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We have developed innovative core-shell hydrogel capsules with a dual-network shell structure designed for precise control of molecular transport in response to external stimuli such as pH and temperature. The capsules were fabricated using a combination of microfluidic electrospray techniques and water-in-water (w/w) core-shell droplets templating. The primary network of the shell, calcium alginate (Ca-Alg), with a pKa around 3.4, exhibits sensitivity to pH. The secondary network of the shell, poly(ethylene glycol) methyl ether methacrylate (PEGMA), undergoes a volume phase transition near 60 °C. These properties enable precise molecular transport control in/out of the capsules by modulating the surface charges through varying pH and modifying pore size through temperature changes. Moreover, the dual-network shell structure not only significantly enhances the mechanical strength of the capsules but also improves their stability under external stimulus, ensuring structural integrity during the transport of molecules. This research lays the groundwork for further investigations into the multimodal stimuli-responsive hydrogel systems to control molecular transport, important in applications such as sensors and reactors for chemical cascade reactions.
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Extracellular matrix (ECM) is essential for tissue development, providing structural support and a microenvironment that is necessary for cells. As tissue engineering advances, there is a growing demand for ECM mimics. Polycaprolactone (PCL) is a commonly used synthetic polymer for ECM mimic materials. However, its biologically inactive surface limits its direct application in tissue engineering. Our study aimed to improve the biocompatibility of PCL by incorporating hemoglobin nanofibrils (HbFs) into PCL using an electrospinning technique. HbFs were formed from bovine hemoglobin (Hb) extracted from industrial byproducts and designed to offer PCL an improved cell adhesion property. The fabricated HbFs@PCL electrospun scaffold exhibits improved fibroblast adherence, proliferation, and deeper fibroblast infiltration into the scaffold compared with the pure PCL scaffold, indicating its potential to be an ECM mimic. This study represents the pioneering utilization of Hb-sourced nanofibrils in the electrospun PCL scaffolds for tissue engineering applications.
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As transient electronics continue to advance, the demand for new materials has given rise to the exploration of conducting polymer (CP)-based electronic materials. The big challenge lies in balancing conductivity while introducing controlled degradable properties into CP-based transient materials. In response to this, we present in this work a concept of using conducting polymers attached to an enzymatically biodegradable biopolymer to create transient polymer electronics materials. Specifically, poly(3-hexyl thiophene) (P3HT) is covalently grafted onto biopolymer gelatin, affording graft copolymer gelatin-graft-poly(3-hexyl thiophene) (termed Gel-g-P3HT). The thin films of Gel-g-P3HT that were produced by optimized processing solvent (THF/H2O cosolvent) showed enhanced π-π stacking domains of P3HT, resulting in semiconducting thin films with good electroactivity. Due to the presence of amide bonds in the gelatin backbone, Gel-g-P3HT underwent degradation over a period of 5 days, resulting in the formation of amphiphilic micellar nanoparticles that are biocompatible and nontoxic. The potential of these conductive and degradable graft copolymers was demonstrated in a pressure sensor. This research paves the way for developing biocompatible and enzymatically degradable polymer materials based on P3HT, enabling the next generation of transient polymer electronics for diverse applications, such as skin, implantable, and environmental electronics.
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Detection of superoxide anion (O2·-) levels holds significant importance for the diagnosis and even clinical treatments of oxidative stress-related diseases. Herein, we prepared a composite electrode material to encapsulate copper-zinc superoxide dismutase (SOD1) for biosensing of O2·-. The sensing material consists of gold nanowires (AuNWs), reduced graphene oxide (rGO), carboxymethyl cellulose (CMC) and PEDOT:PSS. CMC provides abundant -COOH to bind SOD1, with a high adsorption coverage of 1.499 × 10-9 mol cm-2 on the sensor surface. rGO and PEDOT endow the composite with significant conductivity, whereas PSS has antifouling capability. Moreover, AuNWs exhibit excellent electrical conductivity and a high aspect ratio, which promotes electron transfer, and ultimately enhances the catalytic performance of the enzyme. Meanwhile, SOD1(Cu2+) catalyzes the dismutation of O2·- to O2 and H2O2, and H2O2 is then electrochemically oxidized to generate amperometric signals for determination of O2·-. The sensor demonstrates outstanding detection performance for O2·- with a low detection limit of 2.52 nM, and two dynamic ranges (14.30 nM-1.34 µM and 1.34 µM-42.97 µM) with corresponding sensitivity of 0.479 and 0.052 µA µM-1cm-2, respectively. Additionally, the calculated apparent Michaelis constant (Kmapp) of 1.804 µM for SOD1 demonstrates the outstanding catalytic activity and the surface-immobilized enzyme's substrate affinity. Furthermore, the sensor shows the capability to dynamically detect the level of O2·- released from living HepG2 cells. This study provides an inovative design to obtain a biocompatible electrochemical sensing platform with plenty of immobilization sites for biomolecules, large surface area, high conductivity and flexibility.
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Técnicas Biosensibles , Grafito , Superóxidos/química , Carboximetilcelulosa de Sodio , Peróxido de Hidrógeno , Superóxido Dismutasa-1 , Técnicas Biosensibles/métodos , Grafito/química , Superóxido Dismutasa/química , Técnicas ElectroquímicasRESUMEN
Bioelectronic devices are designed to translate biological information into electrical signals and vice versa, thereby bridging the gap between the living biological world and electronic systems. Among different types of bioelectronics devices, wearable and implantable biosensors are particularly important as they offer access to the physiological and biochemical activities of tissues and organs, which is significant in diagnosing and researching various medical conditions. Organic conducting and semiconducting materials, including conducting polymers (CPs) and graphene and carbon nanotubes (CNTs), are some of the most promising candidates for wearable and implantable biosensors. Their unique electrical, electrochemical, and mechanical properties bring new possibilities to bioelectronics that could not be realized by utilizing metals- or silicon-based analogues. The use of organic- and carbon-based conductors in the development of wearable and implantable biosensors has emerged as a rapidly growing research field, with remarkable progress being made in recent years. The use of such materials addresses the issue of mismatched properties between biological tissues and electronic devices, as well as the improvement in the accuracy and fidelity of the transferred information. In this review, we highlight the most recent advances in this field and provide insights into organic and carbon-based (semi)conducting materials' properties and relate these to their applications in wearable/implantable biosensors. We also provide a perspective on the promising potential and exciting future developments of wearable/implantable biosensors.
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Técnicas Biosensibles , Nanotubos de Carbono , Dispositivos Electrónicos Vestibles , Polímeros/química , Nanotubos de Carbono/química , ElectrónicaRESUMEN
With the emergence of deadly viral and bacterial infections, preventing the spread of microorganisms on surfaces has gained ever-increasing importance. This study investigates the potential of solid-state supercapacitors as antibacterial and antiviral devices. We developed a low-cost and flexible carbon cloth supercapacitor (CCSC) with highly efficient antibacterial and antiviral surface properties. The CCSC comprised two parallel layers of carbon cloth (CC) electrodes assembled in a symmetric, electrical double-layer supercapacitor structure that can be charged at low potentials between 1 to 2 V. The optimized CCSC exhibited a capacitance of 4.15 ± 0.3 mF cm-2 at a scan rate of 100 mV s-1, high-rate capability (83% retention of capacitance at 100 mV s-1 compared to its value at 5 mV s-1), and excellent electrochemical stability (97% retention of the initial capacitance after 1000 cycles). Moreover, the CCSC demonstrated outstanding flexibility and retained its full capacitance even when bent at high angles, making it suitable for wearable or flexible devices. Using its stored electrical charge, the charged CCSC disinfects bacteria effectively and neutralizes viruses upon surface contact with the positive and negative electrodes. The charged CCSC device yielded a 6-log CFU reduction of Escherichia coli bacterial inocula and a 5-log PFU reduction of HSV-1 herpes virus. Antibacterial and antiviral carbon cloth supercapacitors represent a promising platform technology for various applications, including electronic textiles and electronic skins, health monitoring or motion sensors, wound dressings, personal protective equipment (e.g., masks) and air filtration systems.
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Antibacterianos , Antivirales , Antivirales/farmacología , Fenómenos Físicos , Movimiento (Física) , Antibacterianos/farmacología , Carbono , Escherichia coliRESUMEN
We developed a heart-on-a-chip platform that integrates highly flexible, vertical, 3D micropillar electrodes for electrophysiological recording and elastic microwires for the tissue's contractile force assessment. The high aspect ratio microelectrodes were 3D-printed into the device using a conductive polymer, poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS). A pair of flexible, quantum dots/thermoplastic elastomer nanocomposite microwires were 3D printed to anchor the tissue and enable continuous contractile force assessment. The 3D microelectrodes and flexible microwires enabled unobstructed human iPSC-based cardiac tissue formation and contraction, suspended above the device surface, under both spontaneous beating and upon pacing with a separate set of integrated carbon electrodes. Recording of extracellular field potentials using the PEDOT:PSS micropillars was demonstrated with and without epinephrine as a model drug, non-invasively, along within situmonitoring of tissue contractile properties and calcium transients. Uniquely, the platform provides integrated profiling of electrical and contractile tissue properties, which is critical for proper evaluation of complex, mechanically and electrically active tissues, such as the heart muscle under both physiological and pathological conditions.
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Elastómeros , Polímeros , Humanos , Microelectrodos , Impresión Tridimensional , Dispositivos Laboratorio en un ChipRESUMEN
This work addresses the challenge of surface modification of porous, electrospun fiber mats containing an insoluble conducting polymer coating. Herein, a novel methodology of grafting a polymer brush onto conducting polymer fiber mats is developed that employs filtering of the polymerization solution through the fiber mat. An electrospun sulfonated polystyrene-poly(ethylene-ran-butylene)-polystyrene (sSEBS) fiber mat is first coated with a layer of conducting copolymer bearing an Atom Transfer Radical Polymerization (ATRP) initiating functionality (PEDOT-Br). The surface-initiated ATRP from the fibers' surface is then carried out to graft a hydrophilic polymer brush (poly(ethylene glycol) methyl ether methacrylate) by means of filtering the polymerization solution through the fiber mat. Scanning electron microscopy (SEM) images reveal a progressive change in the morphology of the fiber mat surface with the increasing volume of the filtrated polymerization solution, while energy dispersive X-ray spectrosdcopy (EDX) spectra show a change in the atomic oxygen to sulfur (O/S) ratio, therefore confirming the successful grafting from the fibers' surface. The conductive fiber mat grafted with hydrophilic brushes shows a 20% reduction in the non-specific adsorption of bovine serum albumin (BSA) compared to a pristine fiber mat. This study is a proof-of-concept for this novel, filtration-based, surface-initiated polymerization methodology.
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The COVID-19 pandemic caused by the virus SARS-CoV-2 was the greatest global threat to human health in the last three years. The most widely used methodologies for the diagnosis of COVID-19 are quantitative reverse transcription polymerase chain reaction (RT-qPCR) and rapid antigen tests (RATs). PCR is time-consuming and requires specialized instrumentation operated by skilled personnel. In contrast, RATs can be used in-home or at point-of-care but are less sensitive, leading to a higher rate of false negative results. In this work, we describe the development of a disposable, electrochemical, and laser-scribed graphene-based biosensor strips for COVID-19 detection that exploits a split-ester bond ligase system (termed 'EsterLigase') for immobilization of a virus-specific nanobody to maintain the out-of-plane orientation of the probe to ensure the efficacy of the probe-target recognition process. An anti-spike VHH E nanobody, genetically fused with the EsterLigase domain, was used as the specific probe for the spike receptor-binding domain (SP-RBD) protein as the target. The recognition between the two was measured by the change in the charge transfer resistance determined by fitting the electrochemical impedance spectroscopy (EIS) spectra. The developed LSG-based biosensor achieved a linear detection range for the SP-RBD from 150 pM to 15 nM with a sensitivity of 0.0866 [log(M)]-1 and a limit of detection (LOD) of 7.68 pM.
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COVID-19 , Grafito , Humanos , SARS-CoV-2 , COVID-19/diagnóstico , Pandemias , Anticuerpos , Rayos LáserRESUMEN
Biofouling on surfaces, caused by the assimilation of proteins, peptides, lipids and microorganisms, leads to contamination, deterioration and failure of biomedical devices and causes implants rejection. To address these issues, various antifouling strategies have been extensively studied, including polyethylene glycol-based polymer brushes. Conducting polymers-based biointerfaces have emerged as advanced surfaces for interfacing biological tissues and organs with electronics. Antifouling of such biointerfaces is a challenge. In this study, we fabricated electrospun fibre mats from sulphonated polystyrene-block-poly(ethylene-ran-butylene)-block-polystyrene (sSEBS), infused with conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) (sSEBS-PEDOT), to produce a conductive (2.06 ± 0.1 S/cm), highly porous, fibre mat that can be used as a biointerface in bioelectronic applications. To afford antifouling, here the poly(oligo (ethylene glycol) methyl ether methacrylate) (POEGMA) brushes were grafted onto the sSEBS-PEDOT conducting fibre mats via surface-initiated atom transfer radical polymerization technique (SI-ATRP). For that, a copolymer of EDOT and an EDOT derivative with SI-ATRP initiating sites, 3,4-ethylenedioxythiophene) methyl 2-bromopropanoate (EDOTBr), was firstly electropolymerized on the sSEBS-PEDOT fibre mat to provide sSEBS-PEDOT/P(EDOT-co-EDOTBr). The POEGMA brushes were grafted from the sSEBS-PEDOT/P(EDOT-co-EDOTBr) and the polymerization kinetics confirmed the successful growth of the brushes. Fibre mats with 10-mers and 30-mers POEGMA brushes were studied for antifouling using a BCA protein assay. The mats with 30-mers grafted brushes exhibited excellent antifouling efficiency, ~82% of proteins repelled, compared to the pristine sSEBS-PEDOT fibre mat. The grafted fibre mats exhibited cell viability >80%, comparable to the standard cell culture plate controls. Such conducting, porous biointerfaces with POEGMA grafted brushes are suitable for applications in various biomedical devices, including biosensors, liquid biopsy, wound healing substrates and drug delivery systems.
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Incrustaciones Biológicas , Polímeros , Polímeros/química , Incrustaciones Biológicas/prevención & control , Poliestirenos , Polietilenglicoles/química , Proteínas/química , Propiedades de SuperficieRESUMEN
Bioelectrical slow waves are fundamental to maintaining the normal motility of the gastrointestinal tract. Slow wave abnormalities are associated with several major digestive disorders. High-resolution electrical mapping arrays have been used to investigate pathological slow wave abnormalities. However, conventional electrode substrate materials are opaque with high mechanical modulus, which leads to non-compliance and sub-par contact with the organ, without additional manipulations. Here we developed highly conformal and transparent conducting polymer electrode arrays using the extrusion wet-printing technique. The performance of electrodes for the electrophysiological recording of the gastric slow wave was validated using in a pig model, against a previously validated reference array over 100 s recording window. The conducting polymer electrodes registered comparable frequency to the reference array ( 3.31±0.20 cpm vs. 3.27±0.07 cpm, p = 0.067), with lower amplitude ( 372±237 vs. ), and signal to noise ratio ( 10.92±7.83 vs. [Formula: see text]). Further adjustments to the deposition parameters and contact material will improve the performance of the conducting polymer array for future experimental applications. Clinical Relevance- These conducting polymer electrodes provide better compliance and minimized mechanical mismatch to the gut tissue thus allowing long-term monitoring and stimulation of the gut. This could be potentially extended to other organs as well.
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Compuestos Bicíclicos Heterocíclicos con Puentes , Polímeros , Animales , Microelectrodos , PorcinosRESUMEN
BACKGROUND: Lipocalins are a large family of proteins, which possess a highly conserved eight-stranded antiparallel beta-barrel structure as distinctive trait. This family includes Major Urinary Proteins (MUPs) from rats and mouse, studied for their role in urinary protein-mediated chemosignalling. Vulpeculin has been identified as the most abundant protein in the urine of the common brushtail possum, Trichosurus vulpecula. On the basis of high similarity with other MUPS, we hypothesised that vulpeculin might have a role in possum chemosignalling and investigated its stability and binding ability. METHODS: We expressed and purified vulpeculin using an E.coli-based system and confirmed correct folding by circular dichroism (CD) spectroscopy. Thermal stability was studied by CD and binding properties were investigated using two optical probes N-phenyl-naphthylamine (NPN) and 8-anilino-1-naphthalene sulphonic acid (ANS). RESULTS: CD revealed a secondary structure typical of a predominantly ß-sheet protein, consistent with the beta barrel structure of the lipocalin family. Vulpeculin showed a high level of thermostability, as assessed by CD, exhibiting a small shift in the secondary structure even at 95 °C. Binding assays indicated that vulpeculin cannot accommodate the NPN ligand but can bind ANS. CONCLUSION: The urinary secretion, high degree of sequence similarity with other lipocalins, its beta sheet structure assessed by CD and potential to bind hydrophobic ligands in the hydrophobic cavity or an external hydrophobic pocket, suggest vulpeculin may be involved in possum chemosignalling. GENERAL SIGNIFICANCE: This work represents a first step towards the further investigation of the newly discovered lipocalin and its role in possum chemosignalling.
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Lipocalinas , Animales , Ligandos , Ratones , Estructura Secundaria de Proteína , RatasRESUMEN
The conformation of a fluorescent polymer, in the solid state or in solution, plays a critical role in the polymer's fluorescent properties. Thus, grafted side chains on a fluorescent polymer can directly influence its optical properties. In this study, the effect of grafted polymeric side chains on the photoluminescent properties of poly(para-phenylene vinylene) (PPV) and poly(para-phenylene ethynylene) (PPE) were investigated. Low- and high-molecular-weight grafts of neutral poly(n-butyl acrylate), cationic poly(trimethylaminoethyl methacrylate) and anionic poly(sulfopropyl acrylate) were grafted onto PPVs and PPEs, and the effect of the grafting on the graft copolymer's absorption and emission wavelengths, the fluorescence intensity and the quantum yield were studied. The results indicate that in the case of the ionic grafts, contrary to the expectations, the polymers have a reduced quantum yield. This contrasts with the copolymers with uncharged side chains (PnBA), where a major increase in the quantum yield is seen for the self-quenching conjugated pristine polymers. These results reinforce that the molecular conformation of the polymer in a solid or solution plays a critical role in fluorescent polymers photoluminescent properties.
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Electrochemical techniques offer great opportunities for the capture of chemical and biological entities from complex mixtures and their subsequent release into clean buffers for analysis. Such methods are clean, robust, rapid, and compatible with a wide range of biological fluids. Here, we designed an electrochemically addressable system, based on a conducting terpolymer [P(EDOT-co-EDOTSAc-co-EDOTEG)] coated onto a carbon cloth substrate, to selectively capture and release biological entities using a simple electrochemical redox process. The conducting terpolymer composition was optimized and the terpolymer-coated carbon cloth was extensively characterized using electrochemical analysis, Raman and Fourier transform-infrared spectroscopy, water contact angle analysis, and scanning electron microscopy. The conductive terpolymer possesses a derivative of EDOT with an acetylthiomethyl moiety (EDOTSAc), which is converted into a "free" thiol that then undergoes reversible oxidation/reduction cycles at +1.0 V and -0.8 V (vs Ag/AgCl), respectively. That redox process enables electrochemical capture and on-demand release. We first demonstrated the successful electrochemical capture/release of a fluorescently labeled IgG antibody. The same capture/release procedure was then applied to release extracellular vesicles (EVs), originating from both MCF7 and SKBR3 breast cancer cell line bioreactors. EVs were captured using the substrate-conjugated HER2 antibody which was purified from commercially available trastuzumab. Capture and release of breast cancer EVs using a trastuzumab-derived HER2 antibody has not been reported before (to the best of our knowledge). A rapid (2 min) release at a low potential (-0.8 V) achieved a high release efficiency (>70%) of the captured, HER2+ve, SKBR3 EVs. The developed system and the electrochemical method are efficient and straightforward and have vast potential for the isolation and concentration of various biological targets from large volumes of biological and other (e.g., environmental) samples.
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Hydrogels are a popular substrate for cell culture due to their mechanical properties closely resembling natural tissue. Stimuli-responsive hydrogels are a good platform for studying cell response to dynamic stimuli. Poly(N-isopropylacrylamide) (pNIPAM) is a thermo-responsive polymer that undergoes a volume-phase transition when heated to 32 °C. Conducting polymers can be incorporated into hydrogels to introduce electrically responsive properties. The conducting polymer, polypyrrole (PPy), has been widely studied as electrochemical actuators due to its electrochemical stability, fast actuation and high strains. We determine the volume-phase transition temperature of pNIPAM hydrogels with PPy electropolymerised with different salts as a film within the hydrogel network. We also investigate the electro-mechanical properties at the transition temperature (32 °C) and physiological temperature (37 °C). We show statistically significant differences in the Young's modulus of the hybrid hydrogel at elevated temperatures upon electrochemical stimulation, with a 5 kPa difference at the transition temperature. Furthermore, we show a three-fold increase in actuation at transition temperature compared to room temperature and physiological temperature, attributed to the movement of ions in/out of the PPy film that induce the volume-phase transition of the pNIPAM hydrogel. Furthermore, cell adhesion to the hybrid hydrogel was demonstrated with mouse articular chondrocytes.
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Hidrogeles , Polímeros , Animales , Técnicas de Cultivo de Célula , Hidrogeles/química , Ratones , Polímeros/química , Pirroles/química , TemperaturaRESUMEN
Cheap artificial light harvesting systems, which competently harvest solar energy and promote efficient energy transfer, are highly sought after in the renewable sector. We report the synthesis of self-assembled peptide-porphyrin fibers (SJ 6) fabricated with iron(III) oxide (Fe3 O4 ) nanoparticles as feasible electron acceptors. Charge-complementarity between the negatively charged peptide (20E) and the protonated Zn-tetraphenyl porphyrin (ZnTPyP) led to an ordered assembly of the ZnTPyP molecules, enabling efficient light harvesting. X-ray diffraction data indicates a more ordered structure in SJ 6 compared to 20E and ZnTPyP. The incorporation of Fe3 O4 nanoparticles into SJ 6 showed significant fluorescence quenching, indicating efficient electron flow from the donor to the acceptor. The SJ 6-nFe3 O4 system performed the light reaction of photosynthesis as confirmed by the reduction of 1 mM NAD+ to 0.180 mM NADH upon exposure to visible light (Xe lamp λ > 420 nm) for 1 h. The photochemical regeneration of NADH using the SJ 6-nFe3 O4 system was coupled to glutamate dehydrogenase-catalyzed conversion of α-ketoglutarate to L-glutamate. These results confirm the successful synthesis of an artificial light harvesting peptide-porphyrin system with Fe3 O4 nanoparticles as promising low-cost electron separators.
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Nanopartículas del Metal , Nanofibras , Porfirinas , Compuestos Férricos , Luz , NAD/química , Óxidos , Péptidos , Porfirinas/químicaRESUMEN
Blood analysis is an established approach to monitor various diseases, ranging from heart defects and diabetes to cancer. Among various tumor markers in the blood, circulating tumor cells (CTCs) have received increasing attention due to the fact that they originate directly from the tumors. Capturing and detecting CTCs represents a promising approach in cancer diagnostics and clinical management of cancers. CTCs in blood progress to self-seeding a tumour or initiating a new lesion mass. Cytosensors are biosensors intended to identify CTCs in a blood sample of cancer patients and provide information about the cancer status. Herein, we firstly discuss different detection methods of state-of-the-art optical cytosensors, including colorimetry, fluorescence, surface plasmon resonance, photoelectrochemistry and electrochemiluminescence. Then we review the significant advances made in implementing biorecognition elements and nanomaterials for the detection of cancer cells. Despite great progress in optical cytosensors, and their integration with smartphones, they have still only been explored to prototype stages. Much more effort is needed to fulfil their potential in modern cancer diagnostics and in monitoring the state of disease for cancer patients.
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Técnicas Biosensibles , Nanoestructuras , Células Neoplásicas Circulantes , Biomarcadores de Tumor/análisis , Técnicas Biosensibles/métodos , Recuento de Células , Humanos , Células Neoplásicas Circulantes/patologíaRESUMEN
Among invasive mammalian predators, rats represent a major threat, endangering ecosystem functioning worldwide. After rat-control operations, detecting their continued presence or reinvasion requires more sensitive and lower cost detection technologies. Here, we develop a new sensing paradigm by using a specific rat urine biomarker (MUP13) to unambiguously signal the presence of rats. As the first step towards a new remote surveillance technology, aptamers were selected to MUP13 using the Flu-Mag SELEX method. Six aptamer candidates were initially screened by dot blot and two of them (Apt-2.5 and Apt-1.4) exhibited high affinity and specificity. Both aptamers were further characterized by bead-based assay to confirm affinity and selectivity. The lead aptamer candidates were then applied to fluorescence anisotropy (FA) and surface plasmon resonance (SPR)-based biosensor platforms, showing dissociation constants in the nanomolar range and high specificity towards their target. The SPR biosensor had limits of detection of 13.8 and 7.5 nM for Apt-2.5 and Apt-1.4, respectively, which are more than three orders of magnitude lower than the physiological concentrations found in rat urine. Selectivity of the aptamers, when comparing with other major urinary proteins, was excellent, indicating strong efficacy in specific detection of rats. In order to validate the aptamer Apt-2.5 for use with real world samples a FA-based assay was performed on a rat urine sample. The assay showed that the aptamer could detect recombinant MUP13 spiked in filtered urine and the natural MUP13 in unfiltered urine, as a first step into translation to real world application. These are the first known assays to detect and quantify a MUP biomarker of rats.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Animales , Ecosistema , Proteínas , Ratas , Técnica SELEX de Producción de Aptámeros , Resonancia por Plasmón de SuperficieRESUMEN
Insulin, a peptide hormone secreted by pancreatic ß cells, affects the development of diabetes and associated complications. Herein, we propose an electrochemical aptasensor for sensitive and selective detection of insulin using laser-scribed graphene electrodes (LSGEs). Before using disposable LSGEs, the development and proof-of-concept sensing experiments were firstly carried out on research-grade glassy carbon electrode (GCE). The aptasensor is based on using Exonuclease I (Exo I) that catalyses the hydrolysis of single-stranded aptamers attached to the electrode surface; however, the hydrolysis does not occur if the insulin is bound to the aptamer. Therefore, the unbound aptamers are cleaved by Exo I while insulin-bound aptamers remain on the electrode surface. In the next step, the gold nanoparticle - aptamer (AuNPs-Apt) probes are introduced to the electrode surface to form a 'sandwich' structure with the insulin on the surface-attached aptamer. The redox probe, methylene blue (MB), intercalates into the aptamers' guanine bases and the sandwich structure of AuNPs-Apt/insulin/surface-bound aptamer amplifies electrochemical signal from MBs. The signal can be well-correlated to the concentrations of insulin. A limit of detection of 22.7 fM was found for the LSGE-based sensors and 9.8 fM for GCE-based sensors used for comparison and initial sensor development. The results demonstrate successful fabrication of the single-use and sensitive LSGEs-based sensors for insulin detection.
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GrafitoRESUMEN
Motility of the stomach is governed by an electrophysiological event termed gastric slow waves. High-resolution (HR) bioelectrical mapping involves placing array of electrodes over the surface of the stomach to record gastric slow waves. Conductive polymer materials have recently been applied to great effect in cardiology and neurophysiology due to its compliant and biocompatible properties. The aim of this study was to quantify the performance of poly(3,4-ethylenedioxythiophene) doped with poly(styrenesulfonate) (PEDOT:PSS) deposited on a flexible print circuit electrode array for gastric slow wave HR mapping. The Au electrodes were coated with PEDOT:PSS at 1 V and different levels of charges (0.3-1.2 mC). HR mapping alongside standard Au electrodes was performed in three anesthetized pigs. Overall, the PEDOT:PSS electrodes detected both antegrade and retrograde slow wave propagations, with comparable frequency, velocity and signal-to-noise ratio to the Au electrodes. Differences between the two electrodes were noted in terms of amplitude and downstroke gradient. The findings of this study will inform designs of future stretchable and implantable HR mapping electrode arrays for gastrointestinal recording and stimulation therapies.