The Effect of IL-6-Primed Platelets on ADAMTS13-Mediated Clearance of Platelet-Bearing ULVWF and Its Mechanism

Inflammation is an essential contributing factor in the development of thrombosis. Using a microfluidic flow chamber, we investigated how the proinflammatory cytokine interleukin 6 (IL-6) affects the cleavage of platelet-bearing ultra-large VWF (ULVWF) by plasma ADAMTS13. We found that IL-6-treated platelets perfused at arteriolar shear stress significantly enhanced the ULVWF-platelet complex formation on activated endothelial cells and suppressed their clearance by ADAMTS13 under flow conditions. We also detected the phosphorylation of the serine/threonine kinase Akt and extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) in platelets treated with IL-6. Treatment of IL-6-primed platelets with either the phosphoinositol-3 kinase (PI3K) inhibitor LY294002 or the mitogen-activated protein kinase kinase (MEK) inhibitor U0126 reduced the ULVWF-platelet complex formation and restored the clearance of the complex by plasma ADAMTS13, compared to IL-6-primed platelets. Furthermore, IL-6 enhanced the phosphorylation of the intracellular adaptor molecule 14-3-3ζ, which regulates VWF binding to the glycoprotein (GP) Ib-IX complex. The 14-3-3 antagonist R18 significantly increased ADAMTS-13 cleavage of ULVWF strings with adherent IL-6-treated platelets. These findings indicate that IL-6 related intracellular signals of platelet is involved in regulating ULVWF-platelet binding and ULVWF cleavage by ADAMTS13.

Modeling of a Graphene Nanoribbon–based Microfluidic Surface Plasmon Resonance Biosensor

A surface plasmon resonance (SPR) biosensor based on a graphene nanoribbon array in a microfluidic flow cell operating in a flow-over format is studied. The optical response of the biosensor is numerically obtained by using rigorous couple wave analysis (RCWA). The performance of the biosensor is described in terms of the limit of detection, which is calculated as a function of key nanoribbon dimensional parameters, such as strip thickness and width, and fill fraction (nanoribbon width to array period ratio). The analysis shows that there are specific values of the fill fraction that optimize, that is, minimize, the limit of detection for particular nanoribbon dimensions. Fabrication issues are also discussed. This study is expected to assist in the design and implementation of SPR biosensors based on nanopatterned 2D materials.

Controlling Droplet Marangoni Flows to Improve Microscopy-Based TB Diagnosis

In developing countries, the most common diagnostic method for tuberculosis (TB) is microscopic examination sputum smears. Current assessment requires time-intensive inspection across the microscope slide area, and this contributes to its poor diagnostic sensitivity of ≈50%. Spatially concentrating TB bacteria in a smaller area is one potential approach to improve visual detection and potentially increase sensitivity. We hypothesized that a combination of magnetic concentration and induced droplet Marangoni flow would spatially concentrate Mycobacterium tuberculosis on the slide surface by preferential deposition of beads and TB–bead complexes in the center of an evaporating droplet. To this end, slide substrate and droplet solvent thermal conductivities and solvent surface tension, variables known to impact microfluidic flow patterns in evaporating droplets, were varied to select the most appropriate slide surface coating. Optimization in a model system used goniometry, optical coherence tomography, and microscope images of the final deposition pattern to observe the droplet flows and maximize central deposition of 1 μm fluorescent polystyrene particles and 200 nm nanoparticles (NPs) in 2 μL droplets. Rain-X® polysiloxane glass coating was identified as the best substrate material, with a PBS-Tween droplet solvent. The use of smaller, 200 nm magnetic NPs instead of larger 1 μm beads allowed for bright field imaging of bacteria. Using these optimized components, we compared standard smear methods to the Marangoni-based spatial concentration system, which was paired with magnetic enrichment using iron oxide NPs, isolating M. bovis BCG (BCG) from samples containing 0 and 103 to 106 bacilli/mL. Compared to standard smear preparation, paired analysis demonstrated a combined volumetric and spatial sample enrichment of 100-fold. With further refinement, this magnetic/Marangoni flow concentration approach is expected to improve whole-pathogen microscopy-based diagnosis of TB and other infectious diseases.

How to Control the Microfluidic Flow and Separate the Magnetic and Non-Magnetic Particles in the Runner of a Disc

Biochips play an important role in both medical and food industry safety testing. Moreover, magnetic activated cell sorting is a well-established technology for biochip development. However, biochips need to be manufactured by precision instruments, resulting in the high cost of biochips. Therefore, this study used magnetic-activation and mechanics theories to create a novel disc that could manipulate the microfluidic flow, mixing, reaction, and separation on the runner of the disc. The goal of the research was to apply in the field of biomedical detection systems to reduce the cost of biochips and simplify the operation process. The simulation and experimental investigation showed that the pattern of the reaction chamber was stomach-shaped and the reservoir chamber was rectangular-shaped on the disc. The microfluid could be controlled to flow to the reaction chamber from the buffer and sample chamber when the disc spun at 175~200 rpm within three minutes. This was defined as the first setting mode. The microfluid could then be controlled to flow to the reservoir chamber from the reaction chamber when the disc spun at 225 rpm within five to ten minutes. This was defined as the second setting mode. This verified that the pattern design of the disc was optimized for control of the microfluid flow, mixing, reaction, and separation in the runner of the disc by different setting modes.

Microfluidic Preparation of Janus Microparticles With Temperature and pH Triggered Degradation Properties

Based on the phase separation phenomenon in micro-droplets, polymer-lipid Janus particles were prepared on a microfluidic flow focusing chip. Phase separation of droplets was caused by solvent volatilization and Janus morphology was formed under the action of interfacial tension. Because phase change from solid to liquid of the lipid hemisphere could be triggered by physiological temperature, the lipid hemisphere could be used for rapid release of drugs. While the polymer we selected was pH sensitive that the polymer hemisphere could degrade under acidic conditions, making it possible to release drugs in a specific pH environment, such as tumor tissues. Janus particles with different structures were obtained by changing the experimental conditions. To widen the application range of the particles, fatty alcohol and fatty acid-based phase change materials were also employed to prepare the particles, such as 1-tetradecanol, 1-hexadecanol and lauric acid. The melting points of these substances are higher than the physiological temperature, which can be applied in fever triggered drug release or in thermotherapy. The introduction of poly (lactic-co-glycolic acid) enabled the formation of multicompartment particles with three distinct materials. With different degradation properties of each compartment, the particles generated in this work may find applications in programmed and sequential drug release triggered by multiple stimuli.