Paper-based multiplex biosensors for inexpensive healthcare diagnostics: a comprehensive review.

Multiplex detection is a smart and an emerging approach in point-of-care testing as it reduces analysis time and testing cost by detecting multiple analytes or biomarkers simultaneously which are crucial for disease detection at an early stage. Application of inexpensive substrate such as paper has immense potential and matter of research interest in the area of point of care testing for multiplexed analysis as it possesses several unique advantages. This study presents the use of paper, strategies adopted to refine the design created on paper and lateral flow strips to enhance the signal, increase the sensitivity and specificity of multiplexed biosensors. An overview of different multiplexed detection studies performed using biological samples has also been reviewed along with the challenges and advantages offered by multiplexed analysis.

Analysis of U87 glioma cells by dielectrophoresis.

Glioblastoma multiforme is the most aggressive and invasive brain cancer consisting of genetically and phenotypically altering glial cells. It has massive heterogeneity due to its highly complex and dynamic microenvironment. Here, electrophysiological properties of U87 human glioma cell line were measured based on a dielectrophoresis phenomenon to quantify the population heterogeneity of glioma cells. Dielectrophoretic forces were generated using a gold-microelectrode array within a microfluidic channel when 3 Vpp and 100, 200, 300, 400, 500 kHz, 1, 2, 5, and 10 MHz frequencies were applied. We analyzed the dielectrophoretic behavior of 500 glioma cells, and revealed that the crossover frequency of glioma cells was around 140 kHz. A quantifying dielectrophoretic movement of the glioma cells exhibited three distinct glioma subpopulations: 50% of the glioma cells experienced strong, 30% of the cells were spread in the microchannel by moderate, and the rest of the cells experienced very weak positive dielectrophoretic forces. Our results demonstrated the dielectrophoretic spectra of U87 glioma cell line. Dielectrophoretic responses of glioma cells linked population heterogeneity to membrane properties of glioma cells rather than their size distribution in the population.

Nondimensional Streaming Dielectrophoresis Number for a System of Continuous Particle Separation.

Cell sorting methods are required in numerous healthcare assays. Although flow cytometry and magnetically actuated sorting are widespread techniques for cell sorting, there is intense research on label-free techniques to reduce the cost and complexity of the process. Among label-free techniques, dielectrophoresis (DEP) offers the capability to separate cells not only on the basis of size but also on their membrane capacitance. This is important because it enables cell discrimination on the basis of specific traits such as viability, identity, fate, and age. StreamingDEP refers to the continuous sorting of cells thanks to the generation of streams of targeted particles by equilibrating the drag and DEP forces acting on targeted particles. In this work, we provide an analytical expression for a streamingDEP number toward enabling the a priori design of DEP devices to agglomerate targeted particles into streams. The nondimensional streamingDEP number (SDN) obtained in this analysis is applied to experiments with 1 µm polystyrene particles and Candida cells. On the basis of these experiments, three characteristic zones are mapped to different values of the SDN: (1) physical capture thanks to DEP for 0 < SDN < 0.6; (2) streaming due to DEP for 0.6 < SDN < 1; (3) elution without experiencing DEP for SDN > 1.

Dielectrophoretic characterization and separation of monocytes and macrophages using 3D carbon-electrodes.

Monocyte heterogeneity and its prevalence are revealed as indicator of several human diseases ranking from cardiovascular diseases to rheumatoid arthritis, chronic kidney diseases, autoimmune multiple sclerosis, and stroke injuries. When monocytes and macrophages are characterized and isolated with preserved genetic, phenotypic and functional properties, they can be used as label-free biomarkers for precise diagnostics and treatment of various diseases. Here, the dielectrophoretic responses of the monocytes and macrophages were examined. We present 3D carbon-electrode dielectrophoresis (carbon-DEP) as a separation tool for U937 monocytes and U937 monocyte-differentiated macrophages. The carbon-electrodes advanced the usability and throughput of DEP separation, presented wider electrochemical stability. Using the 3D carbon-DEP chip, we first identified the selective positive and negative DEP responses and specific crossover frequencies of monocytes and macrophages as their signatures for separation. The crossover frequency of monocytes and macrophages was 17 and 30 kHz, respectively. Next, we separated monocyte and macrophage subpopulations using their specific dielectrophoretic responses. Afterward, we used a fluorescence-activated cell sorter to confirm our results. Finally, we enriched 70% of monocyte cells from the mixed cell population, in other words, concentration of monocyte cells to macrophage cells was five times increased, using the 30-kHz, 10-Vpp electric field and 1 µL/min flow rate.

Dielectrophoretic Separation of Live and Dead Monocytes Using 3D Carbon-Electrodes.

Blood has been the most reliable body fluid commonly used for the diagnosis of diseases. Although there have been promising investigations for the development of novel lab-on-a-chip devices to utilize other body fluids such as urine and sweat samples in diagnosis, their stability remains a problem that limits the reliability and accuracy of readouts. Hence, accurate and quantitative separation and characterization of blood cells are still crucial. The first step in achieving high-resolution characteristics for specific cell subpopulations from the whole blood is the isolation of pure cell populations from a mixture of cell suspensions. Second, live cells need to be purified from dead cells; otherwise, dead cells might introduce biases in the measurements. In addition, the separation and characterization methods being used must preserve the genetic and phenotypic properties of the cells. Among the characterization and separation approaches, dielectrophoresis (DEP) is one of the oldest and most efficient label-free quantification methods, which directly purifies and characterizes cells using their intrinsic, physical properties. In this study, we present the dielectrophoretic separation and characterization of live and dead monocytes using 3D carbon-electrodes. Our approach successfully removed the dead monocytes while preserving the viability of the live monocytes. Therefore, when blood analyses and disease diagnosis are performed with enriched, live monocyte populations, this approach will reduce the dead-cell contamination risk and achieve more reliable and accurate test results.

The ethics of engineered living materials.

Engineered living materials (ELMs) combine living and non-living entities. Their associated ethical concerns must be addressed to promote safety, promote sustainability, and regulate societal impacts. This article identifies key ethical and safety issues by reflecting on fundamental ethical principles. It further discusses a future ethical roadmap for sustainable research in ELMs.

Bio-based epoxy vitrimer with inherent excellent flame retardance and recyclability via molecular design.

The development of biobased fire-safe thermosets with recyclability heralds the switch for a transition towards a circular economy. In this framework, we introduced a novel high-performance bio-epoxy vitrimer (named GVD), which was fabricated by forming a crosslinking network between bio-epoxy glycerol triglycidyl ether (Gte), varying amounts of reactive flame-retardant agent 9,10-dihydro-9-oxa-10-phosphaphenanthrene-10-oxide (DOPO) (0-7 wt%) and a vanillin-based hardener (VA) with imine bonds. For instance, the epoxy vitrimer GVD5, featuring a DOPO content of 5 wt%, achieved a V-0 rating in the vertical burning test (UL-94) and obtained a limiting oxygen index (LOI) value of 31 %, surpassing the performance of pristine epoxy. Furthermore, the peak heat release rate and total heat release of GVD5 were reduced by 38.2 % and 26.3 %, respectively, compared to pristine epoxy. The GVD vitrimers further demonstrated exceptional reprocessability and recyclability, attributed to the presence of dynamic imine bonds within the topological crosslinking network. Remarkably, the epoxy vitrimers maintained the mechanical properties of the parent epoxy. Therefore, this work provides a facile strategy for fabricating high-performance and multi-functional bio-epoxy thermosets.

Investigating Permselectivity in PVDF Mixed Matrix Membranes Using Experimental Optimization, Machine Learning Segmentation, and Statistical Forecasting.

This research examines the correlation between interfacial characteristics and membrane distillation (MD) performance of copper oxide (Cu) nanoparticle-decorated electrospun carbon nanofibers (CNFs) polyvinylidene fluoride (PVDF) mixed matrix membranes. The membranes were fabricated by a bottom-up phase inversion method to incorporate a range of concentrations of CNF and Cu + CNF particles in the polymer matrix to tune the porosity, crystallinity, and wettability of the membranes. The resultant membranes were tested for their application in desalination by comparing the water vapor transport and salt rejection rates in the presence of Cu and CNF. Our results demonstrated a 64% increase in water vapor flux and a salt rejection rate of over 99.8% with just 1 wt % loading of Cu + CNF in the PVDF matrix. This was attributed to enhanced chemical heterogeneity, porosity, hydrophobicity, and crystallinity that was confirmed by electron microscopy, tensiometry, and scattering techniques. A machine learning segmentation model was trained on electron microscopy images to obtain the spatial distribution of pores in the membrane. An Autoregressive Integrated Moving Average with Explanatory Variable (ARIMAX) statistical time series model was trained on MD experimental data obtained for various membranes to forecast the membrane performance over an extended duration.

Microarchitected Compliant Scaffolds of Pyrolytic Carbon for 3D Muscle Cell Growth.

The integration of additive manufacturing technologies with the pyrolysis of polymeric precursors enables the design-controlled fabrication of architected 3D pyrolytic carbon (PyC) structures with complex architectural details. Despite great promise, their use in cellular interaction remains unexplored. This study pioneers the utilization of microarchitected 3D PyC structures as biocompatible scaffolds for the colonization of muscle cells in a 3D environment. PyC scaffolds are fabricated using micro-stereolithography, followed by pyrolysis. Furthermore, an innovative design strategy using revolute joints is employed to obtain novel, compliant structures of architected PyC. The pyrolysis process results in a pyrolysis temperature- and design-geometry-dependent shrinkage of up to 73%, enabling the geometrical features of microarchitected compatible with skeletal muscle cells. The stiffness of architected PyC varies with the pyrolysis temperature, with the highest value of 29.57 ± 0.78 GPa for 900 °C. The PyC scaffolds exhibit excellent biocompatibility and yield 3D cell colonization while culturing skeletal muscle C2C12 cells. They further induce good actin fiber alignment along the compliant PyC construction. However, no conclusive myogenic differentiation is observed here. Nevertheless, these results are highly promising for architected PyC scaffolds as multifunctional tissue implants and encourage more investigations in employing compliant architected PyC structures for high-performance tissue engineering applications.

Enhanced Performance of Laser-Induced Graphene Supercapacitors via Integration with Candle-Soot Nanoparticles.

Laser-induced graphene (LIG) has been emerging as a promising electrode material for supercapacitors due to its cost-effective and straightforward fabrication approach. However, LIG-based supercapacitors still face challenges with limited capacitance and stability. To overcome these limitations, in this work, we present a novel, cost-effective, and facile fabrication approach by integrating LIG materials with candle-soot nanoparticles. The composite electrode is fabricated by laser irradiation on a Kapton sheet to generate LIG material, followed by spray-coating with candle-soot nanoparticles and annealing. Materials characterization reveals that the annealing process enables a robust connection between the nanoparticles and the LIG materials and enhances nanoparticle graphitization. The prepared supercapacitor yields a maximum specific capacitance of 15.1 mF/cm2 at 0.1 mA/cm2, with a maximum energy density of 2.1 µWh/cm2 and a power density of 50 µW/cm2. Notably, the synergistic activity of candle soot and LIG surpasses the performances of previously reported LIG-based supercapacitors. Furthermore, the cyclic stability of the device demonstrates excellent capacitance retention of 80% and Coulombic efficiency of 100% over 10000 cycles.

Fabrication of High Surface Area Microporous ZnO from ZnO/Carbon Sacrificial Composite Monolith Template.

Fabrication of porous materials from the standard sacrificial template method allows metal oxide nanostructures to be produced and have several applications in energy, filtration and constructing sensing devices. However, the low surface area of these nanostructures is a significant drawback for most applications. Here, we report the synthesis of ZnO/carbon composite monoliths in which carbon is used as a sacrificial template to produce zinc oxide (ZnO) porous nanostructures with a high specific surface area. The synthesized porous oxides of ZnO with a specific surface area of 78 m2/g are at least one order of magnitude higher than that of the ZnO nanotubes reported in the literature. The crucial point to achieving this remarkable result was the usage of a novel ZnO/carbon template where the carbon template was removed by simple heating in the air. As a high surface area porous nanostructured ZnO, these synthesized materials can be useful in various applications including catalysis, photocatalysis, separation, sensing, solar energy harvest and Zn-ion battery and as supercapacitors for energy storage.

Evaluating carbon-electrode dielectrophoresis under the ASSURED criteria.

Extreme point-of-care refers to medical testing in unfavorable conditions characterized by a lack of primary resources or infrastructure. As witnessed in the recent past, considerable interest in developing devices and technologies exists for extreme point-of-care applications, for which the World Health Organization has introduced a set of encouraging and regulating guidelines. These are referred to as the ASSURED criteria, an acronym for Affordable (A), Sensitive (S), Specific (S), User friendly (U), Rapid and Robust (R), Equipment-free (E), and Delivered (D). However, the current extreme point of care devices may require an intermediate sample preparation step for performing complex biomedical analysis, including the diagnosis of rare-cell diseases and early-stage detection of sepsis. This article assesses the potential of carbon-electrode dielectrophoresis (CarbonDEP) for sample preparation competent in extreme point-of-care, following the ASSURED criteria. We first discuss the theory and utility of dielectrophoresis (DEP) and the advantages of using carbon microelectrodes for this purpose. We then critically review the literature relevant to the use of CarbonDEP for bioparticle manipulation under the scope of the ASSURED criteria. Lastly, we offer a perspective on the roadmap needed to strengthen the use of CarbonDEP in extreme point-of-care applications.

Carbon-Based Materials for Articular Tissue Engineering: From Innovative Scaffolding Materials toward Engineered Living Carbon.

Carbon materials constitute a growing family of high-performance materials immersed in ongoing scientific technological revolutions. Their biochemical properties are interesting for a wide set of healthcare applications and their biomechanical performance, which can be modulated to mimic most human tissues, make them remarkable candidates for tissue repair and regeneration, especially for articular problems and osteochondral defects involving diverse tissues with very different morphologies and properties. However, more systematic approaches to the engineering design of carbon-based cell niches and scaffolds are needed and relevant challenges should still be overcome through extensive and collaborative research. In consequence, this study presents a comprehensive description of carbon materials and an explanation of their benefits for regenerative medicine, focusing on their rising impact in the area of osteochondral and articular repair and regeneration. Once the state-of-the-art is illustrated, innovative design and fabrication strategies for artificially recreating the cellular microenvironment within complex articular structures are discussed. Together with these modern design and fabrication approaches, current challenges, and research trends for reaching patients and creating social and economic impacts are examined. In a closing perspective, the engineering of living carbon materials is also presented for the first time and the related fundamental breakthroughs ahead are clarified.

Quantifying Deformation and Migration Properties of U87 Glioma Cells Using Dielectrophoretic Forces.

Glioblastoma multiforme is one of the most aggressive malignant primary brain tumors. To design effective treatment strategies, we need to better understand the behavior of glioma cells while maintaining their genetic and phenotypic stability. Here, we investigated the deformation and migration profile of U87 Glioma cells under the influence of dielectrophoretic forces. We fabricated a gold microelectrode array within a microfluidic channel and applied sinusoidal wave AC potential at 3 Vpp, ranging from 30 kHz to 10 MHz frequencies, to generate DEP forces. We followed the dielectrophoretic movement and deformation changes of 100 glioma cells at each frequency. We observed that the mean dielectrophoretic displacements of glioma cells were significantly different at varying frequencies with the maximum and minimum traveling distances of 13.22 µm and 1.37 µm, respectively. The dielectrophoretic deformation indexes of U87 glioma cells altered between 0.027-0.040. It was 0.036 in the absence of dielectrophoretic forces. This approach presents a rapid, robust, and sensitive characterization method for quantifying membrane deformation of glioma cells to determine the state of the cells or efficacy of administrated drugs.

Comparing Carbon Origami from Polyaramid and Cellulose Sheets.

Carbon origami enables the fabrication of lightweight and mechanically stiff 3D complex architectures of carbonaceous materials, which have a high potential to impact a wide range of applications positively. The precursor materials and their inherent microstructure play a crucial role in determining the properties of carbon origami structures. Here, non-porous polyaramid Nomex sheets and macroporous fibril cellulose sheets are explored as the precursor sheets for studying the effect of precursor nature and microstructure on the material and structural properties of the carbon origami structures. The fabrication process involves pre-creasing precursor sheets using a laser engraving process, followed by manual-folding and carbonization. The cellulose precursor experiences a severe structural shrinkage due to its macroporous fibril morphology, compared to the mostly non-porous morphology of Nomex-derived carbon. The morphological differences further yield a higher specific surface area for cellulose-derived carbon. However, Nomex results in more crystalline carbon than cellulose, featuring a turbostratic microstructure like glassy carbon. The combined effect of morphology and glass-like features leads to a high mechanical stiffness of 1.9 ± 0.2 MPa and specific modulus of 2.4 × 104 m2·s-2 for the Nomex-derived carbon Miura-ori structure, which are significantly higher than cellulose-derived carbon Miura-ori (elastic modulus = 504.7 ± 88.2 kPa; specific modulus = 1.2 × 104 m2·s-2) and other carbonaceous origami structures reported in the literature. The results presented here are promising to expand the material library for carbon origami, which will help in the choice of suitable precursor and carbon materials for specific applications.

Electrospun carbon nanofibre-assisted patterning of metal oxide nanostructures.

This work establishes carbon nanofibre-mediated patterning of metal oxide nanostructures, through the combination of electrospinning and vapor-phase transport growth. Electrospinning of a suitable precursor with subsequent carbonization results in the patterning of catalyst gold nanoparticles embedded within carbon nanofibres. During vapor-phase transport growth, these nanofibres allow preferential growth of one-dimensional metal oxide nanostructures, which grow radially outward from the nanofibril axis, yielding a hairy caterpillar-like morphology. The synthesis of metal oxide caterpillars is demonstrated using zinc oxide, indium oxide, and tin oxide. Source and substrate temperatures play the most crucial role in determining the morphology of the metal oxide caterpillars, whereas the distribution of the nanofibres also has a significant impact on the overall morphology. Introducing the current methodology with near-field electrospinning further facilitates user-defined custom patterning of metal oxide caterpillar-like structures.

Carbon fiber/microlattice 3D hybrid architecture as multi-scale scaffold for tissue engineering.

Multiscale 3D carbon architectures are of particular interest in tissue engineering applications, as these structures may allow for three-dimensional cell colonization essential for tissue growth. In this work, carbon fiber/microlattice hybrid architectures are introduced as innovative multi-scale scaffolds for tissue engineering. The microlattice provides the design freedom and structural integrity, whereas the fibrous component creates a cellular microenvironment for cell colonization. The hybrid structures are fabricated by carbonization of stereolithographically 3D printed epoxy microlattice architectures which are pre-filled with cotton fibers within the empty space of the architectures. The cotton filling result in less shrinkage of the architecture during carbonization, as the tight confinement of the fibrous material prevents the free-shrinkage of the microlattices. The hybrid architecture exhibits a compressive strength of 156.9±25.6 kPa, which is significantly higher than an empty carbon microlattice architecture. Furthermore, the hybrid architecture exhibits a flexible behavior up to 30% compressive strain, which is also promising towards soft-tissue regeneration. Osteoblast-like murine MC3T3-E1 cells are cultured within the 3D hybrid structures. Results show that the cells are able to not only proliferate on the carbon microlattice elements as well as along the carbon fibers, but also make connections with each other across the inner pores created by the fibers, leading to a three-dimensional cell colonization. These carbon fiber/microlattice hybrid structures are promising for future fabrication of functionally graded scaffolds for tissue repair applications.

Characterization of the Dielectrophoretic Response of Different Candida Strains Using 3D Carbon Microelectrodes.

Bloodstream infection with Candida fungal cells remains one of the most life-threatening complications among hospitalized patients around the world. Although most of the cases are still due to Candida albicans, the rising incidence of infections caused by other Candida strains that may not respond to traditional anti-fungal treatments merits the development of a method for species-specific isolation of Candida. To this end, here we present the characterization of the dielectrophoresis (DEP) response of Candida albicans, Candida tropicalis and Candida parapsilosis. We complement such characterization with a study of the Candida cells morphology. The Candida strains exhibited subtle differences in their morphology and dimensions. All the Candida strains exhibited positive DEP in the range 10-500 kHz, although the strength of the DEP response was different for each Candida strain at different frequencies. Only Candida tropicalis showed positive DEP at 750 kHz. The current results show potential for manipulation and enrichment of a specific Candida strain at specific DEP conditions towards aiding in the rapid identification of Candida strains to enable the effective and timely treatment of Candida infections.

Facile template-free synthesis of multifunctional 3D cellular carbon from edible rice paper.

Edible rice paper wrapper is found to be an interesting precursor of a porous and light-weight carbon material. During pyrolysis, material samples show significant differences in length change, displaying typical 20-25% shrinking in the in-plane directions, and strongly expanding (up to 500%) across their out-of-plane direction. This results in a template-free synthesis of a 3D network of cellular carbon material. The out-of-plane expansion also allows for fabrication of 3D shapes of cellular carbon material from the 2D precursor. The rice paper derived carbon material features a hierarchical porosity, resulting in a specific surface area ranging from 6 m2 g-1 to 239 m2 g-1 depending on the synthesis temperature. The carbon material has a density of 0.02-0.03 g cm-3, and a higher modulus-density ratio than reported for other cellular carbon materials. It is mechanically stiff and exhibits excellent fire-resistant properties.