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In this work, we simultaneously detected and predicted the concentration levels of serotonin (SE) and dopamine (DA) neurotransmitters (NTs) for in vitro mixtures, with measurements obtained using conventional glassy carbon electrodes (CGCEs) and differential pulse voltammetry (DPV). The NTs were estimated by deconvolving the multiplexed signals of both NTs using Principal Component Analysis with Gaussian Process Regression (PCA-GPR) and Partial Least Squares with Gaussian Process Regression (PLS-GPR), both with exponential–isotropic kernels. The average testing accuracies of estimation using PCA-GPR for DA alone, SE alone and their mixture (DA–SE) were 87.6%, 88.1%, and 96.7%, respectively. Using PLS-GPR, the testing accuracies of estimation for DA alone, SE alone, and their mixture (DA–SE) were 87.3%, 83.8%, and 95.1%, respectively. Furthermore, we explored methods of reducing the procedural complexity in estimating the NTs by finding reduced subsets of features for accurately detecting and predicting their concentrations. The reduced subsets of features found in the oxidation potential windows of the NTs improved the testing accuracy of the estimation of DA–SE to 97.4%. We thus believe that reducing complexity has the potential to increase the detection and prediction accuracies of NT measurements for practical clinical uses such as deep brain stimulation.more » « lessFree, publicly-accessible full text available April 1, 2026
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Reactive oxygen species (ROS) including the superoxide anion (O2•−) are typically studied in cell cultures using fluorescent dyes, which provide only discrete single-point measurements. These methods lack the capabilities for assessing O2•− kinetics and release in a quantitative manner over long monitoring times. Herein, we present the fabrication and application of an electrochemical biosensor that enables real-time continuous monitoring of O2•− release in cell cultures for extended periods (> 8 h) using an O2•− specific microelectrode. To achieve the sensitivity and selectivity requirements for cellular sensing, we developed a biohybrid system consisting of superoxide dismutase (SOD) and Ti3C2Tx MXenes, deposited on a gold microwire electrode (AuME) as O2•− specific materials with catalytic amplification through the synergistic action of the enzyme and the biomimetic MXenes-based structure. The biosensor demonstrated a sensitivity of 18.35 nA/μM with a linear range from 147 to 930 nM in a cell culture medium. To demonstrate its robustness and practicality, we applied the biosensor to monitor O2•− levels in human leukemia monocytic THP-1 cells upon stimulation with lipopolysaccharide (LPS). Using this strategy, we successfully monitored LPS-induced O2•− in THP-1 cells, as well as the quenching effect induced by the ROS scavenger N-acetyl-l-cysteine (NAC). The biosensor is generally useful for exploring the role of oxidative stress and longitudinally monitoring O2•− release in cell cultures, enabling studies of biochemical processes and associated oxidative stress mechanisms in cellular and other biological environments.more » « less
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Dopamine is an essential neurotransmitter for daily cognitive functions controlling many neurophysiological processes including memory, cognition, and physical control. Development of analytical methods and sensors to detect dopamine is important for health monitoring and neurological research. This review provides an overview of recent advances in the development of electrochemical catalytic biosensors based on enzyme and enzyme-mimetic materials and discusses their potential applications for measurements of dopamine in biological fluids. The first part of the review summarizes and critically assesses the different types of enzymes and enzyme mimetic materials that can be used to catalytically convert dopamine, followed by a discussion of the biosensor’s fabrication, key design parameters, and detection mechanism on various electrode platforms ranging from single-use screen-printed electrodes to microneedles and implantable microelectrodes. The second part provides examples of measurements of dopamine in biological samples, including saliva, urine, serum, cell cultures, and brain tissue. We conclude with a summary of advantages and limitations of these devices in the clinical field, and an outlook to future research towards the implementation and broader adoption of electrochemical biosensors in neurophysiology, pharmacology, and the clinical field.more » « less
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Two-dimensional (2D) layered materials that integrate metallic conductivity, catalytic activity and the ability to stabilize biological receptors provide unique capabilities for designing electrochemical biosensors for large-scale detection and diagnostic applications. Herein, we report a multifunctional MXene-based 2D nanostructure decorated with enzyme mimetic cerium oxide nanoparticle (MXCeO2) as a novel platform and catalytic amplifier for electrochemical biosensors, specifically targeting the detection of oxidase enzyme substrates. We demonstrate enhanced catalytic efficiency of the MXCeO2 for the reduction of hydrogen peroxide (H2O2) and its ability to immobilize oxidase enzymes, such as glucose oxidase, lactate oxidase and xanthine oxidase. The designed biosensors exhibit high selectivity, stability, and sensitivity, achieving detection limits of 0.8 μM H2O2, 0.49 μM glucose, 3.6 μM lactate and 1.7 μM hypoxanthine, when the MXCeO2 and their respective enzymes were used. The MXCeO2 was successfully incorporated into a wearable fabric demonstrating high sensitivity for lactate measurements in sweat. The unique combination of MXenes with CeO2 offers excellent conductivity, catalytic efficiency and enhanced enzyme loading, demonstrating potential of the MXCeO2 as a catalytically active material to boost efficiency of oxidase enzyme reactions. This design can be used as a general platform for increasing the sensitivity of enzyme based biosensors and advance the development of electrochemical biosensors for a variety of applications.more » « less
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Cerium oxide nanoparticles (CeNPs) are versatile materials with unique and unusual properties that vary depending on their surface chemistry, size, shape, coating, oxidation states, crystallinity, dopant, structural and surface defects. This review details advances made over the past twenty years in the development of CeNPs and ceria-based nanostructures, the structural determinants affecting their activity, and translation of these distinct features into applications. The two-oxidation states of nanosized CeNPs (Ce3+/Ce4+) coexisting at the nanoscale level, facilitate formation of oxygen vacancies and defect states which confer extremely high reactivity and oxygen buffering capacity, and the ability to act as catalysts for oxidation and reduction reactions. However, the method of synthesis, surface functionalization, surface coating and defects are important factors in determining their properties. This review highlights the key properties of CeNPs, their synthesis, interactions and reaction pathways, and provides examples of emerging applications. Due to their unique properties, CeNPs have become quintessential candidates for catalysis, chemical mechanical planarization (CMP), sensing, biomedical applications and environmental remediation, with tremendous potential to create novel products and translational innovations in a wide range of industries. This review highlights the timely relevance and the transformative potential of these materials in addressing societal challenges and driving technological advancements across these fields.more » « less
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Oxidative stress and excessive accumulation of the superoxide (O2.-) anion are at the genesis of many pathological conditions and the onset of several diseases. The real time monitoring of (O2.-) release is important to assess the extent of oxidative stress in these conditions. Herein, we present the design, fabrication and characterization of a robust (O2.-) biosensor using a simple and straightforward procedure involving deposition of a uniform layer of L-Cysteine on a gold wire electrode to which Cytochrome C (Cyt c) was conjugated. The immobilized layers, studied using conductive Atomic Force Microscopy (c-AFM) revealed a stable and uniformly distributed redox protein on the gold surface, visualized as conductivity and surface topographical plots. The biosensor enabled detection of (O2.-) at an applied potential of 0.15 V with a sensitivity of 42.4 nA/μM and a detection limit of 2.4 nM. Utility of the biosensor was demonstrated in measurements of real time (O2.-) release in activated human blood platelets and skeletal rat limb muscles following ischemia reperfusion injury (IRI), confirming the biosensor's stability and robustness for measurements in complex biological systems. The results demonstrate the ability of these biosensors to monitor real time release of (O2.-) and estimate the extent of oxidative injury in models that could easily be translated to human pathologies.more » « less
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Electrochemical biosensors have the potential to provide rapid and inexpensive diagnostics while moving clinical testing from centralized labs to point-of-care (POC) applications. Conductive materials functionalized with bioreceptors that remain stable and functional for measurements in real-world conditions are essential for the fabrication of electrochemical biosensors, and carbon-based nanomaterials provide the electrical, chemical, structural, and mechanical features that make them suitable for POC devices. This review details the most recent developments in the use of carbon-based nanostructures, with a focus on one-dimensional carbon nanotubes, two-dimensional graphene, and graphene oxide, their interface with biological receptors, deposition on portable, flexible, and wearable substrates, and integration on low-cost platforms for detection of clinical biomarkers. The large-scale manufacturing and implementation of microneedles as implantable and electronic tattoos as wearable devices for on-skin diagnostics, and lab-on-mouth platforms as well as the interface with mobile technologies and their potential implementation for remote POC monitoring and decentralized healthcare through cloud processing and the internet of things (IoT) are discussed with examples of applications. The review concludes with an overview of the regulatory perspectives and future trends, challenges, and opportunities for commercialization and translation of these technologies from the research lab to practice, as useful diagnostic tools for remote monitoring of patient health conditions.more » « less
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Electrochemical sensors are ideally suited for the detection of reactive oxygen and nitrogen species (ROS and RNS) generated during biological processes. This review discusses the latest work in the development of electrochemical microsensors for ROS/RNS and their possible applications for monitoring oxidative stress in biological systems. The performance of recent designs of microelectrodes and electrode materials are discussed along with their functionality in preclinical models of drug efficacy, mitochondrial distress, and endothelial dysfunction. Challenges and opportunities in translating this methodology to study the pathophysiology associated with various diseases are discussed.more » « less
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