Static Conditions Research Articles

Culture insert device with perfusable microchannels enhances in vitro skin model development and barrier function assessment

The ever-stricter regulations on animal experiments in the field of cosmetic testing have prompted a surge in skin-related research with a special focus on recapitulation of the in vivo skin structure in vitro. In vitro human skin models are seen as an important tool for skin research, which in recent years attracted a lot of attention and effort, with researchers moving from the simplest 2-layered models (dermis with epidermis) to models that incorporate other vital skin structures such as hypodermis, vascular structures, and skin appendages. In this study, we designed a microfluidic device with a reverse flange-shaped anchor that allows culturing of an in vitro skin model in a conventional 6-well plate and assessing its barrier function without transferring the skin model to another device or using additional contraptions. Perfusion of the skin model through vascular-like channels improved the morphogenesis of the epidermis compared with skin models cultured under static conditions. This also allowed us to assess the percutaneous penetration of the tested caffeine permeation and vascular absorption, which is one of the key metrics for systemic drug exposure evaluation.

Tuning Electrocatalytic Oxygen Reduction Reaction with Dynamic Control of Electrochemical Interfaces.

Herein, we report the tuning of the activity and selectivity of the oxygen reduction reaction (ORR) through the dynamic regulation of the electrochemical interfaces to surpass the performance of conventional electrocatalysis. This is achieved by applying an oscillating potential between the ORR operating potential and anion adsorbing potential on a gold electrode. Oscillating potential enhances the selectivity for H2O2 by up to 1.35 times compared to the static potential, as confirmed by rotating ring-disk electrode and fluorescence assay measurements. We showed that the enhanced selectivity depends on dynamic adsorption and desorption of anions, and the enhancement occurs in the millisecond time scale or shorter. The transient selectivity to H2O2 can reach ∼97% within the first 5 ms after potential switching. Our results suggest that the dynamic interface can create a transient but unique microenvironment for new reactivity that cannot be reproduced under static conditions, which offers a new dimension in controlling electrocatalysis.

Experimental investigation of dynamic bond behaviors between LRS-FRP and concrete

The dynamic bonding behaviour at the interface between a large-rupture-strain fibre-reinforced polymer (LRS-FRP) and concrete was investigated via drop weight impact tests of notched beam specimens. The main experimental variables included the concrete strength grade, number of LRS-FRP layers, and loading rate. With increasing loading rate, the location of the peeling damage at the interface between the LRS-FRP and concrete shifted from the interior of the concrete layer to the concrete–binder layer interface, binder layer, and even binder layer–FRP interface. The ultimate debonding load of the LRS-FRP strips increased with increasing strain rate, reaching more than 1.7 times the static value at a strain rate of approximately 4/s, while the effective bond length did not show a clear increasing or decreasing tendency with increasing strain rate. For the local bond-slip relationship, both the peak interfacial bond stress and interfacial fracture energy increased with increasing strain rate, and the influences of the concrete strength and number of FRP layers on the strain rate sensitivity were insignificant. Based on the experimental results, improved models to predict the ultimate debonding load and effective bond length at the LRS-FRP–concrete interface under both static and dynamic loading conditions were proposed. Furthermore, dynamic increase factor (DIF) formulas for the peak interfacial bond stress and interfacial fracture energy for bond-slip modelling of the interface between a polyethylene naphthalate (PEN)-FRP and concrete were established.

Development, modeling and analysis of a differential lever-bridge amplification mechanism

The displacement amplification mechanism is commonly utilized in micro/nano-manipulating systems to enhance the output stroke of piezoelectric actuators. Aiming at large amplification ratio, this paper proposes a symmetrical dual-stage amplification mechanism consisting of a differential lever mechanism and a half-bridge mechanism. With consideration of the coupling effect between the dual-stage amplification mechanism, a kinematic modeling method is developed for the symmetrical differential lever-bridge hybrid (SDLBH) mechanism on top of the static balance and constraint conditions of the branch chain. In particular, the force-deformation relationship of the flexible hinge is derived according to the flexibility matrix method. Based on the path constraint conditions of the branch chain, the static equilibrium equation as well as the displacement constraint conditions are obtained. Consequently, the input-output model of the SDLBH mechanism is established, which is capable of accurately predicting the amplification ratio and the mechanical stiffness. Finite element simulations and real time experiments on the SDLBH prototype effectively validate the mechanical performance as well as the accuracy of proposed modeling method. The performance sensitivity of the developed SDLBH mechanism is further analyzed by the Taguchi method, aiming to offer the optimization guideline for real applications.

Interface Engineering to Operate Reversible Protonic Ceramic Electrochemical Cells Below 500 °C

AbstractThe low‐temperature (<500 °C) operation of reversible protonic ceramic electrochemical cells (PCECs) is desirable in achieving efficient and sustainable electricity generation, as well as green hydrogen production. However, significant interfacial resistance, which contributes to both ohmic and polarization resistance, remains a hurdle in lowering the operating temperature. In this study, PrBa0.5Sr0.5Co1.5Fe0.5O5+δ (PBSCF) and BaZr0.4Ce0.4Y0.1Yb0.1O3‐δ (BZCYYb) mono‐grain composite interlayers are introduced, which significantly extend the electrode/electrolyte interface and increase the concentration of vertically aligned oxygen vacancies along the heterointerface. This unique design achieves the lowest ohmic and polarization resistances among previously reported values in solid electrolyte‐based electrochemical cells. As a result, the PCEC can operate at extremely low temperature of 350 °C with an exceptional peak power density of 0.50 W cm−2 in fuel cell mode and current density of 0.25 A cm−2 at 1.3 V in electrolysis cell mode. Furthermore, it demonstrates high energy conversion efficiency and excellent stability under static and dynamic operating conditions.

Activation of heme oxygenase-1 by laminar shear stress ameliorates high glucose-induced endothelial cell and smooth muscle cell dysfunction.

High glucose (HG)-induced endothelial cell (EC) and smooth muscle cell (SMC) dysfunction is critical in diabetes-associated atherosclerosis. However, the roles of heme oxygenase-1 (HO-1), a stress-response protein, in hemodynamic force-generated shear stress and HG-induced metabolic stress remain unclear. This investigation examined the cellular effects and mechanisms of HO-1 under physiologically high shear stress (HSS) in HG-treated ECs and adjacent SMCs. We found that exposure of human aortic ECs to HSS significantly increased HO-1 expression; however, this upregulation appeared to be independent of adenosine monophosphate-activated protein kinase, a regulator of HO-1. Furthermore, HSS inhibited the expression of HG-induced intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and reactive oxygen species (ROS) production in ECs. In an EC/SMC co-culture, compared with static conditions, subjecting ECs close to SMCs to HSS and HG significantly suppressed SMC proliferation while increasing the expression of physiological contractile phenotype markers, such as α-smooth muscle actin and serum response factor. Moreover, HSS and HG decreased the expression of vimentin, an atherogenic synthetic phenotypic marker, in SMCs. Transfecting ECs with HO-1-specific small interfering (si)RNA reversed HSS inhibition on HG-induced inflammation and ROS production in ECs. Similarly, reversed HSS inhibition on HG-induced proliferation and synthetic phenotype formation were observed in co-cultured SMCs. Our findings provide insights into the mechanisms underlying EC-SMC interplay during HG-induced metabolic stress. Strategies to promote HSS in the vessel wall, such as continuous exercise, or the development of HO-1 analogs and mimics of the HSS effect, could provide an effective approach for preventing and treating diabetes-related atherosclerotic vascular complications.

Flexural behavior of reinforced concrete beams strengthened with gradually prestressed near surface mounted carbon fiber-reinforced polymer strips under static and fatigue loading

The near surface mounted (NSM) carbon fiber-reinforced polymer (CFRP) strengthening technique, combined with gradually anchored prestressed technique, is utilized to delay the occurrence of concrete cover separation (CCS) and enhance the ductility of reinforced concrete beams. The load-carrying capacity of fully prestressed beams and gradually prestressed beams are investigated under both static and fatigue loading conditions. The study focused on the effect of gradient prestress on flexural behavior of strengthened beams, analyzed the failure mode, characteristic load, ductility, and stress distribution at CFRP-concrete interface under both prestress and load conditions. Results indicate that gradually prestressed beams outperform fully prestressed ones in restraining crack development, delaying yield of tensile reinforcement, improving bearing capacity and avoiding CCS failure. Bearing capacity was significantly increased by up to 35.48%, while ductility was greatly improved by 100.33% with ultimate deflection for gradually prestressed beams compared to fully prestressed ones. The fatigue life of gradually prestressed beams, which experienced a transition from CCS failure mode to fatigue fracture of tensile reinforcement, was significantly extended. Additionally, their ductility at failure was also greatly enhanced, thus confirming the effectiveness of gradually prestressed NSM CFRP strengthening technique.

Silk Sericin is a Potent, Multifunctional Biomaterial for Endothelialization of Tissue‐Engineered Heart Valves

AbstractEndothelialization is important for offering an anticoagulant surface, which is crucial for the construction of tissue‐engineered heart valves. Silk sericin is extensively investigated in the biomedical field due to its remarkable biological activities and diverse functional groups, favoring chemical modifications for the formation of versatile constructs. Herein, a sericin covalently modified valve (SCMV) is successfully fabricated by coupling maleylated silk sericin (MSS) with thiolated decellularized heart valve (DHV) through a Michael addition reaction. The results demonstrate that SCMV exhibits a smooth surface, higher hydrophilicity, and excellent resistance to platelets adhesion compared to DHV. Additionally, SCMV promotes endothelial cells (ECs) adhesion under both static and flow conditions and enhances their survival in a mouse subcutaneous implant model. The study also discovered that MSS‐stimulated ECs exhibit increased RhoA activation, expression of rho‐associated protein kinase 2 (ROCK2) and phosphorylated myosin light chain 2 (MLC2), actin polymerization, and cell adhesion; whereas actin organization and cell adhesion are significantly reduced by treatment with Y‐27632, a ROCK inhibitor. Furthermore, several integrin subunits, including α1, α2, α3, α5, α6, β1, β3, and β5, are involved in this regulatory process. Collectively, the findings highlight the potential application of silk sericin in promoting endothelialization of DHV while uncovering novel mechanisms underlying its regulatory effect on ECs adhesion.

Fabrication, mechanical, finite element and In vitro evaluation of 3D printed polylactide/biphasic calcium phosphate composite blends

Polylactic acid (PLA) reinforced with biphasic calcium phosphate mixtures were extruded as filaments and subsequently 3D printed into definite shapes in accordance with the ASTM standards. The optimum combination of PLA and BCP to attain a stable 3D printed component has been respectively determined as 80 and 20 wt %. BCP inclusions in the range of 2.5 and 5 wt % in PLA led to a respective upsurge ∼11 % in tensile strength and ∼15 % upsurge in compressive strength than pure PLA. While excess BCP additions led to a decline in the material strength. Further, representative volume element models were developed using the properties of BCP and PLA to build ASTM models for performing Finite element analysis (FEA) under both static and dynamic loading conditions. FEA results specified good uniformity with the experimental mechanical data. In vitro tests ensured the biocompatibility of 3D printed PLA-BCP composites.

Modulation of the Cellular Microenvironment by Mechanical Fluid Shear Stress and Hypoxia Alters the Differentiation Capacity of Skeletal Muscle-Derived Stem Cells

Skeletal muscle-derived stem cells (MDSCs) are the key modulators of muscle regeneration. An inappropriate cellular microenvironment can reduce the regenerative capacity of MDSCs. This study evaluates the effect of microenvironmental alterations on the cell differentiation capacity using either mechanical fluid shear stress (FSS) or hypoxic conditions. C2C12 mouse myoblasts were differentiated under cyclic FSS (CFSS), periodic FSS (PFSS) for one hour, and hypoxia (3% O2) for up to seven days. Cell proliferation and myogenic differentiation capacities were evaluated using cell viability assays, immunohistochemical staining, and morphometric analysis. The expression of MyoD, myogenin, myosin heavy chain, nitric oxide, hypoxia-inducible factor 1 alpha (HIF1α), vascular endothelial growth factor (VEGF) and mammalian target of rapamycin (mTOR) was quantified by means of RT-qPCR. The data showed that FSS conditions altered cell morphology and increased cell viability and cell distribution compared to static conditions. MyoD and myogenin expression was upregulated under both FSS conditions. CFSS induction improved myogenic differentiation parameters including myotube number, size and fusion capacity. Although hypoxia enhanced cell viability compared to normoxia, it reduced differentiation capacity, as indicated by the downregulation of myogenin and mTOR expression, as well as reducing myotube formation. Under hypoxic conditions, increased nitric oxide production and upregulation of VEGF expression were detected for up to 72 h. The data suggest an improved myogenic differentiation capacity under mechanical FSS; in contrast, the cell differentiation capacity was impaired under hypoxic conditions. The data point out that optimizing the biomechanical and oxidative stressors in the cellular microenvironment could improve stem cell transplantation and enhance their regenerative potential in the context of cell-based therapies.

Exploring the electrical robustness of conductive textile fasteners for wearable devices in different human motion conditions

Conventional snap fasteners used in clothing are often used as electrical connectors in e-textile and wearable applications for signal transmission due to their wide availability and ease of use. Nonetheless, limited research exists on the validation of these fasteners, regarding the impact of contact-induced high-amplitude artefacts, especially under motion conditions. In this work, three types of fasteners were used as electromechanical connectors, establishing the interface between a regular sock and an acquisition device. The tested fasteners have different shapes and sizes, as well as have different mechanisms of attachment between the plug and receptacle counterparts. Experimental evaluation was performed under static conditions, slow walking, and rope jumping at a high cadence. The tests were also performed with a test mass of 140 g. Magnetic fasteners presented excellent electromechanical robustness under highly dynamic human movement with and without the additional mass. On the other hand, it was demonstrated that the Spring snap buttons (with a spring-based engaging mechanism) presented a sub-optimal performance under high motion and load conditions, followed by the Prong snap fasteners (without spring), which revealed a high susceptibility to artefacts. Overall, this work provides further evidence on the importance and reliability of clothing fasteners as electrical connectors in wearable systems.

Single molecule studies of dynamic platelet interactions with endothelial cells.

A biotechnological platform consisting of two-color 3D super-resolution readout and a microfluidic system was developed to investigate platelet interaction with a layer of perfused endothelial cells under flow conditions. Platelet activation has been confirmed via CD62P clustering on the membrane and mitochondrial morphology of ECs at the single cell level were examined using 3D two-color single-molecule localization microscopy and classified applying machine learning. To compare binding of activated platelets to intact or stressed ECs, a femtosecond laser was used to induced damage to single ECs within the perfused endothelial layer. We observed that activated platelets bound to the perfused ECs layer preferentially in the proximity to single stressed ECs. Platelets activated under flow were ∼6 times larger compared to activated ones under static conditions. The CD62P expression indicated more CD62P proteins on membrane of dynamically activated platelets, with a tendency to higher densities at the platelet/EC interface. Platelets activated under static conditions showed a less pronounced CD62P top/bottom asymmetry. The clustering of CD62P in the platelet membrane differs depending on the activation conditions. Our results confirm that nanoscopic analysis using two-color 3D super-resolution technology can be used to assess platelet interaction with a stressed endothelium under dynamic conditions.

Alignment of fibrous J-aggregates and the resulting macroscopic optical anisotropy observed in static solution.

J-aggregates, which are supramolecular assemblies that exhibit unique optical properties owing to their excitonic interactions, have potential applications in artificial light-harvesting systems and fluorescence biosensing. Although J-aggregates are formed in solution, in situ observations of their structures and behaviors in solution remain scarce. In this study, we investigated the J-aggregates of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate [DiIC18(3)] in methanol/water (M/W) binary solvents using fluorescence imaging as well as polarized absorption and fluorescence measurements to explore the relationship between their structure and macroscopic optical properties under static conditions. Fluorescence images revealed that the DiIC18(3) J-aggregates have fibrous structures in the M/W = 44/56 (v/v) binary solvent. We measured the polarization-angle dependence of the fluorescence intensity of the fibrous J-aggregates to determine the direction of their transition dipole moment. Furthermore, the J-band absorbance was dependent on the polarization angle of the linearly polarized incident light, even in the absence of an external force such as that generated by a flow or stirring, indicating that the J-aggregates "spontaneously" aligned in solution. We also monitored the time evolution of the degree of alignment of the fibrous J-aggregates, which revealed that the formation and elongation of the fibers induced their alignment, resulting in the observed macroscopic optical anisotropy in solution.

Water-in-Oil Emulsion Temporal and Spatial Evolution at Static Conditions and Its Impact on Pipeline Restart

Summary Understanding emulsion evolution at static conditions is crucial for production operations, such as pipeline operations during the shut-in and restart process and separator optimal design. This study experimentally investigated the temporal and spatial evolution of water-in-oil emulsion properties under static conditions. Numerical simulations were conducted to study their impacts on pipeline restart operations. The experiments were conducted in graduated glass cylinders, with mineral oil and tap water as the testing fluids and Span® 80 as the surfactant. Different water cuts, mixing speeds, and surfactant concentrations were investigated. Along with idle time at static conditions, the mixture demonstrated two layers, namely an upper oil layer and a lower emulsion layer, except for the lowest surfactant concentration that gave a third additional free-water layer at the bottom. Experimental results showed a dramatic increase in viscosity in the emulsion layer with time and depth, which was closely related to the increase in the water volumetric fraction. The increase rate slowed down and plateaued out with time. The increase rate is also related to water cut, mixing speed, and surfactant concentration. Experimental results also show that the relationships between the viscosity and water cut for separated emulsion follow the master curve of viscosity and water cut for homogeneous emulsion. This suggests that one can estimate the viscosity using the master curve given the water volumetric fraction. The numerical simulation was conducted for pipelines with a valley configuration and with the fluid properties obtained from the experimental measurements. It demonstrates that a higher pressure is required to restart the flow to the original flow rate. It also shows that the flow rate may not be able to resume its original value given the same pressure boundaries due to the accumulation of dense emulsion layers in the horizontal and upward inclined sections. For example, for a 16-m pipe, the flow cannot be restarted given the same inlet pressure (100 Pa). It can only resume 4.6% of the original flow rate when the pressure is elevated to 300 Pa.

Functional analysis of postural spinal and pelvic parameters using static and dynamic spinometry

BackgroundSpinometry is a radiation-free method to three-dimensional spine imaging that provides additional information about the functional gait patterns related to the pelvis and lower extremities. This radiation-free technology uses the surface topography of the trunk to analyze surface asymmetry and identify bony landmarks, thereby aiding the assessment of spinal deformity and supporting long-term treatment regimes. Especially reliable dynamic spinometric data for spine and pelvis are necessary to evaluate the management of non-specific back pain. Research aimThis study aims to generate reliable dynamic spinometric data for spine and pelvis parameters that can serve as reference data for future studies and clinical practice. MethodsThis study assessed 366 subjects (185 females) under static and 360 subjects (181 females) under dynamic (walking on a treadmill at 3 km/h and 5 km/h) conditions. The DIERS Formetric 4Dmotion® system uses stripes of light to detect the surface topography of the spine and pelvis and identifies specific landmarks to analyze the spine during standing and walking. ResultsRelevant gender effects were calculated for lordotic angle (ηp2 = 0.22) and pelvic inclination (ηp2 = 0.26). Under static conditions, female subjects showed larger values for both parameters (lordotic angle: 41.6 ± 8.60°; pelvic inclination: 25.5 ± 7.49°). Regarding speed effects, three relevant changes were observed (sagittal imbalance: ηp2 = 0.74, kyphotic angle: ηp2 = 0.13, apical deviation: ηp2 = 0.11). The most considerable changes were observed between static condition and 3 km/h, especially for sagittal imbalance and lordotic angle. For these parameters, relevant effect sizes (d > 0.8) were calculated between static and 3 km/h for males and females. Concerning clinical vertebral parameters, only lordotic angle and pelvic inclination were correlated with each other (r = 0.722). ConclusionThis study generated a gender-specific reference database of asymptomatic individuals for static and dynamic spinometry. It demonstrated that the DIERS Formetric 4Dmotion® system could capture natural changes in static and dynamic situations and catalogue functional adaptations of spino-pelvic statics at different speeds. The lordotic angle is an indirect marker of pelvic inclination, allowing spinometry to identify individuals at risk even under dynamic conditions.

Experimental and Density Functional Theory Study of Inhibitors on Cobalt Corrosion for Chemical Mechanical Planarization Process

In the localized and global chemical mechanical planarization (CMP) process of Co barrier layers, corrosion inhibitors play a crucial role in the removal rate (RR) of Co and the post-polishing surface quality. In this study, quantum chemical calculations were employed to investigate the molecular reactivity of benzotriazole (BTA), 5-methyl benzotriazole (TTA), and 2,2’-{[(methyl-1H-benzotriazol-1-yl)methyl]imino}bis-ethanol (TT-LYK), and their inhibitory performance was predicted to follow the order of TT-LYK > TTA > BTA. The experimental results validated the accuracy of the predicted order under static conditions. However, the performance order of the inhibitors shifted to TTA > BTA > TT-LYK due to varying degrees of damage caused by mechanical friction to the inhibitory film. Nevertheless, all three inhibitors are capable of effectively inhibiting the Co RR to meet industrial requirements. Ultimately, considering the outstanding inhibitory performance of TT-LYK and its ease of removal during post-cleaning processes, TT-LYK is determined as the most promising inhibitor for Co barrier layer CMP.

Study on multi-scale coupled simulation of natural circulation system with 3×3 petal-shaped fuel bundle channel under rolling condition

Petal-shaped fuel rod encompasses a broad application prospect in sea-based nuclear power plants. A multi-scale coupled simulation is conducted based on natural circulation system with 3×3 petal-shaped fuel bundle channel. A multi-scale coupled model considering rolling motion is developed. Its reliability is verified by existing experimental data. The effect of rolling motion on the characteristics of natural circulation system is thoroughly discussed. The results indicate that if the start-up direction of rolling motion is the same as the direction of flow rate, rolling motion will lead to a sharp decrease in flow rate at the initial moment. By increasing rolling amplitude or decreasing rolling period will cause a significant increase in fluctuation amplitude of flow rate. Compared with the results under static condition, the fluid is subjected to the Coriolis force in the non-inertial coordinate system. This research can provide support for the joint optimization of natural circulation system.

Detecting Normal and Cancer Skin Cells via Glycosylation and Adhesion Signatures: A Path to Enhanced Microfluidic Phenotyping

Recruiting circulating cells based on interactions between surface receptors and corresponding ligands holds promise for capturing cells with specific adhesive properties. Our study investigates the adhesion of skin cells to specific lectins, particularly focusing on advancements in lectin-based biosensors with diagnostic potential. We explore whether we can successfully capture normal skin (melanocytes and keratinocytes) and melanoma (WM35, WM115, WM266-4) cells in a low-shear flow environment by coating surfaces with lectins. Specifically, we coated surfaces with Dolichos biflorus (DBA) and Maackia Amurensis (MAL) lectins, which were used to detect and capture specific skin cells from the flow of cell mixture. Alterations in glycan expression (confirmed by fluorescent microscope) demonstrated that DBA binds predominantly to normal skin cells, while MAL interacts strongly with melanoma cells. Assessing adhesion under static and dynamic low-shear conditions (up to 30 mPas) underscores the reliability of DBA and MAL as markers for discriminating cell types. Melanocytes and keratinocytes adhere to DBA-coated surfaces, while melanoma cells prefer MAL-coated surfaces. A comprehensive analysis encompassing cell shape, cytoskeleton, and focal adhesions shows the independence of our approach from the inherent characteristics of cells, thus demonstrating its robustness. Our results carry practical implications for lectin-biosensor designs, emphasizing the significance of glycan-based discrimination of pathologically altered cells. Combined with microfluidics, it demonstrates the value of cell adhesion as a discriminant of cancer-related changes, with potential applications spanning diagnostics, therapeutic interventions, and advanced biomedical technologies.

Collagen IV assembly is influenced by fluid flow in kidney cell-derived matrices

Kidney podocytes and endothelial cells assemble a complex and dynamic basement membrane that is essential for kidney filtration. Whilst many components of this specialised matrix are known, the influence of fluid flow on its assembly and organisation remains poorly understood. Using the coculture of podocytes and glomerular endothelial cells in a low-shear stress, high-flow bioreactor, we investigated the effect of laminar fluid flow on the composition and assembly of cell-derived matrix. With immunofluorescence and matrix image analysis we found flow-mediated remodelling of collagen IV. Using proteomic analysis of the cell-derived matrix we identified changes in both abundance and composition of matrix proteins under flow, including the collagen-modifying enzyme, prolyl 4-hydroxylase (P4HA1). To track collagen IV assembly, we used CRISPR-Cas9 to knock in the luminescent marker HiBiT to the endogenous COL4A2 gene in podocytes. With this system, we found that collagen IV was secreted and accumulated consistently under both static and flow conditions. However knockdown of P4HA1 in podocytes led to a reduction in the secretion of collagen IV and this was more pronounced under flow. Together, this work demonstrates the effect of fluid flow on the composition, modification, and organisation of kidney cell-derived matrix and provides an in vitro system for investigating flow-induced matrix alteration in the context of kidney development and disease.

Methylene blue treatment with zero-valent iron/pyrite/H2O2 system under static and continuous flow conditions: Reaction mechanism and toxicity evaluation

Laboratory batch and continuous flow studies, coupled with surface and toxicity analysis, were performed to evaluate methylene blue treatment by heterogeneous Fenton process using zero-valent iron (ZVI) and pyrite as co-catalyst. The use of pyrite in batch reactors significantly enhanced methylene blue treatment by the ZVI/H2O2 system because of improved iron redox cycling. The continuous-flow experiments revealed that the reactor performance increased in the order of: ZVI/H2O2 < pyrite/H2O2 < ZVI/pyrite/H2O2 under dynamic flow conditions. The methylene blue treatment by the ternary ZVI/pyrite/H2O2 system was described by an initial degradation of methylene blue with *OH radicals, followed by the adsorption and/or co-precipitation of degradation intermediates with some spherical particles. The surface analysis showed that these spherical particles formed, even at pH less than 4. The genotoxicity and cytotoxicity tests performed on mouse embryonic fibroblast (3 T3-L1) and human embryonic kidney (HEK293) cell lines showed that the Fenton treatment of methylene blue using the ZVI/pyrite/H2O2 system resulted in the formation of degradation species with much lower toxicity levels relative to methylene blue. Moreover, the Fenton degradation species of methylene blue significantly enhanced the metabolic activity of several bacterial strains, including E. coli ATCC 8739 and P. aeruginosa PAO1.