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.

Interventional Removal of Travelling Microthrombi Using Targeted Magnetic Microbubble.

Microthrombus is one of the major causes of the sequelae of Corona Virus Disease 2019 (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 min, without damaging the organs. It is a promising method for treating microthrombus issues.

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.

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.

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.

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.

Enhanced Anticoagulation of Hierarchy Liquid Infused Surfaces in Blood Flow.

Liquid infused surfaces (LIS) hold remarkable potential in anticoagulation. However, liquid loss of LIS in the bloodstream remains a challenge toward its clinical application. Here, micronano hierarchy structures are obtained on the titanium alloy substrate by regulating the microspheres' distribution. When the gap between the microspheres is smaller than the diameter of the red blood cell (RBC), the LIS is more stable under the blood wash and presents a better anticoagulation performance. The proper interval is found to prevent the RBCs from entering the gap and remove the liquid on the surface. The retained thickness of the liquid film is measured by the atomic force microscopy (AFM) technique. The LIS is applied on the front guide vane of an artificial heart pump and exhibits significant improvement on anticoagulation in the blood circulation in vitro for 25 h. The techniques and findings can be used to optimize the anticoagulation performance of LIS-related biomedical implant devices.

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.

Nanodroplet-Coated Microbubbles Used in Sonothrombolysis with Two-Step Cavitation Strategy.

Thrombosis is a major cause of morbidity and mortality and sonothrombolysis is a promising method for its treatment. However, the slow diffusion of the thrombolytic agents into the thrombus results in slow recanalization. Here, nanodroplet-coated microbubbles (NCMBs) are designed and fabricated and a two-step cavitation strategy is used to accelerate the thrombolysis. The first cavitation of the NCMBs, cavitation and collapse of the microbubbles induced by low frequency ultrasound, drives the nanodroplets on the shell into the thrombus, while the second cavitation, the phase-change and volume expansion of drug-loaded nanodroplets triggered by high frequency ultrasound, loosens the thrombus by the sono-porosity effect. This two-step cavitation of the NCMBs is verified using a fibrin agarose model, where a rapid diffusion of the thrombolytic agents is observed. Furthermore, the NCMBs reach much higher thrombolysis efficiency in both in vitro and proof-of-concept experiments performed with living mice. The nanodroplet-coated microbubbles are a promising diffusion medicines carrier for efficient drug delivery.

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.

Enhanced thrombolysis by endovascular low-frequency ultrasound with bifunctional microbubbles in venous thrombosis: in vitro and in vivo study.

There is a need to improve the efficacy and safety of endovascular techniques in venous thrombotic diseases, and microbubble enhanced sonothrombolysis is a promising approach. However, whether endovascular low-frequency ultrasound (LFUS) can be utilized in microbubble enhanced sonothrombolysis is unclear. Here, we present a catheter-based thrombolytic system that combines unfocused low-frequency low-intensity ultrasound with novel fibrin-targeted drug-loaded bifunctional microbubbles. We develop an in vitro flow model and an in vivo rabbit inferior vena cava (IVC) thrombosis model to evaluate the safety and efficacy of the thrombolytic system. The results indicate that microbubble enhanced sonothrombolysis with endovascular LFUS treatment for 30 min is equally effective compared to pure pharmacologic treatment. Furthermore, the thrombolytic efficacy of this system is safely and substantially improved by the introduction of a fibrin-targeted drug-loaded bifunctional microbubble with a reduction of the fibrinolytic agent dosage by 60%. The microbubble enhanced endovascular LFUS sonothrombolysis system with excellent thrombolytic efficacy may serve as a new therapeutic approach for venous thrombotic diseases.

Aortic valve opening in mock-loop with continuous-flow left ventricular assist device.

The appropriate opening of aortic valves is crucial for heart failure (HF) patients with left ventricular assist devices (LVADs). Nevertheless, up to the present time, aortic valve monitoring has not been performed in discharged patients. In this study, a mock-loop platform was developed to investigate the aortic valve performance in LVAD patients. An additional sluice valve was placed next to the aortic valve that when the sluice valve is manually closed, the aortic valve will remain closed; when the sluice valve is open, the aortic valve is opened or closed upon the pressures. The results showed that when the LVAD speed was below 2600 rpm, the aortic valve can be intermittently opened, while when the LVAD speed was over 2600 rpm, the aortic valve was persistently closed. The left ventricular end-systolic pressure (LVESP) was found to be an indicator of aortic valve closure that, upon the aortic valve closure LVESP suddenly decreased. The LVESP is suggested for future monitoring the status of the aortic valve for patients with implanted LVADs. The effects of heart failure (HF) degrees, circulation resistance, and aortic compliance on aortic valve closure were further studied. The results revealed that LVAD implantation in patients with early HF degrees will help to avoid persistent aortic valve closure.

Binding properties of flowing fibrin-targeted microbubbles evaluated with a thrombus-embedded microchannel.

Fibrin is found in both arterial and venous thrombi, which provides an important target for thrombus-targeted microbubbles (MBs) used in MB-enhanced ultrasound imaging and sonothrombolysis. A fibrin-targeted peptide, Cys-Arg-Glu-Lys-Ala (CREKA), is used to modify the commercially available SonoVue ultrasound contrast agent using a conjugation method, and the binding capacity and binding strength of the fibrin-targeted CREKA-modified SonoVue MBs are evaluated with a thrombus-embedded microchannel at a typical shear rate range of venous and arterial blood flow. The experimental results indicate that the targeted MBs bind firmly to the thrombus surface when they flow along the microchannel at a wall shear rate of up to 1000 s-1. This work not only provides an effective method for the fabrication of fibrin-targeted MBs based on commercially available SonoVue MBs but also demonstrates an approach for evaluation of the binding properties of flowing targeted MBs under well-controlled flow conditions.

Interventional Microbubble Enhanced Sonothrombolysis on Left Ventricular Assist Devices.

The left ventricular assist device (LVAD) is often used in the treatment of heart failure. However, 4% to 9% implanted LVAD will have thrombosis problem in one year, which is fatal to the patient's life. In this work, an interventional sonothrombolysis (IST) method is developed to realize the thrombolysis on LVAD. A pair of ultrasound transducer rings is installed on the shell of LVAD, and drug-loaded microbubbles are injected into the LVAD through the interventional method. The microbubbles are adhere on the thrombus with the coated thrombus-targeted drugs, and the thrombolytic drugs carried by the bubbles are brought into the thrombus by the cavitation of bubbles under the ultrasound. In a proof-of-concept experiment in a live sheep model, the thrombus on LVAD is dissolved in 30 min, without damages on LVADs and organs. This IST exhibits to be more efficient and safer compared with other thrombolysis methods on LVAD.

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.

Microfluidic Assembly of Microblocks into Interlocked Structures for Enhanced Strength and Toughness.

Compared with monolithic materials, topologically interlocked materials (TIMs) exhibit higher toughness based on their enhanced crack deflection and deformation tolerance. Importantly, by reducing the block size of TIMs, their structural strength can also be improved due to the reduced flexural span. However, the assembly of microscale blocks remains a huge challenge due to the inadequacy of nanoscale self-assembly or macroscale pick-and-place operations. In this work, octahedral microblocks are fabricated and constructed into interlocked structures with different patterns through microfluidic channels with variable cross sections. The pattern of the interlocked panel is demonstrated to affect its strength and toughness. The failure strength and energy absorption of assembled panels significantly exceed that of their monolithic counterpart by ∼33% and ∼19.1 folds, respectively. Generally, the presented microfluidic method provides a unique technique for the assembly of interlocked architecture, which facilitates the design and fabrication of TIMs with highly improved strength and toughness.

Fabrication of sharp-edged 3D microparticles via folded PDMS microfluidic channels.

3D microparticles have promising applications in self-assembly, biomedical engineering, mechanical engineering, etc. The shape of microparticles plays a significant role in their functionalities. Although numerous investigations have been conducted to tailor the shape of microparticles, the diversity is still limited, and it remains a challenge to fabricate 3D microparticles with sharp edges. Here, we present a facile approach that combines a folded PDMS channel and orthogonal projection lithography for shaping sharp-edged 3D microparticles. By adjusting the number and the length of channel sides, both regular and irregular polyhedral cross-sections of the folded channel can be obtained. UV light with diverse patterns is applied vertically as the second shape controlling factor. A variety of 3D microparticles are obtained with sharp edges, which are potential templates for micromachining tools and abrasives. Some sharp-edged microparticles are assembled into 2D and 3D mesoscale structures, which demonstrates their prospective applications in self-assembly, tissue engineering, etc.

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.

Role of glucose in the repair of cell membrane damage during squeeze distortion of erythrocytes in microfluidic capillaries.

The rapid development of portable precision detection methods and the crisis of insufficient blood supply worldwide has led scientists to study mechanical visualization features beyond the biochemical properties of erythrocytes. Combined evaluation of currently known biochemical biomarkers and mechanical morphological biomarkers will become the mainstream of single-cell detection in the future. To explore the mechanical morphology of erythrocytes, a microfluidic capillary system was constructed in vitro, with flow velocity and glucose concentration as the main variables, and the morphology and ability of erythrocytes to recover from deformation as the main objects of analysis. We showed the mechanical distortion of erythrocytes under various experimental conditions. Our results showed that glucose plays important roles in improving the ability of erythrocytes to recover from deformation and in repairing the damage caused to the cell membrane during the repeated squeeze process. These protective effects were also confirmed in in vivo experiments. Our results provide visual detection markers for single-cell chips and may be useful for future studies in cell aging.

Dielectric tetrahedrons as terahertz resonators switched from perfect absorber to reflector.

Tetrahedrons are basic building blocks in natural and artificial materials, while the terahertz response of micro tetrahedrons has been little explored. Here we fabricate subwavelength ceramic tetrahedrons for use in the terahertz frequency range, and find that the three-dimensional geometry significantly affects their terahertz properties. The transmission spectra are independent of the orientation of the tetrahedrons, while the first magnetic resonance disappears in the reflection spectra when an upright tetrahedron is flipped upside down on the metallic substrate, which changes it from a perfect absorber to a perfect reflector. This is attributed to the destructive interference between two magnetic dipoles induced respectively by the incident and the reflected wave. The study brings new insights in the materials design with 3D building blocks to realize more interesting and exotic terahertz properties.