Fatigue of red blood cells under periodic squeezes in ECMO.

Hemolysis usually happens instantly when red blood cells (RBCs) rupture under a high shear stress. However, it is also found to happen gradually in the extracorporeal membrane oxygenation (ECMO) under low but periodic squeezes. In particular, the gradual hemolysis is accompanied by a progressive change in morphology of RBCs. In this work, the gradual hemolysis is studied in a microfluidic device with arrays of narrow gaps the same as the constructions in ECMO. RBCs are seen to deform periodically when they flow through the narrow gaps, which causes the release of adenosine-triphosphate (ATP) from RBCs. The reduced ATP level in the cells leads to the fatigue of RBCs with the progressive changes in morphology and the gradual loss of deformability. An empirical model for the fatigue of RBCs is established under the periodic squeezes with controlled deformation, and it reveals a different way of the hemolysis that is dominated by the squeeze frequency. This finding brings a new insight into the mechanism of hemolysis, and it helps to improve the design of circulatory support devices.

Early-Stage Protein Adsorption Sequence on Blood-Contacting Surfaces: Answer to Vroman's Question.

Plasma protein adsorption on blood-contacting surfaces is the initiating significant event and modulates the subsequent coagulation response. Despite decades of research in this area, Vroman's questions in 1986 "Who gets there first?" and "When does the next protein arrive?" remain unanswered due to the lack of detection techniques with sufficient temporal resolution. In this work, we develop a droplet microfluidic technology to detect protein adsorption sequences on six typical blood-contacting surfaces in milliseconds. Apolipoproteins (Apo) are found to be the first proteins to adsorb onto the surfaces in a plasma droplet, and the specific type of apolipoprotein depends on the surface. Apo CI is the first protein adsorbed on gold, platinum, graphene, stainless steel, and polyvinyl chloride with the adsorption time varying from 0.01 to 1 s, while Apo CIII preferentially reaches the titanium alloy surface within 1 s. Subsequent to the initial adsorption, Apo AI, AII, and other proteins continue to adsorb until albumin arrives. Thus, the adsorption sequence is revealed, and Vroman's questions are answered. Moreover, this finding demonstrates the influence of the initial protein adsorption on subsequent coagulation at the surface, and it offers new insights into the development of anticoagulant surfaces.

Bubble Growth on Hydrophobic Rough Surfaces in the Shear Flow.

It is well known that bubbles will form on a hydrophobic rough surface immersed in water, which can create a surface covered with bubbles and leads to drag reduction. However, it is still not clear how bubbles grow on the surface under flow conditions. In this work, a rotating flow field is created using a parallel-plate setup of a rotational rheometer, and sample surfaces with different roughnesses and wettabilities are examined with different shear rates. The growth of bubbles is exclusively observed on the hydrophobic rough surface, and subsequent drag reduction is also detected simultaneously. The growth of bubbles is attributed to heterogeneous nucleation in the crevices under a local pressure reduction generated by the shear flow. A geometric model is established to describe the profile evolution of the trapped bubble in the crevice based on the contact angle and the pressure balance across the gas-liquid interface, which involves the variations of the Laplace pressure resulting from changes in the local liquid pressure. The growth of bubbles on the hydrophobic rough surface does not need a large decrease of the surrounding pressure or a high moving speed, which will have potential applications in drag reduction under the condition of a moderate shear rate.

The effect of ultrasound-assisted thrombolysis studied in blood-on-a-chip.

BACKGROUND: Thromboembolism, which leads to pulmonary embolism and ischemic stroke, remains one of the main causes of death. Ultrasound-assisted thrombolysis (UAT) is an effective thrombolytic method. However, further studies are required to elucidate the mechanism of ultrasound on arterial and venous thrombi. METHODS: We employed the blood-on-a-chip technology to simulate thrombus formation in coronary stenosis and deep vein valves. Subsequently, UAT was conducted on the chip to assess the impact of ultrasound on thrombolysis under varying flow conditions. Real-time fluorescence was used to assess thrombolysis and drug penetration. Finally, scanning electron microscopy and immunofluorescence were used to determine the effect of ultrasound on fibrinolysis. RESULTS: The study revealed that UAT enhanced the thrombolytic rate by 40% in the coronary stenosis chip and by 10% in the deep venous valves chip. This enhancement is attributed to the disruption of crosslinked fibrin fibers by ultrasound, leading to increased urokinase diffusion within the thrombus and accumulation of plasminogen on the fibrinogen α chain. Moreover, the acceleration of the dissolution rate of thrombi in the venous valve chip by ultrasound was not as significant as that in the coronary stenosis chip. CONCLUSION: These findings highlight the differential impact of ultrasound on thrombolysis under various flow conditions and emphasize the valuable role of the blood-on-a-chip technology in exploring thrombolysis mechanisms.

Flow field around bubbles on formation of air embolism in small vessels.

An air embolism is induced by intravascular bubbles that block the blood flow in vessels, which causes a high risk of pulmonary hypertension and myocardial and cerebral infarction. However, it is still unclear how a moving bubble is stopped in the blood flow to form an air embolism in small vessels. In this work, microfluidic experiments, in vivo and in vitro, are performed in small vessels, where bubbles are seen to deform and stop gradually in the flow. A clot is always found to originate at the tail of a moving bubble, which is attributed to the special flow field around the bubble. As the clot grows, it breaks the lubrication film between the bubble and the channel wall; thus, the friction force is increased to stop the bubble. This study illustrates the stopping process of elongated bubbles in small vessels and brings insight into the formation of air embolism.

LncRNA SNHG12 Promotes Osteoarthritis Progression Through Targeted Down-Regulation of miR-16-5p.

BACKGROUND: In accordance with increasing studies, long non-coding RNAs (LncRNAs) act pivotally in the occurrence as well as development of several human diseases. But how lncRNA SNHG12 acts in osteoarthritis (OA) is still not clear. METHODS: We applied CCK-8 to determine cell viability, along with qRT-PCR to detect mRNA expression. Using luciferase reporter experiment, our team detected the binding relationship between lncRNA SNHG12 along with miR-16-5p. RESULTS: The inflammatory factor IL-1ß induced chondrocytes to express lncRNA SNHG12, and lncRNA SNHG12 expression was up-regulated in OA tissues. Additionally, our personnel proved that IL-1ß inhibited miR-16-5p expression in chondrocytes, which in OA tissues was lower than that in normal tissues. miR-16-5p expression level in the OA patients' tissue was negatively correlated with lncRNA SNHG12 expression. The high-expression lncRNA SNHG12 inhibits chondrocyte proliferation, promoting apoptosis and inflammation as well as extracellular matrix (ECM) degradation. These effects can be reversed by co-transfecting miR-16-5p mimic. In addition, our work revealed that miR-16-5p is a target of lncRNA SNHG12. CONCLUSIONS: lncRNA SNHG12 regulates OA development by inhibiting miR-16-5p expression in chondrocytes. We believe that the lncRNA SNHG12/miR-16-5p axis might be a potential therapeutic and diagnostic target for OA.

Fluorescent reconstitution on deposition of PM2.5 in lung and extrapulmonary organs.

The deposition of PM2.5 (fine particulate matter in air with diameter smaller than 2.5 µm) in lungs is harmful to human health. However, real-time observation on the deposition of particles in the acinar area of the lung is still a challenge in experiments. Here, a fluorescent imaging method is developed to visualize the deposition process with a high temporal and spatial resolution. The observations reveal that the deposition pattern is nonuniform, and the maximum deposition rate in the acinar area differs significantly from the prediction of the widely used average deposition model. The method is also used to find single particles in the kidney and liver, though such particles are commonly believed to be too large to enter the extrapulmonary organs.

Hydrodynamically Formed Uniform Thick Coatings on Microspheres.

Forming uniform thick coatings on microspheres remains a significant challenge in various surface modification and drug delivery applications. In this work, a hydrodynamic method is demonstrated for centering microspheres in droplets with sizes ranging from tens to hundreds of micrometers. The core microspheres stay at the center of the droplets due to the hydrodynamic pressure generated in the surrounding liquid shells, despite the significant density difference between the core microsphere and the liquid shell. Therefore, by using polymerizable liquids that can be solidified thermally or by illumination as the shell layer, core-shell particles with gas, liquid, or solid cores can be surrounded with uniform coatings using the present method.

Size-Dependent Phase Separation in Emulsion Droplets.

Phase separation occurs in emulsion droplets containing poly (ethylene glycol) diacrylate (PEGDA), glycerol, and ethanol to form a glycerol-in-PEGDA structure, and the phase separation process is found to depend on the droplet size. The mechanism of this size-dependent phase separation is dependent on the droplet sizes changing the phase separation time by changing the evaporation speed of mutual solvent ethanol, and the relationship between the separation time T and the droplet diameter D is derived as T≈D2 , which has been validated by experiment results. According to this finding, the structures of the droplets can be designed by applying UV curing at different stages of the phase separation, and the monodispersity of droplets is necessary to achieve polymerized particles with the same structure.

Cleaning of Fluid-Infused Surfaces in Microchannels.

When an immiscible fluid is flowing over a fluid-infused surface with transverse grooves in a microchannel, the infused fluid is either left in or cleaned away from the grooves by the flowing fluid. The cleaning status depends on the geometric parameters of the groove and the contact angle of the flowing fluids. The critical width of the grooves for the infused fluid enclosed in or driven out of the grooves is derived. This study helps to understand the stability of the Cassie status in a low-shear flow where the surface tension plays the key role.

Lactic Dehydrogenase in the In Vitro Evaluation of Hemolytic Properties of Ventricular Assist Device.

Ventricular assist devices (VADs) can effectively improve the survival rate of patients with end-stage heart failure, but the hemolytic complications induced by long-time VAD support have received wide attention recently. The conventional evaluation method of the hemolytic properties of VADs by the indicator of plasma free hemoglobin (PFH) concentration is used but not sensitive enough to meet the needs of the actual examinations. In this study, an experimental method was applied for the evaluation of the injuries and damages caused by VADs to erythrocytes by both indicators of PFH and lactic dehydrogenase (LDH) in the in vitro hemolysis assay of VADs. The changes of LDH and PFH concentrations in plasma with the shear stress and the exposure time under a fixed shear stress were measured and analyzed to investigate the sensitivity and accuracy of the evaluation of erythrocyte damage by LDH. Furthermore, through 24 h in vitro hemolysis tests, the changes of LDH and PFH concentrations in blood samples were measured in combination with the microscopic histological changes and ultrastructural changes of erythrocytes, to assess the possibility of LDH evaluating the hemolysis of VADs. The changes of the concentration of LDH were more sensitive than those of PFH to different shear stress and exposure times, especially lower stress. Meanwhile, in the 24 h in vitro hemolysis assay, the PFH concentration in the blood samples showed no significant changes in the first 8 h, while the LDH concentration increased significantly in the first 3 h, which was consistent with the morphological changes of erythrocytes. Compared with the changes of the PFH concentration, the LDH concentration is sensitive to the damage of erythrocytes caused by VADs. It was considered that LDH could be applied as an additional indicator in the evaluation of erythrocyte damage and hemolytic properties of VADs in combination with the normalized index of hemolysis, for the more accurate assessment of the blood compatibility of VADs.

Mass-Transfer-Induced Multistep Phase Separation in Emulsion Droplets: Toward Self-Assembly Multilayered Emulsions and Onionlike Microspheres.

Mass-transfer-induced multistep phase separation was found in emulsion droplets. The agent system consists of a monomer (ethoxylated trimethylolpropane triacrylate, ETPTA), an oligomer (polyethylene glycol diacrylate, PEGDA 700), and water. The PEGDA in the separated layers offered partial miscibility of all the components throughout the multistep phase-separation procedure, which was terminated by the depletion of PEGDA in the outermost layer. The number of separated portions was determined by the initial PEGDA content, and the initial droplet size influenced the mass-transfer process and consequently determined the sizes of the separated layers. The resultant multilayered emulsions were demonstrated to offer an orderly temperature-responsive release of the inner cores. Moreover, the emulsion droplets can be readily solidified into onionlike microspheres by ultraviolet light curing, providing a new strategy in designing particle structures.

Drag Reduction by Bubble-Covered Surfaces Found in PDMS Microchannel through Depressurization.

Drag reduction was found in polydimethylsiloxane (PDMS) microchannels when the flow was pulled by depressurization at the inlet, and it was attributed to the formation of the bubbles on the PDMS surface. The formed bubbles were examined by atomic force microscopy (AFM), and the resultant effective slip length was measured by microparticle image velocimetry (µPIV). The drag reduction was found to decrease as the bubbles grew and detached from the surface, causing a pulsatile flow in the microchannel.

Ice lubrication for moving heavy stones to the Forbidden City in 15th- and 16th-century China.

Lubrication plays a crucial role in reducing friction for transporting heavy objects, from moving a 60-ton statue in ancient Egypt to relocating a 15,000-ton building in modern society. Although in China spoked wheels appeared ca. 1500 B.C., in the 15th and 16th centuries sliding sledges were still used in transporting huge stones to the Forbidden City in Beijing. We show that an ice lubrication technique of water-lubricated wood-on-ice sliding was used instead of the common ancient approaches, such as wood-on-wood sliding or the use of log rollers. The technique took full advantage of the natural properties of ice, such as sufficient hardness, flatness, and low friction with a water film. This ice-assisted movement is more efficient for such heavy-load and low-speed transportation necessary for the stones of the Forbidden City. The transportation of the huge stones provides an early example of ice lubrication and complements current studies of the high-speed regime relevant to competitive ice sports.

Growth of bubbles on a solid surface in response to a pressure reduction.

A diffusion-controlled method is presented to study the growth of bubbles on a solid surface. The bubbles are nucleated spontaneously on a hydrophobic smooth surface in response to a sudden pressure reduction and then grow with an expanding contact line. The evolution of the bubbles in the early stage is found to grow with a constant bubble radius and a decreasing contact angle, while the bubbles continue their growth with a constant contact angle and an increasing bubble radius after the contact angle reaches its equilibrium value. A total variation of about 60° of the contact angle is observed during the growth of the bubbles with the size scale of 10-100 µm in radius. The growing process is described by the diffusion theory with the validation of the growth constant.

Sonication-microfluidics for fabrication of nanoparticle-stabilized microbubbles.

An approach based upon sonication-microfluidics is presented to fabricate nanoparticle-coated microbubbles. The gas-in-liquid slug flow formed in a microchannel is subjected to ultrasound, leading to cavitation at the gas-liquid interface. Therefore, microbubbles are formed and then stabilized by the nanoparticles contained in the liquid. Compared to the conventional sonication method, this sonication-microfluidics continuous flow approach has unlimited gas nuclei for cavitation that yields continuous production of foam with shorter residence time. By controlling the flow rate ratios of the gas to the liquid, this method also achieves a higher production volume, smaller bubble size, and less waste of the nanoparticles needed to stabilize the microbubbles.

Dual-Frequency Ultrasound Assisted Thrombolysis in Interventional Therapy of Deep Vein Thrombosis.

Deep vein thrombosis (DVT) is one of the main causes of disability and death worldwide. Currently, the treatment of DVT still needs a long time and faces a high risk of major bleeding. It is necessary to find a rapid and safe method for the therapy of DVT. Here, a dual-frequency ultrasound assisted thrombolysis (DF-UAT) is reported for the interventional treatment of DVT. A series of piezoelectric elements are placed in an interventional catheter to emit ultrasound waves with two independent frequencies in turn. The low-frequency ultrasound drives the drug-loaded droplets into the thrombus, while the high-frequency ultrasound causes the cavitation of the droplets in the thrombus. With the joint effect of the enhanced drug diffusion and the cavitation under the dual-frequency ultrasound, the thrombolytic efficacy can be improved. In a proof-of-concept experiment performed with living sheep, the recanalization of the iliac vein is realized in 15 min using the DF-UAT technology. Therefore, the DF-UAT can be one of the most promising methods in the interventional treatment of DVT.

Interventional Removal of Travelling Microthrombi Using Targeted Magnetic Microbubble.

Microthrombus is one of the major causes of the sequelae of COVID-19 and leads to subsequent embolism and necrosis. Due to their small size and irregular movements, the early detection and efficient removal of microthrombi in vivo remain a great challenge. In this work, an interventional method is developed to identify and remove the traveling microthrombi using targeted-magnetic-microbubbles (TMMBs) and an interventional magnetic catheter. The thrombus-targeted drugs are coated on the TMMBs and magnetic nanoparticles are shelled inside, which allow not only targeted adhesion onto the traveling microthrombi, but also the effective capture by the magnetic catheter in the vessel. In the proof-of-concept experiments in the rat models, the concentration of microthrombus is reduced by more than 60% in 3 minutes, without damaging the organs. It is a promising method for treating microthrombus issues. This article is protected by copyright. All rights reserved.

Multifunctional Conductive and Electrogenic Hydrogel Repaired Spinal Cord Injury via Immunoregulation and Enhancement of Neuronal Differentiation.

Spinal cord injury (SCI) is a refractory neurological disorder. Due to the complex pathological processes, especially the secondary inflammatory cascade and the lack of intrinsic regenerative capacity, it is difficult to recover neurological function after SCI. Meanwhile, simulating the conductive microenvironment of the spinal cord reconstructs electrical neural signal transmission interrupted by SCI and facilitates neural repair. Therefore, a double-crosslinked conductive hydrogel (BP@Hydrogel) containing black phosphorus nanoplates (BP) is synthesized. When placed in a rotating magnetic field (RMF), the BP@Hydrogel can generate stable electrical signals and exhibit electrogenic characteristic. In vitro, the BP@Hydrogel shows satisfactory biocompatibility and can alleviate the activation of microglia. When placed in the RMF, it enhances the anti-inflammatory effects. Meanwhile, wireless electrical stimulation promotes the differentiation of neural stem cells (NSCs) into neurons, which is associated with the activation of the PI3K/AKT pathway. In vivo, the BP@Hydrogel is injectable and can elicit behavioral and electrophysiological recovery in complete transected SCI mice by alleviating the inflammation and facilitating endogenous NSCs to form functional neurons and synapses under the RMF. The present research develops a multifunctional conductive and electrogenic hydrogel for SCI repair by targeting multiple mechanisms including immunoregulation and enhancement of neuronal differentiation.

Wall shear gradient dependent thrombosis studied in blood-on-a-chip with stenotic, branched, and valvular constructions.

Thrombosis is the leading cause of death, while the effect of the shear flow on the formation of thrombus in vascular constructions has not been thoroughly understood, and one of the challenges is to observe the origination of thrombus with a controlled flow field. In this work, we use blood-on-a-chip technology to mimic the flow conditions in coronary artery stenosis, neonatal aortic arch, and deep venous valve. The flow field is measured by the microparticle image velocimeter (µPIV). In the experiment, we find that the thrombus often originates at the constructions of stenosis, bifurcation, and the entrance of valve, where the flow stream lines change suddenly, and the maximum wall shear rate gradient appears. Using the blood-on-a-chip technology, the effect of the wall shear rate gradients on the formation of the thrombus has been illustrated, and the blood-on-a-chip is demonstrated to be a perspective tool for further studies on the flow-induced formation of thrombosis.