Vortical Structures Promote Atheroprotective Wall Shear Stress Distributions in a Carotid Artery Bifurcation Model.

Carotid artery diseases, such as atherosclerosis, are a major cause of death in the United States. Wall shear stresses are known to prompt plaque formation, but there is limited understanding of the complex flow structures underlying these stresses and how they differ in a pre-disposed high-risk patient cohort. A 'healthy' and a novel 'pre-disposed' carotid artery bifurcation model was determined based on patient-averaged clinical data, where the 'pre-disposed' model represents a pathological anatomy. Computational fluid dynamic simulations were performed using a physiological flow based on healthy human subjects. A main hairpin vortical structure in the internal carotid artery sinus was observed, which locally increased instantaneous wall shear stress. In the pre-disposed geometry, this vortical structure starts at an earlier instance in the cardiac flow cycle and persists over a much shorter period, where the second half of the cardiac cycle is dominated by perturbed secondary flow structures and vortices. This coincides with weaker favorable axial pressure gradient peaks over the sinus for the 'pre-disposed' geometry. The findings reveal a strong correlation between vortical structures and wall shear stress and imply that an intact internal carotid artery sinus hairpin vortical structure has a physiologically beneficial role by increasing local wall shear stresses. The deterioration of this beneficial vortical structure is expected to play a significant role in atherosclerotic plaque formation.

Shear stress metrics associated with pro-atherogenic high-risk anatomical features in a carotid artery bifurcation model.

BACKGROUND: Diseases associated with atherosclerotic plaques in the carotid artery are a major cause of deaths in the United States. Blood-flow-induced shear-stresses are known to trigger plaque formation. Prior literature suggests that the internal carotid artery sinus is prone to atherosclerosis, but there is limited understanding of why only certain patients are predisposed towards plaque formation. METHODS: We computationally investigate the effect of vessel geometry on wall-shear-stress distribution by comparing flowfields and wall-shear-stress-metrics between a low-risk and a novel predisposed high-risk carotid artery bifurcation anatomy. Both models were developed based on clinical risk estimations and patient-averaged anatomical features. The high-risk geometry has a larger internal carotid artery branching angle and a lower internal-to-carotid-artery-diameter-ratio. A patient-averaged physiological carotid artery inflow waveform is used. FINDINGS: The high-risk geometry experiences stronger flow separation in the sinus. Furthermore, it experiences a more equal flow split at the bifurcation, thereby reducing internal carotid artery flowrate and increasing atherosclerosis-prone low-velocity areas. Lowest time-averaged-wall-shear-stresses are present at the sinus outer wall, where plaques are often found, for both geometries. The high-risk geometry has significantly high, unfavorable oscillatory-shear-index values not found in the low-risk geometry. High oscillatory-shear-index areas are located at the vessels outside walls distal to the bifurcation and on the sinus wall. INTERPRETATION: These results highlight the effectiveness of oscillatory-shear-index, to augment classical time-averaged-wall-shear-stress, in evaluating pro-atherogenic geometry features. Furthermore, the flow split at the bifurcation is a promising clinical indicator for atherosclerosis risk as it can be directly accessed using clinical imaging, whereas shear-stress-metrics cannot.

An in vitro analysis of the effect of geometry-induced flows on endothelial cell behavior in 3D printed small-diameter blood vessels.

Clinical recovery from vascular diseases has increasingly become reliant upon the successful fabrication of artificial blood vessels (BVs) or vascular prostheses due to the shortage of autologous vessels and the high incidence of vessel graft diseases. Even though many attempts at the clinical implementation of large artificial BVs have been reported to be successful, the development of small-diameter BVs remains one of the significant challenges due to the limitation of micro-manufacturing capacity in complexity and reproducibility, as well as the development of thrombosis. The present study aims to develop 3D printed small-diameter artificial BVs that recapitulate the longitudinal geometric elements in the native BVs using biocompatible polylactic acid (PLA). As their intrinsic physical properties are crystallinity dependent, we used two PLA filaments with different crystallinity to investigate the suitability of their physical properties in the micro-manufacturing of BVs. To explore the mechanism of venous thrombosis, our study provided a preliminarily comparative analysis of the effect of geometry-induced flows on the behavior of human endothelial cells (ECs). Our results showed that the adhered healthy ECs in the 3D printed BV exhibited regulated patterns, such as elongated and aligned parallel to the flow direction, as well as geometry-induced EC response mechanisms that are associated with hemodynamic shear stresses. Furthermore, the computational fluid dynamics simulation results provided insightful information to predict velocity profile and wall shear stress distribution in the geometries of BVs in accordance with their spatiotemporally-dependent cell behaviors. Our study demonstrated that 3D printed small-diameter BVs could serve as suitable candidates for fundamental BV studies and hold great potential for clinical applications.

Dual 3D printing for vascularized bone tissue regeneration.

The development of sufficient vascular networks is crucial for the successful fabrication of tissue constructs for regenerative medicine, as vascularization is essential to perform the metabolic functions of tissues, such as nutrient transportation and waste removal. In recent years, efforts to 3D print vascularized bone have gained substantial attention, as bone disorders and defects have a marked impact on the older generations of society. However, conventional and previous 3D printed bone studies have been plagued by the difficulty in obtaining the nanoscale geometrical precision necessary to recapitulate the distinct characteristics of natural bone. Additionally, the process of developing truly biomimetic vascularized bone tissue has been historically complex. In this study, a biomimetic nano-bone tissue construct with a perfusable, endothelialized vessel channel was developed using a combination of simple stereolithography (SLA) and fused deposition modeling (FDM) 3D printing systems. The perfusable vessel channel was created within the SLA printed bone scaffold using an FDM printed polyvinyl alcohol (PVA) sacrificial template. Within the fabricated constructs, bone tissue was formed through the osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs), and distinct capillaries sprouted through the angiogenesis of the endothelialized vessel channel after human umbilical vein endothelial cells (HUVECs) had been perfused throughout. Furthermore, the fabricated constructs were evaluated in physiologically relevant culture conditions to predict tissue development after implantation in the human body. The experimental results revealed that the custom-designed bioreactor with an hMSC-HUVEC co-culture system enhanced the formation of vascular networks and the osteogenic maturation of the constructs for up to 20 days of observation.

Macro-Rheology Characterization of Gill Raker Mucus in the Silver Carp, Hypophthalmichthys molitrix.

The silver carp, Hypophthalmichthys molitrix, is an invasive planktivorous filter feeder fish that infested the natural waterways of the upper Mississippi River basin due to its highly efficient filter feeding mechanism. The characteristic organs called gill rakers (GRs), found in many such filter feeders, facilitate the efficient filtration of food particles such as phytoplankton that are of a few microns in size. The motivation to investigate the rheology of the GR mucus stems from our desire to understand its role in aiding the filter feeding process in the silver carp. The mucus-rich fluid, in a 'thick and sticky' state may facilitate the adhesion of food particulates. The permeation and transport through the GR membrane are facilitated by the action of external shear forces that induce varying shear strain rates. Therefore, mucus rheology can provide a vital clue to the tremendous outcompeting nature of the silver carp within the pool of filter feeding fish. Based on this it was posited that GR mucus may provide an adhesive function to food particles and act as a transport vehicle to assist in the filter feeding process. The main objective of the protocol is to determine the yield stress of the mucus, attributed to the minimum shear stress required to initiate flow at which irreversible plastic deformation is first observed across a structured viscoelastic material. Accordingly, rheological properties of the GR mucus, i.e., viscosity, storage, and loss moduli, were investigated for its non-Newtonian, shear-thinning nature using a rotational rheometer.   A protocol presented here is employed to analyze the rheological properties of mucus extracted from the gill rakers of a silver carp, fished at Hart Creek location of the Missouri River. The protocol aims to develop an effective strategy for rheological testing and material characterization of mucus assumed to be a structured viscoelastic material.

An acoustic source model for asymmetric intraglottal flow with application to reduced-order models of the vocal folds.

The complex three-way interaction between airflow, tissue, and sound, for asymmetric vocal fold vibration, is not well understood. Current modeling efforts are not able to explain clinical observations where drastic differences in sound production are often observed, with no noticeable differences in the vocal fold kinematics. To advance this understanding, an acoustical model for voiced sound generation in the presence of asymmetric intraglottal flows is developed. The source model operates in conjunction with a wave reflection analog propagation scheme and an asymmetric flow description within the glottis. To enable comparison with prior work, the source model is evaluated using a well-studied two-mass vocal fold model. The proposed source model is evaluated through acoustic measures of interest, including radiated sound pressure level, maximum flow declination rate, and spectral tilt, and also via its effects on the vocal fold dynamics. The influence of the model, in comparison to the standard symmetric Bernoulli flow description, results in an increased transfer of energy from the fluid to the vocal folds, increased radiated sound pressure level and maximum flow declination rate, and decreased spectral tilt. These differences are most pronounced for asymmetric vocal fold configurations that mimic unilateral paresis and paralysis, where minor kinematic changes can result in significant acoustic and aerodynamic differences. The results illustrate that fluid effects arising from asymmetric glottal flow can play an important role in the acoustics of pathological voiced speech.

Photolithographic-stereolithographic-tandem fabrication of 4D smart scaffolds for improved stem cell cardiomyogenic differentiation.

4D printing is a highly innovative additive manufacturing process for fabricating smart structures with the ability to transform over time. Significantly different from regular 4D printing techniques, this study focuses on creating novel 4D hierarchical micropatterns using a unique photolithographic-stereolithographic-tandem strategy (PSTS) with smart soybean oil epoxidized acrylate (SOEA) inks for effectively regulating human bone marrow mesenchymal stem cell (hMSC) cardiomyogenic behaviors. The 4D effect refers to autonomous conversion of the surficial-patterned scaffold into a predesigned construct through an external stimulus delivered immediately after printing. Our results show that hMSCs actively grew and were highly aligned along the micropatterns, forming an uninterrupted cellular sheet. The generation of complex patterns was evident by triangular and circular outlines appearing in the scaffolds. This simple, yet efficient, technique was validated by rapid printing of scaffolds with well-defined and consistent micro-surface features. A 4D dynamic shape change transforming a 2-D design into flower-like structures was observed. The printed scaffolds possessed a shape memory effect beyond the 4D features. The advanced 4D dynamic feature may provide seamless integration with damaged tissues or organs, and a proof of concept 4D patch for cardiac regeneration was demonstrated for the first time. The 4D-fabricated cardiac patch showed significant cardiomyogenesis confirmed by immunofluorescence staining and qRT-PCR analysis, indicating its promising potential in future tissue and organ regeneration applications.

PID controller design to generate pulsatile flow rate for in vitro experimental studies of physiological flows.

Producing accurate pulsatile flow rates is essential for many in vitro experimental studies in biofluid dynamics research. A controller system was developed to control a flow loop to produce easily adjustable pulsatile flow rates with sufficient accuracy. An Arduino board is used as a micro-controller to control a pump to produce various pulsatile flow rates, and an open-source proportional-integral-derivative (PID) control algorithm is developed for this purpose. Four non-trivial pulsatile waveforms were produced by the PID controller, as well as an iterative controller, and the performance of both controllers was evaluated. Both the PID and iterative controllers were able to successfully produce slowly-varying signals (single and multi-harmonic low frequency sine waves), but for high frequency signals where the flow has strong acceleration/deceleration (e.g. for physiological waveforms) the iterative controller exhibited significant undershoot. The comparison of PID and iterative controllers suggests that if the desired flow rate is a low frequency, simple waveform then the iterative controller is preferred due to simplicity of implementation. However, if the desired signal is rapidly changing and more complicated then the PID controller achieves better results. This system can be implemented in many flow loops due to its simplicity and low cost, and does not require a mathematical model of the system.

3D printing of novel osteochondral scaffolds with graded microstructure.

Osteochondral tissue has a complex graded structure where biological, physiological, and mechanical properties vary significantly over the full thickness spanning from the subchondral bone region beneath the joint surface to the hyaline cartilage region at the joint surface. This presents a significant challenge for tissue-engineered structures addressing osteochondral defects. Fused deposition modeling (FDM) 3D bioprinters present a unique solution to this problem. The objective of this study is to use FDM-based 3D bioprinting and nanocrystalline hydroxyapatite for improved bone marrow human mesenchymal stem cell (hMSC) adhesion, growth, and osteochondral differentiation. FDM printing parameters can be tuned through computer aided design and computer numerical control software to manipulate scaffold geometries in ways that are beneficial to mechanical performance without hindering cellular behavior. Additionally, the ability to fine-tune 3D printed scaffolds increases further through our investment casting procedure which facilitates the inclusion of nanoparticles with biochemical factors to further elicit desired hMSC differentiation. For this study, FDM was used to print investment-casting molds innovatively designed with varied pore distribution over the full thickness of the scaffold. The mechanical and biological impacts of the varied pore distributions were compared and evaluated to determine the benefits of this physical manipulation. The results indicate that both mechanical properties and cell performance improve in the graded pore structures when compared to homogeneously distributed porous and non-porous structures. Differentiation results indicated successful osteogenic and chondrogenic manipulation in engineered scaffolds.

Experimental Investigation of Secondary Flow Structures Downstream of a Model Type IV Stent Failure in a 180° Curved Artery Test Section.

The arterial network in the human vasculature comprises of ubiquitously present blood vessels with complex geometries (branches, curvatures and tortuosity). Secondary flow structures are vortical flow patterns that occur in curved arteries due to the combined action of centrifugal forces, adverse pressure gradients and inflow characteristics. Such flow morphologies are greatly affected by pulsatility and multiple harmonics of physiological inflow conditions and vary greatly in size-strength-shape characteristics compared to non-physiological (steady and oscillatory) flows (1 - 7). Secondary flow structures may ultimately influence the wall shear stress and exposure time of blood-borne particles toward progression of atherosclerosis, restenosis, sensitization of platelets and thrombosis (4 - 6, 8 - 13). Therefore, the ability to detect and characterize these structures under laboratory-controlled conditions is precursor to further clinical investigations. A common surgical treatment to atherosclerosis is stent implantation, to open up stenosed arteries for unobstructed blood flow. But the concomitant flow perturbations due to stent installations result in multi-scale secondary flow morphologies (4 - 6). Progressively higher order complexities such as asymmetry and loss in coherence can be induced by ensuing stent failures vis-à-vis those under unperturbed flows (5). These stent failures have been classified as "Types I-to-IV" based on failure considerations and clinical severity (14). This study presents a protocol for the experimental investigation of the complex secondary flow structures due to complete transverse stent fracture and linear displacement of fractured parts ("Type IV") in a curved artery model. The experimental method involves the implementation of particle image velocimetry (2C-2D PIV) techniques with an archetypal carotid artery inflow waveform, a refractive index matched blood-analog working fluid for phase-averaged measurements (15 - 18). Quantitative identification of secondary flow structures was achieved using concepts of flow physics, critical point theory and a novel wavelet transform algorithm applied to experimental PIV data (5, 6, 19 - 26).

Investigating the three-dimensional flow separation induced by a model vocal fold polyp.

The fluid-structure energy exchange process for normal speech has been studied extensively, but it is not well understood for pathological conditions. Polyps and nodules, which are geometric abnormalities that form on the medial surface of the vocal folds, can disrupt vocal fold dynamics and thus can have devastating consequences on a patient's ability to communicate. Our laboratory has reported particle image velocimetry (PIV) measurements, within an investigation of a model polyp located on the medial surface of an in vitro driven vocal fold model, which show that such a geometric abnormality considerably disrupts the glottal jet behavior. This flow field adjustment is a likely reason for the severe degradation of the vocal quality in patients with polyps. A more complete understanding of the formation and propagation of vortical structures from a geometric protuberance, such as a vocal fold polyp, and the resulting influence on the aerodynamic loadings that drive the vocal fold dynamics, is necessary for advancing the treatment of this pathological condition. The present investigation concerns the three-dimensional flow separation induced by a wall-mounted prolate hemispheroid with a 2:1 aspect ratio in cross flow, i.e. a model vocal fold polyp, using an oil-film visualization technique. Unsteady, three-dimensional flow separation and its impact of the wall pressure loading are examined using skin friction line visualization and wall pressure measurements.

Response to "Comments on 'A theoretical model of the pressure distributions arising from asymmetric intraglottal flows applied to a two-mass model of the vocal folds'" [J. Acoust. Soc. Am. 130, 389-403 (2011)].

Hirschberg [J. Acoust. Soc. Am. 134, 9-12 (2013)] presents a commentary and criticisms of the viscous flow model presented by Erath et al. [J. Acoust. Soc. Am. 130, 389-403 (2011)] that solves for the asymmetric pressure loading on the vocal fold walls. This pressure loading arises from asymmetric flow attachment to one vocal fold wall when the glottal channel forms a divergent configuration. Hirschberg proposes an alternative model for the asymmetric loading based upon inviscid flow curvature at the glottal inlet. In this manuscript further evidence is provided in support of the model of Erath et al. and the underlying assumptions, and demonstrates that the primary criticisms presented by Hirschberg are unwarranted. The model presented by Hirschberg is compared with the model from the original paper by Erath et al., and it is shown that each model describes different and complementary aspects of divergent glottal flows.

Vortex dynamics and scalar transport in the wake of a bluff body driven through a steady recirculating flow.

The air ventilation system in wide-body aircraft cabins provides passengers with a healthy breathing environment. In recent years, the increase in global air traffic has amplified contamination risks by airborne flu-like diseases and terrorist threats involving the onboard release of noxious materials. In particular, passengers moving through a ventilated cabin may transport infectious pathogens in their wake. This paper presents an experimental investigation of the wake produced by a bluff body driven through a steady recirculating flow. Data were obtained in a water facility using particle image velocimetry and planar laser induced fluorescence. Ventilation attenuated the downward convection of counter-rotating vortices produced near the free-end corners of the body and decoupled the downwash mechanism from forward entrainment, creating stagnant contaminant regions.

Nonlinear vocal fold dynamics resulting from asymmetric fluid loading on a two-mass model of speech.

Nonlinear vocal fold dynamics arising from asymmetric flow formations within the glottis are investigated using a two-mass model of speech with asymmetric vocal fold tensioning, representative of unilateral vocal fold paralysis. A refined theoretical boundary-layer flow solver is implemented to compute the intraglottal pressures, providing a more realistic description of the flow than the standard one-dimensional, inviscid Bernoulli flow solution. Vocal fold dynamics are investigated for subglottal pressures of 0.6 < p(s) < 1.5 kPa and tension asymmetries of 0.5 < Q < 0.8. As tension asymmetries become pronounced the asymmetric flow incites nonlinear behavior in the vocal fold dynamics at subglottal pressures that are associated with normal speech, behavior that is not captured with standard Bernoulli flow solvers. Regions of bifurcation, coexistence of solutions, and chaos are identified.

A theoretical model of the pressure field arising from asymmetric intraglottal flows applied to a two-mass model of the vocal folds.

A theoretical flow solution is presented for predicting the pressure distribution along the vocal fold walls arising from asymmetric flow that forms during the closing phases of speech. The resultant wall jet was analyzed using boundary layer methods in a non-inertial reference frame attached to the moving wall. A solution for the near-wall velocity profiles on the flow wall was developed based on a Falkner-Skan similarity solution and it was demonstrated that the pressure distribution along the flow wall is imposed by the velocity in the inviscid core of the wall jet. The method was validated with experimental velocity data from 7.5 times life-size vocal fold models, acquired for varying flow rates and glottal divergence angles. The solution for the asymmetric pressures was incorporated into a widely used two-mass model of vocal fold oscillation with a coupled acoustical model of sound propagation. Asymmetric pressure loading was found to facilitate glottal closure, which yielded only slightly higher values of maximum flow declination rate and radiated sound, and a small decrease in the slope of the spectral tilt. While the impact on symmetrically tensioned vocal folds was small, results indicate the effect becomes more significant for asymmetrically tensioned vocal folds.

Impact of wall rotation on supraglottal jet stability in voiced speech.

Supraglottal jet variability was investigated in a scaled-up flow facility incorporating driven vocal fold models with and without wall rotation. Principle component analysis was performed on the experimental supraglottal flow fields to ascertain the roll of glottal wall motion on the development of the supraglottal jet. It is shown that intraglottal flow asymmetries that develop due to wall rotation are not the primary mechanism for generating large-scale cycle-to-cycle deflection of the supraglottal jet. However, wall rotation does decrease the energy content of the first mode, redistributing it to the higher modes through an increase in unstructured flow variability.

The occurrence of the Coanda effect in pulsatile flow through static models of the human vocal folds.

Pulsatile flow through a one-sided diffuser and static divergent vocal-fold models is investigated to ascertain the relevance of viscous-driven flow asymmetries in the larynx. The models were 7.5 times real size, and the flow was scaled to match Reynolds and Strouhal numbers, as well as the translaryngeal pressure drop. The Reynolds number varied from 0-2000, for flow oscillation frequencies corresponding to 100 and 150 Hz life-size. Of particular interest was the development of glottal flow skewing by attachment to the bounding walls, or Coanda effect, in a pulsatile flow field, and its impact on speech. The vocal folds form a divergent passage during phases of the phonation cycle when viscous effects such as flow separation are important. It was found that for divergence angles of less than 20 degrees, the attachment of the flow to the vocal-fold walls occurred when the acceleration of the forcing function was zero, and the flow had reached maximum velocity. For a divergence angle of 40 degrees, the fully separated central jet never attached to the vocal-fold walls. Inferences are made regarding the impact of the Coanda effect on the sound source contribution in speech.

Non-uniform flow behavior in a parallel plate flow chamber : alters endothelial cell responses.

Arterial flow characteristics determine vessel health by modulating vascular endothelial cells. One system used to study these interactions is the parallel plate flow chamber. The present in vitro study quantified the uniformity of fluid flow across a parallel plate flow chamber and characterized plate-location dependent endothelial cell gene expression. More specifically, shear stress varied by as much as 11% across the chamber area, which caused non-uniform ecNOS (p < 0.05) and COX-2 (p < 0.05) mRNA expression across the plate area. Results herein suggest that chamber variations may result during construction or assembly, which ultimately affect flow-sensitive cell responses (including mRNA expression). Therefore, these limitations should be considered when reporting endothelial cell responses to fluid flow using parallel plate flow chambers.