Hydrodynamic shear stress' impact on mammalian cell properties and its applications in 3D bioprinting.

As an effective cell assembly method, three-dimensional bioprinting has been widely used in building organ models and tissue repair over the past decade. However, different shear stresses induced throughout the entire printing process can cause complex impacts on cell integrity, including reducing cell viability, provoking morphological changes and altering cellular functionalities. The potential effects that may occur and the conditions under which these effects manifest are not clearly understood. Here, we review systematically how different mammalian cells respond under shear stress. We enumerate available experimental apparatus, and we categorise properties that can be affected under disparate stress patterns. We also summarise cell damaging mathematical models as a predicting reference for the design of bioprinting systems. We concluded that it is essential to quantify specific cell resistance to shear stress for the optimisation of bioprinting systems. Besides, as substantial positive impacts, including inducing cell alignment and promoting cell motility, can be generated by shear stress, we suggest that we find the proper range of shear stress and actively utilise its positive influences in the development of future systems.

Stenosis to stented: decrease in flow disturbances following stent implantation of a diseased arteriovenous fistula.

The arteriovenous fistula (AVF) is commonly faced with stenosis at the juxta-anastomotic (JXA) region of the vein. Implantation of a flexible nitinol stent across the stenosed JXA has led to the retention of functioning AVFs leading to the resulting AVF geometry being distinctly altered, thereby affecting the haemodynamic environment within it. In this study, large eddy simulations of the flow field within a patient-specific AVF geometry before and after stent implantation were conducted to detail the change in flow features. Although the diseased AVF had much lower flow rates, adverse flow features, such as recirculation zones and swirling flow at the anastomosis, and jet flow at the stenosis site were present. Larger velocity fluctuations (leading to higher turbulent kinetic energy) stemming from these flow features were apparent in the diseased AVF compared to the stented AVF. The unsteadiness at the stenosis created large regions of wall shear stress (WSS) fluctuations downstream of the stenosis site that were not as apparent in the stented AVF geometry. The larger pressure drop across the diseased vein, compared to the stented vein, was primarily caused by the constriction at the stenosis, potentially causing the lower flow rate. Furthermore, the WSS fluctuations in the diseased AVF could lead to further disease progression downstream of the stenosis. The change in bulk flow unsteadiness, pressure drop, and WSS behaviour confirms that the haemodynamic environment of the diseased AVF has substantially improved following the flexible stent implantation.

Investigating the hemodynamic implications of triangular cross-sectioned superior sagittal sinus vessels and the errors associated with idealised modelling.

The superior sagittal sinus (SSS) is a blood vessel that is often observed to be approximately triangular in cross-section, due to how the venous wall attaches to the surrounding tissue. Despite this, the vessel has been assumed to be circular, when models are generated without patient-specific data. In this study, the differences between the cerebral hemodynamics of one circular, three triangular and five patient-specific cross-sectional models of a SSS were conducted. The errors associated with using circular cross-sectioned flow extensions were also determined. Computational fluid dynamics (CFD) models were generated from these geometries, with a population mean transient blood flow profile incorporated. The maximal helicity of the fluid flow was found to be elevated in the triangular cross-section, compared to the circular, with a higher wall shear stress (WSS) observed over a smaller, more concentrated region on the posterior sinus wall. The errors associated with using a circular cross-section were detailed, with the cross-sectional area appearing to have a greater influence on the hemodynamic parameters than the triangularity or circularity of the cross-section. This highlighted the importance of exhibiting caution when incorporating idealised modelling, especially when commenting on the true hemodynamics of these models. Errors were also found to be induced when using a circular cross-sectioned flow extension, for a geometry which was non-circular. This study highlights the importance of understanding the human anatomy when modelling blood vessels.

Ultrasonic Traveling Waves for Near-Wall Positioning of Single Microbubbles in a Flowing Channel.

Although microbubbles are used primarily in the medical industry as ultrasonic contrast agents, they can also be manipulated by acoustic waves for targeted drug delivery, sonothrombolysis and sonoporation. Acoustic waves can also potentially remove microbubbles from tubing systems (e.g., in hemodialysis) to prevent the negative effects associated with circulating microbubbles. A deeper understanding of the interactions between the acoustic radiation force, the microbubble and the channel wall could greatly benefit these applications. In this study, single air-filled microbubbles were injected into a flowing (polydimethylsiloxane) channel and monitored by a high-speed camera while passing through a pulsed ultrasonic wave zone (0.5 MHz). This study compared various bubble sizes, flow rates and acoustic pressure amplitudes to better understand the three physical regimes observed: free bubble translation (away from the wall); on-wall translation; and bubble-wall attachment. Comparison with a theoretical model revealed that the acoustic radiation force needs to exceed the combined repulsive forces (shear lift, wall lubrication and repulsive Van der Waal forces) for the dead state of bubble-wall attachment. The bubble dynamics revealed through this investigation provide an opportunity for efficient positioning of microbubbles in a channel flow, for either in vivo manipulation or removal in biological applications.

Long-term Results of Interwoven Nitinol Stents to Treat the Radiocephalic Anastomotic Arteriovenous Fistula Stenosis.

INTRODUCTION: Juxta-anastomotic stenosis (JXAS) is a common problem afflicting the arteriovenous fistula (AVF). This study aimed to evaluate the safety and long-term efficacy of an interwoven nitinol stent (Supera, Abbott Vascular, Santa Clara, CA, USA) in the treatment of radiocephalic AVF JXAS. METHODS: A single-center, retrospective, observational study was conducted of patients with failing AVF due to JXAS treated with an interwoven nitinol stent. End points included JXAS target lesion primary patency, access circuit primary patency, assisted access circuit primary patency, and endovascular intervention rate (EIR). RESULTS: Sixty patients were treated with a Supera stent in the JXAS between February 2014 and March 2020. One patient was excluded (AVF used for illicit drug use), leaving 59 patients (67.8% male, mean age 66.9 ± 11.4 years [range: 40-84]) with typical medical comorbidities. Overall, 45.8% of patients had previous AVF intervention. The stent was inserted with a 100% technical success rate with a mean follow-up of 729.6 ± 456.0 days (range: 5-2182 days). Juxta-anastomotic stenosis target lesion primary patency was 68.2 ± 6.6%, 53.3 ± 7.5%, and 46.2 ± 8.1% at 12, 24, and 36 months, respectively. The EIR was .64 (0-3.29) procedures/patient/year, after which the assisted access circuit primary patency rate was 94.3 ± 3.2% at 12, 24, and 36 month time points. Three thrombosed circuits occurred which were all successfully salvaged with no difference in patency by indication for procedure and no AVFs lost/abandoned out to 3 years. Avoidance of stent post-dilatation and the presence of stent mal-apposition were associated with improved primary patency, and reduced EIR, which may suggest an importance in vessel preparation prior to stent implantation. CONCLUSION: Interwoven nitinol stent treatment of the failing AVF with JXAS results in promising 3 year JXAS patency, with a low rate of endovascular re-intervention for those circuits developing restenosis. All AVFs were maintained over 3 years, demonstrating this treatment allows for long-term radiocephalic AVF vascular access.

Quantified hemodynamic parameters of the venous system in multiple sclerosis: A systematic review.

BACKGROUND: Multiple Sclerosis (MS) is a complex neurodegenerative condition that is influenced by a combination of genetic and environmental factors. Included in these factors is the venous system, however, the extent to which it influences the etiology of MS has yet to be fully characterised. The aim of this review is to critically summarize the literature available concerning the venous system in MS, primarily concerning specific data on the venous pressure and blood flow in this system. METHODS: A systematic review was conducted with the application of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) criteria. The advanced search functions of both the Scopus and PubMed databases were used to conduct the literature search, resulting in 136 unique articles initially identified. Applying relevant exclusion criteria, 22 of the studies were chosen for this review. RESULTS: The selected studies were analysed for venous pressure and blood flow related findings, with 14 studies contributing data on the internal jugular vein (IJV) flow rate, 5 on blood flows of the intracranial venous sinuses, 2 on blood flow pulsatility and 6 supplying information relevant to the venous pressure (3 studies contributed to multiple areas). The general findings of the review included that the IJV flow was not significantly different between MS patients and controls, however, there were variances between stenotic (S) and non-stenotic (NS) MS patients. Due to the limited data in the other two areas defined in this review, further research is required to establish if any variances in MS are present. CONCLUSION: It remains unclear if there are significant differences in many flow variables between MS patients and controls considered in this review. It would be advantageous if future work in this area focused on understanding the hemodynamics of this system, primarily concerning how the flow rate, venous pressure and vascular resistance are related, and any impact that these factors have on the etiology of MS.

A systematic review of three-dimensional printed template-assisted physician-modified stent grafts for fenestrated endovascular aneurysm repair.

OBJECTIVE: Fenestrated endovascular aneurysm repair has yet to gain widespread adoption owing to the technical complexity and increased risk of complications. Three-dimensional (3D) printed templates to guide fenestrated physician-modified stent grafts (PMSGs) are a novel technique that may have the potential to increase the accuracy of fenestration alignment, and to disrupt both the cost and timing of the current commercial fenestrated endograft supply chain. We have conducted a critical appraisal of the emerging literature to assess this. METHODS: A systematic literature search was performed using PubMed and OVID Medline as guided by the PRISMA statement on April 30, 2020. We used "3D printing" and "physician modified" or "surgeon modified" and all related search terms. We identified 50 articles which met our search criteria. None articles were included as being of direct relevance to 3D-printed template-assisted PMSGs for fenestrated endovascular aneurysm repair. Abstracts were screened individually by each investigator to ensure relevance. RESULTS: Nine relevant articles were identified for critical analysis. These included one technical report, five case reports or series, two prospective trials, and one letter to the editor. CONCLUSIONS: These 3D-printed templates are a promising new avenue to assist with the placement of fenestrations in PMSGs, particularly in urgent or emergent cases where custom fenestrated endografts are unavailable, with larger scale studies warranted. Further work to validate the key stages of the template workflow are required, as well as further investigation into the most suitable manufacturing and distribution methods before the mainstream implementation of this novel technique.

The relationship between cerebral blood flow and venous sinus pressure: can hyperemia induce idiopathic intracranial hypertension?

BACKGROUND: It has been shown that idiopathic intracranial hypertension (IIH) in children is associated with cerebral hyperemia, which induces an increase in cerebral venous pressure. The current literature suggests venous pressure scales with blood flow in a linear fashion, however, a linear relationship would not raise the pressure high enough to induce IIH. There is, however, some evidence to suggest that this relationship could be quadratic in nature. The purpose of this paper is to characterize the relationship between cerebral blood flow and the pressure drop across the cerebral venous system. METHODS: 10 CT venogram data sets were collected for this study, with 5 useable geometries created. Computational fluid dynamics (CFD) models were generated using these geometries, with 10 simulations conducted per patient. The flow rates tested ranged from 200 mL/min to 2000 mL/min. 3D pressure and velocity streamline distributions were created and analyzed for each CFD model, with pressure drops across the cerebral venous system determined. The effective and hydraulic diameters were determined at the superior sagittal sinus, transverse sinus and both proximal and distal sigmoid sinuses. RESULTS: A quadratic relationship between blood flow and sinus pressure was found, with correlations of 0.99 or above in all five patients. The presence of vortical blood flow was found to explain this trend, with fluid curl and pressure drop correlations being above 0.97. This suggests that the presence of high blood flow should be considered in the diagnostic workup of IIH. CONCLUSIONS: The cerebral venous sinus blood flow and pressure response relationship are quadratic in nature, with the major cause of this being the degree of rotation induced in the flow. The elevated blood flow found in children with IIH can explain the increased ICP that is found, secondary to the increase in venous pressure that develops.

Hemodialysis taping styles and their effect on reducing the chance of venous needle dislodgement.

INTRODUCTION: Vascular access complications are common among hemodialysis patients, although most are not immediately life-threatening. However, inconsistent taping techniques and incompatible detection mechanisms may lead to venous needle dislodgement (VND), which can lead to catastrophic blood loss. Taping technique does not always meet the recommended best practice and there may be no available protocol for new staff. METHODS: Three commonly used taping patterns (the Chevron, butterfly, and overlapping styles) were tested in a mechanical engineering laboratory to determine the forces that each method was capable of withstanding. RESULTS: While all taping styles were confirmed to have an adhesive force stronger than the inherent force from the venous jet flow of blood, the overlapping style was found to have limited capability beyond this minimum criterion. Both the butterfly and Chevron styles demonstrated excellent holding capability, with elongation of dislodgement particularly noted for the butterfly style, and slightly stronger hold noted for the Chevron style. The Chevron style may be better suited to lateral movements from all directions, due to the taping direction. CONCLUSIONS: We recommend that either the Chevron or butterfly style is used for dialysis needle taping, with the butterfly better suited to home dialysis (where monitors may be used) and the Chevron better suited for in-care patients who may present erratic movements. The overlapping style is not recommended for use.

Arteriovenous fistula maturation and the influence of fluid dynamics.

Arteriovenous fistula creation is the preferred vascular access for haemodialysis therapy, but has a large failure rate in the maturation period. This period generally lasts 6 to 8 weeks after surgical creation, in which the vein and artery undergo extensive vascular remodelling. In this review, we outline proposed mechanisms for both arteriovenous fistula maturation and arteriovenous fistula failure. Clinical, animal and computational studies have not yet shown a definitive link between any metric and disease development, although a number of theories based on wall shear stress metrics have been suggested. Recent work allowing patient-based longitudinal studies may hold the key to understanding arteriovenous fistula maturation processes.

Capillary-assisted microfluidic biosensing platform captures single cell secretion dynamics in nanoliter compartments.

Cancer cells continuously secrete inflammatory biomolecules which play significant roles in disease progression and tumor metastasis toward secondary sites. Despite recent efforts to capture cancer cells' intercellular secretion heterogeneity using microfluidics, the challenges in operation of these systems as well as the complexity of designing a biosensing assay for long-term and real-time measurement of single cell secretions have become grand research barriers. Here, we present a new capillary-based microfluidic biosensing approach to easily and reliably capture ~500 single cells inside isolated dead-end nanoliter compartments using simple pipette injection, and quantify their individual secretion dynamics at the single cell resolution over a long period of culture (~16 h). We first present a detailed investigation of the fluid mechanics underlying the formation of nanoliter compartments in the microfluidic system. Based on the measurement of single cell capture efficiency, we employ a one-step FRET-based biosensor which monitors the single cancer cells' protease activity. The sensor reports the fluorescent signal as a product of amino acid chain cleavage and reduction in its quenching capability. Using the single cell protease secretion data, we identified modes of cell secretion dynamics in our cell sample. While most of the cells had low secretion levels, two other smaller and more aggressive secretion dynamics were cells with secretion modes that include sharp spikes or slow but progressive trend. The method presented here overcomes the difficulties associated with performing single cell secretion assays, enabling a feasible and reliable technique for high throughput measurement of metabolic activities in cancer cells.

Toward Portable Artificial Kidneys: The Role of Advanced Microfluidics and Membrane Technologies in Implantable Systems.

Globally, around 2.6 million people receive renal replacement therapy (RRT), and a further 4.9-9.7 million people need, but do not have access to, RRT [1]. The next generation RRT devices will certainly be in demand due to the increasing occurrence of diabetes, atherosclerosis and the growing population of older citizens. This review provides a comprehensive, yet concise overview of the cleared and remaining hurdles in the development of artificial kidneys to move beyond traditional dialysis technology-the current baseline of renal failure treatment. It compares and contrasts the state-of-the-art in 'cell-based' and 'non-cell-based' approaches. Based on this study, a new engineering perspective on the future of artificial kidneys is described. This review suggests that stem-cell-based artificial kidneys represent a long-term, complete solution but it can take years of development due to the limitations of current cell seeding technology, viability and complicated behaviour control. Alternatively, there is much potential for near- and medium- term solutions with the development of non-cell-based wearable and implantable devices to support current therapies. Based on recent fundamental advances in microfluidics, membranes and related research, it may be possible to integrate these technologies to enable implantable artificial kidneys (iAK) in the near future.

Dynamic Banding (DYBAND) Technique for Symptomatic High-Flow Fistulae.

INTRODUCTION: High flow rates may develop in arteriovenous fistula (AVF), resulting in clinical syndromes of steal, aneurysmal fistula, or high-output cardiac failure. Various techniques with varying success have been advocated to treat this difficult problem. We present a hemodynamically validated novel banding technique. METHODS: We designed a computational fluid dynamic (CFD) model of the native high-flow AVF and tested various juxta-anastomotic venous diameters to determine the effect on AVF blood flow and pressure. We translated this principle in our banding technique, wherein adjustable banding was performed in conjunction with ultrasound-guided brachial artery flow measurement to determine the optimal band diameter. Polyurethane patch was used to fashion the adjustable band. Patient demographics, AVF flow parameters pre- and postintervention, operative intervention, and ultrasound follow-up data were collected prospectively. RESULTS: Our CFD testing demonstrated that the band diameter needed to achieve optimal distal blood pressure and preserve AVF flow depending on blood pressure, end capillary pressure, venous pressure, and vascular diameters. Five patients subsequently underwent dynamic banding of symptomatic high-flow AVF. Mean brachial artery blood flow rates pre- and postbanding were 2964 mL/min (confidence interval [CI]: 1487-4440 mL/min) and 1099 mL/min (CI: 571.7-1627 mL/min), respectively (P = .01). All patients had symptomatic improvement, and at a mean follow-up of 1 year, this benefit was sustained with no AVF thrombosis or loss. CONCLUSION: Adjustable dynamic band using ultrasound-guided brachial artery flow shows promising results in producing accurate AVF blood flow reduction with sustained efficacy in the short term for patients with symptomatic high-flow AVF.

Tracking geometric and hemodynamic alterations of an arteriovenous fistula through patient-specific modelling.

BACKGROUND AND OBJECTIVE: The use of patient-specific CFD modelling for arteriovenous fistulae (AVF) has shown great clinical potential for improving surveillance, yet the use of imaging modes such as MRI and CT for the 3D geometry acquisition presents high costs and exposure risks, preventing regular use. We have developed an ultrasound based procedure to bypass these limitations. METHODS: A scanning procedure and processing pipeline was developed specifically for CFD modelling of AVFs, using a freehand ultrasound setup combining B-mode scanning with 3D probe motion tracking. The scanning procedure involves sweeping along the vasculature to create a high density stack of B-mode frames containing the lumen geometry. This stack is converted into a continuous volume and transient flow waveforms are recorded at the boundaries, synchronised with ECG and automatically digitised, forming realistic boundary conditions for the CFD models. This is demonstrated on a diseased patient-specific AVF. RESULTS: The three scans obtained using this procedure varied in geometry and flow behaviour, with regions of disease located in the first two scans. The outcome of the second procedure seen in the third scan indicated successful restoration with no sites of disease and higher flow. The models gave insight into the lumenal changes in diameter for both the artery and vein segments, as well as characterising hemodynamic behaviours in both the diseased and restored states. Vascular segment resistances obtained from the CFD models indicate a significant reduction once disease was removed, resulting in much higher flows enabling the patient to resume dialysis. CONCLUSION: The methodology described in this study allowed for a multifaceted analysis and high level tracking in terms of both geometry and flow behaviours for a patient case, demonstrating significant clinical utility and practicality, as well as enabling further research into vascular disease progression in AVFs through longitudinal analysis.

Microfluidic Actuation via 3D-Printed Molds toward Multiplex Biosensing of Cell Apoptosis.

Multiplexed analysis of biochemical analytes such as proteins, enzymes, and immune products using a microfluidic device has the potential to cut assay time, reduce sample volume, realize high-throughput, and decrease experimental error without compromising sensitivity. Despite these huge benefits, the need for expensive specialized equipment and the complex photolithography fabrication process for the multiplexed devices have, to date, prevented widespread adoption of microfluidic systems. Here, we present a simple method to fabricate a new microfluidic-based multiplexed biosensing device by taking advantage of 3D-printing. The device is an integration of normally closed (NC) microfluidic valving units which offer superior operational flexibility by using PDMS membrane (E ∼ 1-2 MPa) and require minimized energy input (1-5 kPa). To systematically engineer the device, we first report on the geometrical and operational analysis of a single 3D-printed valving unit. Based on the characterization, we introduce a full prototype multiplexed chip comprising several microfluidic valves. The prototype offers-for the first time in a 3D-printed microfluidic device-the capability of on-demand performce of both a sequential and a parallel biochemical assay. As a proof of concept, our device has been used to simultaneously measure the apoptotic activity of 5 different members of the caspase protease enzyme family. In summary, the 3D-printed valving system showcased in this study overcomes traditional bottlenecks of microfabrication, enabling a new class of sophisticated liquid manipulation required in performing multiplexed sensing for biochemical assays.

Reduction in anastomotic flow disturbance within a modified end-to-side arteriovenous fistula configuration: Results of a computational flow dynamic model.

AIM: We sought to determine if a modification to the peri-anastomotic vein configuration during end-to-side arteriovenous fistula (AVF) creation would achieve a favourable haemodynamic environment compared with the standard acute-angle AVF. METHODS: Computational fluid dynamics (CFD) modelling of two end-to-side AVF geometries (smooth-vein-loop vs. acute-angle) allowed for haemodynamic modelling. Haemodynamic fields at various stages of the pulse cycle were observed and compared across both models. RESULTS: We found a significant reduction in flow disturbance in the modified shape at the peri-anastomotic vein region, a common site for stenosis. The acute anastomotic angle characteristic of a standard-end-to-side AVF was found to cause significant flow disturbance throughout the pulse cycle. CONCLUSION: Computational modelling indicates that improvements in the haemodynamic environment of an AVF are achievable when a gentle change in direction is present in the vein segment. This may lead to improved maturation and AVF patency through reduction in disturbed flow patterns.

Flow-Mediated Drug Transport from Drug-Eluting Stents is Negligible: Numerical and In-vitro Investigations.

Prior numerical studies have shown that the blood flow patterns surrounding drug-eluting stents can enhance drug uptake in stented arteries. However, these studies employed steady-state simulations, wherein flow and drug transport parameters remained constant with respect to time. In the present study, numerical simulations and in-vitro experiments were performed to determine whether luminal blood flow patterns can truly enhance drug uptake in stented arteries. Unlike the aforementioned studies, the time-varying depletion of drug within the stent coating was modelled and the simulation results were validated qualitatively with the in-vitro experiments. The simulations showed that the non-Newtonian properties of blood, its complex near-wall behavior, and the pulsatility of its flow all affect drug uptake only modestly. Furthermore, flow-mediated drug transport was found to be negligible due to the rapid rate at which drug depletes at the stent coating surfaces that are exposed to arterial blood flow. For fluid dynamicists, these results show that steady-state simulations must be avoided when modelling drug transport in stented arteries. For device designers, these results may be used to optimize the shape of drug-eluting stent struts and coatings to improve stent efficacy.

A Novel Microfluidic Device-Based Neurite Outgrowth Inhibition Assay Reveals the Neurite Outgrowth-Promoting Activity of Tropomyosin Tpm3.1 in Hippocampal Neurons.

Overcoming neurite inhibition is integral for restoring neuronal connectivity after CNS injury. Actin dynamics are critical for neurite growth cone formation and extension. The tropomyosin family of proteins is a regarded as master regulator of actin dynamics. This study investigates tropomyosin isoform 3.1 (Tpm3.1) as a potential candidate for overcoming an inhibitory substrate, as it is known to influence neurite branching and outgrowth. We designed a microfluidic device that enables neurons to be grown adjacent to an inhibitory substrate, Nogo-66. Results show that neurons, overexpressing hTpm3.1, have an increased propensity to overcome Nogo-66 inhibition. We propose Tpm3.1 as a potential target for promoting neurite growth in an inhibitory environment in the central nervous system.