Over the past decades, feature-based statistical machine learning and deep neural networks have been extensively utilized for automatic sleep stage classification (ASSC). Feature-based approaches offer clear insights into sleep characteristics and require low computational power but often fail to capture the spatial-temporal context of the data. In contrast, deep neural networks can process raw sleep signals directly and deliver superior performance. However, their overfitting, inconsistent accuracy, and computational cost were the primary drawbacks that limited their end-user acceptance. To address these challenges, we developed a novel neural network model, MLS-Net, which integrates the strengths of neural networks and feature extraction for automated sleep staging in mice. MLS-Net leverages temporal and spectral features from multimodal signals, such as EEG, EMG, and eye movements (EMs), as inputs and incorporates a bidirectional Long Short-Term Memory (bi-LSTM) to effectively capture the spatial-temporal nonlinear characteristics inherent in sleep signals. Our studies demonstrate that MLS-Net achieves an overall classification accuracy of 90.4% and REM state precision of 91.1%, sensitivity of 84.7%, and an F1-Score of 87.5% in mice, outperforming other neural network and feature-based algorithms in our multimodal dataset.
{"title":"MLS-Net: An Automatic Sleep Stage Classifier Utilizing Multimodal Physiological Signals in Mice.","authors":"Chengyong Jiang, Wenbin Xie, Jiadong Zheng, Biao Yan, Junwen Luo, Jiayi Zhang","doi":"10.3390/bios14080406","DOIUrl":"10.3390/bios14080406","url":null,"abstract":"<p><p>Over the past decades, feature-based statistical machine learning and deep neural networks have been extensively utilized for automatic sleep stage classification (ASSC). Feature-based approaches offer clear insights into sleep characteristics and require low computational power but often fail to capture the spatial-temporal context of the data. In contrast, deep neural networks can process raw sleep signals directly and deliver superior performance. However, their overfitting, inconsistent accuracy, and computational cost were the primary drawbacks that limited their end-user acceptance. To address these challenges, we developed a novel neural network model, MLS-Net, which integrates the strengths of neural networks and feature extraction for automated sleep staging in mice. MLS-Net leverages temporal and spectral features from multimodal signals, such as EEG, EMG, and eye movements (EMs), as inputs and incorporates a bidirectional Long Short-Term Memory (bi-LSTM) to effectively capture the spatial-temporal nonlinear characteristics inherent in sleep signals. Our studies demonstrate that MLS-Net achieves an overall classification accuracy of 90.4% and REM state precision of 91.1%, sensitivity of 84.7%, and an F1-Score of 87.5% in mice, outperforming other neural network and feature-based algorithms in our multimodal dataset.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11353116/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142082282","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Here, we report an ultrasoft extra long-lasting, reusable hydrogel-based sensor that enables high-quality electrophysiological recording with low-motion artifacts. The developed sensor can be used and stored in an ambient environment for months before being reused. The developed sensor is made of a self-adhesive electrical-conductivity-enhanced ultrasoft hydrogel mounted in an Ecoflex-based frame. The hydrogel's conductivity was enhanced by incorporating polypyrrole (PPy), resulting in a conductivity of 0.25 S m-1. Young's modulus of the sensor is only 12.9 kPa, and it is stretchable up to 190%. The sensor was successfully used for electrocardiography (ECG) and electromyography (EMG). Our results indicate that using the developed hydrogel-based sensor, the signal-to-noise ratio of recorded electrophysiological signals was improved in comparison to that when medical-grade silver/silver chloride (Ag/AgCl) wet gel electrodes were used (33.55 dB in comparison to 22.16 dB). Due to the ultra-softness, high stretchability, and self-adhesion of the developed sensor, it can conform to the skin and, therefore, shows low susceptibility to motion. In addition, the sensor shows no sign of irritation or allergic reaction, which usually occurs after long-term wearing of medical-grade Ag/AgCl wet gel electrodes on the skin. Further, the sensor is fabricated using a low-cost and scalable fabrication process.
在此,我们报告了一种超软、超长寿命、可重复使用的基于水凝胶的传感器,它能以低运动伪影进行高质量的电生理记录。开发的传感器可在环境中使用和储存数月后再重复使用。开发的传感器由自粘性导电性增强超软水凝胶制成,安装在基于 Ecoflex 的框架中。通过加入聚吡咯(PPy)增强了水凝胶的导电性,使其导电率达到 0.25 S m-1。传感器的杨氏模量仅为 12.9 kPa,可拉伸达 190%。该传感器已成功用于心电图(ECG)和肌电图(EMG)。我们的研究结果表明,与使用医用级银/氯化银(Ag/AgCl)湿凝胶电极相比,使用所开发的水凝胶传感器记录的电生理信号的信噪比有所提高(33.55 dB 比 22.16 dB)。由于所开发的传感器具有超柔软、高伸展性和自粘性,可以贴合皮肤,因此对运动的敏感性较低。此外,该传感器不会出现刺激或过敏反应,而长期在皮肤上佩戴医用级银/氯化银湿凝胶电极通常会出现这种情况。此外,该传感器采用低成本、可扩展的制造工艺制成。
{"title":"Ultrasoft Long-Lasting Reusable Hydrogel-Based Sensor Patch for Biosignal Recording.","authors":"Alexandre Tessier, Shuyun Zhuo, Shideh Kabiri Ameri","doi":"10.3390/bios14080405","DOIUrl":"10.3390/bios14080405","url":null,"abstract":"<p><p>Here, we report an ultrasoft extra long-lasting, reusable hydrogel-based sensor that enables high-quality electrophysiological recording with low-motion artifacts. The developed sensor can be used and stored in an ambient environment for months before being reused. The developed sensor is made of a self-adhesive electrical-conductivity-enhanced ultrasoft hydrogel mounted in an Ecoflex-based frame. The hydrogel's conductivity was enhanced by incorporating polypyrrole (PPy), resulting in a conductivity of 0.25 S m<sup>-1</sup>. Young's modulus of the sensor is only 12.9 kPa, and it is stretchable up to 190%. The sensor was successfully used for electrocardiography (ECG) and electromyography (EMG). Our results indicate that using the developed hydrogel-based sensor, the signal-to-noise ratio of recorded electrophysiological signals was improved in comparison to that when medical-grade silver/silver chloride (Ag/AgCl) wet gel electrodes were used (33.55 dB in comparison to 22.16 dB). Due to the ultra-softness, high stretchability, and self-adhesion of the developed sensor, it can conform to the skin and, therefore, shows low susceptibility to motion. In addition, the sensor shows no sign of irritation or allergic reaction, which usually occurs after long-term wearing of medical-grade Ag/AgCl wet gel electrodes on the skin. Further, the sensor is fabricated using a low-cost and scalable fabrication process.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11352425/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142082318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Chang-Dae Kim, Kyeong-Mo Koo, Hyung-Joo Kim, Tae-Hyung Kim
Challenges in directed differentiation and survival limit the clinical use of stem cells despite their promising therapeutic potential in regenerative medicine. Nanotechnology has emerged as a powerful tool to address these challenges and enable precise control over stem cell fate. In particular, nanomaterials can mimic an extracellular matrix and provide specific cues to guide stem cell differentiation and proliferation in the field of nanotechnology. For instance, recent studies have demonstrated that nanostructured surfaces and scaffolds can enhance stem cell lineage commitment modulated by intracellular regulation and external stimulation, such as reactive oxygen species (ROS) scavenging, autophagy, or electrical stimulation. Furthermore, nanoframework-based and upconversion nanoparticles can be used to deliver bioactive molecules, growth factors, and genetic materials to facilitate stem cell differentiation and tissue regeneration. The increasing use of nanostructures in stem cell research has led to the development of new therapeutic approaches. Therefore, this review provides an overview of recent advances in nanomaterials for modulating stem cell differentiation, including metal-, carbon-, and peptide-based strategies. In addition, we highlight the potential of these nano-enabled technologies for clinical applications of stem cell therapy by focusing on improving the differentiation efficiency and therapeutics. We believe that this review will inspire researchers to intensify their efforts and deepen their understanding, thereby accelerating the development of stem cell differentiation modulation, therapeutic applications in the pharmaceutical industry, and stem cell therapeutics.
{"title":"Recent Advances in Nanomaterials for Modulation of Stem Cell Differentiation and Its Therapeutic Applications.","authors":"Chang-Dae Kim, Kyeong-Mo Koo, Hyung-Joo Kim, Tae-Hyung Kim","doi":"10.3390/bios14080407","DOIUrl":"10.3390/bios14080407","url":null,"abstract":"<p><p>Challenges in directed differentiation and survival limit the clinical use of stem cells despite their promising therapeutic potential in regenerative medicine. Nanotechnology has emerged as a powerful tool to address these challenges and enable precise control over stem cell fate. In particular, nanomaterials can mimic an extracellular matrix and provide specific cues to guide stem cell differentiation and proliferation in the field of nanotechnology. For instance, recent studies have demonstrated that nanostructured surfaces and scaffolds can enhance stem cell lineage commitment modulated by intracellular regulation and external stimulation, such as reactive oxygen species (ROS) scavenging, autophagy, or electrical stimulation. Furthermore, nanoframework-based and upconversion nanoparticles can be used to deliver bioactive molecules, growth factors, and genetic materials to facilitate stem cell differentiation and tissue regeneration. The increasing use of nanostructures in stem cell research has led to the development of new therapeutic approaches. Therefore, this review provides an overview of recent advances in nanomaterials for modulating stem cell differentiation, including metal-, carbon-, and peptide-based strategies. In addition, we highlight the potential of these nano-enabled technologies for clinical applications of stem cell therapy by focusing on improving the differentiation efficiency and therapeutics. We believe that this review will inspire researchers to intensify their efforts and deepen their understanding, thereby accelerating the development of stem cell differentiation modulation, therapeutic applications in the pharmaceutical industry, and stem cell therapeutics.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11352443/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142082314","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The convenient and sensitive detection of metabolites is of great significance for understanding human health status and drug development. Solid-phase electrochemiluminescence (ECL) enzyme electrodes show great potential in metabolite detection based on the enzyme-catalyzed reaction product hydrogen peroxide (H2O2). Herein, a solid-phase ECL enzyme sensor was fabricated based on a confined emitter and an immobilized enzyme using electrostatic nanocage array, constructing a platform for the sensitive detection of cholesterol. The electrostatic cage nanochannel consists of a bipolar and bilayer vertically aligned mesoporous silica film (bp-VMSF). The upper layer of bp-VMSF is an amino-modified, positively charged VMSF (p-VMSF), and the lower layer is a negatively charged VMSF (n-VMSF). The most commonly used ECL probe tris(bipyridine)ruthenium(II) (Ru(bpy)32+) is fixed in n-VMSF by electrostatic adsorption from n-VMSF and electrostatic repulsion from the upper p-VMSF, generating significantly enhanced and stable ECL signals. The successful preparation of the electrostatic cage was characterized by scanning electron microscopy (SEM) and electrochemical methods. After amino groups on the outer surface of bp-VMSF were derivatized with aldehyde, cholesterol oxidase (ChOx) molecules were covalently immobilized. The successful construction of the enzyme electrode was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). When the corresponding enzyme substrate, cholesterol, was present in the solution, the ECL signal of Ru(bpy)32+ was quenched by the enzyme-catalyzed reaction product H2O2, enabling the high-sensitivity detection of cholesterol. The linear range for detecting cholesterol was from 0.05 mM to 5.0 mM, with a limit of detection (LOD) of 1.5 μM.
{"title":"Solid-Phase Electrochemiluminescence Enzyme Electrodes Based on Nanocage Arrays for Highly Sensitive Detection of Cholesterol.","authors":"Xinying Ma, Zhe Zhang, Yanyan Zheng, Jiyang Liu","doi":"10.3390/bios14080403","DOIUrl":"10.3390/bios14080403","url":null,"abstract":"<p><p>The convenient and sensitive detection of metabolites is of great significance for understanding human health status and drug development. Solid-phase electrochemiluminescence (ECL) enzyme electrodes show great potential in metabolite detection based on the enzyme-catalyzed reaction product hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>). Herein, a solid-phase ECL enzyme sensor was fabricated based on a confined emitter and an immobilized enzyme using electrostatic nanocage array, constructing a platform for the sensitive detection of cholesterol. The electrostatic cage nanochannel consists of a bipolar and bilayer vertically aligned mesoporous silica film (bp-VMSF). The upper layer of bp-VMSF is an amino-modified, positively charged VMSF (p-VMSF), and the lower layer is a negatively charged VMSF (n-VMSF). The most commonly used ECL probe tris(bipyridine)ruthenium(II) (Ru(bpy)<sub>3</sub><sup>2+</sup>) is fixed in n-VMSF by electrostatic adsorption from n-VMSF and electrostatic repulsion from the upper p-VMSF, generating significantly enhanced and stable ECL signals. The successful preparation of the electrostatic cage was characterized by scanning electron microscopy (SEM) and electrochemical methods. After amino groups on the outer surface of bp-VMSF were derivatized with aldehyde, cholesterol oxidase (ChOx) molecules were covalently immobilized. The successful construction of the enzyme electrode was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). When the corresponding enzyme substrate, cholesterol, was present in the solution, the ECL signal of Ru(bpy)<sub>3</sub><sup>2+</sup> was quenched by the enzyme-catalyzed reaction product H<sub>2</sub>O<sub>2</sub>, enabling the high-sensitivity detection of cholesterol. The linear range for detecting cholesterol was from 0.05 mM to 5.0 mM, with a limit of detection (LOD) of 1.5 μM.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11353124/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142082316","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Baviththira Suganthan, Ashley M Rogers, Clay S Crippen, Hamid Asadi, Or Zolti, Christine M Szymanski, Ramaraja P Ramasamy
Campylobacter jejuni is a common foodborne pathogen found in poultry that can cause severe life-threatening illnesses in humans. It is important to detect this pathogen in food to manage foodborne outbreaks. This study reports a novel impedimetric phage protein-based biosensor to detect C. jejuni NCTC 11168 at 100 CFU/mL concentrations using a genetically engineered receptor-binding phage protein, FlaGrab, as a bioreceptor. The electrochemical impedance spectroscopy (EIS) technique was employed to measure changes in resistance upon interaction with C. jejuni. The sensitivity of the phage protein-immobilized electrode was assessed using the various concentrations of C. jejuni NCTC 11168 ranging from 102-109 colony forming units (CFU)/mL). The change transfer resistance of the biosensor increased with increasing numbers of C. jejuni NCTC 11168 cells. The detection limit was determined to be approximately 103 CFU/mL in the buffer and 102 CFU/mL in the ex vivo samples. Salmonella enterica subsp. enterica serotype Typhimurium-291RH and Listeria monocytogenes Scott A were used as nontarget bacterial cells to assess the specificity of the developed biosensor. Results showed that the developed biosensor was highly specific toward the target C. jejuni NCTC 11168, as no signal was observed for the nontarget bacterial cells.
{"title":"A Bacteriophage Protein-Based Impedimetric Electrochemical Biosensor for the Detection of <i>Campylobacter jejuni</i>.","authors":"Baviththira Suganthan, Ashley M Rogers, Clay S Crippen, Hamid Asadi, Or Zolti, Christine M Szymanski, Ramaraja P Ramasamy","doi":"10.3390/bios14080402","DOIUrl":"10.3390/bios14080402","url":null,"abstract":"<p><p><i>Campylobacter jejuni</i> is a common foodborne pathogen found in poultry that can cause severe life-threatening illnesses in humans. It is important to detect this pathogen in food to manage foodborne outbreaks. This study reports a novel impedimetric phage protein-based biosensor to detect <i>C. jejuni</i> NCTC 11168 at 100 CFU/mL concentrations using a genetically engineered receptor-binding phage protein, FlaGrab, as a bioreceptor. The electrochemical impedance spectroscopy (EIS) technique was employed to measure changes in resistance upon interaction with <i>C. jejuni</i>. The sensitivity of the phage protein-immobilized electrode was assessed using the various concentrations of <i>C. jejuni</i> NCTC 11168 ranging from 10<sup>2</sup>-10<sup>9</sup> colony forming units (CFU)/mL). The change transfer resistance of the biosensor increased with increasing numbers of <i>C. jejuni</i> NCTC 11168 cells. The detection limit was determined to be approximately 10<sup>3</sup> CFU/mL in the buffer and 10<sup>2</sup> CFU/mL in the ex vivo samples. <i>Salmonella enterica</i> subsp. <i>enterica</i> serotype Typhimurium-291RH and <i>Listeria monocytogenes</i> Scott A were used as nontarget bacterial cells to assess the specificity of the developed biosensor. Results showed that the developed biosensor was highly specific toward the target <i>C. jejuni</i> NCTC 11168, as no signal was observed for the nontarget bacterial cells.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11352438/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142082272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Liliya I Mukhametova, Dmitry O Zherdev, Sergei A Eremin, Anton N Kuznetsov, Viktor I Yudin, Oleg D Sclyarov, Olesia V Babicheva, Anton V Motorygin, Yury E Tsvetkov, Vadim B Krylov, Nikolay E Nifantiev
Brucellosis in animals is an infectious disease caused by bacteria of the genus Brucella. Known methods for diagnosing brucellosis face some challenges, due to the difficulties in isolating and standardizing the natural brucellosis antigen. In this work, we investigated the possibility of using the fluorescence polarization assay (FPA) with synthetic glycoconjugate biosensing tracers to detect antibodies against Brucella as a new methodology for diagnosing brucellosis. Based on the received results, the synthetic fluorescein-labeled trisaccharide tracer is most effective for Brucellosis detection. This tracer is structurally related to the immune determinant fragment of the Brucella LPS buildup of N-formyl-d-perosamine units, connected via α-(1→3)-linkage at the non-reducing end and α-(1→2)-linkage at the reducing end. The sensitivity and specificity in the case of the use of trisaccharide tracer 3b were 71% and 100% (Yuden's method) and 87% and 88% (Euclidean method), respectively, which is comparable with the diagnostic efficiency of traditionally used serological methods, such as the agglutination test (AT), complement fixation test (CFT), and Rose Bengal test (RBT). Given the known advantages of FPA (e.g., speed, compactness of the equipment, and standard reagents) and the increased specificity of the developed test system, it would be appropriate to consider its widespread use for the diagnosis of brucellosis in animals, including rapid testing in the field.
{"title":"Applying a Fluorescence Polarization Assay for Detection of Brucellosis in Animals Using the Fluorescently Labeled Synthetic Oligosaccharides as Biosensing Tracer.","authors":"Liliya I Mukhametova, Dmitry O Zherdev, Sergei A Eremin, Anton N Kuznetsov, Viktor I Yudin, Oleg D Sclyarov, Olesia V Babicheva, Anton V Motorygin, Yury E Tsvetkov, Vadim B Krylov, Nikolay E Nifantiev","doi":"10.3390/bios14080404","DOIUrl":"10.3390/bios14080404","url":null,"abstract":"<p><p>Brucellosis in animals is an infectious disease caused by bacteria of the genus <i>Brucella</i>. Known methods for diagnosing brucellosis face some challenges, due to the difficulties in isolating and standardizing the natural brucellosis antigen. In this work, we investigated the possibility of using the fluorescence polarization assay (FPA) with synthetic glycoconjugate biosensing tracers to detect antibodies against <i>Brucella</i> as a new methodology for diagnosing brucellosis. Based on the received results, the synthetic fluorescein-labeled trisaccharide tracer is most effective for Brucellosis detection. This tracer is structurally related to the immune determinant fragment of the <i>Brucella</i> LPS buildup of N-formyl-d-perosamine units, connected via α-(1→3)-linkage at the non-reducing end and α-(1→2)-linkage at the reducing end. The sensitivity and specificity in the case of the use of trisaccharide tracer <b>3b</b> were 71% and 100% (Yuden's method) and 87% and 88% (Euclidean method), respectively, which is comparable with the diagnostic efficiency of traditionally used serological methods, such as the agglutination test (AT), complement fixation test (CFT), and Rose Bengal test (RBT). Given the known advantages of FPA (e.g., speed, compactness of the equipment, and standard reagents) and the increased specificity of the developed test system, it would be appropriate to consider its widespread use for the diagnosis of brucellosis in animals, including rapid testing in the field.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11352515/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142082274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Juliane Fjelrad Christfort, Morgane Ortis, Hau Van Nguyen, Robert Marsault, Alain Doglio
Static well plates remain the gold standard to study viral infections in vitro, but they cannot accurately mimic dynamic viral infections as they occur in the human body. Therefore, we established a dynamic cell culture platform, based on centrifugal microfluidics, to study viral infections in perfusion. To do so, we used human primary periodontal dental ligament (PDL) cells and herpes simplex virus-1 (HSV-1) as a case study. By microscopy, we confirmed that the PDL cells efficiently attached and grew in the chip. Successful dynamic viral infection of perfused PDL cells was monitored using fluorescent imaging and RT-qPCR-based experiments. Remarkably, viral infection in flow resulted in a gradient of HSV-1-infected cells gradually decreasing from the cell culture chamber entrance towards its end. The perfusion of acyclovir in the chip prevented HSV-1 spreading, demonstrating the usefulness of such a platform for monitoring the effects of antiviral drugs. In addition, the innate antiviral response of PDL cells, measured by interferon gene expression, increased significantly over time in conventional static conditions compared to the perfusion model. These results provide evidence suggesting that dynamic viral infections differ from conventional static infections, which highlights the need for more physiologically relevant in vitro models to study viral infections.
{"title":"Centrifugal Microfluidic Cell Culture Platform for Physiologically Relevant Virus Infection Studies: A Case Study with HSV-1 Infection of Periodontal Cells.","authors":"Juliane Fjelrad Christfort, Morgane Ortis, Hau Van Nguyen, Robert Marsault, Alain Doglio","doi":"10.3390/bios14080401","DOIUrl":"10.3390/bios14080401","url":null,"abstract":"<p><p>Static well plates remain the gold standard to study viral infections in vitro, but they cannot accurately mimic dynamic viral infections as they occur in the human body. Therefore, we established a dynamic cell culture platform, based on centrifugal microfluidics, to study viral infections in perfusion. To do so, we used human primary periodontal dental ligament (PDL) cells and herpes simplex virus-1 (HSV-1) as a case study. By microscopy, we confirmed that the PDL cells efficiently attached and grew in the chip. Successful dynamic viral infection of perfused PDL cells was monitored using fluorescent imaging and RT-qPCR-based experiments. Remarkably, viral infection in flow resulted in a gradient of HSV-1-infected cells gradually decreasing from the cell culture chamber entrance towards its end. The perfusion of acyclovir in the chip prevented HSV-1 spreading, demonstrating the usefulness of such a platform for monitoring the effects of antiviral drugs. In addition, the innate antiviral response of PDL cells, measured by interferon gene expression, increased significantly over time in conventional static conditions compared to the perfusion model. These results provide evidence suggesting that dynamic viral infections differ from conventional static infections, which highlights the need for more physiologically relevant in vitro models to study viral infections.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11352947/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142082277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Taeha Lee, Da Yeon Cheong, Kang Hyun Lee, Jae Hyun You, Jinsung Park, Gyudo Lee
Quantifying the formation and decomposition of amyloid is a crucial issue in the development of new drugs and therapies for treating amyloidosis. The current technologies for grasping amyloid formation and decomposition include fluorescence analysis using thioflavin-T, secondary structure analysis using circular dichroism, and image analysis using atomic force microscopy or transmission electron microscopy. These technologies typically require spectroscopic devices or expensive nanoscale imaging equipment and involve lengthy analysis, which limits the rapid screening of amyloid-degrading drugs. In this study, we introduce a technology for rapidly assessing amyloid decomposition using capillary flow-based paper (CFP). Amyloid solutions exhibit gel-like physical properties due to insoluble denatured polymers, resulting in a shorter flow distance on CFP compared to pure water. Experimental conditions were established to consistently control the flow distance based on a hen-egg-white lysozyme amyloid solution. It was confirmed that as amyloid is decomposed by trypsin, the flow distance increases on the CFP. Our method is highly useful for detecting changes in the gel properties of amyloid solutions within a minute, and we anticipate its use in the rapid, large-scale screening of anti-amyloid agents in the future.
{"title":"Capillary Flow-Based One-Minute Quantification of Amyloid Proteolysis.","authors":"Taeha Lee, Da Yeon Cheong, Kang Hyun Lee, Jae Hyun You, Jinsung Park, Gyudo Lee","doi":"10.3390/bios14080400","DOIUrl":"10.3390/bios14080400","url":null,"abstract":"<p><p>Quantifying the formation and decomposition of amyloid is a crucial issue in the development of new drugs and therapies for treating amyloidosis. The current technologies for grasping amyloid formation and decomposition include fluorescence analysis using thioflavin-T, secondary structure analysis using circular dichroism, and image analysis using atomic force microscopy or transmission electron microscopy. These technologies typically require spectroscopic devices or expensive nanoscale imaging equipment and involve lengthy analysis, which limits the rapid screening of amyloid-degrading drugs. In this study, we introduce a technology for rapidly assessing amyloid decomposition using capillary flow-based paper (CFP). Amyloid solutions exhibit gel-like physical properties due to insoluble denatured polymers, resulting in a shorter flow distance on CFP compared to pure water. Experimental conditions were established to consistently control the flow distance based on a hen-egg-white lysozyme amyloid solution. It was confirmed that as amyloid is decomposed by trypsin, the flow distance increases on the CFP. Our method is highly useful for detecting changes in the gel properties of amyloid solutions within a minute, and we anticipate its use in the rapid, large-scale screening of anti-amyloid agents in the future.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11353070/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142082276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Waleed A El-Said, Tamer S Saleh, Abdullah Saad Al-Bogami, Mohmmad Younus Wani, Jeong-Woo Choi
Pyocyanin is considered a maker of Pseudomonas aeruginosa (P. aeruginosa) infection. Pyocyanin is among the toxins released by the P. aeruginosa bacteria. Therefore, the development of a direct detection of PYO is crucial due to its importance. Among the different optical techniques, the Raman technique showed unique advantages because of its fingerprint data, no sample preparation, and high sensitivity besides its ease of use. Noble metal nanostructures were used to improve the Raman response based on the surface-enhanced Raman scattering (SERS) technique. Anodic metal oxide attracts much interest due to its unique morphology and applications. The porous metal structure provides a large surface area that could be used as a hard template for periodic nanostructure array fabrication. Porous shapes and sizes could be controlled by controlling the anodization parameters, including the anodization voltage, current, temperature, and time, besides the metal purity and the electrolyte type/concentration. The anodization of aluminum foil results in anodic aluminum oxide (AAO) formation with different roughness. Here, we will use the roughness as hotspot centers to enhance the Raman signals. Firstly, a thin film of gold was deposited to develop gold/alumina (Au/AAO) platforms and then applied as SERS-active surfaces. The morphology and roughness of the developed substrates were investigated using scanning electron microscopy (SEM) and atomic force microscopy (AFM) techniques. The Au/AAO substrates were used for monitoring pyocyanin secreted from Pseudomonas aeruginosa microorganisms based on the SERS technique. The results showed that the roughness degree affects the enhancement efficiency of this sensor. The high enhancement was obtained in the case of depositing a 30 nm layer of gold onto the second anodized substrates. The developed sensor showed high sensitivity toward pyocyanin with a limit of detection of 96 nM with a linear response over a dynamic range from 1 µM to 9 µM.
{"title":"Development of Novel Surface-Enhanced Raman Spectroscopy-Based Biosensors by Controlling the Roughness of Gold/Alumina Platforms for Highly Sensitive Detection of Pyocyanin Secreted from <i>Pseudomonas aeruginosa</i>.","authors":"Waleed A El-Said, Tamer S Saleh, Abdullah Saad Al-Bogami, Mohmmad Younus Wani, Jeong-Woo Choi","doi":"10.3390/bios14080399","DOIUrl":"10.3390/bios14080399","url":null,"abstract":"<p><p>Pyocyanin is considered a maker of <i>Pseudomonas aeruginosa</i> (<i>P. aeruginosa</i>) infection. Pyocyanin is among the toxins released by the <i>P. aeruginosa</i> bacteria. Therefore, the development of a direct detection of PYO is crucial due to its importance. Among the different optical techniques, the Raman technique showed unique advantages because of its fingerprint data, no sample preparation, and high sensitivity besides its ease of use. Noble metal nanostructures were used to improve the Raman response based on the surface-enhanced Raman scattering (SERS) technique. Anodic metal oxide attracts much interest due to its unique morphology and applications. The porous metal structure provides a large surface area that could be used as a hard template for periodic nanostructure array fabrication. Porous shapes and sizes could be controlled by controlling the anodization parameters, including the anodization voltage, current, temperature, and time, besides the metal purity and the electrolyte type/concentration. The anodization of aluminum foil results in anodic aluminum oxide (AAO) formation with different roughness. Here, we will use the roughness as hotspot centers to enhance the Raman signals. Firstly, a thin film of gold was deposited to develop gold/alumina (Au/AAO) platforms and then applied as SERS-active surfaces. The morphology and roughness of the developed substrates were investigated using scanning electron microscopy (SEM) and atomic force microscopy (AFM) techniques. The Au/AAO substrates were used for monitoring pyocyanin secreted from <i>Pseudomonas aeruginosa</i> microorganisms based on the SERS technique. The results showed that the roughness degree affects the enhancement efficiency of this sensor. The high enhancement was obtained in the case of depositing a 30 nm layer of gold onto the second anodized substrates. The developed sensor showed high sensitivity toward pyocyanin with a limit of detection of 96 nM with a linear response over a dynamic range from 1 µM to 9 µM.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11352426/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142082278","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Biosensors are used for the specific and sensitive detection of biomolecules. In conventional approaches, the suspected target molecules are bound to selected capture molecules and successful binding is indicated by additional labelling to enable optical readout. This labelling requires additional processing steps tailored to the application. While numerous label-free interaction assays exist, they often compromise on detection characteristics. In this context, we introduce a novel diffractometric biosensor, comprising a diffractive biosensor chip and an associated optical reader assembly. This innovative system can capture an entire assay, detecting various types of molecules in a label-free manner and present the results within in a single, comprehensive image. The applicability of the biosensor is assessed for the detection of viral DNA as well as proteins directly in human plasma, investigating different antigens. In our experiments, we achieve a detection limit of 4.2 pg/mm², which is comparable to other label-free optical biosensors. The simplicity and robustness of the method make it a compelling option for advancing biosensing technologies. This work contributes to the development of an imaging diffractometric biosensor with the potential for multiple applications in molecular interaction analysis.
生物传感器用于对生物分子进行特异而灵敏的检测。在传统方法中,可疑的目标分子与选定的捕获分子结合,并通过额外的标记来表明结合成功,从而实现光学读数。这种标记需要根据应用情况进行额外的处理步骤。虽然存在许多无标记相互作用检测方法,但它们往往在检测特性上大打折扣。在这种情况下,我们推出了一种新型衍射生物传感器,由衍射生物传感器芯片和相关的光学读取器组件组成。这种创新系统可以捕捉整个检测过程,以无标记方式检测各种类型的分子,并将检测结果以单一、全面的图像呈现出来。我们评估了这种生物传感器的适用性,它可以直接检测人体血浆中的病毒 DNA 和蛋白质,并对不同的抗原进行研究。在实验中,我们的检测限达到了 4.2 pg/mm²,与其他无标记光学生物传感器相当。该方法的简便性和稳健性使其成为推动生物传感技术发展的一个令人信服的选择。这项工作有助于开发一种成像衍射生物传感器,它在分子相互作用分析中具有多种应用潜力。
{"title":"Imaging Diffractometric Biosensors for Label-Free, Multi-Molecular Interaction Analysis.","authors":"Cornelia Reuter, Walter Hauswald, Sindy Burgold-Voigt, Uwe Hübner, Ralf Ehricht, Karina Weber, Juergen Popp","doi":"10.3390/bios14080398","DOIUrl":"10.3390/bios14080398","url":null,"abstract":"<p><p>Biosensors are used for the specific and sensitive detection of biomolecules. In conventional approaches, the suspected target molecules are bound to selected capture molecules and successful binding is indicated by additional labelling to enable optical readout. This labelling requires additional processing steps tailored to the application. While numerous label-free interaction assays exist, they often compromise on detection characteristics. In this context, we introduce a novel diffractometric biosensor, comprising a diffractive biosensor chip and an associated optical reader assembly. This innovative system can capture an entire assay, detecting various types of molecules in a label-free manner and present the results within in a single, comprehensive image. The applicability of the biosensor is assessed for the detection of viral DNA as well as proteins directly in human plasma, investigating different antigens. In our experiments, we achieve a detection limit of 4.2 pg/mm², which is comparable to other label-free optical biosensors. The simplicity and robustness of the method make it a compelling option for advancing biosensing technologies. This work contributes to the development of an imaging diffractometric biosensor with the potential for multiple applications in molecular interaction analysis.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9,"publicationDate":"2024-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11352734/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142082281","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}