Biomicrofluidics最新文献

In the field of microfluidics, high-pressure microfluidics technology, which utilizes high driving pressure for microfluidic analysis, is an evolving technology. This technology combines microfluidics and pressurization, where the flow of fluid is controlled by means of high-pressure-driven devices greater than 10 MPa. This paper first reviews the existing high-pressure microfluidics systems and describes their components and applications. Then, it summarizes several materials used in the microfabrication of high-pressure microfluidics chips, reviewing their properties, processing methods, and bonding methods. In addition, advanced laser processing techniques for the microfabrication of high-pressure microfluidics chips are described. Last, the paper examines the analytical detection methods employed in high-pressure microfluidics systems, encompassing optical and electrochemical detection methods. The review of analytical detection methods shows the different functions and application scenarios of high-pressure microfluidics systems. In summary, this study provides an efficient and advanced microfluidics system, which can be widely used in chemical engineering, food industry, and environmental engineering under high pressure conditions.

Monitoring platelet aggregation is crucial for predicting thrombotic diseases and identifying the risk of bleeding or resistance to antiplatelet drugs. This study developed a microfluidic device to measure platelet activation with high sensitivity. By controlling exposure time through repeated reinjections, the device enables the detection of subtle changes in platelet activity influenced by lifestyle factors, such as alcohol consumption. Using computational fluid dynamics simulations, the design was optimized to achieve moderate shear stresses and fabricated with 3D printing. Experimental results revealed that pillars biased to one side partially accelerate the flow and inhibit platelet adhesion. A distinct difference in platelet adhesion was clearly observed before and after alcohol consumption. Despite the high standard deviations in platelet adhesion area, hematocrit, and viscosity after alcohol consumption, the area covered by adhered platelets increased by 3.12 times compared to that before alcohol consumption. This microfluidic chip offers potential for personalized health monitoring by distinguishing platelet variations caused by lifestyle or dietary habits. However, challenges such as reinjection procedures and large sample volumes require further investigation.

Glioblastoma multiforme, the most common type of highly aggressive primary brain tumor, is influenced by complex molecular signaling pathways, where microRNAs (miRNAs) play a critical regulatory role. Originating from glial cells, glioblastoma cells are affected by the physiological direct current electric field (dcEF) in the central nervous system. While dcEF has been shown to affect glioblastoma migration (electrotaxis), the specific impact on glioblastoma intercellular communication and miRNA expression in glioblastoma cells and their exosomes remains unclear. This study aims to fill this gap by investigating the differential expression of microRNAs in glioblastoma cells and exosomes under dcEF stimulation. We have developed a novel, reversibly sealed dcEF stimulation bioreactor that ensures uniform dcEF stimulation across a large cell culture area, specifically targeting glioblastoma cells and primary human astrocytes. Using microarray analysis, we examined differential miRNA profiles in both cellular and exosomal RNAs. Our study identified shared molecular targets and pathways affected by dcEF stimulation. Our findings reveal significant changes in miRNA expression due to dcEF stimulation, with specific miRNAs, such as hsa-miR-4440 being up-regulated and hsa-miR-3201 and hsa-mir-548g being down-regulated. Future research will focus on elucidating the molecular mechanisms of these miRNAs and their potential as diagnostic biomarkers. The developed platform offers high-quality dcEF stimulation and rapid sample recovery, with potential applications in tissue engineering and multi-omics molecular analysis.

Bio-microfluidic technologies offer promising applications in diagnostics and therapy, yet they face significant technical challenges, particularly in the need for external power sources, which limits their practicality and user-friendliness. Recent advancements have explored innovative methods utilizing body fluids, motion, and heat to power these devices, addressing the power supply issue effectively. Among these, body-motion and body-heat-powered systems stand out for their potential to create self-sustaining, wearable, and implantable devices. In this Perspective, we focus on the principles and applications of hydrovoltaic cells, biofuel cells, and piezoelectric and triboelectric nanogenerators. Recent strides in energy conversion efficiency, coupled with the development of biocompatible and durable materials, are driving innovation in bio-integrated electronics. Integration with bio-microfluidic platforms further enhances the linkage to the human body and the potential of these devices for personalized healthcare applications. Ongoing research into these areas promises to deliver sustainable and user-friendly solutions for continuous monitoring, diagnostics, and therapy, potentially revolutionizing the landscape of healthcare delivery.

Chimeric antigen receptor (CAR) T-cell therapy shows unprecedented efficacy for cancer treatment, particularly in treating patients with various blood cancers, most notably B-cell acute lymphoblastic leukemia. In recent years, CAR T-cell therapies have been investigated for treating other hematologic malignancies and solid tumors. Despite the remarkable success of CAR T-cell therapy, cytokine release syndrome (CRS) is an unexpected side effect that is potentially life-threatening. Our aim is to reduce pro-inflammatory cytokine release associated with CRS by controlling CAR surface density on CAR T cells. We show that CAR expression density can be titrated on the surface of primary T cells using an acoustic-electric microfluidic platform. The platform performs dosage-controlled delivery by uniformly mixing and shearing cells, delivering approximately the same amount of CAR gene coding mRNA into each T cell.

Platelet transfusion is a lifesaving therapy intended to prevent and treat bleeding. However, in addition to platelets, a typical unit also contains a large volume of supernatant that accumulates multiple pro-inflammatory contaminants, including residual leukocytes, microaggregates, microparticles, antibodies, and cytokines. Infusion of this supernatant is responsible for virtually all adverse reactions to platelet transfusions. Conventional methods for removing residual leukocytes (leukoreduction) and reducing the volume of transfused supernatant (volume reduction) struggle to mitigate these risks holistically. Leukoreduction filters can remove leukocytes and microaggregates but fail to reduce supernatant volume, whereas centrifugation can reduce volume, but it is ineffective against larger contaminants and damages platelets. Additionally, platelet purification based on these methods is often too logistically complex, time-consuming, and labor-intensive to implement routinely. Emerging microfluidic technologies offer promising alternatives through passive separation mechanisms that enable cell separation with minimal damage and drastically reduced instrumentation size and facility requirements. This review examines recent innovations in microfluidic cell separation that can be used for leukoreduction and volume reduction of platelets. It begins by defining the performance requirements that any separation method must meet to successfully replace conventional methods currently used to perform these tasks. Standard performance metrics are described, including leukocyte depletion efficiency, degree of volume reduction, processing throughput, and platelet recovery. Finally, the review outlines the primary challenges that must be overcome to enable simple-to-use, disposable microfluidic devices capable of both reducing the platelet unit volume and removing pro-inflammatory contaminants, while preserving most functional platelets for transfusion.

Drug delivery technologies, which are a crucial area of research in the field of cell biology, aim to actively or passively deliver drugs to target cells to enhance therapeutic efficacy and minimize off-target effects. In recent years, with advances in drug development, particularly, the increasing demand for macromolecular drugs (e.g., proteins and nucleic acids), novel drug delivery technologies and intracellular cargo delivery systems have emerged as promising tools for cell and gene therapy. These systems include various viral- and chemical-mediated methods as well as physical delivery strategies. Physical methods, such as electroporation and microinjection, have shown promise in early studies but have not been widely adopted due to concerns regarding efficiency and cellular viability. Recently, microfluidic technologies have provided new opportunities for cargo delivery by allowing for precise control of fluid dynamic parameters to achieve efficient and safe penetration of cell membranes, as well as for foreign material transport. Microfluidics-based mechanical delivery methods utilize biophysical phenomena, such as cell constriction and fluid shear, and are associated with high throughput and high transfection efficiency. In this review, we summarize the latest advancements in microfluidic mechanical delivery technologies, and we discuss constriction- and fluid shear-induced delivery strategies. Furthermore, we explore the potential application of artificial intelligence in optimizing cargo delivery technologies, aiming to provide theoretical support and practical guidance for the future development of novel cellular drug delivery technologies.

Accurate detection of pathogenic nucleic acids is crucial for early diagnosis, effective treatment, and containment of infectious diseases. It facilitates the timely identification of pathogens, aids in monitoring disease outbreaks, and helps prevent the spread of infections within healthcare settings and communities. We developed a multi-layered, paper-based microfluidic and miniaturized electrophoresis system for rapid nucleic acid extraction, separation, amplification, and detection, designed for resource-limited settings. Constructed from acrylic, transparency film, pressure-sensitive adhesion, and Whatman paper using a CO₂ laser, the setup simplifies traditional methods and eliminates the need for complex equipment. DNA extraction and purification are achieved using Zweifach-Fung bifurcation and Fahraeus effect principles, with detection via a hydrogel-assisted colorimetric isothermal reverse transcriptase-loop-mediated isothermal amplification technique. The system accurately identified the SARS-CoV-2 N-gene and β-actin human gene, validated by a compact electrophoresis setup. In clinical validation with 12 patient specimens, the system demonstrated a positive predictive agreement of 83.0% and a negative predictive agreement of 100%. The system achieves a limit of detection of 1 copy/μl and can potentially transform nucleic acid detection assays in healthcare settings. This study addresses key challenges in nucleic acid detection, such as ensuring sample quality and quantity, reducing reliance on sophisticated equipment, preventing contamination, simplifying procedures, and providing rapid and accurate diagnostics for emerging pathogens.

Rheotaxis is one of the major migratory mechanisms used in autonomous swimmers such as sperms and bacteria. Here, we present a microfluidic chip using joint rheotaxis and boundary-following behavior that selects sperms based on the motility and persistence. The proposed device consists of a channel decorated with diamond-shaped pillars that create spots of increased velocity field and shear rate. These spots are supposed as hydrodynamic barriers that impede the passage of less motile sperms through the channels, while highly motile sperms were able to overcome the generated barrier and swim through the structures. The proposed device was able to populate the chamber with sorted sperms that were fully viable and motile. The experimental results validated the separation of highly motile sperms with enhanced motility parameters compared with the initial sample. Our device was able to improve linear straight velocity, curvilinear velocity, and average path velocity of the sorted population surpassing 35%, compared with the raw semen. The processing time was also reduced to 20 min.

Single-cell printing technology has arisen as a potent instrument for investigating cell biology and disease pathophysiology. Nonetheless, current single-cell printing methodologies are hindered by restricted throughput, a limited field of view, and diminished efficiency. We present an innovative single-cell printing chip that utilizes thermal inkjet technology for single-cell printing, therefore addressing these constraints. We have accomplished high-throughput, wide-field, and efficient single-cell printing by merging a high-density thermal foam-based inkjet nozzle array on a chip with high-speed cameras and computer vision technologies for optical image capture and single-cell identification training. We have shown the efficacy and adaptability of the printing chip by printing various concentrations of Chinese hamster ovary cells and human embryonic kidney 293 cells. The printing of a single 96-well plate is accomplished in 2-3 min, facilitating one-time loading and uninterrupted multi-plate paving. Our thermal bubble single-cell printing chip serves as a viable platform for high-throughput single-cell analysis applications.