrast to control devices without PMMA, PMMA-containing devices gave rise to decreases in oxygen and pH levels as well as an increased consumption of glucose after two days of culture, indicating a rapid metabolic switch of the tumor cells under hypoxic conditions towards increased glycolysis. This platform will open new avenues for testing anti-cancer therapies targeting hypoxic areas. Full article

In this work, a disposable passive microfluidic device for cell culturing that does not require any additional/external pressure sources is introduced. By regulating the height of fluidic columns and the aperture and closure of the source wells, the device can provide different media and/or drug flows, thereby allowing different flow patterns with respect to time. The device is made of two Polymethylmethacrylate (PMMA) layers fabricated by micro-milling and solvent assisted bonding and allows us to ensure a flow rate of 18.6 μL/h - 7%/day, due to a decrease of the fluid height while the liquid is driven from the reservoirs into the channels. Simulations and experiments were conducted to characterize flows and diffusion in the culture chamber. Melanoma tumor cells were used to test the device and carry out cell culturing experiments for 48 h. Moreover, HeLa, Jurkat, A549 and HEK293T cell lines were cultivated successfully inside the microfluidic device for 72 h. Full article

To develop proper drug formulations and to optimize the delivery of their active ingredients through the dermal barrier, the Franz diffusion cell system is the most widely used in vitro/ex vivo technique. However, different providers and manufacturers make various types of this equipment (horizontal, vertical, static, flow-through, smaller and larger chambers, etc.) with high variability and not fully comparable and consistent data. Furthermore, a high amount of test drug formulations and large size of diffusion skin surface and membranes are important requirements for the application of these methods. The aim of our study was to develop a novel Microfluidic Diffusion Chamber device and compare it with the traditional techniques. Here the design, fabrication, and a pilot testing of a microfluidic skin-on-a chip device are described. Based on this chip, further developments can also be implemented for industrial purposes to assist the characterization and optimization of drug formulations, dermal pharmacokinetics, and pharmacodynamic studies. The advantages of our device, beside the low costs, are the small drug and skin consumption, low sample volumes, dynamic arrangement with continuous flow mimicking the dermal circulation, as well as rapid and reproducible results. Full article

Microfluidic DNA biochips capable of detecting specific DNA sequences are useful in medical diagnostics, drug discovery, food safety monitoring and agriculture. They are used as miniaturized platforms for analysis of nucleic acids-based biomarkers. Binding kinetics between immobilized single stranded DNA on the surface and its complementary strand present in the sample are of interest. To achieve optimal sensitivity with minimum sample size and rapid hybridization, ability to predict the kinetics of hybridization based on the thermodynamic characteristics of the probe is crucial. In this study, a computer aided numerical model for the design and optimization of a flow-through biochip was developed using a finite element technique packaged software tool (FEMLAB; package included in COMSOL Multiphysics) to simulate the transport of DNA through a microfluidic chamber to the reaction surface. The model accounts for fluid flow, convection and diffusion in the channel and on the reaction surface. Concentration, association rate constant, dissociation rate constant, recirculation flow rate, and temperature were key parameters affecting the rate of hybridization. The model predicted the kinetic profile and signal intensities of eighteen 20-mer probes targeting vancomycin resistance genes (VRGs). Predicted signal intensities and hybridization kinetics strongly correlated with experimental data in the biochip (R² = 0.8131). Full article

This paper presents an innovative mixing technology for centrifugal microfluidic platforms actuated using a specially designed flyball governor. The multilayer microfluidic disc was fabricated using a polydimethylsiloxane (PDMS) replica molding process with a soft lithography technique. The operational principle is based on the interaction between the elastic covering membrane and an actuator pin installed on the flyball governor system. The flyball governor was used as the transducer to convert the rotary motion into a reciprocating linear motion of the pin pressing against the covering membrane of the mixer chamber. When the rotation speed of the microfluidic disc was periodically altered, the mixing chamber was compressed and released accordingly. In this way, enhanced active mixing can be achieved with much better efficiency in comparison with diffusive mixing. Full article