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A special issue of Biosensors (ISSN 2079-6374). This special issue belongs to the section "Biosensors and Healthcare".

Deadline for manuscript submissions: closed (31 July 2024) | Viewed by 25605

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Dr. Georgia Moschopoulou

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School of Food, Biotechnology and Development (TBA), Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece
Interests: cell-based biosensors; cell and neuronal differentiation; real-time monitoring systems; high-throughput screening or diagnostics systems
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Cells as biorecognition elements have many advantages, such as sensitivity and rapid response to various stimuli. During the last decade, researchers have achieved progress in improving the selectivity of cells against desired target analytes as well as their integration into biosensors, which have led to their use in many applications. For this reason, cell-based biosensors represent one of the most advanced and, at the same time, challenging scientific and technological domains in analytical and diagnostic sciences. The most popular application is in toxicology research, but due to selectivity improvement, they are also used as analytical devices for specific molecules. This Biosensors Special Issue on “Cell-Based Biosensors for Rapid Detection and Monitoring” is intended to be a timely and comprehensive issue on very recent and emerging technologies in the fascinating field of cell-based biosensors for the rapid detection of molecules such as biomarkers, environmental pollutants, etc. and/or monitoring of cell physiology in response to pharmacological or environmental stimuli. Topics include but are not restricted to cell-based methodological approaches, synthetic cell manufacturing (targeted genome editing, genetic circuits, membrane engineering, etc.), integration of cell-based biosensors into platforms, and current applications and perspectives for cell-based biosensors and analytical devices for toxicology and drug research, such as lab-on-a-chip or organ-like cultures. Research papers, short communications, and reviews are all welcome.

Dr. Georgia Moschopoulou
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Biosensors is an international peer-reviewed open access monthly journal published by MDPI.

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Keywords

analytical devices
bioelectric
cell-based biosensors
enviromental pollutants
food safety
impedimetric biosensors
medical diagnostics
microbial fuel cells
toxicology
synthetic cells

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Related Special Issue

Cell-Based Biosensors for Rapid Detection and Monitoring (2nd Edition) in Biosensors

Published Papers (11 papers)

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Research

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12 pages, 1852 KiB

Open AccessArticle

Development of an Escherichia coli Cell-Based Biosensor for Aspirin Monitoring by Genetic Engineering of MarR

by Yeonhong Kim, Yangwon Jeon, Kyeoungseok Song, Haekang Ji, Soon-Jin Hwang and Youngdae Yoon

Biosensors 2024, 14(11), 547; https://doi.org/10.3390/bios14110547 - 12 Nov 2024

Viewed by 578

Abstract

Multiple antibiotic resistance regulators (MarRs) control the transcription of genes in the mar operon of Escherichia coli in the presence of salicylic acid (SA). The interaction with SA induces conformational changes in the MarR released from the promoter of the mar operon, turning on transcription. We constructed an SA-specific E. coli cell-based biosensor by fusing the promoter of the mar operon (P_marO) and the gene that encodes an enhanced green fluorescent protein (egfp). Because SA and aspirin are structurally similar, a biosensor for monitoring aspirin can be obtained by genetically engineering MarR to be aspirin (ASP)-responsive. To shift the selectivity of MarR toward ASP, we changed the residues around the ligand-binding sites by site-directed mutagenesis. We examined the effects of genetic engineering on MarR by introducing MarRs with P_marO-egfp into E. coli. Among the tested mutants, MarR T72A improved the ASP responses by approximately 3 times compared to the wild-type MarR, while still showing an SA response. Although the MarR T72A biosensor exhibited mutual interference between SA and ASP, it accurately determined the ASP concentration in spiked water and medicine samples with over 90% accuracy. While the ASP biosensors still require improvement, our results provide valuable insights for developing E. coli cell-based biosensors for ASP and transcription factor-based biosensors in general. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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14 pages, 2828 KiB

Open AccessArticle

Development of a Portable Cell-Based Biosensor for the Ultra-Rapid Screening for Boscalid Residues in Lettuce

by Georgia Moschopoulou, Vasileios Tsekouras, Josep V. Mercader, Antonio Abad-Fuentes and Spyridon Kintzios

Biosensors 2024, 14(6), 311; https://doi.org/10.3390/bios14060311 - 18 Jun 2024

Viewed by 1024

Abstract

Fungal plant pathogens have posed a significant threat to crop production. However, the large-scale application of pesticides is associated with possible risks for human health and the environment. Boscalid is a widely used fungicide, consistently implemented for the management of significant plant pathogens. Conventionally, the detection and determination of boscalid residues is based on chromatographic separations. In the present study, a Bioelectric Recognition Assay (BERA)-based experimental approach combined with MIME technology was used, where changes in the electric properties of the membrane-engineering cells with anti-boscalid antibodies were recorded in response to the presence of boscalid at different concentrations based on the maximum residue level (MRL) for lettuce. The membrane-engineering Vero cells with 0.5 μg/mL of antibody in their surface were selected as the best cell line in combination with the lowest antibody concentration. Furthermore, the biosensor was tested against another fungicide in order to prove its selectivity. Finally, the BERA cell-based biosensor was able to detect the boscalid residue, below and above the MRL, in spiked lettuce leaf extracts in an entirely distinct and reproducible manner. This study indicates that the BERA-based biosensor, after further development and optimization, could be used for the routine, high-throughput detection of boscalid residue in lettuce, and not only that. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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16 pages, 6741 KiB

Open AccessArticle

Targeted Formation of Biofilms on the Surface of Graphite Electrodes as an Effective Approach to the Development of Biosensors for Early Warning Systems

by Anna Kharkova, Roman Perchikov, Saniyat Kurbanalieva, Kristina Osina, Nadezhda Popova, Andrey Machulin, Olga Kamanina, Evgeniya Saverina, Ivan Saltanov, Sergey Melenkov, Denis Butusov and Vyacheslav Arlyapov

Biosensors 2024, 14(5), 239; https://doi.org/10.3390/bios14050239 - 9 May 2024

Cited by 2 | Viewed by 2041

Abstract

Biofilms based on bacteria Pseudomonas veronii (Ps. veronii) and Escherichia coli (E. coli) and yeast Saccharomyces cerevisiae (S. cerevisiae) were used for novel biosensor creation for rapid biochemical oxygen demand (BOD) monitoring. Based on the electrochemical measurement results, it was shown that the endogenous mediator in the matrix of E. coli and Ps. veronii biofilms and ferrocene form a two-mediator system that improves electron transport in the system. Biofilms based on Ps. veronii and E. coli had a high biotechnological potential for BOD assessment; bioreceptors based on such biofilms had high sensitivity (the lower limits of detectable BOD₅ concentrations were 0.61 (Ps. veronii) and 0.87 (E. coli) mg/dm³) and high efficiency of analysis (a measurement time 5–10 min). The maximum biosensor response based on bacterial biofilms has been observed in the pH range of 6.6–7.2. The greatest protective effect was found for biofilms based on E. coli, which has high long-term stability (151 days for Ps. veronii and 163 days for E. coli). The results of the BOD₅ analysis of water samples obtained using the developed biosensors had a high correlation with the results of the standard 5-day method (R² = 0.9820, number of tested samples is 10 for Ps. veronii, and R² = 0.9862, number of tested samples is 10 for E. coli). Thus, biosensors based on Ps. veronii biofilms and E. coli biofilms could be a novel analytical system to give early warnings of pollution. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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16 pages, 5676 KiB

Open AccessArticle

Highly Sensitive and Linear Resonator-Based Biosensor for White Blood Cell Counting: Feasible Measurement Method and Intrinsic Mechanism Exploration

by Yi-Ke Wang, Bo-Wen Shi, Jun-Ming Zhao, Yan-Xiong Wang, Yan-Feng Jiang, Gang-Long Yang, Xiao-Dong Gao and Tian Qiang

Biosensors 2024, 14(4), 180; https://doi.org/10.3390/bios14040180 - 7 Apr 2024

Viewed by 1569

Abstract

Since different quantities of white blood cells (WBCs) in solution possess an adaptive osmotic pressure of cells, the WBCs themselves and in solution have similar concentrations, resulting in them having similar dielectric properties. Therefore, a microwave sensor could have difficulty in sensing the quantity variation when WBCs are in solution. This paper presents a highly sensitive, linear permittivity-inspired microwave biosensor for WBCs, counting through the evaporation method. Such a measurement method is proposed to record measurements after the cell solution is dripped onto the chip and is completely evaporated naturally. The proposed biosensor consists of an air-bridged asymmetric differential inductor and a centrally located circular fork-finger capacitor fabricated on a GaAs substrate using integrated passive fabrication technology. It is optimized to feature a larger sensitive area and improved Q-factor, which increases the effective area of interaction between cells and the electromagnetic field and facilitates the detection of their changes in number. The sensing relies on the dielectric properties of the cells and the change in the dielectric constant for different concentrations, and the change in resonance properties, which mainly represents the frequency shift, corresponds to the macroscopic change in the concentration of the cells. The microwave biosensors are used to measure biological samples with concentrations ranging from 0.25 × 10⁶ to 8 × 10⁶ cells per mL in a temperature (26.00 ± 0.40 °C) and humidity (54.40 ± 3.90 RH%) environment. The measurement results show a high sensitivity of 25.06 Hz/cells·mL⁻¹ with a highly linear response of r² = 0.99748. In addition, a mathematical modeling of individual cells in suspension is performed to estimate the dielectric constant of individual cells and further explain the working mechanism of the proposed microwave biosensor. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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15 pages, 4971 KiB

Open AccessArticle

Using Rapid Prototyping to Develop a Cell-Based Platform with Electrical Impedance Sensor Membranes for In Vitro RPMI2650 Nasal Nanotoxicology Monitoring

by Mateo Gabriel Vasconez Martinez, Eva I. Reihs, Helene M. Stuetz, Astrid Hafner, Konstanze Brandauer, Florian Selinger, Patrick Schuller, Neus Bastus, Victor Puntes, Johannes Frank, Wolfgang Tomischko, Martin Frauenlob, Peter Ertl, Christian Resch, Gerald Bauer, Guenter Povoden and Mario Rothbauer

Biosensors 2024, 14(2), 107; https://doi.org/10.3390/bios14020107 - 18 Feb 2024

Cited by 2 | Viewed by 2199

Abstract

Due to advances in additive manufacturing and prototyping, affordable and rapid microfluidic sensor-integrated assays can be fabricated using additive manufacturing, xurography and electrode shadow masking to create versatile platform technologies aimed toward qualitative assessment of acute cytotoxic or cytolytic events using stand-alone biochip platforms in the context of environmental risk assessment. In the current study, we established a nasal mucosa biosensing platform using RPMI2650 mucosa cells inside a membrane-integrated impedance-sensing biochip using exclusively rapid prototyping technologies. In a final proof-of-concept, we applied this biosensing platform to create human cell models of nasal mucosa for monitoring the acute cytotoxic effect of zinc oxide reference nanoparticles. Our data generated with the biochip platform successfully monitored the acute toxicity and cytolytic activity of 6 mM zinc oxide nanoparticles, which was non-invasively monitored as a negative impedance slope on nasal epithelial models, demonstrating the feasibility of rapid prototyping technologies such as additive manufacturing and xurography for cell-based platform development. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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12 pages, 3133 KiB

Open AccessCommunication

Capacitance Contribution of NIH/3T3 Cells Existing on and between Electrodes of an Impedance Biosensor

by Yeeun Kim, Dahyun Kang, Seokgyu Kim, Eunchae Hong and Moongyu Jang

Biosensors 2023, 13(11), 970; https://doi.org/10.3390/bios13110970 - 6 Nov 2023

Viewed by 1812

Abstract

In this study, an impedance biosensor capable of real-time monitoring of the growth and drug reactions using NIH/3T3 cells was fabricated through a semiconductor process. With the fabricated impedance biosensor, the cell growth and drug reaction states are monitored in real-time, showing the validness of the developed biosensor. By using the developed impedance biosensor, we have investigated the capacitance contribution of NIH/3T3 cells existing on electrodes and between electrodes. To compare the capacitance value contributions of the cells on and between electrodes, wide- and narrow-gap electrode patterns are manufactured with 3.7 and 0.3 mm electrode gap spacings, respectively. From the detailed analysis, the capacitance contributions of NIH/3T3 cells existing on electrodes are estimated around less than 20 percent compared to the cells existing between electrodes. In other words, a minimized electrode area with maximized electrode spacing is the promising impedance biosensor design guide for accurate cell capacitance measurements. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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15 pages, 2973 KiB

Open AccessArticle

Predictive Cell Culture Time Evolution Based on Electric Models

by Juan Alfonso Serrano, Pablo Pérez, Paula Daza, Gloria Huertas and Alberto Yúfera

Biosensors 2023, 13(6), 668; https://doi.org/10.3390/bios13060668 - 20 Jun 2023

Cited by 2 | Viewed by 1309

Abstract

Obtaining cell concentration measurements from a culture assay by using bioimpedance is a very useful method that can be used to translate impedances to cell concentration values. The purpose of this study was to find a method to obtain the cell concentration values of a given cell culture assay in real time by using an oscillator as the measurement circuit. From a basic cell–electrode model, enhanced models of a cell culture immersed in a saline solution (culture medium) were derived. These models were used as part of a fitting routine to estimate the cell concentration in a cell culture in real time by using the oscillation frequency and amplitude delivered by the measurement circuits proposed by previous authors. Using real experimental data (the frequency and amplitude of oscillations) that were obtained by connecting the cell culture to an oscillator as the load, the fitting routine was simulated, and real-time data of the cell concentration were obtained. These results were compared to concentration data that were obtained by using traditional optical methods for counting. In addition, the error that we obtained was divided and analyzed in two parts: the first part of the experiment (when the few cells were adapting to the culture medium) and the second part of the experiment (when the cells exponentially grew until they completely covered the well). Low error values were obtained during the growth phase of the cell culture (the relevant phase); therefore, the results obtained were considered promising and show that the fitting routine is valid and that the cell concentration can be measured in real time by using an oscillator. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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14 pages, 2840 KiB

Open AccessArticle

Bioluminescent RIPoptosome Assay for FADD/RIPK1 Interaction Based on Split Luciferase Assay in a Human Neuroblastoma Cell Line SH-SY5Y

by Parisa Ghanavatian, Hossein Salehi-Sedeh, Farangis Ataei and Saman Hosseinkhani

Biosensors 2023, 13(2), 297; https://doi.org/10.3390/bios13020297 - 20 Feb 2023

Cited by 3 | Viewed by 3223

Abstract

Different programed cell death (PCD) modalities involve protein–protein interactions in large complexes. Tumor necrosis factor α (TNFα) stimulated assembly of receptor-interacting protein kinase 1 (RIPK1)/Fas-associated death domain (FADD) interaction forms Ripoptosome complex that may cause either apoptosis or necroptosis. The present study addresses the interaction of RIPK1 and FADD in TNFα signaling by fusion of C-terminal (CLuc) and N-terminal (NLuc) luciferase fragments to RIPK1-CLuc (R1C) or FADD-NLuc (FN) in a caspase 8 negative neuroblastic SH-SY5Y cell line, respectively. In addition, based on our findings, an RIPK1 mutant (R1C K612R) had less interaction with FN, resulting in increasing cell viability. Moreover, presence of a caspase inhibitor (zVAD.fmk) increases luciferase activity compared to Smac mimetic BV6 (B), TNFα -induced (T) and non-induced cell. Furthermore, etoposide decreased luciferase activity, but dexamethasone was not effective in SH-SY5Y. This reporter assay might be used to evaluate basic aspects of this interaction as well as for screening of necroptosis and apoptosis targeting drugs with potential therapeutic application. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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13 pages, 1509 KiB

Open AccessArticle

A Luciferase Mutant with Improved Brightness and Stability for Whole-Cell Bioluminescent Biosensors and In Vitro Biosensing

by Maria Maddalena Calabretta, Denise Gregucci, Héctor Martínez-Pérez-Cejuela and Elisa Michelini

Biosensors 2022, 12(9), 742; https://doi.org/10.3390/bios12090742 - 9 Sep 2022

Cited by 16 | Viewed by 4786

Abstract

The availability of new bioluminescent proteins with tuned properties, both in terms of emission wavelength, kinetics and protein stability, is highly valuable in the bioanalytical field, with the potential to improve the sensitivity and analytical performance of the currently used methods for ATP detection, whole-cell biosensors, and viability assays among others. We present a new luciferase mutant, called BgLuc, suitable for developing whole-cell biosensors and in vitro biosensors characterized by a bioluminescence maximum of 548 nm, narrow emission bandwidth, favorable kinetic properties, and excellent pH- and thermo-stabilities at 37 and 45 °C and pH from 5.0 to 8.0. We assessed the suitability of this new luciferase for whole-cell biosensing with a cell-based bioreporter assay for Nuclear Factor-kappa B (NF-kB) signal transduction pathway using 2D and 3D human embryonic kidney (HEK293T) cells, and for ATP detection with the purified enzyme. In both cases the luciferase showed suitable for sensitive detection of the target analytes, with better or similar performance than the commercial counterparts. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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Review

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30 pages, 7707 KiB

Open AccessReview

Recent Advances in Electrochemical Detection of Cell Energy Metabolism

by Kyeong-Mo Koo, Chang-Dae Kim and Tae-Hyung Kim

Biosensors 2024, 14(1), 46; https://doi.org/10.3390/bios14010046 - 15 Jan 2024

Viewed by 3581

Abstract

Cell energy metabolism is a complex and multifaceted process by which some of the most important nutrients, particularly glucose and other sugars, are transformed into energy. This complexity is a result of dynamic interactions between multiple components, including ions, metabolic intermediates, and products that arise from biochemical reactions, such as glycolysis and mitochondrial oxidative phosphorylation (OXPHOS), the two main metabolic pathways that provide adenosine triphosphate (ATP), the main source of chemical energy driving various physiological activities. Impaired cell energy metabolism and perturbations or dysfunctions in associated metabolites are frequently implicated in numerous diseases, such as diabetes, cancer, and neurodegenerative and cardiovascular disorders. As a result, altered metabolites hold value as potential disease biomarkers. Electrochemical biosensors are attractive devices for the early diagnosis of many diseases and disorders based on biomarkers due to their advantages of efficiency, simplicity, low cost, high sensitivity, and high selectivity in the detection of anomalies in cellular energy metabolism, including key metabolites involved in glycolysis and mitochondrial processes, such as glucose, lactate, nicotinamide adenine dinucleotide (NADH), reactive oxygen species (ROS), glutamate, and ATP, both in vivo and in vitro. This paper offers a detailed examination of electrochemical biosensors for the detection of glycolytic and mitochondrial metabolites, along with their many applications in cell chips and wearable sensors. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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12 pages, 663 KiB

Open AccessReview

Three-Dimensional Printing and Its Potential to Develop Sensors for Cancer with Improved Performance

by João B. M. Rocha Neto, Juliana Coatrini Soares, Guilherme A. Longhitano, Andrey Coatrini-Soares, Hernandes F. Carvalho, Osvaldo N. Oliveira, Jr., Marisa M. Beppu and Jorge V. L. da Silva

Biosensors 2022, 12(9), 685; https://doi.org/10.3390/bios12090685 - 26 Aug 2022

Cited by 6 | Viewed by 2285

Abstract

Cancer is the second leading cause of death globally and early diagnosis is the best strategy to reduce mortality risk. Biosensors to detect cancer biomarkers are based on various principles of detection, including electrochemical, optical, electrical, and mechanical measurements. Despite the advances in the identification of biomarkers and the conventional 2D manufacturing processes, detection methods for cancers still require improvements in terms of selectivity and sensitivity, especially for point-of-care diagnosis. Three-dimensional printing may offer the features to produce complex geometries in the design of high-precision, low-cost sensors. Three-dimensional printing, also known as additive manufacturing, allows for the production of sensitive, user-friendly, and semi-automated sensors, whose composition, geometry, and functionality can be controlled. This paper reviews the recent use of 3D printing in biosensors for cancer diagnosis, highlighting the main advantages and advances achieved with this technology. Additionally, the challenges in 3D printing technology for the mass production of high-performance biosensors for cancer diagnosis are addressed. Full article

(This article belongs to the Special Issue Cell-Based Biosensors for Rapid Detection and Monitoring)

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