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Simple and stable generation of monodispersed droplets with volume from picolitre to nanoliter is one of the key factors in high-throughput quantitative microreactors for chemical and biomedical applications. In this work, an efficient method that could realize simple manipulating microflow with a broad operation window for preparing monodispersed droplets with controllable diameter is developed. The microfluidic device is constructed by inserting a capillary with an oblique angle (α) into the continuous phase, named a floating capillary-based open microfluidic device (FCOMD). The transition of droplet-generating mode between dripping and jetting can be achieved by changing capillary number and α. A computational model based on the volume-of-fluid/continuum-surface-force method to explain the controllability of α on the droplet formation regime and droplet breakage, verifying the synergistic effect of ΔP and Fb, facilitates the droplet pinching. A descending order of Pn of capillary with different α is that 45° > 30° > 15° > 60° > 75°, leading to the same order of generated droplet's D. When compared with the traditional capillary co-flow device, the generating throughput of the integrated FCOMD obtained by integrating different numbers of capillaries is at least ten times. Moreover, water in oil, oil in water double-emulsion, colloidal dispersed droplets, and liquid crystal droplets with diameters ranging from 25 to 800 μm are prepared on-demand by the FCOMD, indicating the universality of the microfluidic device. Thus, the FCOMD shows the features of simplicity, practicability, and flexibility, offering valuable guidance for generating controllable droplets with wide size change and showing a great potential application in material science, foods, pharmaceuticals, and cosmetics. [ABSTRACT FROM AUTHOR]

Unlike the conventional electroosmotic flow (EOF) driven by direct current and alternating current electric fields, this study investigates the pulse EOF of Newtonian fluids through a parallel plate microchannel actuated by pulse electric fields. Specifically, the pulses considered encompass triangular and half-sinusoidal pulse waves. By applying the Laplace transform method and the residual theorem, the analytical solutions for the velocity and volumetric flow rate of the pulse EOF associated with these two pulse waves are derived, respectively. The influence of pulse width a ¯ and electrokinetic width K on velocity is further considered, while the volumetric flow rate as a function of time t ¯ and electrokinetic width K is examined separately. A comparison of the volumetric flow rates related to these two pulse waves under varying parameters is also conducted. The research findings indicate that irrespective of the pulse wave, a broader pulse width results in a prolonged period and increased amplitude of the velocity profile. Elevating the electrokinetic width yields higher near-wall velocities, with negligible effect on near-center velocities. It is noteworthy that regardless of the electrokinetic width, the near-wall velocity exceeds that of the near-center during the first half-cycle, while the situation reverses during the second half-cycle. The volumetric flow rate varies periodically with time, initially surging rapidly with electrokinetic width before gradually stabilizing at a constant level. More interestingly, independent of pulse width and electrokinetic width, the volumetric flow rates linked to the half-sinusoidal pulse wave consistently surpass those of the triangular pulse wave. For any pulse width, the volumetric flow rates corresponding to the two pulse waves grow with higher electrokinetic widths, especially prominent at alternating intervals of the two half-cycles within a complete cycle. These findings have important implications for improving the design and optimization of microfluidic devices in engineering and biomedical applications utilizing pulse EOF. [ABSTRACT FROM AUTHOR]

The power-law fluid flow in tree-like self-similar branching networks is prevalent throughout the natural world and also finds numerous applications in technology such as oil recovery and microfluidic devices. We investigate analysis of optimal power-law fluid flow conditions and the optimal structures within tree-like branching networks, focusing on maximizing flow conductance under the constraint of the network tube's volume and the surface area. The study considered fully developed laminar power-law fluid flow regimes without considering any losses in the network system. A key observation was the sensitivity of the dimensionless effective flow conductance to the network's geometrical parameters. We found that the maximum flow conductance occurs when a dimensionless radius ratio β ∗ satisfies the equation β ∗ = N − 1 / 3 and β ∗ = N − (n + 1) / (3 n + 2) under constrained tube-volume and surface-area, respectively. Here, N represents the bifurcation number of branches splitting at each junction, and n is the fluid power-law index. We further find that this optimal condition occurs when pressure drops are equipartition across each branching level. We validated our results with various experimental results and theories under limiting conditions. Further, Hess–Murray's law is justified and extended for the shear-thinning and shear-thickening fluid flows for an arbitrary number of branches N. Further, in this study, we also derive the relationships between the geometrical and flow characteristics of the parent and daughter tubes as well as the generalized scaling laws at the optimal conditions for the other essential parameters such as tube-wall stresses, length ratios, mean velocities, tube-volume, and surface-area of the tube distributing within the networks. We find that the fluid power-law index n does not influence the constrained tube-volume scaling at the optimal conditions; however, the scaling laws vary with n under the constrained tube's surface area. These findings offer valuable design principles for developing efficient transport and flow systems. [ABSTRACT FROM AUTHOR]

3D printing techniques allow for the precise placement of living cells, biological substances, and biochemical components, establishing themselves as a promising approach in bioengineering. Recently, 3D printing has been applied to develop human-relevant in vitro cancer models with highly controlled complexity and as a potential method for drug screening and disease modeling. Compared to 2D culture, 3D-printed in vitro cancer models more closely replicate the in vivo microenvironment. Additionally, they offer a reduction in the complexity and ethical issues associated with using in vivo animal models. This focused review discusses the relevance of 3D printing technologies and the applied cells and materials used in cutting-edge in vitro cancer models and microfluidic device systems. Future prospective solutions were discussed to establish 3D-printed in vitro models as reliable tools for drug screening and understanding cancer disease mechanisms. [ABSTRACT FROM AUTHOR]

Accurate rotation of microparticles is of great significance in micro-rotors, multi-angle microscopic observation, microbial three-dimensional phenotyping, and microsystem assembly. However, most methods can only rotate a single object, thus limiting the throughput. In this study, we realized the simultaneous rotation of many trapped and aligned subwavelength glass cylinders inside an evanescent wave field excited by a resonant phononic crystal plate. The unique feature of the rotation lies in its periodic distribution as well as the rotation axis being perpendicular to the acoustic axis. The rotary power originates from viscous torque generated by the evanescent wave-induced near-boundary acoustic streaming's asymmetry distribution on the trapped cylinder. Furthermore, the three-dimensional topographies of rotated cylinders can be reconstructed from the microscopic images under different rotating angles. Our findings can pave the way toward developing simple, disposable, and scalable microfluidic devices for massive subwavelength acoustic rotation by carefully designing acoustic metamaterials. [ABSTRACT FROM AUTHOR]

This paper focuses on the study of Blasius–Rayleigh–Stokes nanofluid dusty flow with generalized Fourier and Fick laws under the influence of a transitive magnetic field. The Blasius–Rayleigh–Stokes nanofluid dusty flow has significant applications in various fields, including the design of heat exchangers, microfluidic devices and industrial processes involving the transportation of suspended particles in fluids. This paper considers the impact of melting heat transfer at the surface boundary. By employing relevant transformations, the mathematical model is transformed into a system of self-similar equations. The solution to this set of highly nonlinear equations is obtained using the bvp4c numerical method in combination with the response surface methodology (RSM) statistical approach. The results are presented through graphical illustrations and numerically calculated tabulated values. It is observed that the fitted model for the skin friction coefficient Re x 1 ∕ 2 C f optimal. Moreover, the model parameters Pr , Q, β v , M and L are linearly and quadratically significant in explaining the variation presented for skin friction coefficient Re x 1 ∕ 2 C f . This indicates the response skin friction coefficient Re x 1 ∕ 2 C f is minimized to 0.005 at Pr = 2. 2 0 2 7 , Q = 0. 2 5 2 , β v = 0. 2 2 , M = 0. 2 1 7 and L = 0. 2 5 2 and is maximized to 1.387 at Pr = 2. 0 6 6 , Q = 0. 2 5 4 , β v = 0. 0 2 1 , M = 0. 0 5 6 and L = 0. 2 5 2. [ABSTRACT FROM AUTHOR]

Cell-based assays are widely exploited for drug screening and biosensing, providing useful information about bioactivity of target analytes and complex biological samples. It is well recognized that 3D cell models are required to achieve highly valuable information, also from the perspective of replacing animal models. However, bioassays relying on 3D cell models are generally highly demanding in terms of facilities, equipment, and skilled personnel requirements. To reduce cost, increase sustainability, and provide a flexible 3D cell-based platform for bioassays, we here report a novel approach based on a 3D-printed microtissue device. To assess the suitability of this strategy for reporter gene technology, we selected to monitor two molecular pathways which were of interest in several applications, hypoxia signaling and the p53 pathway. The investigation of such pathways is highly relevant in fields spanning from drug screening to bioactivity monitoring for industrial by-product valorization. Microtissues of human hepatocarcinoma (HepG2) and human embryonic kidney (Hek293T) cell lines were obtained with a low-cost and sustainable chip platform and bioassays were developed to monitor the hypoxia-inducible factors (HIFs) and the p53 tumor suppressor pathway. HepG2 and Hek293T 3D cell models were genetically engineered to express the Luc2P from Photinus pyralis firefly either under the regulation of p53 or HIF response elements. The bioassays allowed quantitative assessment of hypoxia and tumoral activity with 1,10-phenanthroline for HIF and with doxorubicin for p53 pathway activation, respectively, showing good potential for applications of this sustainable and low-cost 3D-printed microfluidic platform for bioactivity analyses, drug screening, and precision medicine. [ABSTRACT FROM AUTHOR]

When exposed to an alternating current (AC) electric field, a polarized microparticle is moved by the interaction between the voltage-induced dipoles and the AC electric field under dielectrophoresis (DEP). The DEP force is widely used for manipulation of microparticles in diverse practical applications such as 3D manipulation, sorting, transfer, and separation of various particles such as living cells. In this study, we propose the integration of surface-enhanced Raman spectroscopy (SERS), an extremely sensitive and versatile technique based on the Raman scattering of molecules supported by nanostructured materials, with DEP using a microfluidic device. The microfluidic device combines microelectrodes with gold nanohole arrays to characterize the electrophysiological and biochemical properties of biological cells. The movement of particles, which varies depending on the electrical properties such as conductivity and permittivity of particles, can be manipulated by the cross-frequency change. For proof of concept, Raman spectroscopy using the DEP–SERS integration was performed for polystyrene beads and biological cells and resulted in an improved signal-to-noise ratio by determining the direction of the DEP force applied to the cells with respect to the applied AC power and collecting them on the nanohole arrays. The result illustrates the potential of the concept for simultaneously examining the electrical and biochemical properties of diverse chemical and biological microparticles in the microfluidic environment. [ABSTRACT FROM AUTHOR]

Gynecological cancers, including cervical, endometrial, and ovarian cancer, contribute to a significant portion of female cancer-related deaths. Despite advancements in cancer detection, these diseases continue to pose challenges due to limited cost-effective screening methods and late-stage diagnoses. This review paper focuses on the utilization of microfluidic devices (MFDs) as a cost-effective tool for diagnosing and screening gynecological cancers. MFDs are portable instruments capable of sample separation, extraction, dilution, mixing, and biomarker detection. Their compact size and efficiency make them advantageous for comprehensive sample analysis. The emergence of microfluidic point-of-care devices offers potential for developing biomarker-based screening technologies and facilitating early detection of gynecological cancers. This paper aims to consolidate the knowledge and findings surrounding the utilization of MFDs in gynecological cancer research by summarizing the current literatures that exist between 2006 and 2023. The review of previous research in this area will contribute to a comprehensive understanding of the recent state of utilizing MFDs for gynecological cancer screening and detection as it will shed light on the advancements made thus far and provide insights into the future prospects and potential directions of research in this field. [ABSTRACT FROM AUTHOR]

Cell-based therapies hold great potential for cancer immunotherapy. This approach is based on manipulation of dendritic cells to activate immune system against specific cancer antigens. For the development of an effective cell vaccine platform, gene transfer, and cell fusion have been used for modification of dendritic or tumor cells to express immune (co)stimulatory signals and to load dendritic cells with tumor antigens. Both, gene transfer and cell fusion can be achieved by single technique, a cell membrane electroporation. The cell membrane exposed to external electric field becomes temporarily permeable, enabling introduction of genetic material, and also fusogenic, enabling the fusion of cells in the close contact. We tested the feasability of combining gene electrotransfer and electrofusion into a single-step technique and evaluated the effects of electroporation buffer, pulse parameters, and cell membrane fluidity for single or combined method of gene delivery or cell fusdion. We determined the percentage of fused cells expressing green fluorescence protein (GFP) in a murine cell model of melanoma B16F1, cell line used in our previous studies. Our results suggest that gene electrotransfer and cell electrofusion can be applied in a single step. The percentage of viable hybrid cells expressing GFP depends on electric pulse parameters and the composition of the electroporation buffer. Furthermore, our results suggest that cell membrane fluidity is not related to the efficiency of the gene electrotransfer and electrofusion. The protocol is compatible with microfluidic devices, however further optimization of electric pulse parameters and buffers is still needed. [ABSTRACT FROM AUTHOR]

Continuous monitoring of pathogens finds applications in environmental, medical, and food industry settings. Quartz crystal microbalance (QCM) is one of the promising methods for real-time detection of bacteria and viruses. QCM is a technology that utilizes piezoelectric principles to measure mass and is commonly used in detecting the mass of chemicals adhering to a surface. Due to its high sensitivity and rapid detection times, QCM biosensors have attracted considerable attention as a potential method for detecting infections early and tracking the course of diseases, making it a promising tool for global public health professionals in the fight against infectious diseases. This review first provides an overview of the QCM biosensing method, including its principle of operation, various recognition elements used in biosensor creation, and its limitations and then summarizes notable examples of QCM biosensors for pathogens, focusing on microfluidic magnetic separation techniques as a promising tool in the pretreatment of samples. The review explores the use of QCM sensors in detecting pathogens in various samples, such as food, wastewater, and biological samples. The review also discusses the use of magnetic nanoparticles for sample preparation in QCM biosensors and their integration into microfluidic devices for automated detection of pathogens and highlights the importance of accurate and sensitive detection methods for early diagnosis of infections and the need for point-of-care approaches to simplify and reduce the cost of operation. [ABSTRACT FROM AUTHOR]

Femtosecond laser processing has become a critical technique for the microfabrication of hard and brittle materials, particularly in microfluidic device applications. This study focuses on the fabrication of microchannels with controllable cross-sectional profiles in silica glass, a material known for its excellent physical and chemical properties. Through a combination of experimental research and theoretical analysis, the surface generation mechanisms governing microchannel morphology are investigated, alongside the influence of various processing parameters on the surface roughness at the microchannel bottom. A comprehensive optimization method is developed to control sidewall taper and surface roughness by adjusting laser scanning paths and modes. Utilizing a composite scanning approach, the study achieves near-rectangular microchannels with average sidewall taper angles below 5° and surface roughness (Sa) of 2.53 μm. These results provide a new strategy for precise control of microchannel morphology in silica glass, offering significant potential to enhance the efficiency and precision of microfluidic device fabrication, with broad applications in both industrial and research settings. [ABSTRACT FROM AUTHOR]

Conventional testing of novel contrast agents for magnetic resonance imaging (MRI) involves cell and animal studies. However, 2D cultures lack dynamic flow and in vivo MRI is limited by regulatory approval of long‐term anesthesia use. Microfluidic tumor models (MTMs) offer a cost‐effective, reproducible, and high throughput platform for bridging cell and animal models. Yet, MRI of microfluidic devices is challenging, due to small fluid volumes generating low sensitivity. For the first time, an MRI of MTMs was performed at low field strength (1 T) using conventional imaging equipment without microcoils. To enable longitudinal MRI, we developed (1) CHAMP‐3 (controlled heating apparatus for microfluidics and portability) which heats MTMs during MRI scans and (2) an MRI‐compatible temperature monitoring system. CHAMP‐3 maintained chip surface temperature at ~37°C and the media inside at ~35.5°C. Enhanced T1‐weighted MRI contrast was achieved in 3D MTMs with free manganese (Mn2+) solutions and Mn2+ labeled tumor cells. [ABSTRACT FROM AUTHOR]

Renal cell carcinoma (RCC) is one of the deadliest malignant tumors. Despite significant advances in RCC treatment over the past decade, complete remission is rarely achieved. Consequently, there is an urgent need to explore and develop new therapies to improve the survival rates and quality of life for patients. In recent years, the development of tumor organoid technology has attracted widespread attention as it can more accurately simulate the spatial structure and physiological characteristics of tumors within the human body. In this review, we summarize the main methods currently used to construct kidney cancer organoids, as well as their various biological and clinical applications. Furthermore, combining organoids with other technologies, such as co-culture techniques and microfluidic technologies, can further develop organoids and address their limitations, creating more practical models. This approach summarizes the interactions between different tissues or organs during tumor progression. Finally, we also provide an outlook on the construction and application of kidney cancer organoids. These rapidly evolving kidney cancer organoids may soon become a focal point in the development of in vitro clinical models and therapeutic research for kidney cancer. [ABSTRACT FROM AUTHOR]

In vitro neuronal culture is an important research platform in cellular and network neuroscience. However, neurons cultured on a homogeneous scaffold form dense, randomly connected networks and display excessively synchronized activity; this phenomenon has limited their applications in network‐level studies, such as studies of neuronal ensembles, or coordinated activity by a group of neurons. Herein, polydimethylsiloxane‐based microfluidic devices are developed to create small neuronal networks exhibiting a hierarchically modular structure resembling the connectivity observed in the mammalian cortex. The strength of intermodular coupling is manipulated by varying the width and height of the microchannels that connect the modules. Neuronal activity recording via calcium imaging shows that the spontaneous activity in networks with smaller microchannels (2.2–5.5 µm2) has lower synchrony and exhibits a threefold variety of neuronal ensembles. Optogenetic stimulation demonstrates that a reduction in intermodular coupling enriches evoked neuronal activity patterns and that repeated stimulation induces plasticity in neuronal ensembles in these networks. These findings suggest that cell engineering technologies based on microfluidic devices enable in vitro reconstruction of the intricate dynamics of neuronal ensembles, thus providing a robust platform for studying neuronal ensembles in a well‐defined physicochemical environment. [ABSTRACT FROM AUTHOR]

Electrically controlled droplet manipulation is attractive for lots of applications ranging from material synthesis and drug delivery. However, the present techniques using bulky and expensive power supplies to generate high‐voltage electrical signals are easy to cause electrode electrolysis, bubble formation in buffer solution, and are incapable for point‐of‐care testing. Herein, a novel and portable electrical method for self‐powered emulsion droplet core coalescence and release based on triboelectric nanogenerator (TENG) is presented. The electrical signal with a 2.25 kV open‐voltage and a 35 µA short‐circuit current can be conveniently generated via frictional electrification effect, and the harmful electrochemical reaction in solutions can be avoided. The high transient pulse voltage and stable sinusoidal signal between two electrodes in microfluidic device can generate a strong electric field, which causes apparent Maxwell electrical stresses at droplet interfaces and subsequently promotes the core coalescence and release of double‐emulsions. By adjusting the voltage magnitude, the core coalescence and release behaviors of double‐emulsion droplets can be controllably achieved, followed by the synthesis copper‐based metal–organic framework (Cu‐MOF) nanoparticles. Thus, this portable, effective and self‐powered droplet manipulation technique can be attractive for those droplet‐based applications. [ABSTRACT FROM AUTHOR]

Inflammatory microenvironments often become acidic (pH < 7.4) due to tissue oxygen deprivation and lactate release in glycolysis by activated immune cells. Although neutrophils are known to accumulate in such microenvironments, the effects of pH on their migration are not fully understood. Here, we first investigated the pH control around cultured cells with a microfluidic device, which was equipped with two gas channels above three parallel media channels. By supplying gas mixtures with predefined carbon dioxide (CO2) concentrations to the gas channels, the gas exchange adjusted the dissolved CO2 and affected the chemical equilibrium of sodium hydrogen carbonate in the cell culture medium. A pH gradient from 8.3 to 6.8 was generated along the media channels when gas mixtures containing 1% and 50% CO2 were supplied to the left and right gas channels, respectively. Neutrophil-like differentiated human promyelocytic leukemia cells (HL-60) were then seeded to the fibronectin-coated media channels and their migratory behaviors were quantified while varying the pH. The cell migration became more active and faster under high pH than under low pH conditions. However, no directional migration along the pH gradient was detected during the three-hour observation. Thus, the microfluidic device is useful to elucidate pH-dependent cellular dynamics. [ABSTRACT FROM AUTHOR]

Pathogenic bacteria are commonly found in food, water, and soil, posing significant public health challenges globally. Therefore, early, rapid, and highly sensitive strategies for monitoring the bacterial proliferation are crucial for ensuring public health, medical diagnosis, and food safety. Compared to traditional techniques, microfluidic platforms provide powerful detective tools characterized by high integration, high throughput, ease of operation, low reagent consumption, and high sensitivity. Driven by substantial commercial demand, research and development in microfluidic‐based rapid detection methods and technologies has progressed significantly derived by the interdisciplinary integration of multiple disciplines. In this review, progress in clinical detection of pathogenic bacteria with microfluidic biosensors, including microfluidic devices for point‐of‐care (POC) testing, is summarized. Strategies for pathogenic bacteria detection, containing their advantages and disadvantages are discussed in detail. Advanced platforms for capturing and detecting pathogenic bacteria, such as microchannels, microarrays, digital microfluidics (DMF) and paper‐based platforms, are highlighted. The accomplishments and shortcomings of these microfluidic devices are also summarized. Additionally, case studies of biosensor‑based microfluidic devices used for detecting diseases caused by bacterial imbalances are listed. Finally, possible research perspectives for further development in highly effective biosensor‑based microfluidics for clinical detection of pathogenic bacteria are proposed. [ABSTRACT FROM AUTHOR]

A significant challenge in the treatment of central nervous system (CNS) disorders is represented by the presence of the blood--brain barrier (BBB), a highly selective membrane that regulates molecular transport and restricts the passage of pathogens and therapeutic compounds. Traditional in vivo models are constrained by high costs, lengthy experimental timelines, ethical concerns, and interspecies variations. In vitro models, particularly microfluidic BBB-on-a-chip devices, have been developed to address these limitations. These advanced models aim to more accurately replicate human BBB conditions by incorporating human cells and physiological flow dynamics. In this framework, here we developed an innovative microfluidic system that integrates thin-film electrodes for non-invasive, real-time monitoring of BBB integrity using electrochemical impedance spectroscopy (EIS). EIS measurements showed frequencydependent impedance changes, indicating BBB integrity and distinguishing well-formed from non-mature barriers. The data from EIS monitoring was confirmed by permeability assays performed with a fluorescence tracer. The model incorporates human endothelial cells in a vessel-like arrangement to mimic the vascular component and three-dimensional cell distribution of human astrocytes and microglia to simulate the parenchymal compartment. By modeling the BBB-on-a-chip with an equivalent circuit, a more accurate trans-endothelial electrical resistance (TEER) value was extracted. The device demonstrated successful BBB formation and maturation, confirmed through live/dead assays, immunofluorescence and permeability assays. Computational fluid dynamics (CFD) simulations confirmed that the device mimics in vivo shear stress conditions. Drug crossing assessment was performed with two chemotherapy drugs: doxorubicin, with a known poor BBB penetration, and temozolomide, conversely a specific drug for CNS disorders and able to cross the BBB, to validate the model predictive capability for drug crossing behavior. The proposed sensorized microfluidic device represents a significant advancement in BBB modeling, offering a versatile platform for CNS drug development, disease modeling, and personalized medicine. [ABSTRACT FROM AUTHOR]

Detecting glucose levels is crucial for diabetes patients as it enables timely and effective management, preventing complications and promoting overall health. In this endeavor, we have designed a novel, affordable point-of-care diagnostic device utilizing microfluidic principles, a smartphone camera, and established laboratory colorimetric methods for accurate glucose estimation. Our proposed microfluidic device comprises layers of adhesive poly-vinyl films stacked on a poly methyl methacrylate (PMMA) base sheet, with micro-channel contours precision-cut using a cutting printer. Employing the gold standard glucose-oxidase/peroxidase reaction on this microfluidic platform, we achieve enzymatic glucose determination. The resulting colored complex, formed by phenol and 4-aminoantipyrine in the presence of hydrogen peroxide generated during glucose oxidation, is captured at various glucose concentrations using a smartphone camera. Raw images are processed and utilized as input data for a 2-D convolutional neural network (CNN) deep learning classifier, demonstrating an impressive 95% overall accuracy against new images. The glucose predictions done by CNN are compared with ISO 15197:2013/2015 gold standard norms. Furthermore, the classifier exhibits outstanding precision, recall, and F1 score of 94%, 93%, and 93%, respectively, as validated through our study, showcasing its exceptional predictive capability. Next, a user-friendly smartphone application named "GLUCOLENS AI" was developed to capture images, perform image processing, and communicate with cloud server containing the CNN classifier. The developed CNN model can be successfully used as a pre-trained model for future glucose concentration predictions. [ABSTRACT FROM AUTHOR]

Simple Summary: Interactions between breast cancer (BCa) cells and carcinoma-associated fibroblasts (CAFs) in the tumor microenvironment (TME) are critical for cancer development and progression and can mitigate responses to therapies. Historically, it has been difficult to replicate in vivo long-term interactions between BCa cells and CAFs for in vitro studies and to define the precise roles of CAFs in BCa cell progression. Our novel microfluidic-capable culture devices called TAME (tissue architecture and microenvironment engineering) devices enable us to study cell-cell interactions of human breast cancer (BCa) cells and human CAFs through their secretome in 3D cultures for extended periods, up to and beyond 70 days. Using these systems, we observed that CAFs enhance BCa cell progression to an invasive phenotype. Moreover, secretome-mediated reciprocal interactions of BCa cells and CAFs promote their migration toward each other, suggesting that targeting signaling pathways to mediate interactions between BCa cells and CAFs could be a potential therapeutic approach for the prevention of BCa progression. Background/Objectives: Carcinoma-associated fibroblasts (CAFs), a prominent cell type in the tumor microenvironment (TME), significantly contributes to cancer progression through interactions with cancer cells and other TME components. Consequently, targeting signaling pathways driven by CAFs has potential to yield new therapeutic approaches to inhibit cancer progression. However, the mechanisms underlying their long-term interactions with cancer cells in vivo remains poorly understood. Methods: To address this, we developed a three-dimensional (3D) parallel coculture model of human triple-negative breast cancer (TNBC) cells and CAFs using our innovative TAME devices. This model allowed for the analysis of TNBC paracrine interactions via their secretome over extended culture periods (at least 70 days). Results: Using TNBC cell lines (MDA-MB-231, MCF10.DCIS, and HCC70), we found that TNBC spheroids in 3D parallel cocultures with CAFs exhibited more pronounced invasive finger-like outgrowths than those in cocultures of TNBC cells and normal fibroblasts (NFs) over a period of 50–70 days. We also established that the CAF-derived secretome affects TNBC migration towards the CAF secretome region. Additionally, we observed a preferential migration of CAFs, but not NFs, toward TNBC spheroids. Conclusions: Overall, our results suggest that paracrine interactions between TNBC cells and CAFs enhance TNBC invasive phenotypes and promote reciprocal migration. [ABSTRACT FROM AUTHOR]

Polydimethylsiloxane (PDMS) is the most studied organosilicon polymer, finding broad applications in soft lithography of microfluidic devices, wearable electronics insulation, medical tubing, shielding coatings, and sealant elements. However, their cross‐linked forms swell severely in nonpolar oils due to the comparable solubility parameters. Here, an oil‐stable cross‐linked PDMS discovered by grafting flexible PDMS chains onto its polymeric networks is reported, which enables surface‐exposed chains with high dynamics. It is found that the synergetic effect of mobile nanopore filling and quasi‐liquid lubrication of PDMS brushes endows the cross‐linked matrix with a reduced oil absorption ratio of up to 91.7% without altering its surface composition. Moreover, stretching polymeric networks can be re‐arranged with an adequate grafting of PDMS chains, further enhancing the antipenetration performance. This discovery of the oil‐stable surface configuration of cross‐linked PDMS breaks the constraint of siloxane elastomers not being applicable in nonpolar soluble liquid, opening a new era of research in organosilicon chemistry. [ABSTRACT FROM AUTHOR]

Electrode fabrication in microfluidic devices is often avoided due to the smaller footprint of the platform, associated complexity in the process, and absence of a well-established method. In this article a glass microchip with chemically modified and vapor deposited gold internal electrode was fabricated and used for biomolecule immobilization and electrochemical applications. The silanol (-OH) on glass surface of the microdevice was modified with (3-mercaptopropyl)trimethoxysilane prior to gold deposition to achieve a strong attachment of the metal film using the gold-thiol binding. The practical utility of the microdevice with gold surface was demonstrated by performing quantitative enzyme linked immunosorbent assay (ELISA) of human interferon gamma (IFN-γ) antigen, yielding a limit of detection (LOD) of 0.15 pg/mL compared to 4 pg/mL in the microplate assay. Additionally, the functionality of the internal electrode was established by performing cyclic voltammetry (CV) and linear sweep voltammetry (LSV) of potassium ferricyanide (III) in potassium chloride (KCl) solution using the proposed gold patterned electrode in the microchip, offering limits of detection of 0.14 and 0.11 mM for CV and LSV respectively. Both the ELISA and electrochemical assays were validated by generating calibration curves and using a control sample for the electroanalytical sensing applications. The measurements yielded experimental results that agreed with control concentration with 7% relative standard deviation (RSD). The promising reproducibility of the electrode in fabrication with no/minimum failure rate and its outstanding performance in immunosensor applications as well as an electron exchange surface makes our technique of gold electrode fabrication in microchips a state of the art. [ABSTRACT FROM AUTHOR]

Emerging diagnostic scenarios, such as population surveillance by pooled testing and on‐site rapid diagnosis, highlight the importance of advanced microfluidic systems for in vitro diagnostics. However, the widespread adoption of microfluidic technology faces challenges due to the lack of standardized design paradigms, posing difficulties in managing macro‐micro fluidic interfaces, reagent storage, and complex macrofluidic operations. This paper introduces a novel modular‐based mesoscopic design paradigm, featuring a core "needle‐plug/piston" structure with versatile variants for complex fluidic operations. These structures can be easily coupled with various microfluidic platforms to achieve truly self‐contained microsystems. Incorporated into a "3D extensible" design architecture, the mesoscopic design meets the demands of function integration, macrofluid manipulations, and flexible throughputs for point‐of‐care nucleic acid testing. Using this approach, an ultra‐sensitive nucleic acid detection system is developed with a limit of detection of ten copies of SARS‐CoV‐2 per mL. This system efficiently conducts large‐scale pooled testing from 50 pharyngeal swabs in a tube with an uncompromised sensitivity, enabling a truly "sample‐in‐answer‐out" microsystem with exceptional performance. [ABSTRACT FROM AUTHOR]

The importance of microfluidics research is growing, especially in the fields of chemistry, biology or medicine. This is coupled with a growing demand for specialized capillary equipment that allows advanced research at the microscale. Conventional methods of manufacturing such devices are expensive, time-consuming and do not guarantee good results. An alternative to these methods is the use of 3D printing technology. Despite the existence of numerous works presenting the possibilities of 3D printing in the context of creating microfluidic devices, there is a lack of comprehensive works presenting qualitative analysis of printed objects. This paper presents a method of producing microcapillary structures for microfluidics research with the help of 3D printing using stereolithography technology. The quality requirements that the printed objects should meet are defined and all stages of production are characterized. A qualitative analysis of the obtained objects was carried out, taking into account both the influence of individual printing parameters and print processing methods. The results of microfluidic tests using printed objects are also presented. This work is aimed at providing specific knowledge that allows the manufacture of precision devices for microfluidics purposes at low cost. [ABSTRACT FROM AUTHOR]

This review delves into the pivotal role of circulating tumor cells (CTCs) in colorectal cancer (CRC) metastasis, focusing on their biological properties, interactions with the immune system, advanced detection techniques, and clinical implications. We explored how metastasis-competent CTCs evade immune surveillance and proliferate, utilizing cutting-edge detection and isolation technologies, such as microfluidic devices and immunological assays, to enhance sensitivity and specificity. The review highlights the significant impact of CTC interactions with immune cells on tumor progression and patient outcomes. It discusses the application of these findings in clinical settings, including non-invasive liquid biopsies for early diagnosis, prognosis, and treatment monitoring. Despite advancements, challenges remain, such as the need for standardized methods to consistently capture and analyze CTCs. Addressing these challenges through further molecular and cellular research on CTCs could lead to improved interventions and outcomes for CRC patients, underscoring the importance of unraveling the complex dynamics of CTCs in cancer progression. [ABSTRACT FROM AUTHOR]

The "Lab on a Chip" journal, published by The Royal Society of Chemistry, features various papers on micro- and nanoscale devices and applications. The current issue includes topics such as surface modification of paper-based microfluidic devices, reversible bonding in thermoplastic elastomer microfluidic platforms, and multiplexed ultrasensitive detection of cancer biomarkers. Other papers cover areas like neuroplasticity investigations, miRNA detection in skin interstitial fluid, and artificial intelligence performance in testing microfluidics for point-of-care applications. The journal also explores topics such as magnetophoresis in millifluidic systems, dynamic sampling from ex vivo adipose tissue, and inertial co-focusing of heterogeneous particles in hybrid microfluidic channels. [Extracted from the article]

Pathologies in adipose (fat) tissue function are linked with human diseases such as diabetes, obesity, metabolic syndrome, and cancer. Dynamic, rapid release of metabolites has been observed in adipocyte cells and tissue, yet higher temporal resolution is needed to adequately study this process. In this work, a microfluidic device with precise and regular valve-automated droplet sampling, termed a microfluidic analog-to-digital converter (μADC), was used to sample secretions from ∼0.75 mm diameter adipose explants from mice, and on-chip salt water electrodes were used to merge sampled droplets with reagent droplets from two different fluorometric coupled enzyme assays. By integrating sampling and assays on-chip, either glycerol or non-esterified fatty acids (NEFA), or both, were quantified optically within merged 12 nanoliter droplets using a fluorescence microscope with as high as 20 second temporal resolution. Limits of detection were 6 μM for glycerol (70 fmol) and 0.9 μM for NEFA (10 fmol). Multiple ex vivo adipose tissue explants were analyzed with this system, all showing clear increases in lipolytic function after switching from feeding to fasting conditions. Enabled by high temporal resolution, lipolytic oscillations of both glycerol and NEFA were observed for the first time in the range of 0.2 to 1.6 min−1. Continuous wavelet transform (CWT) spectrograms and burst analyses (0.1 to 4.0 pmol bursts) revealed complex dynamics, with multiplexed assays (duplex for glycerol and NEFA) from the same explants showing mostly discordant bursts. These data support separate mechanisms of NEFA and glycerol release, although the connection to intracellular metabolic oscillations remains unknown. Overall, this device allowed automated and highly precise temporal sampling of tissue explants at high resolution and programmable downstream merging with multiple assay reagents, revealing unique biological information. Such device features should be applicable to various other tissue or spheroid types and to other assay formats. [ABSTRACT FROM AUTHOR]

We report on the use of a microfluidic antisolvent crystallization method to investigate the effect of solution volume on the chiral symmetry breaking (CSB) phenomena of enantiomeric sodium chlorate crystals. The utilization of a microfluidic device is capable of periodically producing emulsion droplets of uniform size and facilitates the quantitative analysis and visualization of crystallization phenomena occurring within the individual emulsions immersed in an oil continuous medium (i.e., dodecane). To promote nucleation and crystallization, a small amount of an antisolvent (i.e., ethanol) is introduced into the continuous phase. We observe that 100% CSB occurs within a certain critical emulsion volume. Beyond this critical volume, the probability of forming two different enantiomeric crystal particles increases. This solution volume-dependent CSB phenomenon can be attributed to the rapid depletion of surrounding molecules by spontaneous crystal growth after the formation of the initial nucleus within the critical volume, thereby suppressing further primary nucleation. [ABSTRACT FROM AUTHOR]

Transplantable ready-made microvessels have therapeutic potential for tissue regeneration and cell replacement therapy. Inspired by the natural rapid angiogenic sprouting of microvessels in vivo, engineered injectable 3D microvessel networks are created using thermoplastic elastomer (TPE) microfluidic devices. The TPE material used here is flexible, optically transparent, and can be robustly yet reversibly bonded to a variety of plastic substrates, making it a versatile choice for microfluidic device fabrication because it overcomes the weak self-adhesion properties and limited manufacturing options of poly(dimethylsiloxane) (PDMS). By leveraging the reversible bonding characteristics of TPE material templates, we present their utility as an organ-on-a-chip platform for forming and handling microvessel networks, and demonstrate their potential for animal-free tissue generation and transplantation in clinical applications. We first show that TPE-based devices have nearly 6-fold higher bonding strength during the cell culture step compared to PDMS-based devices while simultaneously maintaining a full reversible bond to (PS) culture plates, which are widely used for biological cell studies. We also demonstrate the successful generation of perfusable and interconnected 3D microvessel networks using TPE–PS microfluidic devices on both single and multi-vessel loading platforms. Importantly, after removing the TPE slab, microvessel networks remain intact on the PS substrate without any structural damage and can be effectively harvested following gel digestion. The TPE-based organ-on-a-chip platform offers substantial advantages by facilitating the harvesting procedure and maintaining the integrity of microfluidic-engineered microvessels for transplant. To the best of our knowledge, our TPE-based reversible bonding approach marks the first confirmation of successful retrieval of organ-specific vessel segments from the reversibly-bonded TPE microfluidic platform. We anticipate that the method will find applications in organ-on-a-chip and microphysiological system research, particularly in tissue analysis and vessel engraftment, where flexible and reversible bonding can be utilized. [ABSTRACT FROM AUTHOR]

Paper-based microfluidic devices offer an ideal platform for biological and environmental detection because they are low-cost, small, disposable, and fill by natural capillary action. In this tutorial review, we discuss the surface modification of paper-based microfluidic devices with functional polymers using the initiated chemical vapor deposition (iCVD) process. The iCVD process is solventless and therefore ideal for coating cellulose paper because there are no surface tension effects or solvent compatibility issues. The process can also be scaled up for roll-to-roll manufacturing. The chemical functionality of the iCVD coating can be tuned by varying the monomer and the structure of the coating can be tuned by varying the processing parameters. [ABSTRACT FROM AUTHOR]

Digital nucleic acid quantification has emerged as an exceptionally valuable technique in molecular diagnosis, capturing significant attention in the biomedical field. This study introduces a novel multi‐volume digital loop‐mediated isothermal amplification (dLAMP) chip that offers an extensive detection range and user‐friendly operation. The chip consists of 1024 large chambers with the volume of 7.2 nL and 6400 small chambers with the volume of 0.064 nL. The ingenious combination of chambers with volume difference beyond a 100‐fold on the single chip enables a broad dynamic range exceeding 105 for nucleic acid quantification. Furthermore, this chip incorporates an automated strategy, enabling the rapid loading and partitioning of the reaction mixture into subvolumes within 4 min. In order to evaluate the quantitative capability of the multi‐volume dLAMP chip, a 10‐fold serial dilution of the β‐actin DNA template is measured. Additionally, the chip is tested for β‐actin DNA concentration in plasma samples from colorectal cancer patients and healthy individuals. By providing enhanced automation, wide dynamic range, and improved accuracy, this chip paves the way for broader applications and the advancement of biomedical research fields. [ABSTRACT FROM AUTHOR]

Unidirectional liquid transport, a special self‐propelled wetting phenomenon observed in natural structures like the Nepenthes pitcher plant peristome, Araucaria leaves, and Crassula muscosa shoots, has inspired scientists to develop various novel microfluidic devices for liquid collection, physical/chemical reactions, and irrigation. Recently, the concept of capillary transistors is proposed to enable a programmable transport area and a significant increase in unidirectional capillary height, which is expected to greatly expand the applications of conventional microfluidic chips. In this work, using black resin as the three‐dimensional (3D) printing material, we construct microfluidic chips with capillary transistors for photothermal‐responsive unidirectional liquid transport. The transistors consist of asymmetric overhanging structures with connected overhangs, allowing for fast, long‐distance, and large‐area unidirectional liquid transport, with a unidirectional capillary height exceeding 90.0 mm—more than double the values reported in most previous studies. Furthermore, smart control of capillary height is achieved by applying external photothermal stimuli to the capillary transistors, demonstrating their potential applications in photothermal‐responsive unidirectional liquid transport and mixing in 3D space. It is envisioned that additional functions such as liquid patterning, desalination, and biochemical microreactions would be developed by engineering capillary transistors and their responsiveness. [ABSTRACT FROM AUTHOR]

An incompressible boundary‐layer third‐grade nanomaterial stagnated flow confined by stretchy regime is formulated. The laminar magnetized thermal and solutal flow features the aspects of Brownian diffusion, generalized thermal‐flux, thermophoresis, and generalized solutal‐flux. The dimensionalized third‐grade steady‐state model is obtained by implementing appropriate variables. The subsequent nonlinear mathematical model is analytically computed by utilizing homotopy algorithm. The graphics demonstration of nondimensional fields (i.e., velocity, concentration, and temperature) for distinct physical variables is exhibited. The outcomes for the skin‐friction coefficient are calculated and confirmed to be precise. The analysis reveals that augmented values of material parameters escalate nanomaterial velocity while the Hartman number exhibits lower velocity. The temperature field displays a decaying trend for Prandtl number and thermal relaxation variable while contrary effects for nanoscale parameters (i.e., thermophoresis diffusive and Brownian diffusive) are reported. Besides, the nanomaterial concentration shows a decreasing trend with respect to the Schmidt number, Brownian motion, and solutal‐relaxation variable while thermophoresis parameter exhibits opposite effects. Furthermore, this study could be valuable for the design of various lab‐on‐a‐chip and thermal microfluidic devices in the field of biomedicine. Besides contributing to the creation of new devices, they may also be advantageous for performing genetic testing and developing innovative tools. [ABSTRACT FROM AUTHOR]

Shear stress plays a crucial role in modulating cell adhesion and signaling. We present a microfluidic shear stress generator used to investigate the adhesion dynamics of Dictyostelium discoideum, an amoeba cell model organism with well-characterized adhesion properties. We applied shear stress and tracked cell adhesion, motility, and detachment using time-lapse videomicroscopy. In the precise shear conditions generated on-chip, our results show cell migration patterns are influenced by shear stress, with cells displaying an adaptive response to shear forces as they alter their adhesion and motility behavior. Additionally, we observed that DH1-10 wild-type D. discoideum cells exhibit stronger adhesion and resistance to shear-induced detachment compared to phg2 adhesion-defective mutant cells. We also highlight the influence of cell density on detachment kinetics. [ABSTRACT FROM AUTHOR]

Human norovirus (HuNoV) is recognized as the leading causative agent of foodborne outbreaks of epidemic gastroenteritis. Consequently, there is a high demand for developing point-of-care testing for HuNoV. We developed an origami microfluidic device that facilitates rapid detection of murine norovirus 1 (MNV-1), a surrogate for HuNoV, encompassing the entire process from sample preparation to result visualization. This process includes RNA absorption via a paper strip, RNA amplification using recombinase polymerase amplification (RPA), and a lateral flow assay for signal readout. The on-chip detection of MNV-1 was completed within 35 min, demonstrating 100% specificity to MNV-1 in our settings. The detection limit of this microfluidic device for MNV-1 was 200 PFU/mL, comparable to the in-tube RPA reaction. It also successfully detected MNV-1 in lettuce and raspberries at concentrations of 170 PFU/g and 230 PFU/g, respectively, without requiring extra concentration steps. This device demonstrates high compatibility with isothermal nucleic acid amplification and holds significant potential for detecting foodborne viruses in agri-food products in remote and resourcelimited settings. IMPORTANCE HuNoV belongs to the family of Caliciviridae and is a leading cause of acute gastroenteritis that can be transmitted through contaminated foods. HuNoV causes around one out of five cases of acute gastroenteritis that lead to diarrhea and vomiting, placing a substantial burden on the healthcare system worldwide. HuNoV outbreaks can occur when food is contaminated at the source (e.g., wild mussels exposed to polluted water), on farms (e.g., during crop cultivation, harvesting, or livestock handling), during packaging, or at catered events. The research outcomes of this study expand the approaches of HuNoV testing, adding value to the framework for routine testing of food products. This microfluidic device can facilitate the monitoring of HuNoV outbreaks, reduce the economic loss of the agri-food industry, and enhance food safety. [ABSTRACT FROM AUTHOR]

Technologies for rapid and high-throughput separation of rare cells from large populations of other types of cells have recently attracted much attention in the field of bioengineering. Among the various cell separation technologies proposed in the past, dielectrophoresis has shown particular promise because of its preciseness of manipulation and noninvasiveness to cells. However, one drawback of dielectrophoresis devices is that their application of high voltage generates Joule heat that exposes the cells within the device to high temperatures. To further explore this problem, this study investigated the temperature field in a previously developed cell separation device in detail. The temperature rise at the bottom of the microfluidic channel in the device was measured using a micro-LIF method. Moreover, the thermofluidic behavior of the cell separation device was numerically investigated by adopting a heat generation model that takes the electric-field-dependent heat generation term into account in the energy equation. Under the operating conditions of the previously developed cell separation device, the experimentally obtained temperature rise in the device was approximately 20 °C, and the numerical simulation results generally agreed well. Next, parametric calculations were performed with changes in the flow rate of the cell sample solution and the solution conductivity, and a temperature increase of more than 40 °C was predicted. The results demonstrated that an increase in temperature within the cell separation device may have a significant impact on the physiological functions of the cells, depending on the operating conditions of the device. [ABSTRACT FROM AUTHOR]

STUDY QUESTION Can a functional in vitro model, containing the main cellular components of the uterine wall, be generated from cells derived from patient tissues? SUMMARY ANSWER We present a three-dimensional (3D) physiologically relevant, organ-on-a-chip model of the uterine wall containing primary endometrial and myometrial cellular participants, generated from human uterine tissue. WHAT IS KNOWN ALREADY As a highly dynamic reproductive organ, the human uterus plays fundamental physiological roles in menstruation and childbirth. The endometrial–myometrial junction (EMJ) defines the interface between the inner mucosal layer (endometrium) and outer smooth muscle zone (myometrium) that comprises the uterine wall. The EMJ is implicit in several uterine pathologies of unknown aetiology, including adenomyosis and abnormally invasive placenta; however, despite this, no patient-derived in vitro models of the uterine wall containing all EMJ participants currently exist. STUDY DESIGN, SIZE, DURATION We employed microfluidic technology to characterize multiple miniaturized models of the uterine wall. Protocols were tested that included variations in the seeding order of endometrial and myometrial fractions, and the addition of a low viscosity extracellular matrix to influence cell behaviour. Ultimately, functional hormone responses of patient-derived uterine wall models were assessed. PARTICIPANTS/MATERIALS, SETTING, METHODS Endometrial (n = 9) and myometrial biopsies (n = 4) were enzymatically dissociated to create epithelial, stromal and myometrial cellular fractions. Cell suspensions were seeded into non-adhesive poly(dimethylsiloxane) microfluidic devices containing 5 × 5 microwell arrays. The fate of individual cell types was monitored in real-time using fluorescent tracers, and cell phenotype was characterized by immunocytochemistry. Model functionality was assessed by measuring Ca2+ responses to agonist stimulation, and both insulin-like growth factor binding protein 1 (IGFBP-1) and osteopontin secretion in response to hormone stimulation. MAIN RESULTS AND THE ROLE OF CHANCE When subjected to microfluidic culture in isolation, endometrial stromal cells and smooth muscle myocytes formed compact spheroids, whilst epithelial cells produced diffuse aggregates. Tri-cultures were established by sequential seeding of individual or combined cell fractions at various ratios. Regardless of the protocol, epithelial cells localized to the outer periphery of tri-culture spheroids, which varied in morphology across the protocols. Incorporation of 5% [v/v] Matrigel® improved the reproducibility of 3D aggregates which exhibited robust self-assembly of a stromal/smooth muscle core encased in epithelium. Exposure of tri-cultures to oestradiol, medroxyprogesterone acetate and cyclic adenosine monophosphate (cAMP) increased secretion of IGFBP-1, which indicates stromal decidualization, and enhanced epithelial cell osteopontin secretion. Stimulation with endothelin-1 induced Ca2+ signalling in myocytes. LIMITATIONS, REASONS FOR CAUTION Endometrial and myometrial tissue was collected from relatively few donors. Myometrial tissue was collected from pregnant donors, which may have influenced the myocyte phenotype. Furthermore, endometrial tissue sampling was from women not having a hysterectomy, thus may not include the deeper basalis region, which may limit the physiological mimicry of the final models. WIDER IMPLICATIONS OF THE FINDINGS Our novel approach to modelling the uterine wall in 3D captures all of the main cell types in a medium-throughput system, enabling the screening of hundreds of cultures in parallel from a single biopsy. This system shows great promise for examining the cellular interplay between physiological cues and EMJ pathologies, such as the impact of uterine peristalsis and cyclical hormones on the pathogenesis of adenomyosis. STUDY FUNDING/COMPETING INTEREST(S) C.B. was supported by an Organ-on-a-Chip Technologies Network Pump Priming Project grant. C.J.H. was supported by a Wellbeing of Women project grant (RG2137), SRI/Bayer and Wellcome Trust IFFS3. D.K.H. was supported by a Wellbeing of Women project grant (RG2137) and MRC clinical research training fellowship (MR/V007238/1). M.Z. is Director and Co-Founder of ScreenIn3D Limited. The other authors declare no conflict of interest. TRIAL REGISTRATION NUMBER N/A. [ABSTRACT FROM AUTHOR]

Microfluidic devices have been the subject of considerable attention in recent years. The development of novel microfluidic devices, their evaluation, and their validation requires simulations. While common methods based on Computational Fluid Dynamics (CFD) can be time-consuming, 1D simulation provides an appealing alternative that leads to efficient results with reasonable quality. Current 1D simulation tools cover specific microfluidic applications; however, these tools are still rare and not widely adopted. There is a need for a more versatile and adaptable tool that covers novel applications, like mixing and the addition of membranes, and allows easy extension, resulting in one comprehensive 1D simulation tool for microfluidic devices. In this work, we present an open-source, modular, and extendable 1D simulation approach for microfluidic devices, which is available as an open-source software package at https://github.com/cda-tum/mmft-modular-1D-simulator. To this end, we propose an implementation that consists of a base module (providing the core functionality) that can be extended with dedicated application-specific modules (providing dedicated support for common microfluidic applications such as mixing, droplets, membranes, etc.). Case studies show that this indeed allows to efficiently simulate a broad spectrum of microfluidic applications in a quality that matches previous results or even fabricated devices. [ABSTRACT FROM AUTHOR]

The demand for rapid, high-quality, and controlled mixing at the microscale has led to the development of various types of micromixers. Micromixers are commonly categorised as active, or passive based on whether they utilise external energy to enhance mixing. Passive micromixers utilise a complex geometry to enhance the diffusion coefficient at lower Reynolds numbers and induce chaotic advection at higher Reynolds numbers for effectively mixing fluids without external energy. Active micromixers, on the other hand, achieve precise, fast, and controllable mixing by employing external energy sources such as pressure, electric, magnetic, or acoustic fields. Some active methods such as magnetic field-driven micromixers need fluids with specific properties. Others, such as acoustic field-driven micromixers apply to various types of fluids. Bubbles can be used as membranes or stirrers in microfluidic devices for both passive and active micromixers. They are easy to use, compatible with microfluidic systems, low cost, and effective. Improvements in manufacturing methods, notably, 3D printing have emerged as promising methods for the development of new micromixer designs. In this paper, a wide range of micromixer types is reviewed and the main mechanism for enhanced mixing is investigated. This study aims to guide researchers proposing innovative designs. Furthermore, it is shown that combining different methods can lead to the development of more effective micromixers, promising further advancements in microscale mixing technology. [ABSTRACT FROM AUTHOR]

The practice of conventional cell culture techniques affects the quality of biological research in mechanobiology by not replicating the cell's microenvironment adequately. Microfluidic devices resembling 3D in vivo cell culture offer viable alternatives for mechanobiology studies and other applications including cell screening, cell separation, and point-of-care diagnostics. This review highlights recent advances in additive manufacturing (AM) to fabricate microfluidic and millifluidic devices through fast design iterations and cost reduction compared with conventional device manufacturing. Key advances in AM technologies such as fused deposition modeling (FDM), stereolithography apparatus (SLA), and digital light processing (DLP) have allowed the direct manufacture of complex microfluidic devices with master molds and sacrificial structures using multiple material components. Currently, available 3D printed devices for the precise fabrication of milli- and microfluidic devices, essential to mimicking the cellular microenvironment, while economically reducing production costs by eliminating expensive clean-room environments and reducing material waste are highlighted. [ABSTRACT FROM AUTHOR]

The aim of this research is to validate a new prototyping method for microfluidic devices which could compete with the widely used polydimethylsiloxane soft lithography. This alternative methodology uses laser‐etched glass and pre‐cured silicone as main materials. A dual approach is chosen for validation, testing prototypes of a mixing device well known in the scientific literature. Specifically the focus is to evaluate the mixing efficiency (ME) of two fluids flowing through the device. After fine‐tuning the fabrication process, the ME is experimentally evaluated, and the results are compared with a 3D computational fluid dynamic analysis. [ABSTRACT FROM AUTHOR]

The present study explores the application of X-ray scattering, using synchrotron radiation, to assess the diffusive transport of nanomedicines in tumor on a chip devices fabricated by 3D stereolithography using a resin with high optical and X-ray transmittance. Unlike conventional methods that require fluorescent labeling of nanoparticles, potentially altering their in vitro and in vivo behavior, this approach enables the investigation of the transport properties for unlabeled nanoparticles. In particular, the results presented confirm the influence of the porosity of the extracellular matrix-like microenvironment, specifically Matrigel, on the diffusive transport of oligonucleotide-functionalized gold nanoparticles. The analysis of the scattering patterns allows to create 2D maps showing the nanoparticle distribution with high spatial resolution. The proposed approach demonstrates the potential for studying other factors involved in nanoparticle diffusion processes. By implementing X-ray scattering to track unmodified nanomedicines within extracellular matrix-like microenvironments, increasingly accurate models for evaluating and predicting therapeutics transport behaviors can be developed. [ABSTRACT FROM AUTHOR]

Nanocarriers are essential for targeted and enhanced drug delivery. However, the use of toxic solvent and the complex of chemical functionalization have seriously limited their clinical translation. Here, a one‐step strategy is developed to direct the self‐assembly of biocompatible core‐shell nanocapsules with tunable surface functionality by microfluidics. Upon rapid mixing and solvent exchange of ethanol with water in a microfluidic device, drug, oil, polymer, and polymer‐PEG‐function co‐precipitate and self‐assemble into core‐shell nanocapsules with desired surface functionality as driven by energy minimization. The prepared PCL‐PEG‐FA core‐shell nanocapsules demonstrate a high encapsulation efficiency and loading capacity and exhibit excellent tumor‐targeting drug delivery performances both in vitro and in vivo via the binding of functional group on the nanocapsule surface with corresponding receptor on the tumor cell membrane. The immune activation in the tumor microenvironment is investigated in detail by flow cytometry and multiplex immunohistochemistry staining to reveal the underlying mechanism for enhanced anti‐tumor performances. The developed strategy is green, facile, versatile, scalable and offers a promising platform for the design of advanced nanocarriers with hierarchical structure and desired function for enhanced drug delivery. [ABSTRACT FROM AUTHOR]

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the loss of motoneurons (MNs), and despite progress, there is no effective treatment. A large body of evidence shows that astrocytes expressing ALS-linked mutant proteins cause non-cell autonomous toxicity of MNs. Although MNs innervate muscle fibers and ALS is characterized by the early disruption of the neuromuscular junction (NMJ) and axon degeneration, there are controversies about whether muscle contributes to non-cell-autonomous toxicity to MNs. In this study, we generated primary skeletal myotubes from myoblasts derived from ALS mice expressing human mutant SOD1G93A (termed hereafter mutSOD1). Characterization revealed that mutSOD1 skeletal myotubes display intrinsic phenotypic and functional differences compared to control myotubes generated from non-transgenic (NTg) littermates. Next, we analyzed whether ALS myotubes exert non-cell-autonomous toxicity to MNs. We report that conditioned media from mutSOD1 myotubes (mutSOD1-MCM), but not from control myotubes (NTg-MCM), induced robust death of primary MNs in mixed spinal cord cultures and compartmentalized microfluidic chambers. Our study further revealed that applying mutSOD1-MCM to the MN axonal side in microfluidic devices rapidly reduces mitochondrial axonal transport while increasing Ca2 + transients and reactive oxygen species (i.e., H2O2). These results indicate that soluble factor(s) released by mutSOD1 myotubes cause MN axonopathy that leads to lethal pathogenic changes. [ABSTRACT FROM AUTHOR]

Currently, despite the vast amounts of time and money invested in cancer treatment, cancer remains one of the primary threats to human life. The primary factor contributing to the low treatment efficacy is cancer heterogeneity. The unclear molecular mechanisms underlying tumorigenesis, coupled with the complexity of human physiology, and the inability of animal models to accurately replicate the human tumor microenvironment, pose significant hurdles in the development of novel cancer therapies. Tumor-microenvironment-on-chip (TMOC) represents a research platform that integrates three-dimensional cell culture with microfluidic systems, simulating the essential components and physiological traits of the in vivo tumor microenvironment. It offers a dynamic setting within the chip system to study tumor progression, potentially heralding a breakthrough in cancer research. In this review, we will summarize the current advancements in this platform, encompassing various types of TMOCs and their applications in different types of cancer. From our perspective, the TMOC platform necessitates enhanced integration with tissue engineering techniques and microphysiological environments before it can evolve into a more refined preclinical model for cancer research. [ABSTRACT FROM AUTHOR]

AbstractRestoring cartilage to healthy state is challenging due to low cell density and hence low regenerative capacity. The current platforms are not compatible with clinical translation and require dedicated handling of trained personnel. However, by engineering and implanting cell microaggregates in higher concentrations, efficient formation of new cartilage can be achieved, even in the absence of exogenous growth factors. Therefore, one-step surgeries are preferable for novel treatments and we need cell laden microgels allowing the formation of microaggregaets in vivo. Injectability is a key parameter for in situ forming the shape and minimally invasive clinical applications. Hydrogels as bioinks can restore damaged tissues to their primary shape. Chitosan is a polysaccharide derived from chitin with abundant usage in tissue engineering. This review highlights the use of chitosan as an injectable hydrogel for osteochondral defects. Several studies focused on encapsulating mesenchymal stem cells within chitosan hydrogels have been categorized and incorporating microfluidic devices has been identified in the forefront to form microgels. Additionally, the printability is another convenience of chitosan for using in 3D printing for cartilage tissue engineering which is described in this review. [ABSTRACT FROM AUTHOR]

Sperm capture techniques that immobilize sperm to halt their motility are essential for the long-term analysis of individual sperm. These techniques are beneficial in assisted reproductive technologies such as intracytoplasmic sperm injection (ICSI) by allowing selective screening of sperm. However, there is a notable lack of high-throughput and non-destructive sperm capture methods that allow the flagellum to beat freely, which is crucial for accurately reflecting the behavior of unfettered, freely swimming sperm. To bridge this gap, we introduce a novel microfluidic device specifically engineered to capture sperm without restricting flagellar motion. The design utilizes sperm's innate boundary-following behavior in both 3D and 2D environments to direct them into a capture zone. Once captured, the sperm head is restrained while the flagellum remains free to exhibit natural beating patterns. Utilizing this device, we explore the effects of hyperactivating agents, temperature, and their combined influence on the dynamics of bovine sperm flagella. The unrestricted flagellar motion offered by our device yields two prominent advantages: it mirrors the flagellar behavior of free-swimming sperm, ensuring research findings are consistent with natural sperm activity, and it prevents imaging overlap between the flagellum and the capture structures, simplifying the automation of flagellar tracking and analysis. This technological advancement facilitates the collection of waveform parameters along the entire flagellum, addressing inconsistencies that have arisen in previous research due to differing measurement sites, and enabling precise extraction of sperm behavioral properties. [ABSTRACT FROM AUTHOR]

Fluorescence polarization (FP) assays are widely used to quantify biomolecules, and their combination with microfluidic devices has the potential for application in onsite analysis. However, the hydrophobic surface of polydimethylsiloxane (PDMS)-based microfluidic devices and the amphiphilicity of the blocking agents can cause the nonspecific adsorption of biomolecules, which in turn reduces the sensitivity of the FP assay. To address this, we demonstrated an FP assay with improved sensitivity in microfluidic devices using a polyethylene glycol-based surface modification to avoid the use of blocking agents. We evaluated the effectiveness of the modification in inhibiting nonspecific protein adsorption and demonstrated the improved sensitivity of the FP immunoassay (FPIA). Our study addressed the lack of sensitivity of FP assays in microfluidic devices, particularly for the quantification of low-abundance analytes. [ABSTRACT FROM AUTHOR]