Integrated Cardiomyocyte-Based Biosensing Platform for Electroporation-Triggered Intracellular Recording in Parallel with Delivery Efficiency Evaluation.

Electroporation is a proven technique that can record action potential of cardiomyocytes and serve for biomolecular delivery. To ensure high cell viability, micro-nanodevices cooperating with low-voltage electroporation are frequently utilized in research, and the effectiveness of delivery for intracellular access is typically assessed using an optical imaging approach like flow cytometry. However, the efficiency of in situ biomedical studies is hampered by the intricacy of these analytical approaches. Here, we develop an integrated cardiomyocyte-based biosensing platform to effectively record action potential and evaluate the electroporation quality in terms of viability, delivery efficiency, and mortality. The ITO-MEA device of the platform possesses sensing/stimulating electrodes which combines with the self-developed system to achieve intracellular action potential recording and delivery by electroporation trigger. Moreover, the image acquisition processing system analyzes various parameters effectively to assess delivery performance. Therefore, this platform has the potential for drug delivery therapy and pathology research for cardiology.

Scalable and Robust Hollow Nanopillar Electrode for Enhanced Intracellular Action Potential Recording.

Electrophysiology is a unique biomarker of the electrogenic cells that can perform a disease investigation or drug assessment. In the recent decade, vertical nanoelectrode arrays can successfully achieve a high-quality intracellular electrophysiological study in electrogenic cells and their networks. However, a high success rate and high-quality and long-term intracellular recording using low-cost nanostructures is still a considerable challenge. Herein, we develop a scalable and robust hollow nanopillar electrode to achieve enhanced intracellular recording of cardiomyocytes. The template-based synthesis of vertical hollow nanopillars is compatible with large-scale and efficient microfabrication processes and is convenient to regulate the geometry of hollow nanopillars. Compared with the conventional same-size planar electrode, the regulating height of a hollow nanopillar can achieve high-quality and prolonged intracellular recordings, which can improve the cell-electrode interface for tight coupling and effective electroporation. It is demonstrated that the geometry regulation of a nanostructure is a powerful strategy to enhance intracellular recording.

Dynamic and Label-Free Sensing of Cardiomyocyte Responses to Nanosized Vesicles for Cardiac Oxidative Stress Injury Therapy.

Cardiac oxidative stress is a significant phenotype of myocardial infarction disease, a leading cause of global health threat. There is an urgent need to develop innovative therapies. Nanosized extracellular vesicle (nEV)-based therapy shows promise, yet real-time monitoring of cardiomyocyte responses to nEVs remains a challenge. In this study, a dynamic and label-free cardiomyocyte biosensing system using microelectrode arrays (MEAs) was constructed. Cardiomyocytes were cultured on MEA devices for electrophysiological signal detection and treated with nEVs from E. coli, gardenia, HEK293 cells, and mesenchymal stem cells (MSC), respectively. E. coli-nEVs and gardenia-nEVs induced severe paroxysmal fibrillation, revealing distinct biochemical communication compared to MSC-nEVs. Principal component analysis identified variations and correlations between nEV types. MSC-nEVs enhanced recovery without inducing arrhythmias in a H2O2-induced oxidative stress injury model. This study establishes a fundamental platform for assessing biochemical communication between nEVs and cardiomyocytes, offering new avenues for understanding nEVs' functions in the cardiovascular system.

Optimizing the Cell-Nanostructure Interface: Nanoconcave/Nanoconvex Device for Intracellular Recording of Cardiomyocytes.

Nanostructures are powerful components for the development of high-performance nanodevices. Revealing and understanding the cell-nanostructure interface are essential for improving and guiding nanodevice design for investigations of cell physiology. For intracellular electrophysiological detection, the cell-nanostructure interface significantly affects the quality of recorded intracellular action potentials and the application of nanodevices in cardiology research and pharmacological screening. Most of the current investigations of biointerfaces focus on nanovertical structures, and few involve nanoconcave structures. Here, we design both nanoconvex and nanoconcave devices to perform intracellular electrophysiological recordings. The amplitude, signal-to-noise ratio, duration, and repeatability of the recorded intracellular electrophysiological signals provide a multifaceted characterization of the cell-nanostructure interface. We demonstrate that devices based on both convex and concave nanostructures can create tight coupling, which facilitates high-quality and stable intracellular recordings and paves the way for precise electrophysiological study.

Porous Polyethylene Terephthalate Nanotemplate Electrodes for Sensitive Intracellular Recording of Action Potentials.

New strategies for intracellular electrophysiology break the spatiotemporal limitation of the action potential and lead a notable advance in the investigation of electrically excitable cells and their network. Although successful applications of intracellular recording have been achieved by 3D micro/nanodevices, complex micro/nanofabrication processes preclude the progress of extensive applications. We address this challenge by introducing porous polyethylene terephthalate (PET) membrane to develop a new type of nanotemplate electrode. This nanotemplate electrode is manufactured following a fabrication process on a porous PET membrane by atomic layer deposition. The 3D nanotemplate electrodes afford intracellular access to cardiomyocytes to report intracellular-like action potentials. These controllable nanotemplate electrodes exhibit sensitive and prolonged intracellular recordings of action potentials compared with free-growing 3D nanoelectrodes. This study indicates that the optimized structure of the nanoelectrode significantly promotes the performance of intracellular recording to assess electrophysiology in the fields of cardiology and neuroscience at an action potential level.

Scalable Nanotrap Matrix Enhanced Electroporation for Intracellular Recording of Action Potential.

Electrophysiological recording, as a long-sought objective, plays a crucial role in fundamental biomedical research and practical clinical applications. The challenge in developing electrophysiological detection platforms is to combine simplicity, stability, and sensitivity in the same device. In this study, we develop a nanotrapped microelectrode based on a porous PET membrane, which is compatible with large-scale microtechnologies. The nanotraps can promote the protrusion of the local cell membrane in the hollow center and offer a unique nanoedge structure for tight sealing and effective electroporation. We demonstrate that scalable nanotraps can enhance cell-electrode coupling and perform high-quality intracellular recording. Further, the nanoedge-enhanced electroporation and minimally invasive nanotrapped recordings afford much longer intracellular access of over 100 min and permit consecutive electroporation events in a short period of time. This study suggests that the geometry-regulating strategy of the cell-electrode nanointerface could significantly improve the intracellular recording performance of a nanopatterned electrode.

Double-Drum Test Bench for Variable Load Transfer Simulation by Electromechanical Inertia Compensation.

To improve the accuracy and actual road equivalence of vehicle performance testing using test benches, a double-drum test bench that meets the test requirements of vehicle control system prototypes and in-use vehicles was designed. Dynamic models of the single-wheel test bench and the vehicle test bench were established, and mechanisms were theoretically analyzed for single-wheel variable adhesion and vehicle load transfer for equivalent testing using the variable placement angle. The mechanism of electromechanical inertia compensation was studied to realize stepless simulation of vehicle inertia and simulate dynamic load while braking. The simulation model of the vehicle test bench system was established based on MATLAB/Simulink. Simulations were carried out to verify the anti-lock braking system (ABS) performance test functionality of the test bench under high adhesion, bisectional, and low adhesion conditions. Referring to the simulation conditions, ABS tests under actual test bench and road conditions were carried out. Results demonstrated that the mechanism of variable load transfer simulation by electromechanical inertia compensation improves the equivalent accuracy compared to that of its road test equivalent, verifying the feasibility of the simulation mechanism. This study could help further improve the accuracy and reduce the cost of vehicle performance testing, thus greatly benefitting the vehicle development and testing industry.

Automated Vocal Analysis of Children With Hearing Loss and Their Typical and Atypical Peers.

OBJECTIVES: This study investigated automatic assessment of vocal development in children with hearing loss compared with children who are typically developing, have language delays, and have autism spectrum disorder. Statistical models are examined for performance in a classification model and to predict age within the four groups of children. DESIGN: The vocal analysis system analyzed 1913 whole-day, naturalistic acoustic recordings from 273 toddlers and preschoolers comprising children who were typically developing, hard of hearing, language delayed, or autistic. RESULTS: Samples from children who were hard of hearing patterned more similarly to those of typically developing children than to the language delayed or autistic samples. The statistical models were able to classify children from the four groups examined and estimate developmental age based on automated vocal analysis. CONCLUSIONS: This work shows a broad similarity between children with hearing loss and typically developing children, although children with hearing loss show some delay in their production of speech. Automatic acoustic analysis can now be used to quantitatively compare vocal development in children with and without speech-related disorders. The work may serve to better distinguish among various developmental disorders and ultimately contribute to improved intervention.

[Relevant studies on effect of Fuzheng Sanjie recipe in regulating immune microenvironment remodeling of TAMs in Lewis lung cancer mice].

OBJECTIVE: To study the effect of Fuzheng Sanjie recipe in regulating tumor-associated macrophages (TAMs) in Lewis lung cancer mice. METHOD: Efforts were made to establish the Lewis lung cancer mouse model, weigh tumors and calculate the anti-tumor rate. The immunohistochemical method was used to examine the infiltration degree of CD68 + in tumor tissues in each group. ELISA was used to examine the content of IFN-γ, TGF-ß, IL-4, IL-13, IL-6, IL-10, IL-12, TNF-α in mice serum. RESULT: Compared with the tumor-bearing model group, all of the other groups showed higher tumor inhibition rates, i. e. 50.28% for the DDP group, 34.37% for the TCM-preventing group and 66.76% for the Chinese and western medicine group, with statistical difference (P < 0.05), but without statistical difference in the infiltration degree of CD68+. The expressions of the IFN-γ, IL-6, IL-12 in tumor-bearing groups were lower than that in the blank control group, but with higher contents of IL-4, IL-13, TGF-ß. Intervened with different drugs, there were significant differences in content among some relevant cytokines (P < 0.05), as well as statistical differences among the TCM prevention group, the Chinese and western medicine group and the tumor-bearing control group (P <0. 05) , but without statistical difference in TNF-α and IL-10 content from the tumor-bearing control group (P < 0.05). CONCLUSION: Fuzheng Sanjie recipe could reverse the immune remodeling effect and control the tumor growth by down-regulating the expressions of IL-4, IL-13, TGF-α in lung cancer immune microenvironment and up-regulating the expression of IFN-γ.

Language ENvironment Analysis Language and Autism Screen and the Child Development Inventory Social Subscale as a possible autism screen for children who are deaf or hard of hearing.

The Language ENvironment Analysis Language and Autism Screen (LLAS) is an automated vocal production analysis that has been shown to be a valid screener for autism in hearing children between the ages of 24 to 48 months of age. Although there is reportedly a higher incidence of autism among children who are deaf or hard of hearing, the diagnosis of autism is usually later than that in children with hearing. None of the traditional screening instruments have been used with children with hearing loss. Data about the utility of LLAS with children who are deaf or hard of hearing will be presented and discussed. Though more data will be needed, an LLAS at-risk flag in conjunction with the Social Quotient from the Child Development Inventory holds significant promise for a screen for children who are deaf or hard of hearing.

Elevating intracellular action potential recording in cardiomyocytes: A precision-enhanced and biosafe single-pulse electroporation system.

Action potentials play a pivotal role in diverse cardiovascular physiological mechanisms. A comprehensive understanding of these intricate mechanisms necessitates a high-fidelity intracellular electrophysiological investigative approach. The amalgamation of micro-/nano-electrode arrays and electroporation confers substantial advantages in terms of high-resolution intracellular recording capabilities. Nonetheless, electroporation systems typically lack precise control, and commonly employed electroporation modes, involving tailored sequences, may escalate cellular damage and perturbation of normal physiological functions due to the multiple or higher-intensity electrical pulses. In this study, we developed an innovative electrophysiological biosensing system customized to facilitate precise single-pulse electroporation. This advancement serves to achieve optimal and uninterrupted intracellular action potential recording within cardiomyocytes. The refinement of the single-pulse electroporation technique is realized through the integration of the electroporation and assessment biosensing system, thereby ensuring a consistent and reliable means of achieving stable intracellular access. Our investigation has unveiled that the optimized single-pulse electroporation technique not only maintains robust biosafety standards but also enables the continuous capture of intracellular electrophysiological signals across an expansive three-day period. The universality of this biosensing system, adaptable to various micro/nano devices, furnishes real-time analysis and feedback concerning electroporation efficacy, guaranteeing the sustained, secure, and high-fidelity acquisition of intracellular data, thereby propelling the field of cardiovascular electrophysiological research.

Micronano Synergetic Three-Dimensional Bioelectronics: A Revolutionary Breakthrough Platform for Cardiac Electrophysiology.

Cardiovascular diseases (CVDs) are the leading cause of mortality and therefore pose a significant threat to human health. Cardiac electrophysiology plays a crucial role in the investigation and treatment of CVDs, including arrhythmia. The long-term and accurate detection of electrophysiological activity in cardiomyocytes is essential for advancing cardiology and pharmacology. Regarding the electrophysiological study of cardiac cells, many micronano bioelectric devices and systems have been developed. Such bioelectronic devices possess unique geometric structures of electrodes that enhance quality of electrophysiological signal recording. Though planar multielectrode/multitransistors are widely used for simultaneous multichannel measurement of cell electrophysiological signals, their use for extracellular electrophysiological recording exhibits low signal strength and quality. However, the integration of three-dimensional (3D) multielectrode/multitransistor arrays that use advanced penetration strategies can achieve high-quality intracellular signal recording. This review provides an overview of the manufacturing, geometric structure, and penetration paradigms of 3D micronano devices, as well as their applications for precise drug screening and biomimetic disease modeling. Furthermore, this review also summarizes the current challenges and outlines future directions for the preparation and application of micronano bioelectronic devices, with an aim to promote the development of intracellular electrophysiological platforms and thereby meet the demands of emerging clinical applications.

Fabrication and Characterization of Silicon-Based Antimonene Thin Film via Electron Beam Evaporation.

Antimonene has attracted much attention due to its excellent characteristics of high carrier mobility, thermoelectric properties and high stability. It has great application prospects in Q-switched lasers, laser protection and spintronics. At present, the epitaxy growth of antimonene mainly depends on molecular beam epitaxy. We have successfully prepared antimonene films on silicon, germanium/silicon substrates for the first time using electron beam evaporation coating and studied the effects of the deposition rate and substrate on the preparation of antimonene; film characterization was performed via confocal microprobe Raman spectroscopy, via X-ray diffraction and using a scanning electron microscope. Raman spectroscopy showed that different deposition rates can lead to the formation of different structures of antimonene, such as α phase and ß phase. At the same time, it was found that the growth of antimonene is also affected by different substrates and ion beams.

Three-Dimensional Cardiomyocyte-Nanobiosensing System for Specific Recognition of Drug Subgroups.

Abnormal cardiac electrophysiological activities significantly contribute to the incidence of cardiovascular diseases. Therefore, it is crucial to recognize effective drugs, which require an accurate, stable, and sensitive platform. Although conventional extracellular recordings offer a non-invasive and label-free manner to monitor the electrophysiological state of cardiomyocytes, the misrepresented and low-quality extracellular action potentials are difficult to provide accurate and high-content information for drug screening. This study presents the development of a three-dimensional cardiomyocyte-nanobiosensing system that can specifically recognize drug subgroups. The nanopillar-based electrode is manufactured by template synthesis and standard microfabrication technology on a porous polyethylene terephthalate membrane. Based on the cardiomyocyte-nanopillar interface, high-quality intracellular action potentials can be recorded by the minimally invasive electroporation. We validate the performance of a cardiomyocyte-nanopillar-based intracellular electrophysiological biosensing platform by two subclasses of sodium channel blockers, quinidine and lidocaine. The recorded intracellular action potentials accurately reveal the subtle differences between these drugs. Our study indicates that high-content intracellular recordings utilizing nanopillar-based biosensing can provide a promising platform for the electrophysiological and pharmacological investigation of cardiovascular diseases.

Integrated Au-Nanoroded Biosensing and Regulating Platform for Photothermal Therapy of Bradyarrhythmia.

Bradyarrhythmia is a kind of cardiovascular disease caused by dysregulation of cardiomyocytes, which seriously threatens human life. Currently, treatment strategies of bradyarrhythmia mainly include drug therapy, surgery, or implantable cardioverter defibrillators, but these strategies are limited by drug side effect, surgical trauma, and instability of implanted devices. Here, we developed an integrated Au-nanoroded biosensing and regulating platform to investigate the photothermal therapy of cardiac bradyarrhythmia in vitro. Au-nanoroded electrode array can simultaneously accumulate energy from the photothermal regulation and monitor the electrophsiological state to restore normal rhythm of cardiomyocytes in real time. To treat the cardiomyocytes cultured on Au-nanoroded device by near-infrared (NIR) laser irradiation, cardiomyocytes return to normal for long term after irradiation of suitable NIR energy and maintenance. Compared with the conventional strategies, the photothermal strategy is more effective and convenient to regulate the cardiomyocytes. Furthermore, mRNA sequencing shows that the differential expression genes in cardiomyocytes are significantly increased after photothermal strategy, which are involved in the regulation of the heart rate, cardiac conduction, and ion transport. This work establishes a promising integrated biosensing and regulating platform for photothermal therapy of bradyarrhythmia in vitro and provides reliable evidence of photothermal regulation on cardiomyocytes for cardiological clinical studies.

Universal and Sensitive Drug Assessment Biosensing Platform Using Optimal Mechanical Beating Detection of Single Cardiomyocyte.

The preclinical assessment of efficacy and safety is essential for cardiovascular drug development in order to guarantee effective prevention and treatment of cardiovascular disease and avoid human health endangerment and a huge waste of resources. Rhythmic mechanical beating as one of the crucial cardiomyocyte properties has been exploited to establish a drug assessment biosensing platform. However, the conventional label-free biosensing platforms are difficult to perform high-throughput and high-resolution mechanical beating detection for a single cardiomyocyte, while label-based strategies are limited by pharmacologically adverse effects and phototoxicity. Herein, we propose a biosensing platform involving the multichannel electrode array device and the universal mechanical beating detection system. The platform can determine the optimal characteristic working frequency of different devices and dynamically interrogate the viability of multisite single cardiomyocytes to establish the optimized cell-based model for sensitive drug assessment. The subtle changes of mechanical beating signals induced by cardiovascular drugs can be detected by the platform, thereby demonstrating its high performance in pharmacological assessment. The universal and sensitive drug assessment biosensing platform is believed to be widely applied in cardiology investigating and preclinical drug screening.

A dynamic and quantitative biosensing assessment for electroporated membrane evolution of cardiomyocytes.

The electrophysiological study is an essential approach to perform the biology and basic medicine research. To achieve the intracellular electrophysiological investigation, electroporation is introduced as an effective and convenient strategy to achieve the intracellular access of electrogenic cells and obtain high-fidelity action potentials. However, seldom platform could provide a quantitative and dynamic strategy to assess the electroporation-induced membrane perforation and recovery during intracellular electrophysiological investigation. Here we develop a high-throughput, sensitive, and stable biosensing platform to assess the evolution of electroporated cell membrane dynamically and quantitatively based on the recorded intracellular electrophysiological signals of cardiomyocytes. Following the electroporation, the extracellular action potentials transiently convert to the intracellular action potentials, whose amplitude rapidly increases to the maximum and then gradually decays. The intracellular action potentials finally convert back to the extracellular action potentials. This biosensing platform can dynamically explore and characterize the evolution procedures of perforation, stabilization, and resealing of the cell membrane by intracellular recordings. Moreover, the effect of electroporation voltages on the cell membrane is segmentally and quantitatively analyzed, demonstrating that a higher electroporation voltage induced a longer resealing time within the safe range of electroporation voltage. We believed that this dynamic and quantitative electroporated membrane evolution biosensing assessment platform will be a promising tool to pave a new avenue to bridge the electrophysiology and electroporated membrane evolution.

A universal, multimodal cell-based biosensing platform for optimal intracellular action potential recording.

Intracellular recording of action potentials is an essential mean for studying disease mechanisms, and for electrophysiological studies, particularly in excitable cells as cardiomyocytes or neurons. Current strategies to obtain intracellular recordings include three-dimensional (3D) nanoelectrodes that can effectively penetrate the cell membrane and achieve high-quality intracellular recordings in a minimally invasive manner, or transient electroporation of the membrane that can yield temporary intracellular access. However, the former strategy requires a complicated and costly fabrication process, and the latter strategy suffers from high dependency on the method of application of electroporation, yielding inconsistent, suboptimal recordings. These factors hinder the high throughput use of these strategies in electrophysiological studies. In this work, we propose an advanced cell-based biosensing platform that relies on electroporation to produce consistent, high-quality intracellular recordings. The suggested universal system can be integrated with any electrode array, and it enables tunable electroporation with controllable pulse parameters, while the recorded potentials can be analyzed in real time to provide instantaneous feedback on the electroporation effectiveness. This integrated system enables the user to perform electroporation, record and assess the obtained signals in a facile manner, to ultimately achieve stable, reliable, intracellular recording. Moreover, the proposed platform relies on microelectrode arrays which are suited for large-scale production, and additional modules that are low-cost. Using this platform, we demonstrate the tuning of electroporation pulse width, pulse number, and amplitude, to achieve effective electroporation and high-quality intracellular recordings. This integrated platform has the potential to enable larger scale, repeatable, convenient, and low-cost electrophysiological studies.

High-throughput rhythmic regulation of cardiomyocytes by integrated electrical stimulation and video-based automated analysis biosensing platform.

In cardiac tissue engineering, electric stimulation is an efficient approach to improve the formation of cardiac tissue from individual cardiomyocyte. The regulation conditions of electric stimulation should be screened in an efficient way. However, the lack of high-throughput and large-scale assessment platforms limited the effectively screen the regulation conditions. Here, we develop a high-throughput integrated electrical stimulation system to rhythmically regulate the cardiomyocytes in situ. The state of regulated cardiomyocytes is characterized by a video-based automated biosensing system to analyze the beating of cardiomyocytes. Electrical stimulation conditions are optimized to regulate the cardiomyocyte state in vitro to replace the complex bioactive molecules and materials. By the video analysis, the accurate beating rate and regularity of cardiomyocyte can be determined. The results show that electrical stimulation frequency is a significant factor to regulate the cardiomyocyte beating. The electrical stimulation with a frequency of 3 Hz can effectively regulate the primary rat cardiomyocytes with normal rhythm. This high-throughput electrical stimulation and a video-based automated biosensing system will be a promising and powerful tool to effectively optimize the regulation conditions of cardiomyocyte in vitro, and possess broad application prospects in cardiac tissue engineering and pharmaceutical industry.

Research Progress of Wide Tunable Bragg Grating External Cavity Semiconductor Lasers.

In this paper, we review the progress of wide tunable Bragg grating external cavity semiconductor lasers (BG-ECSLs). We concentrate on BG-ECSLs based on the wide tunable range for multicomponent detection. Wide tunable BG-ECSLs have many important applications, such as wavelength-division multiplexing (WDM) systems, coherent optical communications, gas detection and atom cooling. Wide tunability, narrow linewidth and a high side-mode suppression ratio BG-ECSLs have attracted much attention for their merits. In this paper, three main structures for achieving widely tunable, narrow linewidth, high side-mode suppression ratio BG-ECSLs are reviewed and compared in detail, such as the volume Bragg grating (VBG) structure, fiber Bragg grating (FBG) structure and waveguide Bragg grating (WBG) structure of ECSLs. The advantages and disadvantages of different structures of BG-ECSLs are analyzed. The results show that WBG-ECSLs are a potential way to realize the integration, small size, wide tuning range, stable spectral output and high side-mode suppression ratio laser output. Therefore, the use of WBG as optical feedback elements is still the mainstream direction of BG-ECSLs, and BG-ECSLs offer a further new option for multicomponent detection and multi-atoms cooling.