Real-Time Monitoring of Cellular Bioenergetics with a Multianalyte Screen-Printed Electrode

McKenzie, Jennifer R.; Cognata, Andrew C.; Davis, Anna N; Wikswo, John P.; Cliffel, David E.

doi:10.1021/acs.analchem.5b01533

Cited by 32 publications

(41 citation statements)

References 34 publications

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“…This may be due to a combination of the storage method (wet storage), the use of Nafion (antifouling), and the enzyme entrapment technique (crosslinking), which all stabilize enzyme sensors. 6,17 The dynamic and durable nature of this sensor make it well suited for integration into the μCA to measure cellular microphysiometry.…”

Section: Discussionmentioning

confidence: 99%

“…17 The electrode array features five electrodes; three platinum disk electrodes and two band electrodes. The three platinum disk electrodes (A = 1.8 mm 2 ) were used for enzymatic detection of glutamate.…”

Section: Methodsmentioning

confidence: 99%

“…Microclinical analyzer fabrication.-The μCA bioreactor was designed in-house and fabricated by Vanderbilt Institute for Integrative Biosystems Research and Education (VIIBRE) at Vanderbilt. 17 The screws, magnets, O-rings, membranes, nuts, and ferrules can be purchased commercially. The screen-printed electrode was designed in-house and fabricated by Pine Research.…”

Section: Methodsmentioning

confidence: 99%

“…5,23 This versatile platform can easily integrate cells or other enzyme sensors to monitor cellular bioenergetics in real-time. 17 In addition to increasing the analytical power, microfluidic systems also help recapitulate physiological conditions. 24 Microfluidic systems can add flow, and, as a result, mechanical forces.…”

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confidence: 99%

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Electrochemical Microphysiometry Detects Cellular Glutamate Uptake

Miller

McClain

Cliffel

2018

J. Electrochem. Soc.

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Glutamate is the principal excitatory amino acid in the vertebrate nervous system and is responsible for learning and memory. Understanding of these complex biological processes can be gained through experimentally accessible systems of glutamate detection. In this work, the microclinical analyzer (μCA) was used with a sensitive and stable glutamate sensor and model neuronal cells for quantitative glutamate detection under physiological relevant shear. Glutamate was detected by immobilized glutamate oxidase on a screen-printed electrode array. The sensor's linear range spanned glutamate's physiological to pathophysiological concentration range, and the biologically relevant sub-second to month temporal range. After 11 hours of use, the sensor retained 91 ± 1% of its signal, and it was able to be stored for a month without a significant decrease. When model neuronal cells were integrated into the μCA bioreactor and exposed to glutamate, they initially took up 210 ± 100 μmoles of glutamate, which increased to 390 ± 50 μmoles during their second exposure. These data suggest that the neurotransmitter uptake systems were functional and may be upregulated. The dynamic and durable μCA platform offers an experimentally accessible system of glutamate detection that can be used to monitor glutamate metabolism and signaling. Glutamate is one of the 20 canonical amino acids that together provide the structural and enzymatic foundation of proteins. Alone, glutamate plays a very different role and is itself an excitatory signaling molecule widely distributed throughout the central nervous system. In fact, glutamate is the most prevalent neurotransmitter and its function is essential for proper neurocognition, learning, and memory. Overactivation of glutamate receptors causes excitotoxicity, a pathological process whereby neurons are damaged or killed, which may result in neurodegeneration.1,2 Because of its central role in metabolic and cognitive processes, a strong effort has been put toward developing methods to detect glutamate. [2][3][4][5] Accurate detection of glutamate can be accomplished using many techniques, including spectrometry, spectroscopy, and electrochemistry. The benefits of spectrometry and spectroscopy include sensitivity and selectivity. However, mass spectrometry requires chromatographic separation or vacuum preparation, which decreases the temporal resolution of the system. 4 Although spectroscopic techniques, such as the iGluSNFR, 2 offer impressive temporal resolution, they require optical transparency thereby limiting the scope of samples that can be analyzed. In contrast, electrochemical sensors require almost no sample manipulation and can be placed directly in the area of interest, allowing, for example, detection of glutamate and dopamine signaling in the brain.6-8 Electrochemical sensors are also versatile: They can be made on the nanoscale, 1 have been 3D-printed, 9 and can be inexpensive.10,11 The highly translatable nature of electrochemical techniques has already been demonstrated with the a...

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Electrochemical Microphysiometry Detects Cellular Glutamate Uptake

Miller

McClain

Cliffel

2018

J. Electrochem. Soc.

Self Cite

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“…Several efforts have been initiated toward the accomplishment of this ambitious goal. For example, prototype microphysiometers have been developed for on-line measurement of glucose and lactate (19,35), oxygen (35), transepithelial electrical resistance (24,36,37), ions (38), extracellular acidification rate (pH), and protein biomarkers (39) in organ-on-a-chip systems (40)(41)(42)(43)(44)(45)(46). However, these sensing units were either built upon a single chip and limited in system-level integration and full automation (19), or were designed for measurements over relatively short periods of time in the range of minutes to hours.…”

Section: Significancementioning

confidence: 99%

Multisensor-integrated organs-on-chips platform for automated and continual in situ monitoring of organoid behaviors

Zhang

Aleman

Shin

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

624

571

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Organ-on-a-chip systems are miniaturized microfluidic 3D human tissue and organ models designed to recapitulate the important biological and physiological parameters of their in vivo counterparts. They have recently emerged as a viable platform for personalized medicine and drug screening. These in vitro models, featuring biomimetic compositions, architectures, and functions, are expected to replace the conventional planar, static cell cultures and bridge the gap between the currently used preclinical animal models and the human body. Multiple organoid models may be further connected together through the microfluidics in a similar manner in which they are arranged in vivo, providing the capability to analyze multiorgan interactions. Although a wide variety of human organ-on-a-chip models have been created, there are limited efforts on the integration of multisensor systems. However, in situ continual measuring is critical in precise assessment of the microenvironment parameters and the dynamic responses of the organs to pharmaceutical compounds over extended periods of time. In addition, automated and noninvasive capability is strongly desired for long-term monitoring. Here, we report a fully integrated modular physical, biochemical, and optical sensing platform through a fluidics-routing breadboard, which operates organ-on-a-chip units in a continual, dynamic, and automated manner. We believe that this platform technology has paved a potential avenue to promote the performance of current organ-on-a-chip models in drug screening by integrating a multitude of real-time sensors to achieve automated in situ monitoring of biophysical and biochemical parameters.organ-on-a-chip | microbioreactor | electrochemical biosensor | physical sensor | drug screening D rug discovery is a lengthy process associated with tremendous cost and high failure rate (1). On average, less than 1 in 10 drug candidates are eventually approved by the Food and Drug Administration (2), during which successful transition ratios to next phases are roughly 65, 32, and 60% for phase I, phase II, and phase III, respectively (3). Among the primary causes of failure, nonclinical/ clinical safety (>50%) and efficacy (>10%) stand out in the front, more than all other factors (e.g., strategic, commercial, operational) combined (3, 4). These two major causes not only contribute to the low success rates during the drug development but also lead to the withdrawal of approved drugs from the market. Among all of the drug attritions, it is estimated that safety liabilities related to the cardiovascular system account for 45%, whereas 32% are due to hepatotoxicity (5, 6). These high failure/attrition ratios of drugs SignificanceMonitoring human organ-on-a-chip systems presents a significant challenge, where the capability of in situ continual monitoring of organ behaviors and their responses to pharmaceutical compounds over extended periods of time is critical in understanding the dynamics of drug effects and therefore accurate prediction of human organ reacti...

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Printed Microfluidics

Dixon

Lamanna

Wheeler

2017

Adv Funct Materials

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Microfluidics has become an important tool that is useful for a wide range of applications. A drawback for microfluidics is that many of the techniques that are commonly used to fabricate devices are not widely accessible, not scalable to high‐volume manufacturing processes, or both. Recently, a number of printing strategies that were originally developed for other applications have been applied to microfluidic device fabrication. These techniques, which include inkjet printing (IJP), screen printing (SP), and solid wax printing (SWP), are proposed to have a transformative effect on the field. Here microfluidics and printing, are introduced and a list of favorite examples is provided that highlights the accessibility and scalability that the combination is bringing to the field.

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Real-Time Monitoring of Cellular Bioenergetics with a Multianalyte Screen-Printed Electrode

Cited by 32 publications

References 34 publications

Electrochemical Microphysiometry Detects Cellular Glutamate Uptake

Electrochemical Microphysiometry Detects Cellular Glutamate Uptake

Multisensor-integrated organs-on-chips platform for automated and continual in situ monitoring of organoid behaviors

Printed Microfluidics

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