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Platelet-Rich Plasma, also known as PRP, is an autologous biologic product used in medicine as a treatment for tissue repair. Nowadays, the majority of PRP obtention methods enrich only platelets, not considering extraplatelet biomolecules, which take part in several cell processes. In the present work, a novel PRP preparation method was developed to obtain a PRP rich in both platelet and plasma extraplatelet molecules. The method is based on the evaporation of the water of the plasma using a rotary evaporator. With this new methodology an increase in plasmatic growth factors and, as a consequence, a better dermal fibroblast cell viability was achieved, compared to a standard PRP formulation. This novel PRP product obtained with this new methodology showed promising results in vitro as an improved PRP treatment in future application. [ABSTRACT FROM AUTHOR]

The Dean Flow, a physics phenomenon that accounts for the impact of channel curvature on fluid dynamics, has great potential to be used in microfluidic synthesis of nanoparticles. This study explores the impact of the Dean Flow on the synthesis of ZIF‐8 particles. Several variables that influence the Dean Equation (the mathematical expression of Dean Flow) are tested to validate the applicability of this expression in microfluidic synthesis, including the flow rate, radius of curvature, channel cross sectional area, and reagent concentration. It is demonstrated that the current standard of reporting, providing only the flow rate and crucially not the radius of curvature, is an incomplete description that will invariably lead to irreproducible syntheses across different laboratories. An alternative standard of reporting is presented and it is demonstrated how the sleek and simple math of the Dean Equation can be used to precisely tune the final dimensions of high quality, monodisperse ZIF‐8 nanoparticles between 40 and 700 nm. [ABSTRACT FROM AUTHOR]

Remotely handled miniaturized colorimetric sensor platforms with multiple functionalities are able to mimic conventional laboratory operations with reduced assets. The integration of a magnetic phase in a miniaturized hydrogel sensor system allows the manipulation of the sensor, under a magnetic field, in a programmable manner. However, dark color interferences from conventional magnetic phases (Fe3O4, Fe2O3, and Fe micro/nanoparticles) affect the signal readout, hindering the colorimetric response. Therefore, a novel Janus bead configuration is introduced and tested by the localized incorporation of ferromagnetic iron microparticles into a TiO2 nanotubes/alginate hydrogel bead biosystem, under an applied magnetic field. The so‐called hydrogel Janus bead shows both, magnetic translocation and biosensing properties in the same bead. These beads are used for the direct colorimetric analysis of biomarkers in blood. The surface of the Janus bead prevents the biofouling of red blood cells, keeping the sensor surface clean for accurate optical colorimetric analysis. Moreover, the external magnetic manipulation of the bead permits precise control of the time and position of the bead in the sample. Therefore, multiple functionalities are presented by a single magnetic TiO2 nanotubes/alginate Janus bead such as actuation, low biofouling, and sensing, positioning this technology as a truly Lab‐in‐a‐Bead System. [ABSTRACT FROM AUTHOR]

In recent years, innovative cell-based biosensing systems have been developed, showing impact in healthcare and life science research. Now, there is a need to design mass-production processes to enable their commercialization and reach society. However, current protocols for their fabrication employ materials that are not optimal for industrial production, and their preparation requires several chemical coating steps, resulting in cumbersome protocols. We have developed a simplified two-step method for generating controlled cell patterns on PMMA, a durable and transparent material frequently employed in the mass manufacturing of microfluidic devices. It involves air plasma and microcontact printing. This approach allows the formation of well-defined cell arrays on PMMA without the need for blocking agents to define the patterns. Patterns of various adherent cell types in dozens of individual cell cultures, allowing the regulation of cell–material and cell–cell interactions, were developed. These cell patterns were integrated into a microfluidic device, and their viability for more than 20 h under controlled flow conditions was demonstrated. This work demonstrated the potential to adapt polymeric cytophobic materials to simple fabrication protocols of cell-based microsystems, leveraging the possibilities for commercialization. [ABSTRACT FROM AUTHOR]

Platelet-rich plasma (PRP) is an autologous biologic product used in several fields of medicine for tissue repair due to the regenerative capacity of the biomolecules of its formulation. PRP consists of a plasma with a platelet concentration higher than basal levels but with basal levels of any biomolecules present out of the platelets. Plasma contains extraplatelet biomolecules known to enhance its regenerative properties. Therefore, a PRP containing not only a higher concentration of platelets but also a higher concentration of extraplatelet biomolecules that could have a stronger regenerative performance than a standard PRP. Considering this, the aim of this work is to develop a new method to obtain PRP enriched in both platelet and extraplatelet molecules. The method is based on the absorption of the water of the plasma using hydroxyethyl acrylamide (HEAA)-based hydrogels. A plasma fraction obtained from blood, containing the basal levels of platelets and proteins, was placed in contact with the HEAA hydrogel powder to absorb half the volume of the water. The resulting plasma was characterized, and its bioactivity was analyzed in vitro. The novel PRP (nPRP) showed a platelet concentration and platelet derived growth factor (PDGF) levels similar to the standard PRP (sPRP), but the concentration of the extraplatelet growth factors IGF-1 (p < 0.0001) and HGF (p < 0.001) were significantly increased. Additionally, the cells exposed to the nPRP showed increased cell viability than those exposed to a sPRP in human dermal fibroblasts (p < 0.001) and primary chondrocytes (p < 0.01). In conclusion, this novel absorption-based method produces a PRP with novel characteristics compared to the standard PRPs, with promising in vitro results that could potentially trigger improved tissue regeneration capacity. [ABSTRACT FROM AUTHOR]

Platelet-Rich Plasma (PRP) is an autologous biological product which, due to its regenerative capacity, is currently used in different fields of medicine. This biological treatment has proven to be effective in numerous research studies due to its high content of growth factors released by platelets. However, the current systems used to obtain PRP do not enrich the growth factors and cytokines outside platelets. Considering this, the present work aims to develop a new technique by which all the biomolecules present in plasma are enriched. Thus, a new method based on ultrafiltration has been developed for the obtaining of the novel PRP. By this method, ultrafiltration of the plasma water is carried out using a 3KDa filtering unit. The results showed that the technique was able to concentrate extraplatelet factors, such as IGF-1 and HGF, in contrast with conventional plasmas. Thus, the cultured cells responded with increased viability to this new PRP. These results could provide a new approach to the treatment of injuries requiring regenerative medicine, potentially improving the outcomes of the conventional PRPs. [ABSTRACT FROM AUTHOR]

This work presents a new architecture concept for microfluidic devices, which combines the conventional 3D printing fabrication process with the stable and precise integration of polymeric functional materials in small footprints within the microchannels in well-defined locations. The approach solves the assembly errors that normally occur during the integration of functional and/or sensing materials in hybrid microfluidic devices. The method was demonstrated by embedding four pH-sensitive ionogel microstructures along the main microfluidic channel of a complex 3D printed microfluidic device. The results showed that this microfluidic architecture, comprising the internal integration of sensing microstructures of diverse chemical compositions, highly enhanced the adhesion force between the microstructures and the 3D printed microfluidic device that contains them. In addition, the performance of this novel 3D printed pH sensor device was investigated using image analysis of the pH colour variations obtained from photos taken with a conventional camera. The device presented accurate and repetitive pH responses in the 2 to 12 pH range without showing any type of device deterioration or lack of performance over time. [ABSTRACT FROM AUTHOR]

Glucose is an analyte of great importance, both in the clinical and sports fields. Since blood is the gold standard biofluid used for the analytical determination of glucose, there is high interest in finding alternative non-invasive biofluids, such as sweat, for its determination. In this research, we present an alginate-based bead-like biosystem integrated with an enzymatic assay for the determination of glucose in sweat. The system was calibrated and verified in artificial sweat, and a linear calibration range was obtained for glucose of 10–1000 µM. The colorimetric determination was investigated, and the analysis was carried out both in the black and white and in the Red:Green:Blue color code. A limit of detection and quantification of 3.8 µM and 12.7 µM, respectively, were obtained for glucose determination. The biosystem was also applied with real sweat, using a prototype of a microfluidic device platform as a proof of concept. This research demonstrated the potential of alginate hydrogels as scaffolds for the fabrication of biosystems and their possible integration in microfluidic devices. These results are intended to bring awareness of sweat as a complementary tool for standard analytical diagnosis. [ABSTRACT FROM AUTHOR]

Additive manufacturing technology is an emerging method for rapid prototyping, which enables the creation of complex geometries by one-step fabrication processes through a layer-by-layer approach. The simplified fabrication achieved with this methodology opens the way towards a more efficient industrial production, with applications in a great number of fields such as biomedical devices. In biomedicine, blood is the gold-standard biofluid for clinical analysis. However, blood cells generate analytical interferences in many test procedures; hence, it is important to separate plasma from blood cells before analytical testing of blood samples. In this research, a custom-made resin formulation combined with a high-resolution 3D printing methodology were used to achieve a methodology for the fast prototype optimization of an operative plasma separation modular device. Through an iterative process, 17 different prototypes were designed and fabricated with printing times ranging from 5 to 12 min. The final device was evaluated through colorimetric analysis, validating this fabrication approach for the qualitative assessment of plasma separation from whole blood. The 3D printing method used here demonstrates the great contribution that this microfluidic technology will bring to the plasma separation biomedical devices market. [ABSTRACT FROM AUTHOR]

The accumulation and deposition of undesired air bubbles on the surface of microfluidic channels and on underwater functional systems is a major drawback, which worsens their performance. Present de-bubbling strategies are usually complex, and they require advanced tools or tedious operation protocols. In this study, we propose a flexible and easily adaptable strategy to efficiently collect, transport and evacuate the underwater deposited air bubbles by a remote stimulus using superhydrophobic ferromagnetic particles. The low surface energy of the superhydrophobic ferromagnetic particles assures the trapping of underwater air bubbles due to their underwater superaerophilicity, while the high magnetisation of the superhydrophobic ferromagnetic particles facilitates the manipulation of the formed particle-bubble system using an external magnetic field, induced by a simple permanent magnet. The properties of the system were characterised for different bubble-deposited surfaces. Remarkably, a ∼99% air bubble removal efficiency was observed in microfluidic channels, providing an easy and inexpensive strategy to address the air bubble accumulation issue faced by the microfluidic technology without integrating any advanced tools or protocols in the device. [ABSTRACT FROM AUTHOR]

The manipulation of droplets via a magnetic field forms the basis of a fascinating technology that is currently in development. Often, the movement of droplets with magnets involves adding magnetic particles in or around the droplet; alternatively, magneto responsive surfaces may also be used. This work, presents and characterizes experimentally the formation and properties of a tunable superparamagnetic ring (tSPRing), which precisely adjusts itself around a water droplet, due to liquid–liquid interaction, and enables the physical manipulation of droplets. The ring is made of an oil‐based ferrofluid, a stable suspension of ferromagnetic particles in an oily phase. It appears spontaneously due to the oil–water interfacial interaction under the influence of a magnetic field. The ferrofluid–water interaction resembles a cupcake assembly, with the surrounding ring only at the base of the droplet. The ring is analogous to a soft matter ring magnet, showing dipole repulsive forces, which stabilizes the droplets on a surface. It enables robust, controllable, and programmable manipulation of enclosed water droplets. This work opens the door to new applications in open surface upside or upside‐down microfluidics and lays the groundwork for new studies on tunable interfaces between two immiscible liquids. [ABSTRACT FROM AUTHOR]

The effect of cell–cell contact on gene transfection is mainly unknown. Usually, transfection is carried out in batch cell cultures without control over cellular interactions, and efficiency analysis relies on complex and expensive protocols commonly involving flow cytometry as the final analytical step. Novel platforms and cell patterning are being studied to control cellular interactions and improve quantification methods. In this study, we report the use of surface patterning of fibronectin for the generation of two types of mesenchymal stromal cell patterns: single‐cell patterns without cell‐to‐cell contact, and small cell‐colony patterns. Both scenarios allowed the integration of the full transfection process and the continuous monitoring of thousands of individualized events by fluorescence microscopy. Our results showed that cell‐to‐cell contact clearly affected the transfection, as single cells presented a maximum transfection peak 6 h earlier and had a 10% higher transfection efficiency than cells with cell‐to‐cell contact. [ABSTRACT FROM AUTHOR]

Lipid membrane nanotubes (NTs) are a widespread template for in vitro studies of cellular processes happening at high membrane curvature. Traditionally NTs are manufactured one by one, using sophisticated membrane micromanipulations, while simplified methods for controlled batch production of NTs are in growing demand. Here we propose a lab-on-a-chip (LOC) approach to the simultaneous formation of multiple NTs with length and radius controlled by the chip design. The NTs form upon rolling silica microbeads covered by lipid lamellas over the pillars of a polymer micropillar array. The array's design and surface chemistry set the geometry of the resulting free-standing NTs. The integration of the array inside a microfluidic chamber further enables fast and turbulence-free addition of components, such as proteins, to multiple preformed NTs. This LOC approach to NT production is compatible with the use of high power objectives of a fluorescence microscope, making real-time quantification of the different modes of the protein activity in a single experiment possible. [ABSTRACT FROM AUTHOR]

Self-assembled monolayers (SAMs) on glass were used as a platform to sequentially deposit fluorophores and small molecules for ion sensing. It was shown that sequential deposition of a flurophore and a binding group onto quartz results in a complete sensing system for cations and anions with no need to independently design and synthesize receptors.

A great breadth of questions remains in cellular biology. Some questions cannot be answered using traditional analytical techniques and so demand the development of new tools for research. In the near future, the development of highly integrated microfluidic analytical platforms will enable the acquisition of unknown biological data. These microfluidic systems must allow cell culture under controlled microenvironment and high throughput analysis. For this purpose, the integration of a variable number of newly developed micro- and nano-technologies, which enable control of topography and surface chemistry, soluble factors, mechanical forces and cell-cell contacts, as well as technology for monitoring cell phenotype and genotype with high spatial and temporal resolution will be necessary. These multifunctional devices must be accompanied by appropriate data analysis and management of the expected large datasets generated. The knowledge gained with these platforms has the potential to improve predictive models of the behavior of cells, impacting directly in better therapies for disease treatment. In this review, we give an overview of the microtechnology toolbox available for the design of high throughput microfluidic platforms for cell analysis.We discuss current microtechnologies for cell microenvironment control, different methodologies to create large arrays of cellular systems and finally techniques for monitoring cells in microfluidic devices. [ABSTRACT FROM AUTHOR]

The combination of cell microenvironment control and real-time monitoring of cell signaling events can provide key biological information. Through precise multipatterning of gold nanoparticles (GNPs) around cells, sensing and actuating elements can be introduced in the cells' microenviroment, providing a powerful substrate for cell studies. In this work, a combination of techniques are implemented to engineer complex substrates for cell studies. Alternating GNPs and bioactive areas are created with micrometer separation by means of a combination of vacumm soft-lithography of GNPs and protein microcontract printing. Instead of conventional microfluidics that need syringe pumps to flow liquid in the microchannels, degas driven flow is used to fill dead-end channels with GNP solutions, rendering the fabrication process straightforward and accessible. This new combined technique is called Printing and Vacuum lithography (PnV lithography). By using different GNPs with various organic coating ligands, different macroscale patterns are obtained, such as wires, supercrystals, and uniformly spread nanoparticle layers that can find different applications depending on the need of the user. The application of the system is tested to pattern a range of mammalian cell lines and obtain readouts on cell viability, cell morphology, and the presence of cell adhesive proteins. [ABSTRACT FROM AUTHOR]

We report a microfluidic chip-based hydrodynamic focusing approach that minimizes sample volume for the analysis of cell–surface interactions under controlled fluid-shear conditions. Assays of statistically meaningful numbers of translocating platelets interacting with immobilized von Willebrand factor at arterial shear rates (∼1500 s ^-1) are demonstrated. By controlling spatial disposition and relative flow rates of two contacting fluid streams, e.g., sample (blood) and aqueous buffer, on-chip hydrodynamic focusing guides the cell-containing stream across the protein surface as a thin fluid layer, consuming ∼50 μL of undiluted whole blood for a 2-min platelet assay. Control of wall shear stress is independent of sample consumption for a given flow time. The device design implements a mass-manufacturable fabrication approach. Fluorescent labeling of cells enables readout using standard microscopy tools. Customized image-analysis software rapidly quantifies cellular surface coverage and aggregate size distributions as a function of time during blood-flow analyses, facilitating assessment of drug treatment efficacy or diagnosis of disease state. [ABSTRACT FROM PUBLISHER]

Lactate is present in sweat at high concentrations, being a metabolite of high interest in sport science and medicine. Therefore, the potential to determine lactate concentrations in physiological fluids, at the point of need with minimal invasiveness, is very valuable. In this work, the synthesis and performance of an alginate bead biosystem was investigated. Artificial sweat with different lactate concentrations was used as a proof of concept. The lactate detection was based on a colorimetric assay and an image analysis method using lactate oxidase, horseradish peroxidase and tetramethyl benzidine as the reaction mix. Lactate in artificial sweat was detected with a R² = 0.9907 in a linear range from 10 mM to 100 mM, with a limit of detection of 6.4 mM and a limit of quantification of 21.2 mM. Real sweat samples were used as a proof of concept to test the performance of the biosystem, obtaining a lactate concentration of 48 ± 3 mM. This novel sensing configuration, using alginate beads, gives a fast and reliable method for lactate sensing, which could be integrated into more complex analytical systems. [ABSTRACT FROM AUTHOR]

The main problem for the expansion of the use of microfluidic paper-based analytical devices and, thus, their mass production is their inherent lack of fluid flow control due to its uncontrolled fabrication protocols. To address this issue, the first step is the generation of uniform and reliable microfluidic channels. The most common paper microfluidic fabrication method is wax printing, which consists of two parts, printing and heating, where heating is a critical step for the fabrication of reproducible device dimensions. In order to bring paper-based devices to success, it is essential to optimize the fabrication process in order to always get a reproducible device. Therefore, the optimization of the heating process and the analysis of the parameters that could affect the final dimensions of the device, such as its shape, the width of the wax barrier and the internal area of the device, were performed. Moreover, we present a method to predict reproducible devices with controlled working areas in a simple manner. [ABSTRACT FROM AUTHOR]

Nowadays, great efforts are made in developing new technology for rapid detection of specific DNA sequences, for environmental monitoring, forensic analysis and rapid biomedical diagnosis applications. Microfluidic paper-based analytical devices are suitable platforms for the development of point of care devices, due to their simple fabrication protocols, ease of use and low cost. Herein, a methodology for in situ detection of single strand DNA by using a colorimetric assay based on the formation of a DNAzyme within a paper substrate was developed. A DNAzyme that could only be formed in the presence of a specific sequence of the Y human amelogenin gene was designed. The performance of the DNAzyme was followed colorimetrically first in solution and then in paper substrates. The reaction was found to be specific to the Y fragment selected as analyte. The DNAzyme reaction on paper enabled the unequivocal colorimetric identification of the Y single strand DNA fragment both qualitatively, with the naked eye (143 ng), and quantitatively by image analysis (45.7 ng). As a proof of concept, a microfluidic paper-based device, pre-loaded with all DNAzyme reagents, was characterized and implemented for the simultaneous detection of X and Y single strand DNA fragments. Image 1 • Optimized methodology for in situ DNA detection in a paper device format. • Detection based on a DNAzyme using Y human amelogenin fragment. • Presents a non-enzymatic, colorimetric and rapid way to detect DNA sequences. • Detection: quantitatively by calculating the grey intensity values. • Detection: qualitatively by the presence of a green color "coffee ring". [ABSTRACT FROM AUTHOR]