Search Results (42)

Breast cancer remains the most common malignancy in women, yet, many patients fail to achieve full remission despite significant advancements. This is largely due to tumour heterogeneity and the limitations of current experimental models in accurately replicating the complexity of in vivo tumour environment. In this study, we present a compartmentalised alginate hydrogel platform as an innovative in vitro tool for three-dimensional breast cancer cell culture. To mimic the heterogeneity of tumour tissues, we developed a core–shell structure (3.5% alginate core and 2% alginate shell) that mimic the stiffer, denser internal tumour matrix. The human triple-negative breast cancer cell line (MDA-MB-231) was embedded in core–shell alginate gels to assess viability, proliferation and hypoxic activity. Over one week, good cells proliferation and viability was observed, especially in the softer shell. Interestingly, cells within the stiffer core were more positive to hypoxic marker expression (HIF-1α) than those embedded in the shell, confirming the presence of a hypoxic niche, as observed in vivo. When cultured in the MIVO^® milli fluidic organ-on-chip resembling the physiological fluid flow conditions, cancer cells viability became comparable between core and shell hydrogel area, emphasising the importance of the fluid flow in nutrients diffusion within three-dimensional matrixes. Cisplatin chemotherapy treatment further highlighted these differences: under static conditions, cancer cell death was prominent in the softer shell, whereas cells in the stiffer core remained resistant to cisplatin. Conversely, drug diffusion was more homogeneous in the core–shell structured treated in the organ-on-chip, leading to a uniform reduction in cell viability. These findings suggest that integrating a compartmentalised core–shell cell laden alginate model with the millifluidic organ on chip offers a more physiologically relevant experimental approach to deepening cancer cell behaviour and drug response. Full article

Silver nanowires (AgNWs) have garnered significant attention in nanotechnology due to their unique mechanical and electrical properties and versatile applications. This review explores the synthesis of AgNWs, with a specific focus on the utilization of millifluidic flow reactors (MFRs) as a promising platform for controlled and efficient production. It begins by elucidating the exceptional characteristics and relevance of AgNWs in various technological domains and then delves into the principles and advantages of MFRs by showcasing their pivotal role in enhancing the precision and scalability of nanowire synthesis. Within this review, an overview of the diverse synthetic methods employed for AgNW production using MFRs is provided. Special attention is given to the intricate parameters and factors influencing synthesis and how MFRs offer superior control over these critical variables. Recent advances in this field are highlighted, revealing innovative strategies and promising developments that have emerged. As with any burgeoning field, challenges are expected, so future directions are explored, offering insights into the current limitations and opportunities for further exploration. In conclusion, this review consolidates the state-of-the-art knowledge in AgNW synthesis and emphasizes the critical role of MFRs in shaping the future of nanomaterial production and nanomanufacturing. Full article

Background: Hypertension is a major risk factor for cerebrovascular diseases due to its damaging effects on the blood–brain barrier (BBB) and associated pathologies. Oxidative stress-induced endothelial damage plays a critical role in BBB disruption, potentially leading to cognitive impairment and neurodegeneration. In this study, we investigated the protective effects of two essential trace elements, zinc (Zn) and selenium (Se), against oxidative stress in human brain endothelial cells (HBCE5i) exposed to hypertensive shear stress. Using an innovative millifluidic system (LiveBox2), which allows for the precise simulation of continuous flow conditions, we replicated the hemodynamic forces associated with hypertension. Methods: Cells were treated with ZnCl₂ (5–50 µM) or Na₂SeO₃ (50–500 nM) at concentrations selected based on previous studies and confirmed by cytotoxicity assays. Results: Our results demonstrated that shear stress significantly altered the localization of the tight junction protein zonula occludens-1 (ZO-1) and induced the nuclear translocation of the transcription factor NRF2, a hallmark of oxidative stress. Importantly, treatment with 10 µM ZnCl₂ preserved ZO-1 membrane localization and prevented NRF2 translocation, as confirmed by quantitative image analysis. In contrast, Na₂SeO₃ did not provide comparable protection, although modest improvements in ZO-1 localization were observed in some replicates. Discussion: We discuss potential reasons for selenium’s limited efficacy, including differences in bioavailability and cellular uptake. Our findings underscore zinc’s promising role as a neurovascular protector and suggest that further investigation into more complex in vitro models and in vivo studies is warranted. Full article

Drug-coated balloons (DCBs) are designed to deliver an anti-proliferative drug to the stenotic vessel to combat restenosis after an angioplasty treatment. However, significant drug loss can occur during device navigation toward the lesion site, thus reducing the delivery efficiency and increasing the off-target drug loss. In this framework, this study aimed to design a novel in vitro setup to estimate the drug loss due to blood flow–coating interaction during tracking. The system consists of a millifluidic chamber, able to host small drug-coated flat patches representative of DCBs, connected at the inlet to a syringe pump able to provide an ad hoc flow and, at the outlet, to a vial collecting the testing fluid with possible drug removed from the specimen. Unlike other studies, the device presented here uniquely evaluates flow-related drug loss from smaller-scale DCB samples, making it a precise, easy-to-use, and efficient assessment tool. In order to define proper boundary conditions for these washing off tests, computational fluid dynamics (CFD) models of a DCB in an idealized vessel were developed to estimate the wall shear stresses (WSSs) experienced in vivo by the device when inserted into leg arteries. From these simulations, different target WSSs were identified as of interest to be replicated in the in vitro setup. A combined analytical–CFD approach was followed to design the testing system and set the flow rates to be imposed to generate the desired WSSs. Finally, a proof-of-concept study was performed by testing eight coated flat specimens and analyzing drug content via high-performance liquid chromatography (HPLC). Results indicated different amounts of drug loss according to the different imposed WSSs and confirmed the suitability of the designed system to assess the washing off resistance of different drug coatings for angioplasty balloons. Full article

On-chip microfluidics are advanced in vitro models that simulate lung tissue’s native 3D environment more closely than static 2D models to investigate the complex lung architecture and multifactorial processes that lead to pulmonary disease. Current microfluidic systems can be restrictive in the quantities of biological sample that can be retrieved from a single micro-channel, such as RNA, protein, and supernatant. Here, we describe a newly developed large-scale airway-on-chip model that employs a surface area for a cell culture wider than that in currently available systems. This enables the collection of samples comparable in volume to traditional cell culture systems, making the device applicable to any workflow utilizing these static systems (RNA isolation, ELISA, etc.). With our construction method, this larger culture area allows for easier handling, the potential for a wide range of exposures, as well as the collection of low-quantity samples (e.g., volatiles or mitochondrial RNA). The model consists of two large polydimethylsiloxane (PDMS) cell culture chambers under an independent flow of medium or air, separated by a semi-permeable polyethylene (PET) cell culture membrane (23 μm thick, 0.4 μm pore size). Each chamber carries a 5 × 18 mm, 90 mm² (92 mm² with tapered chamber inlets) surface area that can contain up to 1–2 × 10⁴ adherent structural lung cells and can be utilized for close contact co-culture studies of different lung cell types, including airway epithelial cells, fibroblasts, smooth muscle cells, and endothelial cells. The parallel bi-chambered design of the chip allows for epithelial cells to be cultured at the air–liquid interface (ALI) and differentiation into a dense, multi-layered, pseudostratified epithelium under biological flow rates. This millifluidic airway-on-chip advances the field by providing a readily reproducible, easily adjustable, and cost-effective large-scale fluidic 3D airway cell culture platform. Full article

The creation of highly specific particle systems in the nano- and micrometer size range is a challenging task. The demand for particle systems with narrowly distributed properties is increasing in many applications, especially for use in high-tech products. Conventional separation techniques often reach their limits in the micrometer size range or become (labor-)intensive, which makes them economically or ecologically unsustainable. In addition, sorting based on several properties is rarely feasible in just one separator. Dielectrophoretic processes can be a viable option for complex sorting tasks like this, given their ability to address several particle properties and their high degree of selectivity. In this paper, we summarize the progress of a project in which the capability of dielectrophoretic methods for multidimensional sorting of microparticles was investigated. We were able to develop an operation mode for multidimensional sorting of microparticles using dielectrophoresis as well as a scalable electrically switchable filter. This creates a basis for high-throughput and multi-target sorting of technical microparticles using dielectrophoretic processes. Full article

Microfluidic devices (µFDs) have been explored extensively in drug screening and studying cellular processes such as migration and metastasis. However, the fabrication and implementation of microfluidic devices pose cost and logistical challenges that limit wider-spread adoption. Despite these challenges, light-based 3D printing offers a potential alternative to device fabrication. This study reports on the development of millifluidic devices (MiFDs) for disease modeling and elucidates the methods and implications of the design, production, and testing of 3D-printed MiFDs. It further details how such millifluidic devices can be cost-efficiently and effortlessly produced. The MiFD was developed through an iterative process with analytical tests (flow tests, leak tests, cytotoxicity assays, and microscopic analyses), driving design evolution and determination of the suitability of the devices for disease modeling and cancer research. The design evolution also considered flow within tissues and replicates interstitial flow between the main flow path and the modules designed to house and support organ-mimicking cancer cell spheroids. Although the primary stereolithographic (SLA) resin used in this study showed cytotoxic potential despite its biocompatibility certifications, the MiFDs possessed essential attributes for cell culturing. In summary, SLA 3D printing enables the production of MiFDs as a cost-effective, rapid prototyping alternative to standard µFD fabrication for investigating disease-related processes. Full article

The detection of magnetic nanoparticles in a liquid medium and the quantification of their concentration have the potential to improve the efficiency of several relevant applications in different fields, including medicine, environmental remediation, and mechanical engineering. To this end, sensors based on the magneto-impedance effect have attracted much attention due to their high sensitivity to the stray magnetic field generated by magnetic nanoparticles, their simple fabrication process, and their relatively low cost. To improve the sensitivity of these sensors, a multidisciplinary approach is required to study a wide range of soft magnetic materials as sensing elements and to customize the magnetic properties of nanoparticles. The combination of magneto-impedance sensors with ad hoc microfluidic systems favors the design of integrated portable devices with high specificity towards magnetic ferrofluids, allowing the use of very small sample volumes and making measurements faster and more reliable. In this work, a magneto-impedance sensor based on an amorphous Fe_73.5Nb₃Cu₁Si_13.5B₉ wire as the sensing element is integrated into a customized millifluidic chip. The sensor detects the presence of magnetic nanoparticles in the ferrofluid and distinguishes the different stray fields generated by single-domain superparamagnetic iron oxide nanoparticles or magnetically blocked Co-ferrite nanoparticles. Full article

The luminal surface of the intestinal epithelium is protected by a vital mucus layer, which is essential for lubrication, hydration, and fostering symbiotic bacterial relationships. Replicating and studying this complex mucus structure in vitro presents considerable challenges. To address this, we developed a hydrogel-integrated millifluidic tissue chamber capable of applying precise apical shear stress to intestinal models cultured on flat or 3D structured hydrogel scaffolds with adjustable stiffness. The chamber is designed to accommodate nine hydrogel scaffolds, 3D-printed as flat disks with a storage modulus matching the physiological range of intestinal tissue stiffness (~3.7 kPa) from bioactive decellularized and methacrylated small intestinal submucosa (dSIS-MA). Computational fluid dynamics simulations were conducted to confirm a laminar flow profile for both flat and 3D villi-comprising scaffolds in the physiologically relevant regime. The system was initially validated with HT29-MTX seeded hydrogel scaffolds, demonstrating accelerated differentiation, increased mucus production, and enhanced 3D organization under shear stress. These characteristic intestinal tissue features are essential for advanced in vitro models as they critically contribute to a functional barrier. Subsequently, the chamber was challenged with human intestinal stem cells (ISCs) from the terminal ileum. Our findings indicate that biomimicking hydrogel scaffolds, in combination with physiological shear stress, promote multi-lineage differentiation, as evidenced by a gene and protein expression analysis of basic markers and the 3D structural organization of ISCs in the absence of chemical differentiation triggers. The quantitative analysis of the alkaline phosphatase (ALP) activity and secreted mucus demonstrates the functional differentiation of the cells into enterocyte and goblet cell lineages. The millifluidic system, which has been developed and optimized for performance and cost efficiency, enables the creation and modulation of advanced intestinal models under biomimicking conditions, including tunable matrix stiffness and varying fluid shear stresses. Moreover, the readily accessible and scalable mucus-producing cellular tissue models permit comprehensive mucus analysis and the investigation of pathogen interactions and penetration, thereby offering the potential to advance our understanding of intestinal mucus in health and disease. Full article

The development of highly selective separation processes is a focus of current research. In 2016, the German Science Foundation funded a priority program SPP 2045 “MehrDimPart—highly specific multidimensional fractionation of fine particles with technical relevance” that aims to develop new or enhance existing approaches for the separation of nano- and micrometer-sized particles. Dielectrophoretic separators achieve highly selective separations of (bio-)particles in microfluidic devices or can handle large quantities when non-selective separation is sufficient. Recently, separator designs were developed that aim to combine a high throughput and high selectivity. Here, we summarize the development from a microfluidic fast chromatographic separation via frequency modulated dielectrophoretic particle chromatography (DPC) toward a macrofluidic high throughput separation. Further, we provide a starting point for future work by providing new experimental data demonstrating for the first time the trapping of 200 nm polystyrene particles in a dielectrophoretic high-throughput separator that uses printed circuit boards as alternatives for expensive electrode arrays. Full article

This study attempts to address the challenge of accurately measuring the degradation of biodegradable hydrogels, which are frequently employed in drug delivery for controlled and sustained release. The traditional method utilizes a mass-loss approach, which is cumbersome and time consuming. The aim of this study was to develop an innovative screening platform using a millifluidic device coupled with automated image analysis to measure the degradation of Gelatin methacrylate (GelMA) and the subsequent release of an entrapped wetting agent, polyvinyl alcohol (PVA). Gel samples were placed within circular wells on a custom millifluidic chip and stained with a red dye for enhanced visualization. A camera module captured time-lapse images of the gels throughout their degradation. An image-analysis algorithm was used to translate the image data into degradation rates. Simultaneously, the eluate from the chip was collected to quantify the amount of GelMA degraded and PVA released at various time points. The visual method was validated by comparing it with the mass-loss approach (R = 0.91), as well as the amount of GelMA eluted (R = 0.97). The degradation of the GelMA gels was also facilitated with matrix metalloproteinases 9. Notably, as the gels degraded, there was an increase in the amount of PVA released. Overall, these results support the use of the screening platform to assess hydrogel degradation and the subsequent release of entrapped therapeutic compounds. Full article

Flow focusing is an important hydrodynamic technique for cytometric analysis, enabling the rapid study of cellular samples to identify a variety of biological processes. To date, the majority of flow-focusing devices are fabricated using conventional photolithography or flame processing of glass capillaries. This article presents a suite of low-cost, millifluidic, flow-focusing devices that were fabricated using a desktop sterolithgraphy (SLA) 3D printer. The suite of SLA printing strategies consists of a monolithic SLA method and a hybrid molding process. In the monolithic SLA approach, 1.3 mm square millifluidic channels were printed as a single piece. The printed device does not require any post processing, such as bonding or surface polishing for optical access. The hybrid molding approach consists of printing a mold using the SLA 3D printer. The mold is treated to an extended UV exposure and oven baked before using PDMS as the molding material for the channel. To demonstrate the viability of these channels, we performed a series of experiments using several flow-rate ratios to show the range of focusing widths that can be achieved in these devices. The experiments are validated using a numerical model developed in ANSYS. Full article

In recent years, measuring and monitoring analyte concentrations continuously, frequently, and periodically has been a vital necessity for certain individuals. We developed a cotton-based millifluidic fabric-based electrochemical device (mFED) to monitor glucose continuously and evaluate the effects of mechanical deformation on the device’s electrochemical performance. The mFED was fabricated using stencil printing (thick film method) for patterning the electrodes and wax-patterning to make the reaction zone. The analytical performance of the device was carried out using the chronoamperometry method at a detection potential of −0.2 V. The mFED has a linear working range of 0–20 mM of glucose, with LOD and LOQ of 0.98 mM and 3.26 mM. The 3D mFED shows the potential to be integrated as a wearable sensor that can continuously measure glucose under mechanical deformation. Full article

Bacteria primarily live in structured environments, such as colonies and biofilms, attached to surfaces or growing within soft tissues. They are engaged in local competitive and cooperative interactions impacting our health and well-being, for example, by affecting population-level drug resistance. Our knowledge of bacterial competition and cooperation within soft matrices is incomplete, partly because we lack high-throughput tools to quantitatively study their interactions. Here, we introduce a method to generate a large amount of agarose microbeads that mimic the natural culture conditions experienced by bacteria to co-encapsulate two strains of fluorescence-labeled Escherichia coli. Focusing specifically on low bacterial inoculum (1–100 cells/capsule), we demonstrate a study on the formation of colonies of both strains within these 3D scaffolds and follow their growth kinetics and interaction using fluorescence microscopy in highly replicated experiments. We confirmed that the average final colony size is inversely proportional to the inoculum size in this semi-solid environment as a result of limited available resources. Furthermore, the colony shape and fluorescence intensity per colony are distinctly different in monoculture and co-culture. The experimental observations in mono- and co-culture are compared with predictions from a simple growth model. We suggest that our high throughput and small footprint microbead system is an excellent platform for future investigation of competitive and cooperative interactions in bacterial communities under diverse conditions, including antibiotics stress. Full article

In conventional assisted reproductive technologies (ARTs), oocytes are in vitro cultured in static conditions. Instead, dynamic systems could better mimic the physiological in vivo environment. In this study, a millifluidic in vitro oocyte maturation (mIVM) system, in a transparent bioreactor integrated with 3D printed supports, was investigated and modeled thanks to computational fluid dynamic (CFD) and oxygen convection-reaction-diffusion (CRD) models. Cumulus-oocyte complexes (COCs) from slaughtered lambs were cultured for 24 h under static (controls) or dynamic IVM in absence (native) or presence of 3D-printed devices with different shapes and assembly modes, with/without alginate filling. Nuclear chromatin configuration, mitochondria distribution patterns, and activity of in vitro matured oocytes were assessed. The native dynamic mIVM significantly reduced the maturation rate compared to the static group (p < 0.001) and metaphase II (MII) oocytes showed impaired mitochondria distribution (p < 0.05) and activity (p < 0.001). When COCs were included in a combination of concave+ring support, particularly with alginate filling, oocyte maturation and mitochondria pattern were preserved, and bioenergetic/oxidative status was improved (p < 0.05) compared to controls. Results were supported by computational models demonstrating that, in mIVM in biocompatible inserts, COCs were protected from shear stresses while ensuring physiological oxygen diffusion replicating the one occurring in vivo from capillaries. Full article