Nucleic acid aptamer-based electrochemical sensor for the detection of serum P-tau231 and the instant screening test of Alzheimer's disease.

The instant screening of patients with a tendency towards developing Alzheimer's disease (AD) is significant for providing preventive measures and treatment. However, the current imaging-based technology cannot meet the requirements in the early stage. Developing biosensor-based liquid biopsy technology could be overcoming this bottleneck problem. Herein, we developed a simple, low-cost, and sensitive electrochemical aptamer biosensor for detecting phosphorylated tau protein threonine 231 (P-tau231), the earliest and one of the most efficacious abnormally elevated biomarkers of AD. Gold nanoparticles (AuNPs) were electrochemically synthesized on a glassy carbon electrode as the transducer, exhibiting excellent conductivity, and were applied to amplify the electrochemical signal. A nucleic acid aptamer was designed as the receptor to capture the P-tau231 protein, specifically through the formation of an aptamer-antigen complex. The proposed biosensor showed excellent sensitivity in detecting P-tau 231, with a broad linear detection range from 10 to 107 pg/mL and a limit of detection (LOD) of 2.31 pg/mL. The recoveries of the biosensor in human serum ranged from 97.59 to 103.26%, demonstrating that the biosensor could be used in complex practical samples. In addition, the results showed that the developed biosensor has good repeatability, reproducibility, and stability, which provides a novel method for the early screening of AD.

Advances in Engineered Nanoparticles for the Treatment of Ischemic Stroke by Enhancing Angiogenesis.

Angiogenesis, or the formation of new blood vessels, is a natural defensive mechanism that aids in the restoration of oxygen and nutrition delivery to injured brain tissue after an ischemic stroke. Angiogenesis, by increasing vessel development, may maintain brain perfusion, enabling neuronal survival, brain plasticity, and neurologic recovery. Induction of angiogenesis and the formation of new vessels aid in neurorepair processes such as neurogenesis and synaptogenesis. Advanced nano drug delivery systems hold promise for treatment stroke by facilitating efficient transportation across the the blood-brain barrier and maintaining optimal drug concentrations. Nanoparticle has recently been shown to greatly boost angiogenesis and decrease vascular permeability, as well as improve neuroplasticity and neurological recovery after ischemic stroke. We describe current breakthroughs in the development of nanoparticle-based treatments for better angiogenesis therapy for ischemic stroke employing polymeric nanoparticles, liposomes, inorganic nanoparticles, and biomimetic nanoparticles in this study. We outline new nanoparticles in detail, review the hurdles and strategies for conveying nanoparticle to lesions, and demonstrate the most recent advances in nanoparticle in angiogenesis for stroke treatment.

Rapid Evaluation of Antibacterial Carbohydrates on a Microfluidic Chip Integrated with the Impedimetric Neoglycoprotein Biosensor.

The colonization of some bacteria to their host cell is mediated by selective adhesion between adhesin and glycan. The evaluation of antiadhesive carbohydrates in vitro has great significance in discovering new antibacterial drugs. In this paper, a microfluidic chip integrated with impedimetric neoglycoprotein biosensors was developed to evaluate the antibacterial effect of carbohydrates. Mannosylated bovine serum albumin (Man-BSA) was taken as the neoglycoprotein and immobilized on the microelectrode-modified gold nanoparticles (Au NPs) to form a bionic glycoprotein nanosensing surface (Man-BSA/Au NPs). Salmonella typhimurium (S. typhimurium) was selected as a bacteria model owing to its selective adhesion to the mannose. Electrochemical impedance spectroscopy (EIS) was used to characterize the adhesion capacity of S. typhimurium to the Man-BSA/Au NPs and evaluate the antiadhesive efficacy of nine different carbohydrates. It was illustrated that the 4-methoxyphenyl-α-D-pyran mannoside (Phenyl-Man) and mannan peptide (Mannatide) showed excellent antiadhesive efficacy, with IC50 values of 0.086 mM and 0.094 mM, respectively. The microfluidic device developed in this study can be tested in multiple channels. Compared with traditional methods for evaluating the antibacterial drug in vitro, it has the advantages of being fast, convenient, and cost-effective.

In-situ synthesized V2CTx MXene-based immune tag for the electrochemical detection of Interleukin 6 (IL-6) from breast cancer cells.

Interleukin-6 (IL-6) is a proinflammatory cytokine with a critical role in immune regulation and treatment of many diseases, including breast cancer. Herein, we developed a novel V2CTx MXene-based immunosensor for rapid and accurate IL-6 detection. The chosen substrate was V2CTx, a 2-dimensional (2D) MXene nanomaterial with excellent electronic properties. Prussian blue (Fe4[Fe(CN)6]3), used for its electrochemical properties, and spindle-shaped gold nanoparticles (Au SSNPs), used to combine with antibodies, were in-situ synthesized on the surface of the MXene. The in-situ synthesis ensures a firm chemical connection compared to other tags formed by a less stable physical absorption. Inspired by a sandwich ELISA test, the modified V2CTx tag was captured by the electrode surface with cysteamine to detect the analyte, IL-6, after being attached with a capture antibody (cAb). Benefiting from an increased surface area, an enhanced charge transfer rate, and a firm connection of the tag, this biosensor exhibited excellent analytical performance. The high sensitivity, high selectivity, and wide detection range covering the IL-6 level of both healthy individuals and breast cancer patients were obtained to meet clinical demands. Herein, this V2CTx MXene-based immunosensor is a potential therapeutic and diagnostic point-of-care alternative to routine ELISA IL-6 detection methods.

3D co-culture of macrophages and fibroblasts in a sessile drop array for unveiling the role of macrophages in skin wound-healing.

Three-dimensional (3D) heterotypic multicellular spheroid models play important roles in researches of the proliferation and remodeling phases in wound healing. This study aimed to develop a sessile drop array to cultivate 3D spheroids and simulate wound healing stage in vitro using NIH-3T3 fibroblasts and M2-type macrophages. By the aid of the offset of surface tension and gravity, the sessile drop array is able to transfer cell suspensions to spheroids via the superhydrophobic surface of each microwell. Meanwhile, each microwell has a cylinder hole at its bottom that provides adequate oxygen to the spheroid. It demonstrated that the NIH-3T3 fibroblast spheroid and the 3T3 fibroblast/M2-type macrophage heterotypic multicellular spheroid can form and maintain physiological activities within nine days. In order to further investigate the structure without destroying the entire spheroid, we reconstructed its 3D morphology using transparent processing technology and the Z-stack function of confocal microscopy. Additionally, a nano antibody-based 3D immunostaining assay was used to analyze the proliferation and differentiation characteristics of these cells. It found that M2-type macrophages were capable of promoting the differentiation of 3T3 fibroblast spheroid. In this study, a novel, inexpensive platform was constructed for developing spheroids, as well as a 3D immunofluorescence method for investigating the macrophage-associated wound healing microenvironment.

A novel alternative strategy for monitoring and insight into liver fibrosis progression: The combination of surface-enhanced Raman spectroscopy (SERS) and gut microbiota.

Nowadays, the studies on the interaction and relationship between the intestinal microorganisms and liver diseases are increasing. However, it is still a huge challenge for the in-depth investigation and dynamic monitoring of such a complex network. Herein, a significant discovery was made. A strong association between gut microbial structural and functional genomics and SERS spectra of hepatocytes were revealed. Based on the study of gut microbes and SERS spectra, complementary information could be provided for the mechanism analysis of related diseases. Liver fibrosis, a chronic liver disease that lack specific cure was thus comprehensive studied. Liver targeting gold nanoparticle dimers were prepared as the SERS tags, and abundant SERS peak signals were acquired. Meanwhile, the gut microbiomes were also comparative studied. The changes of carbohydrates and lipids in liver cells were observed at the early stages of liver fibrosis, and TLR4 (toll-like receptors 4) was activated to elicit immune responses. Then again, oxidative stress, endotoxin and serum inflammatory factors were the major observations at the late stages. The SERS signals and the microbiome analysis were well confirmed and complemented each other, which suggested that the detection strategy could be another valuable method for the "gut-liver axis" study.

Bacterial identification and adhesive strength evaluation based on a mannose biosensor with dual-mode detection.

A biosensor integrated with mannose nano-surface was developed for the identification and adhesive strength evaluation of bacteria. Different bacteria were studied on the designed surface by both electrochemical impedance spectroscopy (EIS) and surface enhanced Raman spectroscopy (SERS). S. typhimurium and E. coli JM109 (type 1 pili) were found to be captured by the mannose nano-surface. SERS spectra were used to identify the species of captured bacteria by combing with partial least squares discriminant analysis (PLS-DA). Meanwhile, binding affinities of the two captured bacteria to mannose nano-surface were obtained by EIS measurements and Frumkin isotherm model analysis, which were 6.859 × 1023 M-1 and 2.054 × 1017 M-1 respectively. A higher binding affinity indicates a stronger adhesive strength. Hence the results show the S. typhimurium has a stronger adhesive strength to mannose. Normalized impedance change (NIC) was proved to have a positive relevant relationship with binding affinities, which could be used as an indicator for the adhesive strength of bacteria. It was demonstrated that 100% accuracy of bacteria species discrimination and good consistency of NIC and adhesive strength for blind samples. The developed biosensor can provide both qualitative and quantitative information of selective recognition between bacteria and mannose.

Laser-induced graphene (LIG)-driven medical sensors for health monitoring and diseases diagnosis.

Laser-induced graphene (LIG) is a class of three-dimensional (3D) porous carbon nanomaterial. It can be prepared by direct laser writing on some polymer materials in the air. Because of its features of simplicity, fast production, and excellent physicochemical properties, it was widely used in medical sensing devices. This minireview gives an overview of the characteristics of LIG and LIG-driven sensors. Various methods for preparing graphene were compared and discussed. The applications of the LIG in biochemical sensors for ions, small molecules, microRNA, protein, and cell detection were highlighted. LIG-based physical physiological sensors and wearable electronics for medical applications were also included. Finally, our insights into current challenges and prospects for LIG-based medical sensing devices were presented.

A Microfluidic Cell Co-Culture Chip for the Monitoring of Interactions between Macrophages and Fibroblasts.

Macrophages and fibroblasts are two types of important cells in wound healing. The development of novel platforms for studying the interrelationship between these two cells is crucial for the exploration of wound-healing mechanisms and drug development. In this study, a microfluidic chip composed of two layers was designed for the co-culturing of these two cells. An air valve was employed to isolate fibroblasts to simulate the wound-healing microenvironment. The confluence rate of fibroblasts in the co-culture system with different macrophages was explored to reflect the role of different macrophages in wound healing. It was demonstrated that M2-type macrophages could promote the activation and migration of fibroblasts and it can be inferred that they could promote the wound-healing process. The proposed microfluidic co-culture system was designed for non-contact cell-cell interactions, which has potential significance for the study of cell-cell interactions in biological processes such as wound healing, tumor microenvironment, and embryonic development.

Effect of vimentin on cell migration in collagen-coated microchannels: A mimetic physiological confined environment.

Cancer cell migration through tissue pores and tracks into the bloodstream is a critical biological step for cancer metastasis. Although in vivo studies have shown that expression of vimentin can induce invasive cell lines, its role in cell cytoskeleton reorganization and cell motility under in vitro physical confinement remains unknown. Here, a microfluidic device with cell culture chamber and collagen-coated microchannels was developed as an in vitro model for physiological confinement environments. Using this microchannel assay, we demonstrated that the knockdown of vimentin decreases 3T3 fibroblast cell directional migration speed in confined microchannels. Additionally, as cells form dynamic membranes that define the leading edge of motile cells, different leading edge morphologies of 3T3 fibroblast and 3T3 vimentin knockdown cells were observed. The leading edge morphology change under confinement can be explained by the effect of vimentin on cytoskeletal organization and focal adhesion. The microfluidic device integrated with a time-lapse microscope provided a new approach to study the effect of vimentin on cell adhesion, migration, and invasiveness.

Advancing Biosensors with Machine Learning.

Chemometrics play a critical role in biosensors-based detection, analysis, and diagnosis. Nowadays, as a branch of artificial intelligence (AI), machine learning (ML) have achieved impressive advances. However, novel advanced ML methods, especially deep learning, which is famous for image analysis, facial recognition, and speech recognition, has remained relatively elusive to the biosensor community. Herein, how ML can be beneficial to biosensors is systematically discussed. The advantages and drawbacks of most popular ML algorithms are summarized on the basis of sensing data analysis. Specially, deep learning methods such as convolutional neural network (CNN) and recurrent neural network (RNN) are emphasized. Diverse ML-assisted electrochemical biosensors, wearable electronics, SERS and other spectra-based biosensors, fluorescence biosensors and colorimetric biosensors are comprehensively discussed. Furthermore, biosensor networks and multibiosensor data fusion are introduced. This review will nicely bridge ML with biosensors, and greatly expand chemometrics for detection, analysis, and diagnosis.

Batch fabrication of electrochemical sensors on a glycol-modified polyethylene terephthalate-based microfluidic device.

Developing low-cost methods for the fabrication of electrochemical microfluidic devices is urgently needed for transferring such devices from fundamental research to daily-life technology. Herein, glycol-modified polyethylene terephthalate (PETG)-based microfluidic devices with embedded channels and gold film electrode (GFE) are developed by a one-step, low-cost, straightforward, and mass-producible method, and are sealed by a reversible hydrophilic tape-based mechanism. Easily accessible poly (methyl methacrylate) (PMMA), polyethylene terephthalate (polyester, PET), and PETG are explored as substrate options for fabricating electrochemical sensors. The results demonstrated that PETG can be an excellent substrate for fabricating the electrode. The electrochemical stability and morphology of the device are investigated. Both redox ions ([Fe(CN)6]3-/4-) and redox organic compounds (dopamine) are used as model analytes to prove the electrochemical performance of the device. The PETG-based microfluidic devices integrated with electrochemical sensors can be used as alternative electrochemical devices for the detection of biological and chemical analytes. Meanwhile, batch-fabricated flexible electrochemical sensors based on PETG film and their electrochemical performance are reported.

Diagnostic methods and potential portable biosensors for coronavirus disease 2019.

Timely detection and diagnosis are urgently needed to guide epidemiological measures, infection control, antiviral treatment, and vaccine research. In this review, biomarkers/indicators for diagnosis of coronavirus disease 2019 (COVID-19) or detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the environment are summarized and discussed. It is concluded that the detection methods targeting antibodies are not suitable for screening of early and asymptomatic cases since most patients had an antibody response at about 10 days after onset of symptoms. However, antibody detection methods can be combined with quantitative real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) to significantly improve the sensitivity and specificity of diagnosis, and boost vaccine research. Fast, sensitive and accurate detection methods targeting antigens need to be developed urgently. Various specimens for diagnosis or detection are compared and analyzed. Among them, deep throat saliva and induced sputum are desired for RT-qPCR test or other early detection technologies. Chest computerized tomography (CT) scan, RT-qPCR, lateral flow immunochromatographic strip (LFICS) for diagnosis of COVID-19 are summarized and compared. Specially, potential electrochemical (EC) biosensor, surface enhanced Raman scattering (SERS)-based biosensor, field-effect transistor (FET)-based biosensor, surface plasmon resonance (SPR)-based biosensor and artificial intelligence (AI) assisted diagnosis of COVID-19 are emphasized. Finally, some commercialized portable detection device, current challenges and future directions are discussed.

Molecularly Imprinted Polymers and Surface Imprinted Polymers Based Electrochemical Biosensor for Infectious Diseases.

Owing to their merits of simple, fast, sensitive, and low cost, electrochemical biosensors have been widely used for the diagnosis of infectious diseases. As a critical element, the receptor determines the selectivity, stability, and accuracy of the electrochemical biosensors. Molecularly imprinted polymers (MIPs) and surface imprinted polymers (SIPs) have great potential to be robust artificial receptors. Therefore, extensive studies have been reported to develop MIPs/SIPs for the detection of infectious diseases with high selectivity and reliability. In this review, we discuss mechanisms of recognition events between imprinted polymers with different biomarkers, such as signaling molecules, microbial toxins, viruses, and bacterial and fungal cells. Then, various preparation methods of MIPs/SIPs for electrochemical biosensors are summarized. Especially, the methods of electropolymerization and micro-contact imprinting are emphasized. Furthermore, applications of MIPs/SIPs based electrochemical biosensors for infectious disease detection are highlighted. At last, challenges and perspectives are discussed.

CuO/Cu composite nanospheres on a TiO2 nanotube array for amperometric sensing of glucose.

A non-enzymatic glucose sensor based on the use of CuO-Cu nanospheres placed on a TiO2 nanotube (TNT) array with excellent performance is described. The electrode was fabricated by coating the CuO-Cu nanospheres onto the TNT array through electrochemical deposition. The CuO-Cu nanospheres with a diameter of ~200 nm are well dispersed on the TNT surface, which warrants smooth interaction and a 3D nanostructure with high uniformity. The modified electrode was then used for amperometric determination of glucose in 0.1 M NaOH solution. Figures of merit include (a) a typical working voltage of 0.65 V (vs. Ag/AgCl). (b) a linear range as wide as from 0.2-90 mM, (c) good sensitivity (234 µA mM-1 cm-2), and a 19 nM lower detection limit. The sensor is selective over ascorbic acid (AA), dopamine (DA), uric acid (UA), lactose, sucrose, and fructose. Graphical abstract.

Monitoring of bacteria biofilms forming process by in-situ impedimetric biosensor chip.

A biosensor chip integrated interdigital microelectrodes was proposed and applied to monitor the formation process of Salmonella and E. coli biofilms in this paper. The biosensor chip was composed of a glass substrate with interdigital microelectrodes and PDMS layer with micro cavities. The electrochemical impedance spectroscopy (EIS) of Salmonella and E. coli biofilms was measured by the biosensor chip using alternating voltage of 100 mv in the frequency range from 1 Hz to 100 kHz for 48 h. It was illustrated that the changes of impedance spectroscopy of biofilms occurred with culture time. Furthermore, impedance spectroscopy of biofilms was fitted by an equivalent circuit model including the biofilms capacitance (Cb) and the biofilms resistance (Rb) parameters. The results indicated that the Cb presented a tendency to decrease first and then rise with culture time, while the Rb was in the opposite direction. These changing trends were consistent with the formation process of biofilms that bacteria adhered to electrodes surface, and then formed mature biofilms, finally escaped from biofilms. In addition, it was also demonstrated that the changing trends of Cb and Rb with culture time were quite different between Salmonella and E. coli. The results obtained by impedance detection were in accordance with the results of using crystal violet staining to analyze biofilms formation process, under the same conditions for bacterial culture. The biosensor chip provided a promising platform for further study of biofilms owing to its unique advantages of real time, continuity, and non-invasion for bacteria biofilms detection and in-situ monitoring.

[Detection of biofilms formation by electrochemical impedance spectroscopy].

Biofilms are microbial communities composed of bacteria cells and self-produced extracellular polymeric substances and lead to antibiotic resistance, forming a great threat in the fields of medicine, food industries and so on. The formation of biofilms is a multi-stage dynamic process regulated by multiple mechanisms. Recently, researchers paid much more attention to the related research areas of biofilms, especially the detection methods of biofilms. This paper introduced the characteristics and formation process of biofilms as well as the effects of quorum sensing on biofilms development. Meanwhile, we discussed the detection methods of biofilms, especially electrochemical impedance spectroscopy (EIS) in detection of biofilms in detail. Furthermore, we reviewed and predicted that microfluidic chips based EIS served as an in-situ and online monitoring method for biofilms growth.

Label-free impedimetric glycan biosensor for quantitative evaluation interactions between pathogenic bacteria and mannose.

In order to understanding the pathogenic mechanism of infectious diseases, it was important to study the selective recognition and interaction between pathogenic bacteria and host cells. In this paper, a novel electrochemical impedance biosensor was proposed, in which the Man/MUA-MH/Au sensing surface (Man: mannose; MUA: 11-mercapto eleven acid; MH: 6-mercapto hexanol) was fabricated and was of good biologically active and stability. The capture capacity of the designed sensing surface for S. typhimurium ATCC14028, E. coli JM109 and E. coli DH5α were characterized by Electrochemical impedance spectroscopy (EIS). According to Randless equivalent circuit and the Frumkin isotherm model, electron transfer impedance (Ret) was obtained and the binding affinity of the three bacteria and Man was calculated. It was shown that the sensing surface had a better binding affinity for S. typhimurium ATCC14028 with KADS(S.T.) = 2.16 × 106 CFU/mL. The impedance normalized value NIC (S.T.-Man) was of a good linear relationship with the logarithm of bacterial concentration (R2 = 0.96) in the range of 50-1000 CFU/mL. The detection limit was 50 CFU/mL. Meanwhile, the E. coli JM109 which expresses type 1 fimbriae was also adsorbed on the designed sensing surface with KADS(JM109) = 5.84 × 103 CFU/mL. It was illustrated that the novel electrochemical impedance biosensor could be more rapid and reliable for studying interactions between pathogen and glycan, and it was also perspective for a new point-of-care diagnostic tool for evaluating the pathogenicity bacteria.

An integrated microsystem with dielectrophoresis enrichment and impedance detection for detection of Escherichia coli.

An integrated microsystem device with matched interdigitated microelectrode chip was fabricated for enrichment and detection of Escherichia coli O157:H7. The microsystem has integrated with positive dielectrophoresis (pDEP) enrichment and in situ impedance detection, whose total volume is only 3.0 × 10-3 m3, and could provide impedance testing voltages of 0 ~ 10 V, detection frequencies of 1 KHz ~ 1 MHz, DEP excitation signals with amplitude of 0 ~ 10 Vpp and frequencies of 1KHz ~ 1 MHz, which fully meets the demands of pDEP enrichment and impedance detection for bacteria. The microfluidic chip with interdigitated microelectrodes was manufactured by microfabrication methods. The interdigital microelectrode array has sufficient contact area with a bacterial suspension to improve enrichment efficiency and detection sensitivity. Bacteria in the interdigital microelectrode area of the microfluidic chip were firstly captured and enriched by pDEP. Then, in situ impedance detection of the enriched bacteria was realized by switching test conditions. Using the self-assembly microsystem, a novel quantitative detection method was established and demonstrated to detect Escherichia coli O157:H7. Experimental results showed that the detection limits of Escherichia coli O157:H7 was 5 × 104 cfu mL-1, and testing time was only 6 min under the optimized detection voltage of 100 mV and frequency of 500 KHz. The method was successfully used to detect Escherichia coli O157:H7 in synthetic chicken synthetic samples.