Anastasia Desyatova

Patterning mechanisms of cytoskeletal and cell wall systems during leaf trichome morphogenesis

The plant actin cytoskeleton is an unstable network of filaments that influences polarized growth through poorly understood mechanisms. Here, we used a combination of live cell imaging and finite element computational modelling of Arabidopsis trichome morphogenesis to determine how the actin and microtubule cytoskeletons cooperate to pattern the cell wall and growth. The actin-related protein (ARP)2/3 complex generates an actin meshwork that operates within a tip-localized, microtubule-depleted zone to modulate cell wall anisotropy locally. The actin meshwork also positions an actin bundle network that organizes organelle flow patterns. This activity is required to maintain cell wall thickness gradients that enable tip-biased diffuse growth. These newly discovered couplings between cytoskeletal patterns and wall textures provide important insights into the cellular mechanism of growth control in plants.

Effects of age on the physiological and mechanical characteristics of human femoropopliteal arteries

Surgical and interventional therapies for peripheral artery disease (PAD) are notorious for high rates of failure. Interactions between the artery and repair materials play an important role, but comprehensive data describing the physiological and mechanical characteristics of human femoropopliteal arteries are not available. Fresh femoropopliteal arteries were obtained from 70 human subjects (13-79 years old), and in situ vs. excised arterial lengths were measured. Circumferential and longitudinal opening angles were determined for proximal superficial femoral, proximal popliteal and distal popliteal arteries. Mechanical properties were assessed by multi-ratio planar biaxial extension, and experimental data were used to calculate physiological stresses and stretches, in situ axial force and anisotropy. Verhoeff-Van Gieson-stained axial and transverse arterial sections were used for histological analysis. Most specimens demonstrated nonlinear deformations and were more compliant longitudinally than circumferentially. In situ axial pre-stretch decreased 0.088 per decade of life. In situ axial force and axial stress also decreased with age, but circumferential physiological stress remained constant. Physiological circumferential stretch decreased 55-75% after 45 years of age. Histology demonstrated a thickened external elastic lamina with longitudinally oriented elastin that was denser in smaller, younger arteries. Axial elastin likely regulates axial pre-stretch to help accommodate the complex deformations required of the artery wall during locomotion. Degradation and fragmentation of elastin as a consequence of age, cyclic mechanical stress and atherosclerotic arterial disease may contribute to decreased in situ axial pre-stretch, predisposing to more severe kinking of the artery during limb flexion and loss of energy-efficient arterial function.

Three-dimensional bending, torsion and axial compression of the femoropopliteal artery during limb flexion

High failure rates of femoropopliteal artery reconstruction are commonly attributed to complex 3D arterial deformations that occur with limb movement. The purpose of this study was to develop a method for accurate assessment of these deformations. Custom-made stainless-steel markers were deployed into 5 in situ cadaveric femoropopliteal arteries using fluoroscopy. Thin-section CT images were acquired with each limb in the straight and acutely bent states. Image segmentation and 3D reconstruction allowed comparison of the relative locations of each intra-arterial marker position for determination of the arterys bending, torsion and axial compression. After imaging, each artery was excised for histological analysis using Verhoeff-Van Gieson staining. Femoropopliteal arteries deformed non-uniformly with highly localized deformations in the proximal superficial femoral artery, and between the adductor hiatus and distal popliteal artery. The largest bending (11±3-6±1 mm radius of curvature), twisting (28±9-77±27°/cm) and axial compression (19±10-30±8%) were registered at the adductor hiatus and the below knee popliteal artery. These deformations were 3.7, 19 and 2.5 fold more severe than values currently reported in the literature. Histology demonstrated a distinct sub-adventitial layer of longitudinally oriented elastin fibers with intimal thickening in the segments with the largest deformations. This endovascular intra-arterial marker technique can quantify the non-uniform 3D deformations of the femoropopliteal artery during knee flexion without disturbing surrounding structures. We demonstrate that 3D arterial bending, torsion and compression in the flexed lower limb are highly localized and are substantially more severe than previously reported.

In vivo three-dimensional blood velocity profile shapes in the human common, internal, and external carotid arteries

OBJECTIVE True understanding of carotid bifurcation pathophysiology requires a detailed knowledge of the hemodynamic conditions within the arteries. Data on carotid artery hemodynamics are usually based on simplified, computer-based, or in vitro experimental models, most of which assume that the velocity profiles are axially symmetric away from the carotid bulb. Modeling accuracy and, more importantly, our understanding of the pathophysiology of carotid bifurcation disease could be considerably improved by more precise knowledge of the in vivo flow properties within the human carotid artery. The purpose of this work was to determine the three-dimensional pulsatile velocity profiles of human carotid arteries. METHODS Flow velocities were measured over the cardiac cycle using duplex ultrasonography, before and after endarterectomy, in the surgically exposed common (CCA), internal (ICA), and external (ECA) carotid arteries (n = 16) proximal and distal to the stenosis/endarterectomy zone. These measurements were linked to a standardized grid across the flow lumina of the CCA, ICA, and ECA. The individual velocities were then used to build mean three-dimensional pulsatile velocity profiles for each of the carotid artery branches. RESULTS Pulsatile velocity profiles in all arteries were asymmetric about the arterial centerline. Posterior velocities were higher than anterior velocities in all arteries. In the CCA and ECA, velocities were higher laterally, while in the ICA, velocities were higher medially. Pre- and postendarterectomy velocity profiles were significantly different. After endarterectomy, velocity values increased in the common and internal and decreased in the external carotid artery. CONCLUSIONS The in vivo hemodynamics of the human carotid artery are different from those used in most current computer-based and in vitro models. The new information on three-dimensional blood velocity profiles can be used to design models that more closely replicate the actual hemodynamic conditions within the carotid bifurcation. Such models can be used to further improve our understanding of the pathophysiologic processes leading to stroke and for the rational design of medical and interventional therapies.

IN SITU LONGITUDINAL PRE-STRETCH IN THE HUMAN FEMOROPOPLITEAL ARTERY

UNLABELLED In situ longitudinal (axial) pre-stretch (LPS) plays a fundamental role in the mechanics of the femoropopliteal artery (FPA). It conserves energy during pulsation and prevents buckling of the artery during limb movement. We investigated how LPS is affected by demographics and risk factors, and how these patient characteristics associate with the structural and physiologic features of the FPA. LPS was measured in n=148 fresh human FPAs (14-80 years old). Mechanical properties were characterized with biaxial extension and histopathological characteristics were quantified with Verhoeff-Van Gieson Staining. Constitutive modeling was used to calculate physiological stresses and stretches which were then analyzed in the context of demographics, risk factors and structural characteristics. Age had the strongest negative effect (r=-0.812, p<0.01) on LPS and could alone explain 66% of LPS variability. Male gender, higher body mass index, hypertension, diabetes, coronary artery disease, dyslipidemia and tobacco use had negative effects on LPS, but only the effect of tobacco was not associated with aging. FPAs with less pre-stretch had thicker medial layers, but thinner intramural elastic fibers with less dense and more fragmented external elastic laminae. Elastin degradation was associated with decreased physiological tethering force and longitudinal stress, while circumferential stress remained constant. FPA wall pathology was negatively associated with LPS (r=-0.553, p<0.01), but the effect was due primarily to aging. LPS in the FPA may serve as an energy reserve for adaptive remodeling. Reduction of LPS due to degradation and fragmentation of intramural longitudinal elastin during aging can be accelerated in tobacco users. STATEMENT OF SIGNIFICANCE This work studies in situ longitudinal pre-stretch (LPS) in the human femoropopliteal artery. LPS has a fundamental role in arterial mechanics, but is rather poorly studied due to lack of direct in vivo measurement method. We have investigated LPS in the n=148 human femoropopliteal arteries in the context of subject demographics and risk factors, and structural and physiologic characteristics of the artery. Our results demonstrate that LPS reduces with age due to degradation and fragmentation of intramural elastin. LPS may serve as an energy reserve for adaptive remodeling, and reduction of LPS can be accelerated in tobacco users.

Finite Element Model of the Patched Human Carotid

Introduction: The hemodynamic effects of carotid artery patching are not well known. Our objective was to develop a fluid-solid finite element model of the endarterectomized and patched carotid artery. Methods: Hyperelastic materials parameters were determined from studies of 8 cadaveric carotids. Blood flow characteristics were based on intraoperative data from a patient undergoing endarterectomy. Wall shear stress, cyclic strain and effective stress were computed as hemodynamic parameters with known association with endothelial injury, neointimal hyperplasia abd atherogenesis. Results: Low wall shear stress, high cyclic strain and high effective stress were identified diffusely in the carotid bulb, at the margins around the patch and in the flow divider. Conclusion: Endarterectomy and Polytetrafluoroethylene patching produce considerable abnormalities in the hemodynamics of the repaired carotid. Advanced mechanical modeling can be used to evaluate different carotid revascularization approaches to obtain optimized biomechanical and hemodynamic results for the care of patients with carotid bifurcation disease.

Morphometric roadmaps to improve accurate device delivery for fluoroscopy-free resuscitative endovascular balloon occlusion of the aorta.

BACKGROUND Uncontrolled hemorrhage from vessel injuries within the torso remains a significant source of prehospital trauma mortality. Resuscitative endovascular balloon occlusion of the aorta can effectively control noncompressible hemorrhage, but this minimally invasive technique relies heavily on imaging not available in the field. Our goal was to develop morphometric roadmaps to enhance the safety and accuracy of fluoroscopy-free endovascular navigation of hemorrhage control devices. METHODS Three-dimensional reconstructions of computed tomographic angiography scans from 122 trauma patients (mean [SD] age, 47 [24] years; range 5–93 years; 64 males; 58 females) were used to measure centerline distances from femoral artery access sites to the major aortic branch artery origins. Morphometric roadmap equations were created using multiple linear regression analysis to predict distances to the origins of the major arteries in the chest, abdomen, and pelvis using torso length, demographics, and risk factors as independent variables. A 40-mm-long occlusion balloon was then virtually deployed targeting Zones 1 and 3 of the aorta using these equations. Balloon placement accuracy was determined by comparing predicted versus actual measured distances to the target zone locations within the aortas from the database. RESULTS Torso length and age were the strongest predictors of centerline distances from femoral artery access sites to the major artery origins. Male sex contributed to longer distances, while diabetes and smoking were associated with shorter distances. Hypertension, dyslipidemia, and coronary artery disease had no effect. With the use of morphometric roadmaps, virtual occlusion balloon placement accuracy was 100% for Zone 3 of the aorta, compared with 87% accuracy when using torso length alone. CONCLUSION Morphometric roadmaps demonstrate a potential for improving the safety and accuracy of fluoroscopy-free aortic occlusion balloon delivery. Continued development of minimally invasive hemorrhage control techniques holds promise to improve prehospital mortality for patients with noncompressible exsanguinating torso injuries. LEVEL OF EVIDENCE Therapeutic study, level IV; diagnostic study, level III.

Limb flexion-induced twist and associated intramural stresses in the human femoropopliteal artery

High failure rates of femoropopliteal artery (FPA) interventions are often attributed to severe mechanical deformations that occur with limb movement. Torsion of the FPA likely plays a significant role, but is poorly characterized and the associated intramural stresses are currently unknown. FPA torsion in the walking, sitting and gardening postures was characterized in n = 28 in situ FPAs using intra-arterial markers. Principal mechanical stresses and strains were quantified in the superficial femoral artery (SFA), adductor hiatus segment (AH) and the popliteal artery (PA) using analytical modelling. The FPA experienced significant torsion during limb flexion that was most severe in the gardening posture. The associated mechanical stresses were non-uniformly distributed along the length of the artery, increasing distally and achieving maximum values in the PA. Maximum twist in the SFA ranged 10–13° cm−1, at the AH 8–16° cm−1, and in the PA 14–26° cm−1 in the walking, sitting and gardening postures. Maximum principal stresses were 30–35 kPa in the SFA, 27–37 kPa at the AH and 39–43 kPa in the PA. Understanding torsional deformations and intramural stresses in the FPA can assist with device selection for peripheral arterial disease interventions and may help guide the development of devices with improved characteristics.

Constitutive modeling of human femoropopliteal artery biaxial stiffening due to aging and diabetes

Atherosclerotic obstructive disease of the femoropopliteal artery (Peripheral Arterial Disease, PAD) is notorious for high treatment failure rates. Older age and diabetes mellitus (DM) are among the major risk factors for PAD, and both are associated with increased arterial stiffness. Our goal was to develop a constitutive model describing multiaxial arterial stiffening, and use it to portray aging of normal and diabetic human femoropopliteal arteries (FPA). Fresh human FPAs (n=744) were obtained from 13-82-year-old donors. Arteries were tested using planar biaxial extension, and their behavior was modeled with a constitutive relation that included stiffening functions of age. FPA diameter, wall thickness, circumferential, and longitudinal opening angles increased with age, while longitudinal pre-stretch decreased. Diameter and circumferential opening angle did not change with age in subjects with DM. Younger FPAs were more compliant longitudinally but became more isotropic with age. Arteries with DM stiffened significantly faster in the circumferential direction than arteries without DM. Constitutive model accurately portrayed orthotropic stiffening with age of both normal and diabetic arteries. Constitutive description of FPA aging contributes to understanding of arterial pathophysiology and can help improve fidelity of computational models investigating device-artery interaction in PAD repair by providing more personalized arterial properties. STATEMENT OF SIGNIFICANCE We have analyzed n=744 human femoropopliteal artery (FPA) specimens using biaxial tensile testing to derive constitutive description of FPA aging in diabetic and non-diabetic subjects. The proposed model allows determination of FPA mechanical properties for subjects of any given age in the range of 13-82years. These results contribute to understanding of FPA pathophysiology and can help improve fidelity of computational models investigating device-artery interaction in peripheral arterial disease repair by providing more personalized arterial properties. In addition, they can guide the development of new materials tunable to diabetic and non-diabetic arteries.

Comparison of femoropopliteal artery stents under axial and radial compression, axial tension, bending, and torsion deformations

High failure rates of Peripheral Arterial Disease (PAD) stenting appear to be associated with the inability of certain stent designs to accommodate severe biomechanical environment of the femoropopliteal artery (FPA) that bends, twists, and axially compresses during limb flexion. Twelve Nitinol stents (Absolute Pro, Supera, Lifestent, Innova, Zilver, Smart Control, Smart Flex, EverFlex, Viabahn, Tigris, Misago, and Complete SE) were quasi-statically tested under bench-top axial and radial compression, axial tension, bending, and torsional deformations. Stents were compared in terms of force-strain behavior, stiffness, and geometrical shape under each deformation mode. Tigris was the least stiff stent under axial compression (6.6N/m axial stiffness) and bending (0.1N/m) deformations, while Smart Control was the stiffest (575.3N/m and 105.4N/m, respectively). Under radial compression Complete SE was the stiffest (892.8N/m), while Smart Control had the lowest radial stiffness (211.0N/m). Viabahn and Supera had the lowest and highest torsional stiffness (2.2μNm/° and 959.2μNm/°), respectively. None of the 12 PAD stents demonstrated superior characteristics under all deformation modes and many experienced global buckling and diameter pinching. Though it is yet to be determined which of these deformation modes might have greater clinical impact, results of the current analysis may help guide development of new stents with improved mechanical characteristics.