Immobilisation of Neuro-2a cells on electrodes and electrochemical detection of MTT formazan crystals to assess their viability

Alkassar, Mounira; Leonardo, Sandra; Diogène, Jorge; Campàs, Mònica

doi:10.1016/j.bioelechem.2022.108274

Cited by 8 publications

(5 citation statements)

References 49 publications

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“…The adsorption of cells on sensing materials can be enhanced through the modification of these surfaces with components that emulate the extracellular matrix environment [71]. For example, polymers such as polyaniline, poly-L-lysine, and poly(3,4-ethylenedioxythiophene) have been proven to be effective for the adhesion of Neuro-2A cells on carbon electrodes [72]. Another factor that needs to be considered in the immobilization of cells is signal transduction.…”

Section: Cell-based Biosensorsmentioning

confidence: 99%

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Detection of Ciguatoxins and Tetrodotoxins in Seafood with Biosensors and Other Smart Bioanalytical Systems

Reverté

Alkassar

Diogène

et al. 2023

Foods

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The emergence of marine toxins such as ciguatoxins (CTXs) and tetrodotoxins (TTXs) in non-endemic regions may pose a serious food safety threat and public health concern if proper control measures are not applied. This article provides an overview of the main biorecognition molecules used for the detection of CTXs and TTXs and the different assay configurations and transduction strategies explored in the development of biosensors and other biotechnological tools for these marine toxins. The advantages and limitations of the systems based on cells, receptors, antibodies, and aptamers are described, and new challenges in marine toxin detection are identified. The validation of these smart bioanalytical systems through analysis of samples and comparison with other techniques is also rationally discussed. These tools have already been demonstrated to be useful in the detection and quantification of CTXs and TTXs, and are, therefore, highly promising for their implementation in research activities and monitoring programs.

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Section: Cell-based Biosensorsmentioning

confidence: 99%

“…Another type of CBB is the strategy used by Alkassar and co-authors, where cell viability is recorded with electrochemical techniques [72]. As explained above, in metabolic active cells, the MTT dye is transformed to a formazan product with redox properties.…”

Section: Cell-based Biosensorsmentioning

confidence: 99%

Detection of Ciguatoxins and Tetrodotoxins in Seafood with Biosensors and Other Smart Bioanalytical Systems

Reverté

Alkassar

Diogène

et al. 2023

Foods

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“…The mouse bioassay (MBA), which has been used for a long time for CTXs’ detection, has become obsolete owing to its insufficient detection capability and ethical concerns [ 31 ]. Therefore, alternative methods, such as liquid chromatography with tandem mass spectrometry (LC-MS/MS) [ 32 , 33 ], capillary LC-HRMS [ 25 ], sandwich enzyme-linked immunosorbent assay (ELISA) [ 34 ], receptor binding assay (RBA) [ 32 , 35 , 36 , 37 ], neuroblastoma cell-based assay (N2a assay) [ 32 , 36 , 38 , 39 ], and even portable biosensors [ 40 , 41 , 42 , 43 ] have been developed in recent years. The RBA and N2a assays, which respond indistinguishably to all CTXs, complement LC-MS/MS and sandwich ELISA, which quantify the congeners individually.…”

Section: Introductionmentioning

confidence: 99%

Sensitive Detection of Ciguatoxins Using a Neuroblastoma Cell-Based Assay with Voltage-Gated Potassium Channel Inhibitors

Yokozeki,

Kawabata,

Fujita

et al. 2024

Toxins

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Ciguatoxins (CTXs) are neurotoxins responsible for ciguatera poisoning (CP), which affects more than 50,000 people worldwide annually. The development of analytical methods to prevent CP is a pressing global issue, and the N2a assay is one of the most promising methods for detecting CTXs. CTXs are highly toxic, and an action level of 0.01 μg CTX1B equivalent (eq)/kg in fish has been proposed. It is desirable to further increase the detection sensitivity of CTXs in the N2a assay to detect such low concentrations reliably. The opening of voltage-gated sodium channels (NaV channels) and blocking of voltage-gated potassium channels (KV channels) are thought to be involved in the toxicity of CTXs. Therefore, in this study, we developed an assay that could detect CTXs with higher sensitivity than conventional N2a assays, using KV channel inhibitors as sensitizing reagents for N2a cells. The addition of the KV channel inhibitors 4-aminopyridine and tetraethylammonium chloride to N2a cells, in addition to the traditional sensitizing reagents ouabain and veratridine, increased the sensitivity of N2a cells to CTXs by up to approximately 4-fold. This is also the first study to demonstrate the influence of KV channels on the toxicity of CTXs in a cell-based assay.

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“…Typically, neuroblastoma cell lines have been used to detect toxins of marine origin [19,20]. This is due to the presence in the cells of several targets that are normally affected by marine toxins and that may cause a loss in function or cell death, the endpoint of which is usually measured in many methods [21][22][23]. Among them, Neuro-2a cells are a particular type of neuroblastoma from mice that has been used for TTX detection [24,25].…”

Section: Introductionmentioning

confidence: 99%

Automated Patch Clamp for the Detection of Tetrodotoxin in Pufferfish Samples

Campàs,

Reverté,

Tudó

et al. 2024

Marine Drugs

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Tetrodotoxin (TTX) is a marine toxin responsible for many intoxications around the world. Its presence in some pufferfish species and, as recently reported, in shellfish, poses a serious health concern. Although TTX is not routinely monitored, there is a need for fast, sensitive, reliable, and simple methods for its detection and quantification. In this work, we describe the use of an automated patch clamp (APC) system with Neuro-2a cells for the determination of TTX contents in pufferfish samples. The cells showed an IC50 of 6.4 nM for TTX and were not affected by the presence of muscle, skin, liver, and gonad tissues of a Sphoeroides pachygaster specimen (TTX-free) when analysed at 10 mg/mL. The LOD achieved with this technique was 0.05 mg TTX equiv./kg, which is far below the Japanese regulatory limit of 2 mg TTX equiv./kg. The APC system was applied to the analysis of extracts of a Lagocephalus sceleratus specimen, showing TTX contents that followed the trend of gonads > liver > skin > muscle. The APC system, providing an in vitro toxicological approach, offers the advantages of being sensitive, rapid, and reliable for the detection of TTX-like compounds in seafood.

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Immobilisation of Neuro-2a cells on electrodes and electrochemical detection of MTT formazan crystals to assess their viability

Cited by 8 publications

References 49 publications

Detection of Ciguatoxins and Tetrodotoxins in Seafood with Biosensors and Other Smart Bioanalytical Systems

Detection of Ciguatoxins and Tetrodotoxins in Seafood with Biosensors and Other Smart Bioanalytical Systems

Sensitive Detection of Ciguatoxins Using a Neuroblastoma Cell-Based Assay with Voltage-Gated Potassium Channel Inhibitors

Automated Patch Clamp for the Detection of Tetrodotoxin in Pufferfish Samples

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