Numerical and experimental investigation of the deviation of microparticles inside the microchannel using the vortices caused by the ICEK phenomenon.

One field of study in microfluidics is the control, trapping, and separation of microparticles suspended in fluid. Some of its applications are related to cell handling, virus detection, and so on. One of the new methods in this field is using ICEK phenomena and dielectrophoresis forces. In the present study, considering the ICEK phenomena, the microparticles inside the fluid are deviated in the desired ratio using a novel ICEK microchip. The deviation is such that after the microparticles reach the floating electrode, they are trapped in the ICEK flow vortex and deviated through a secondary channel that was placed crosswise and noncoplanar above the main channel. For simulation verification, an experimental test is done. The method used for making two noncoplanar channels and separating the particles in the desired ratio with a simple ICEK microchip is an innovation of the present study. Moreover, the adjustment of the percentage of separation of microparticles by adjusting the parameters of the applied voltage and fluid inlet velocity is one of the other innovations of the present experimental study. We observed that for input velocities of 150-1200 µm/s with applied voltages of 10-33 V, 100% of the particles can be directed toward the secondary-channel.

Effects of 3D electrodes arrangement in a novel AC electroosmotic micropump: Numerical modeling and experimental validation.

To date, a comprehensive systematic optimization framework, capable of accurately predicting an efficient electrode geometry, is not available. Here, different geometries, including 3D step electrodes, have been designed in order to fabricate AC electroosmosis micropumps. It is essential to optimize both geometrical parameters of electrode, such as width and height of steps on each base electrode and their location in one pair, the size of each base electrode (symmetric or asymmetric), the gap of electrode pairs, and nongeometrical parameters such as fluid flow in a channel and electrical characteristics (e.g., frequency and voltage). The governing equations comprising of electric domain and fluid domain have been coupled using finite element method. The developed model was employed to investigate the effect of electrode geometric parameters on electroosmotic slip velocity and its subsequent effect on pressure and flow rate. Numerical simulation indicates that the optimal performance can be achieved using a design with varying step height and displacement, at a given voltage (2.5 V) and frequency (1 kHz). Finally, in order to validate the numerical simulation, the optimal microchip was fabricated using a combination of photolithography, electroplating, and a polydimethylsiloxane microchannel. Our results indicate that our micropump is capable of generating a pressure, velocity, and flow rate of 74.2 Pa, 1.76 mm/s, and 14.8 µl/min, respectively. This result reveals that our proposed geometry outperforms the state-of-the-art micropumps previously reported in the literature by improving the fluid velocity by 32%, with 80% less electrodes per unit length, and whereas the channel length is ∼80% shorter.

Hydrodynamic cavitation for scalable exfoliation of few-layered graphene nanosheets.

A scalable manufacturing method for the production of biocompatible fewlayered graphene nanosheets is developed using hydrodynamic cavitation. Scalable exfoliation is induced by employing hydrodynamic cavitation and a serum albumin protein. Unlike acoustic cavitation, the primary means of bubble collapse in hydrodynamic cavitation is caused laterally, thereby separating two adjacent flakes through a shear effect. In this process, bovine serum albumin, a known protein, was employed to act as an effective exfoliation agent and provide desired stability by preventing restacking of the graphene layers. This method was used to study the effect of time of graphene exfoliation in a novel hydrodynamic cavitation system. The fabricated products were characterized using Raman spectroscopy, Transmission electron microscopy, Fourier transform infrared spectroscopy and differential scanning calorimetry. The results showed that with increasing the time of exfoliation, the number of graphene layers decreased based on theI2D/IGratio but disorder increased based on theID/IGratio. At 3 h, theI2D/IGratio was at 0.39 and theID/IGratio was 0.25, while at 6 h theI2D/IGratio was 0.35 andID/IGratio was 0.29. The results of the theoretical and computational analysis this research outlines are needed to obtain an effective cavitation model that can be used to potentially improve graphene synthesis and quality. The captured images of bubble propagation in the solution imply that this fluidic phenomenon could assist the graphene exfoliation. To prove this, a simple cavitation model using a needle valve was designed. The needle valve cavitation setup was able to identify that cavitation assists in graphene exfoliation and this was proved using the graphene characterization data. Based on these findings, the simulation models were designed in ANSYS and COMSOL. Specifically, through the ANSYS simulation, we were able to calculate cavitation numbers for specific flow rates and fluid temperatures.

Advancement of Sensor Integrated Organ-on-Chip Devices.

Organ-on-chip devices have provided the pharmaceutical and tissue engineering worlds much hope since they arrived and began to grow in sophistication. However, limitations for their applicability were soon realized as they lacked real-time monitoring and sensing capabilities. The users of these devices relied solely on endpoint analysis for the results of their tests, which created a chasm in the understanding of life between the lab the natural world. However, this gap is being bridged with sensors that are integrated into organ-on-chip devices. This review goes in-depth on different sensing methods, giving examples for various research on mechanical, electrical resistance, and bead-based sensors, and the prospects of each. Furthermore, the review covers works conducted that use specific sensors for oxygen, and various metabolites to characterize cellular behavior and response in real-time. Together, the outline of these works gives a thorough analysis of the design methodology and sophistication of the current sensor integrated organ-on-chips.

Capacitance of Flexible Polymer/Graphene Microstructures with High Mechanical Strength.

Carbon-modified fibrous structures with high biocompatibility have attracted much attention due to their low cost, sustainability, abundance, and excellent electrical properties. However, some carbon-based materials possess low specific capacitance and electrochemical performance, which pose significant challenges in developing electronic microdevices. In this study, we report a microfluidic-based technique of manufacturing alginate hollow microfibers incorporated by water dispersed modified graphene (bovine serum albumin-graphene). These architectures successfully exhibited enhanced conductivity ∼20 times higher than alginate hollow microfibers without any significant change in the inner dimension of the hollow region (220.0 ± 10.0 µm) compared with pure alginate hollow microfibers. In the presence of graphene, higher specific surface permeability, active ion adsorption sites, and shorter pathways were created. These continuous ion transport networks resulted in improved electrochemical performance. The desired electrochemical properties of the microfibers make alginate/graphene hollow fibers an excellent choice for further use in the development of flexible capacitors with the potential to be used in smart health electronics.

Hydrodynamic Assembly of Astrocyte Cells in Conductive Hollow Microfibers.

The manufacturing of 3D cell scaffoldings provides advantages for modeling diseases and injuries as it enables the creation of physiologically relevant platforms. A triple-flow microfluidic device is developed to rapidly fabricate alginate/graphene hollow microfibers based on the gelation of alginate induced with CaCl2 . This five-channel microdevice actualizes continuous mild fabrication of hollow fibers under an optimized flow rate ratio of 300:200:100 µL min-1 . The polymer solution is 2.5% alginate in 0.1% graphene and a 30% polyethylene glycol solution is used as the sheath and core solutions. The biocompatibility of these conductive microfibers by encapsulating mouse astrocyte cells (C8D1A) within the scaffolds is investigated. The cells can successfully survive both the manufacturing process and prolonged encapsulation for up to 8 days, where there is between 18-53% of live cells on both the alginate microfibers and alginate/graphene microfibers. These unique 3D hollow scaffolds can significantly enhance the available surface area for nutrient transport to the cells. In addition, these conductive hollow scaffolds illustrate unique advantages such as 0.728 cm3  gr-1 porosity and two times more electrical conductivity in comparison to alginate scaffolds. The results confirm the potential of these scaffolds as a microenvironment that supports cell growth.

Graphene Microelectrodes for Real-Time Impedance Spectroscopy of Neural Cells.

Understanding the changes in the electrochemical properties of neural cells upon exposure to stress factors imparts vital information about the conditions prior to their death. This study presents a graphene-based biosensor for real-time monitoring of N27 rat dopaminergic neural cells which characterizes cell adhesion and cytotoxicity factors through impedance spectroscopy. The aim was to monitor the growth of the entire cell network via a nonmetallic flexible electrode array. Therefore, a water-based graphene solution was formulized as a conductive ink, 3D-printed into a flexible substrate through an electrohydrodynamic approach, resulting in electrodes with a conductivity of 6750 s/m. The presented high-throughput method enabled microscale monitoring of the entire cell network via the design of PDMS-based growth channels. The electrical resistance of the cell network was measured continuously along with their network density, constituting a mean density of 1890 cell/mm2 at full cell confluency. The results demonstrate the applicability of the impedance-based sensing of the cell network for rapid screening of the cytotoxic elements, and the real-time effect of UV exposure on dopaminergic neural cells was reported as an immediate application of the device.

Transport of Maternally Administered Pharmaceutical Agents Across the Placental Barrier In Vitro.

To understand the transport of pharmaceutical agents and their effects on developing fetus, we have created a placental microsystem that mimics structural phenotypes and physiological characteristic of a placental barrier. We have shown the formation of a continuous network of epithelial adherens junctions and endothelial cell-cell junctions confirming the integrity of the placental barrier. More importantly, the formation of elongated microvilli under dynamic flow condition is demonstrated. Fluid shear stress acts as a mechanical cue triggering the microvilli formation. Pharmaceutical agents were administered to the maternal channel, and the concentration of pharmaceutical agents in fetal channel for coculture and control models were evaluated. In fetal channel, the coculture model exhibited about 2.5 and 2.2% of the maternal initial concentration for naltrexone and 6ß-naltrexol, respectively. In acellular model, fetal channel showed about 10.5 and 10.3% of the maternal initial concentration for naltrexone and 6ß-naltrexol, respectively. Gene expressions of epithelial cells after direct administration of naltrexone and 6ß-naltrexol to the maternal channel and endothelial cells after exposure due to transport through placental barrier are also reported.

Minute-sensitive real-time monitoring of neural cells through printed graphene microelectrodes.

Real-time and high-throughput cytometric monitoring of neural cells exposed to injury mechanisms is invaluable for in-vitro studies. Electrical impedance spectroscopy via microelectrode arrays is a label-free technique for monitoring of neural growth and their detachment upon death. In this method, the interface material plays a vital role to provide desirable attachment cues for the cell network. Thus, here we demonstrate the electrohydrodynamic patterning of aqueous graphene for microelectrode fabrication. We investigated whether the wrinkled surface morphology of the electrodes fabricated by this deposition method expands their electroactive surface area and thus enables a rapid response time. The nano-scale quality of the graphene lattice is characterized by Raman spectroscopy and Transmittance electron microscopy. N27 rat dopaminergic neural cells were cultured on the chips and the surface morphology of the microelectrodes during cellular growth was investigated by Scanning electrode spectroscopy. Attachment of the neural population on the graphene microelectrodes was parametrized and the change in the impedance spectrum of this cell population was quantified at 10 Hz to 10 kHz frequencies along with the change in TUBB3 gene expression. The viability test of the cell population on the biosensor demonstrated no significant difference in comparison to the control, and a cell density of 2289 cell/mm2 was achieved. As a proof of concept, the confluent N27 cell population was exposed to UV and its cytotoxic impact on neural detachment and lift-off was monitored. The multiplexed detection of cellular activity was reported with a temporal resolution of one minute.

Microfluidic Seeding of Cells on the Inner Surface of Alginate Hollow Microfibers.

Mimicking microvascular tissue microenvironment in vitro calls for a cytocompatible technique of manufacturing biocompatible hollow microfibers suitable for cell-encapsulation/seeding in and around them. The techniques reported to date either have a limit on the microfiber dimensions or undergo a complex manufacturing process. Here, a microfluidic-based method for cell seeding inside alginate hollow microfibers is designed whereby mouse astrocytes (C8-D1A) are passively seeded on the inner surface of these hollow microfibers. Collagen I and poly-d-lysine, as cell attachment additives, are tested to assess cell adhesion and viability; the results are compared with nonadditive-based hollow microfibers (BARE). The BARE furnishes better cell attachment and higher cell viability immediately after manufacturing, and an increasing trend in the cell viability is observed between Day 0 and Day 2. Swelling analysis using percentage initial weight and width is performed on BARE microfibers furnishing a maximum of 124.1% and 106.1%, respectively. Degradation analysis using weight observed a 62% loss after 3 days, with 46% occurring in the first 12 h. In the frequency sweep test performed, the storage modulus (G') remains comparatively higher than the loss modulus (G″) in the frequency range 0-20 Hz, indicating high elastic behavior of the hollow microfibers.

Machine learning-assisted E-jet printing for manufacturing of organic flexible electronics.

Electrohydrodynamic-jet (E-jet) printing technique enables the high-resolution printing of complex soft electronic devices. As such, it has an unmatched potential for becoming the conventional technique for printing soft electronic devices. In this study, the electrical conductivity of the E-jet printed circuits was studied as a function of key printing parameters (nozzle speed, ink flow rate, and voltage). The collected experimental dataset was then used to train a machine learning algorithm to establish models capable of predicting the characteristics of the printed circuits in real-time. A decision tree was applied to the data set and resulted in an accuracy of 0.72, and further evaluations showed that pruning the tree increased the accuracy while sensitivity decreased in the highly pruned trees. The k-fold cross-validation (CV) method was used in model selection to test the ability of the model to get trained on data. The accuracy of CV method was the highest for random forest at 0.83 and K-NN model (k = 10) at 0.82. Precision parameters were compared to evaluate the supervised classification models. According to F-measure values, the K-NN model (k = 10) and random forest are the best methods to classify the conductivity of electrodes.

Recent Advances in Microfluidically Spun Microfibers for Tissue Engineering and Drug Delivery Applications.

In recent years, the unique and tunable properties of microfluidically spun microfibers have led to tremendous advancements for the field of biomedical engineering, which have been applied to areas such as tissue engineering, wound dressing, and drug delivery, as well as cell encapsulation and cell seeding. In this article, we analyze the most recent advances in microfluidics and microfluidically spun microfibers, with an emphasis on biomedical applications. We explore in detail these new and innovative experiments, how microfibers are made, the experimental purpose of making microfibers, and the future work that can be done as a result of these new types of microfibers. We also focus on the applications of various materials used to fabricate microfibers, as well as their many promises and limitations.

Targeted Microfluidic Manufacturing to Mimic Biological Microenvironments: Cell-Encapsulated Hollow Fibers.

At present, the blood-brain barrier (BBB) poses a challenge for treating a wide range of central nervous system disorders; reliable BBB models are still needed to understand and manipulate the transfer of molecules into the brain, thereby improving the efficiency of treatments. In this study, hollow, cell-laden microfibers are fabricated and investigated as a starting point for generating BBB models. The genetic effects of the manufacturing process are analyzed to understand the implications of encapsulating cells in this manner. These fibers are created using different manufacturing parameters to understand the effects on wall thickness and overall diameter. Then, dopaminergic rat cells are encapsulated into hollow fibers, which maintained at least 60% live cells throughout the three-day observation period. Lastly, genetic changes tyrosine hydroxylase (TH) and tubulin beta 3 class III (TUBB-3) are investigated to elucidate the effects on cell health and behavior; while the TH levels in encapsulated cells were similar to control cells, showing similar levels of TH synthesis, TUBB-3 was downregulated, indicating lower amounts of cellular neurogenesis.

Protein-assisted scalable mechanochemical exfoliation of few-layer biocompatible graphene nanosheets.

A facile method to produce few-layer graphene (FLG) nanosheets is developed using protein-assisted mechanical exfoliation. The predominant shear forces that are generated in a planetary ball mill facilitate the exfoliation of graphene layers from graphite flakes. The process employs a commonly known protein, bovine serum albumin (BSA), which not only acts as an effective exfoliation agent but also provides stability by preventing restacking of the graphene layers. The latter is demonstrated by the excellent long-term dispersibility of exfoliated graphene in an aqueous BSA solution, which exemplifies a common biological medium. The development of such potentially scalable and toxin-free methods is critical for producing cost-effective biocompatible graphene, enabling numerous possible biomedical and biological applications. A methodical study was performed to identify the effect of time and varying concentrations of BSA towards graphene exfoliation. The fabricated product has been characterized using Raman spectroscopy, powder X-ray diffraction, transmission electron microscopy and scanning electron microscopy. The BSA-FLG dispersion was then placed in media containing Astrocyte cells to check for cytotoxicity. It was found that lower concentrations of BSA-FLG dispersion had only minute cytotoxic effects on the Astrocyte cells.

Behavior of Neural Cells Post Manufacturing and After Prolonged Encapsulation within Conductive Graphene-Laden Alginate Microfibers.

Engineering conductive 3D cell scaffoldings offer advantages toward the creation of physiologically relevant platforms with integrated real-time sensing capabilities. Dopaminergic neural cells are encapsulated into graphene-laden alginate microfibers using a microfluidic approach, which is unmatched for creating highly-tunable microfibers. Incorporating graphene increases the conductivity of the alginate microfibers by 148%, creating a similar conductivity to native brain tissue. The cell encapsulation procedure has an efficiency of 50%, and of those cells, ≈30% remain for the entire 6-day observation period. To understand how the microfluidic encapsulation affects cell genetics, tyrosine hydroxylase, tubulin beta 3 class 3, interleukin 1 beta, and tumor necrosis factor alfa are analyzed primarily with real-time reverse transcription-quantitative polymerase chain reaction and secondarily with enzyme-linked immunosorbent assay, immediately after manufacturing, after encapsulation in polymer matrix for 6 days, and after encapsulation in the graphene-polymer composite for 6 days. Preliminary data shows that the manufacturing process and combination with alginate matrix affect the expression of the studied genes immediately after manufacturing. In addition, the introduction of graphene further changes gene expressions. Long-term encapsulation of neural cells in alginate and 6-day exposure to graphene also leads to changes in gene expressions.

Manufacturing of poly(ethylene glycol diacrylate)-based hollow microvessels using microfluidics.

The microvasculature is a vital organ that distributes nutrients within tissues, and collects waste products from them, and which defines the environmental conditions in both normal and disease situations. Here, a microfluidic chip was developed for the fabrication of poly(ethylene glycol diacrylate) (PEGDA)-based hollow self-standing microvessels having inner dimensions ranging from 15 µm to 73 µm and displaying biocompatibility/cytocompatibility. Macromer solutions were hydrodynamically focused into a single microchannel to form a concentric flow regime, and were subsequently solidified through photopolymerization. This approach uniquely allowed the fabrication of hollow microvessels having a defined structure and integrity suitable for cell culturing.

High-Yield Production of Aqueous Graphene for Electrohydrodynamic Drop-on-Demand Printing of Biocompatible Conductive Patterns.

Presented here is a scalable and aqueous phase exfoliation of graphite to high yield and quality of few layer graphene (FLG) using Bovine Serum Albomine (BSA) and wet ball milling. The produced graphene ink is tailored for printable and flexible electronics, having shown promising results in terms of electrical conductivity and temporal stability. Shear force generated by steel balls which resulted in 2-3 layer defect-free graphene platelets with an average size of hundreds of nm, and with a concentration of about 5.1 mg/mL characterized by Raman spectroscopy, atomic force microscopy (AFM), transmittance electron microscopy (TEM) and UV-vis spectroscopy. Further, a conductive ink was prepared and printed on flexible substrate (Polyimide) with controlled resolution. Scanning electron microscopy (SEM) and Profilometry revealed the effect of thermal annealing on the prints to concede consistent morphological characteristics. The resulted sheet resistance was measured to be R s   =   36.75   Ω / sqr for prints as long as 100 mm. Printable inks were produced in volumes ranging from 20 mL to 1 L, with potential to facilitate large scale production of graphene for applications in biosensors, as well as flexible and printable electronics.

Characterization of Astrocytic Response after Experiencing Cavitation In Vitro.

When a traumatic brain injury (TBI) occurs, low-pressure regions inside the skull can cause vapor contents in the cerebral spinal fluid (CSF) to expand and collapse, a phenomenon known as cavitation. When these microbubbles (MBs) collapse, shock waves are radiated outward and are known to damage surrounding materials in other applications, like the steel foundation of boat propellers, so it is alarming to realize the damage that cavitation inflicts on vulnerable brain tissue. Using cell-laden microfibers, the longitudinal morphological response that mouse astrocytes have to surrounding cavitation in vitro is visually analyzed. Astrocytic damage is evident immediately after cavitation when compared to a control sample, as their processes retract. Forty-eight hours later, the astrocytes appeared to spread across the fibers, as normal. This study also analyzes the gene expression changes that occur post-cavitation via quantitative polymerase chain reaction (qPCR) methods. After cavitation a number of pro-inflammatory genes are upregulated, including TNFα, IL-1ß, C1q, Serping1, NOS1, IL-6, and JMJD3. Taken together, these results confirm that surrounding cavitation is detrimental to astrocytic function, and yield opportunities to further the understanding of how protective headgear can minimize or eliminate the occurrence of cavitation.

Recovery of Encapsulated Adult Neural Progenitor Cells from Microfluidic-Spun Hydrogel Fibers Enhances Proliferation and Neuronal Differentiation.

Because of the limitations imposed by traditional two-dimensional (2D) cultures, biomaterials have become a major focus in neural and tissue engineering to study cell behavior in vitro. 2D systems fail to account for interactions between cells and the surrounding environment; these cell-matrix interactions are important to guide cell differentiation and influence cell behavior such as adhesion and migration. Biomaterials provide a unique approach to help mimic the native microenvironment in vivo. In this study, a novel microfluidic technique is used to encapsulate adult rat hippocampal stem/progenitor cells (AHPCs) within alginate-based fibrous hydrogels. To our knowledge, this is the first study to encapsulate AHPCs within a fibrous hydrogel. Alginate-based hydrogels were cultured for 4 days in vitro and recovered to investigate the effects of a 3D environment on the stem cell fate. Post recovery, cells were cultured for an additional 24 or 72 h in vitro before fixing cells to determine if proliferation and neuronal differentiation were impacted after encapsulation. The results indicate that the 3D environment created within a hydrogel is one factor promoting AHPC proliferation and neuronal differentiation (19.1 and 13.5%, respectively); however, this effect is acute. By 72 h post recovery, cells had similar levels of proliferation and neuronal differentiation (10.3 and 8.3%, respectively) compared to the control conditions. Fibrous hydrogels may better mimic the natural micro-environment present in vivo and be used to encapsulate AHPCs, enhancing cell proliferation and selective differentiation. Understanding cell behavior within 3D scaffolds may lead to the development of directed therapies for central nervous system repair and rescue.

Drug transport across the human placenta: review of placenta-on-a-chip and previous approaches.

In the past few decades, the placenta became a very controversial topic that has had many researchers and pharmacists discussing the significance of the effects of pharmaceutical drug intake and how it is a possible leading cause towards birth defects. The creation of an in vitro microengineered model of the placenta can be used to replicate the interactions between the mother and fetus, specifically pharmaceutical drug intake reactions. As the field of nanotechnology significantly continues growing, nanotechnology will become more apparent in the study of medicine and other scientific disciplines, specifically microengineering applications. This review is based on past and current research that compares the feasibility and testing of the placenta-on-a-chip microengineered model to the previous and underdeveloped in vivo and ex vivo approaches. The testing of the practicality and effectiveness of the in vitro, in vivo and ex vivo models requires the experimentation of prominent pharmaceutical drugs that most mothers consume during pregnancy. In this case, these drugs need to be studied and tested more often. However, there are challenges associated with the in vitro, in vivo and ex vivo processes when developing a practical placental model, which are discussed in further detail.