Coupling of nanostraws with diverse physicochemical perforation strategies for intracellular DNA delivery.

Effective intracellular DNA transfection is imperative for cell-based therapy and gene therapy. Conventional gene transfection methods, including biochemical carriers, physical electroporation and microinjection, face challenges such as cell type dependency, low efficiency, safety concerns, and technical complexity. Nanoneedle arrays have emerged as a promising avenue for improving cellular nucleic acid delivery through direct penetration of the cell membrane, bypassing endocytosis and endosome escape processes. Nanostraws (NS), characterized by their hollow tubular structure, offer the advantage of flexible solution delivery compared to solid nanoneedles. However, NS struggle to stably self-penetrate the cell membrane, resulting in limited delivery efficiency. Coupling with extra physiochemical perforation strategies is a viable approach to improve their performance. This study systematically compared the efficiency of NS coupled with polyethylenimine (PEI) chemical modification, mechanical force, photothermal effect, and electric field on cell membrane perforation and DNA transfection. The results indicate that coupling NS with PEI modification, mechanical force, photothermal effects provide limited enhancement effects. In contrast, NS-electric field coupling significantly improves intracellular DNA transfection efficiency. This work demonstrates that NS serve as a versatile platform capable of integrating various physicochemical strategies, while electric field coupling stands out as a form worthy of primary consideration for efficient DNA transfection.

Technological Advances of Wearable Device for Continuous Monitoring of In Vivo Glucose.

Managing diabetes is a chronic challenge today, requiring monitoring and timely insulin injections to maintain stable blood glucose levels. Traditional clinical testing relies on fingertip or venous blood collection, which has facilitated the emergence of continuous glucose monitoring (CGM) technology to address data limitations. Continuous glucose monitoring technology is recognized for tracking long-term blood glucose fluctuations, and its development, particularly in wearable devices, has given rise to compact and portable continuous glucose monitoring devices, which facilitates the measurement of blood glucose and adjustment of medication. This review introduces the development of wearable CGM-based technologies, including noninvasive methods using body fluids and invasive methods using implantable electrodes. The advantages and disadvantages of these approaches are discussed as well as the use of microneedle arrays in minimally invasive CGM. Microneedle arrays allow for painless transdermal puncture and are expected to facilitate the development of wearable CGM devices. Finally, we discuss the challenges and opportunities and look forward to the biomedical applications and future directions of wearable CGM-based technologies in biological research.

Integrated electronic/fluidic microneedle system for glucose sensing and insulin delivery.

Background: Precise and dynamic blood glucose regulation is paramount for both diagnosing and managing diabetes. Continuous glucose monitoring (CGM) coupled with insulin pumps forms an artificial pancreas, enabling closed-loop control of blood glucose levels. Indeed, this integration necessitates advanced micro-nano fabrication techniques to miniaturize and combine sensing and delivery modules on a single electrode. While microneedle technology can mitigate discomfort, concerns remain regarding infection risk and potential sensitivity limitations due to their short needle length. Methods: This study presents the development of an integrated electronic/fluidic microneedle patch (IEFMN) designed for both glucose sensing and insulin delivery. The use of minimally invasive microneedles mitigates nerve contact and reduces infection risks. The incorporation of wired enzymes addresses the issue of "oxygen deprivation" during glucose detection by decreasing the reliance on oxygen. The glucose-sensing electrodes employ wired enzyme functionalization to achieve lower operating voltages and enhanced resilience to sensor interference. The hollow microneedles' inner channel facilitates precise drug delivery for blood glucose regulation. Results: Our IEFMN-based system demonstrated high sensitivity, selectivity, and a wide response range in glucose detection at relatively low voltages. This effectively reduced interference from both external and internal active substances. The microneedle array ensured painless and minimally invasive skin penetration, while wired enzyme functionalization not only lowered sensing potential but also improved glucose detection accuracy. In vivo, experiments conducted in rats showed that the device could track subcutaneous glucose fluctuations in real-time and deliver insulin to regulate blood glucose levels. Conclusions: Our work suggests that the IEFMN-based system, developed for glucose sensing and insulin delivery, exhibits good performance during in vivo glucose detection and drug delivery. It holds the potential to contribute to real-time, intelligent, and controllable diabetes management.

Machine learning-coupled tactile recognition with high spatiotemporal resolution based on cross-striped nanocarbon piezoresistive sensor array.

Flexible pressure sensor arrays have been playing important roles in various applications of human-machine interface, including robotic tactile sensing, electronic skin, prosthetics, and human-machine interaction. However, it remains challenging to simultaneously achieve high spatial and temporal resolution in developing pressure sensor arrays for tactile sensing with robust function to achieve precise signal recognition. This work presents the development of a flexible high spatiotemporal piezoresistive sensor array (PRSA) by coupling with machine learning algorithms to enhance tactile recognition. The sensor employs cross-striped nanocarbon-polymer composite as an active layer, though screen printing manufacture processes. A miniaturized signal readout circuit and transmission board is developed to achieve high-speed acquisition of distributed pressure signals from the PRSA. Test results indicate that the developed PRSA platform simultaneously possesses the characteristics of high spatial resolution up to 1.5 mm, fast temporal resolution of about 5 ms, and long-term durability with a variation of less than 2%. The PRSA platform also exhibits excellent performance in real-time visualization of multi-point touch, mapping embossed shapes, and tracking motion trajectory. To test the performance of PRSA in recognizing different shapes, we acquired pressure images by pressing the finger-type device coated with PRSA film on different embossed shapes and implementing the T-distributed Stochastic Neighbor Embedding model to visualize the distinction between images of different shapes. Then we adopted a one-layer neural network to quantify the discernibility between images of different shapes. The analysis results show that the PRSA could capture the embossed shapes clearly by one contact with high discernibility up to 98.9%. Collectively, the PRSA as a promising platform demonstrates its promising potential for robotic tactile sensing.

Personalized Machine Learning-Coupled Nanopillar Triboelectric Pulse Sensor for Cuffless Blood Pressure Continuous Monitoring.

A wearable system that can continuously track the fluctuation of blood pressure (BP) based on pulse signals is highly desirable for the treatments of cardiovascular diseases, yet the sensitivity, reliability, and accuracy remain challenging. Since the correlations of pulse waveforms to BP are highly individualized due to the diversity of the patients' physiological characteristics, wearable sensors based on universal designs and algorithms often fail to derive BP accurately when applied on individual patients. Herein, a wearable triboelectric pulse sensor based on a biomimetic nanopillar layer was developed and coupled with Personalized Machine Learning (ML) to provide accurate and continuous monitoring of BP. Flexible conductive nanopillars as the triboelectric layer were fabricated through soft lithography replication of a cicada wing, which could effectively enhance the sensor's output performance to detect weak signal characteristics of pulse waveform for BP derivation. The sensors were coupled with a personalized Partial Least-Squares Regression (PLSR) ML to derive unknown BP based on individual pulse characteristics with reasonable accuracy, avoiding the issue of individual variability that was encountered by General PLSR ML or formula algorithms. The cuffless and intelligent design endow this ML-sensor as a highly promising platform for the care and treatments of hypertensive patients.

Nanopore Electroporation Device for DNA Transfection into Various Spreading and Nonadherent Cell Types.

Cell transfection plays a crucial role in the study of gene function and regulation of gene expression. The existing gene transfection methods, such as chemical carriers, viruses, electroporation, and microinjection, suffer from limitations, including cell type dependence, reliance on cellular endocytosis, low efficiency, safety concerns, and technical complexity. Nanopore-coupled electroporation offers a promising approach to localizing electric fields for efficient cell membrane perforation and nucleic acid transfection. However, the applicability of nanopore electroporation technology across different cell types lacks a systematic investigation. In this study, we explore the potential of nanopore electroporation for transfecting DNA plasmids into various cell types. Our nanopore electroporation device employs track-etched membranes as the core component. We find that nanopore electroporation efficiently transfects adherent cells, including well-spreading epithelial-like HeLa cells, cardiomyocyte-like HL-1 cells, and dendritic-cell-like DC2.4 cells. However, it shows a limited transfection efficiency in weakly spreading macrophages (RAW264.7) and suspension cells (Jurkat). To gain insights into these observations, we develop a COMSOL model, revealing that nanopore electroporation better localizes the electric field on adherent and well-spreading cells, promoting favorable membrane poration conditions. Our findings provide valuable references for advancing nanopore electroporation as a high-throughput, safe, and efficient gene transfection platform.

Capacitive-piezoresistive hybrid flexible pressure sensor based on conductive micropillar arrays with high sensitivity over a wide dynamic range.

Flexible pressure sensors are the foundation of wearable/implantable biosensing and human-machine interfaces, and mainly comprise piezoresistive-, capacitive-, piezoelectric-, and triboelectric-type sensors. As each type of sensor exhibits different electro-mechanical behaviors, it is challenging to detect various physiological mechanical signals that cover a large pressure range using a given sensor configuration, or even a single type of sensor. Here, we report a capacitive-piezoresistive hybrid flexible pressure sensor based on face-to-face-mounted conductive micropillar arrays as a solution to this challenge. The sensor exhibited high sensitivity over a wide dynamic range of five orders of magnitude, which covers almost the full range of physiological mechanical signals. A process for fabricating large-scale and morphologically homogeneous conductive micropillar arrays was first developed and refined. This track-etched-membrane-based process provides a facile, cost-effective, and highly flexible way to precisely adjust the morphology, modulus, and conductivity of the micropillars according to the application requirements. Subsequently, conductive-micropillar-array-based pressure sensors (MAPS) were developed and optimized to attain all-round sensing performance. The pillar contact behaviors generated significant variations in both the capacitance and resistance of the MAPS in the low-pressure regime (10-4-0.2 kPa), providing high sensitivity in both the capacitive and piezoresistive working modes. The vertical contact, bending and thickening of the pillars under medium pressure (0.2-16 kPa) led to a continuous linear response in both modes. Configuration and optimization enabled the MAPS to detect acoustic pressure (<1 Pa), milligram weights, soft touch (<1 kPa), arterial pulses (1-16 kPa preload), joint motions and plantar pressure (∼100 kPa), and the hybrid sensing mode allowed the MAPS to work in a desirable way. In this work, the piezoresistive mode was mainly employed for a higher accuracy and sampling rate, and can apparently simplify IC design for wearable applications. The circuit converts the resistive variations into electrical signals via the voltage division method and directly reads out the signals after further amplification, filtering and transmission. The improved facile and highly adjustable fabrication process, as well as the flexible hybrid sensing strategy, will benefit the unified design, batch production, quantifiable optimization, and functional diversity of wearable/implantable bioelectronics.