Biomechanics and inflammation in atherosclerotic plaque erosion and plaque rupture: implications for cardiovascular events in women.

OBJECTIVE: Although plaque erosion causes approximately 40% of all coronary thrombi and disproportionally affects women more than men, its mechanism is not well understood. The role of tissue mechanics in plaque rupture and regulation of mechanosensitive inflammatory proteins is well established, but their role in plaque erosion is unknown. Given obvious differences in morphology between plaque erosion and rupture, we hypothesized that inflammation in general as well as the association between local mechanical strain and inflammation known to exist in plaque rupture may not occur in plaque erosion. Therefore, our objective was to determine if similar mechanisms underlie plaque rupture and plaque erosion. METHODS AND RESULTS: We studied a total of 74 human coronary plaque specimens obtained at autopsy. Using lesion-specific computer modeling of solid mechanics, we calculated the stress and strain distribution for each plaque and determined if there were any relationships with markers of inflammation. Consistent with previous studies, inflammatory markers were positively associated with increasing strain in specimens with rupture and thin-cap fibroatheromas. Conversely, overall staining for inflammatory markers and apoptosis were significantly lower in erosion, and there was no relationship with mechanical strain. Samples with plaque erosion most closely resembled those with the stable phenotype of thick-cap fibroatheromas. CONCLUSIONS: In contrast to classic plaque rupture, plaque erosion was not associated with markers of inflammation and mechanical strain. These data suggest that plaque erosion is a distinct pathophysiological process with a different etiology and therefore raises the possibility that a different therapeutic approach may be required to prevent plaque erosion.

Calculation of the outcomes of remodeling of arteries subjected to sustained hypertension using a 3D two-layered model.

Arteries manifest a remodeling response to long-term alterations in arterial pressure and blood flow by changing geometry, structure, and composition through processes driven by perturbations of the local stresses in the vascular wall from their baseline values. The objective of this study is twofold--to develop a general method for calculating the remodeling responses of an artery considered as a two-layered tube; and to provide results for adaptive and maladaptive remodeling of a coronary artery. By formulating an inverse problem of vascular mechanics, the geometrical dimensions and mechanical properties of an artery are calculated from a prescribed deformed configuration, stress field, structural stiffness, and applied load. As an illustrative example we consider a human LAD coronary artery in both a perfect and incomplete adaptive response to a sustained step-wise change in pressure and a maladaptive response due to impaired remodeling of adventitia. The results obtained show that adventitia plays an important role in vascular mechanics when an artery is subjected to high arterial pressure. In addition to its well-known short term function of preventing over-inflation of an artery, it seems reasonable to accept that the manner by which adventitia remodels in response to a chronic increase in pressure is essential for preserving normal arterial function or may lead to an increased risk of developing vascular disorders.

Biomechanical modeling and morphology analysis indicates plaque rupture due to mechanical failure unlikely in atherosclerosis-prone mice.

Spontaneous plaque rupture in mouse models of atherosclerosis is controversial, although numerous studies have discussed so-called "vulnerable plaque" phenotypes in mice. We compared the morphology and biomechanics of two acute and one chronic murine model of atherosclerosis to human coronaries of the thin-cap fibroatheroma (TCFA) phenotype. Our acute models were apolipoprotein E-deficient (ApoE(-/-)) and LDL receptor-deficient (LDLr(-/-)) mice, both fed a high-fat diet for 8 wk with simultaneous infusion of angiotensin II (ANG II), and our chronic mouse model was the apolipoprotein E-deficient strain fed a regular chow diet for 1 yr. We found that the mouse plaques from all three models exhibited significant morphological differences from human TCFA plaques, including the plaque burden, plaque thickness, eccentricity, and amount of the vessel wall covered by lesion as well as significant differences in the relative composition of plaques. These morphological differences suggested that the distribution of solid mechanical stresses in the walls may differ as well. Using a finite-element analysis computational solid mechanics model, we computed the relative distribution of stresses in the walls of murine and human plaques and found that although human TCFA plaques have the highest stresses in the thin fibrous cap, murine lesions do not have such stress distributions. Instead, local maxima of stresses were on the media and adventitia, away from the plaque. Our results suggest that if plaque rupture is possible in mice, it may be driven by a different mechanism than mechanics.

Matrix metalloproteinase-2 and -9 are associated with high stresses predicted using a nonlinear heterogeneous model of arteries.

Arteries adapt to their mechanical environment by undergoing remodeling of the structural scaffold via the action of matrix metalloproteinases (MMPs). Cell culture studies have shown that stretching vascular smooth muscle cells (VSMCs) positively correlates to the production of MMP-2 and -9. In tissue level studies, the expressions and activations of MMP-2 and -9 are generally higher in the outer media. However, homogeneous mechanical models of arteries predict lower stress and strain in the outer media, which appear inconsistent with experimental findings. The effects of heterogeneity may be important to our understanding of VSMC function since arteries exhibit structural heterogeneity across the wall. We hypothesized that local stresses, computed using a heterogeneous mechanical model of arteries, positively correlate to the levels of MMP-2 and -9 in situ. We developed a model of the arterial wall accounting for nonlinearity, residual strain, anisotropy, and structural heterogeneity. The distributions of elastin and collagen fibers in situ, measured in the media of porcine carotid arteries, showed significant nonuniformities. Anisotropy was represented by the direction of collagen fibers measured by the helical angle of VSMC nuclei. The points at which the collagen fibers became load bearing were computed, assuming a uniform fiber strain and orientation under physiological loading conditions, an assumption motivated by morphological measurements. The distributions of circumferential stresses, computed using both heterogeneous and homogeneous models, were correlated to the distributions of expressions and activations of MMP-2 and -9 in porcine common carotid arteries incubated in an ex vivo perfusion organ culture system under physiological conditions for 48 h. While strains computed using incompressibility were identical in both models, the heterogeneous model, unlike the homogeneous model, predicted higher circumferential stresses in the outer layer correlated to the expressions and activations of MMP-2 and -9. This implies that localized remodeling occurs in the areas of high stress and agrees with results from cell culture studies. The results support the role of mechanical stress in vascular remodeling and the importance of structural heterogeneity in understanding mechanobiological responses.

Morphologic adaptation of arterial endothelial cells to longitudinal stretch in organ culture.

Arteries in vivo are subjected to large longitudinal stretch, which changes significantly due to vascular disease and surgery. However, little is known about the effect of longitudinal stretch on arterial endothelium. The aim of this study was to determine the morphologic adaptation of arterial endothelial cells (ECs) to elevated axial stretch. Porcine carotid arteries were stretched 20% more than their in vivo length while being maintained at physiological pressure and flow rate in an organ culture system. The ECs were elongated with the application of the axial stretch (aspect ratio 2.81+/-0.25 versus 3.65+/-0.38, n=8, p<0.001). The elongation was slightly decreased after three days and the ECs recovered their normal shape after seven days, as measured by the shape index and aspect ratio (0.55+/-0.03 versus 0.56+/-0.04, and 2.93+/-0.28 versus 2.88+/-0.20, respectively, n=5). Cell proliferation was increased in the intima of stretched arteries in three days as compared to control arteries but showed no difference after seven days in organ culture. These results demonstrate that the ECs adapt to axial stretch and maintain their normal shape.

Arteries respond to independent control of circumferential and shear stress in organ culture.

Arteries respond to changes in global mechanical parameters (pressure, flow rate, and longitudinal stretching) by remodeling to restore local parameters (circumferential stress, shear stress, and axial strain) to baseline levels. Because a change in a single global parameter results in changes of multiple local parameters, the effects of individual local parameters on remodeling remain unknown. This study uses a novel approach to study remodeling in organ culture based on independent control of local mechanical parameters. The approach is illustrated by studying the short term effects of circumferential and shear stress on remodeling-related biological markers. Porcine carotid arteries were cultured for 3 days at a circumferential stress of 50 or 150 kPa or, in separate experiments, a shear stress of 0.75 or 2.25 Pa. At high circumferential stress, matrix synthesis, smooth muscle cell proliferation, and cell death are significantly greater, but matrix metalloproteinase-2 (MMP-2) and pro-MMP-2 activity are significantly less. In contrast, biological markers measured were unaffected by shear stress. Applications of the proposed approach for improved understanding of remodeling, optimizing mechanical conditioning of tissue engineered arteries, and selection of experimentally motivated growth laws are discussed.

Effect of low shear stress on permeability and occludin expression in porcine artery endothelial cells.

INTRODUCTION: Although both fluid shear stress and mass transport of atherogenic substances into the vascular wall are known to be important factors in atherogenesis, there has been little research on the effect of shear stress on vascular permeability. Therefore, the effects of shear stress on the permeability of arteries and the expression of the endothelial cell tight junction molecule occludin, an important regulator of vascular permeability, were investigated. METHODS: Porcine carotid arteries were perfusion cultured ex vivo with low (1.5 dyne/cm(2)) or physiologic (15 dyne/cm(2)) shear stress and 100 mmHg pressure for 24 hours. Subsequently, 20 nm gold particles in solution were infused into the lumen of vessels at 100 mmHg for 30 minutes. Frozen sections were then cut and stained for gold particles. Image analysis was used to determine the density of the particles in the vessel walls. The expression of endothelial cell occludin mRNA and protein were determined using reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively. RESULTS: Permeability results showed a 2.8-fold increase in the apparent permeability of vessels cultured with low versus physiologic shear stress. RT-PCR and Western blotting results showed significant decreases in occludin mRNA and protein expression at 12 and 24 hours in vessels cultured with low versus physiologic shear stress. CONCLUSIONS: These results demonstrate that low shear stress increases vascular permeability in porcine carotid arteries, possibly owing to decreased occludin expression. These results may have implications in the preferential formation of atherosclerotic vascular disease adjacent to branches and bifurcations where low mean shear stresses may occur.

Correction of distortion of histologic sections of arteries.

Histologic sections of arteries can be used to generate three-dimensional (3D) geometric models and identify structural constituents. However, geometric distortions are introduced by fixation, embedding and sectioning; distortions which can, for example, lead to errors in stresses predicted by finite element models. We developed a method to measure and correct for distortions caused by acrylic processing and applied it to intact, healthy porcine coronary arteries. Micro-computed tomography was used to image arteries in the fresh and embedded states. Tissue blocks were sectioned, stained and imaged using a light microscope. Each section contained four registration marks used to determine strains introduced by sectioning and staining. Using these three image sets, 3D geometric models were generated and distortions were measured. Fixation, processing, and embedding resulted in shrinkage of 6.4+/-2.3% axially and 35.4+/-5.0% in mean cross-sectional area (n=5). Shrinkage in a cross section was well characterized by a uniform, equibiaxial strain. Sectioning and staining resulted in additional compressive strains in the sectioning direction of 0.067+/-0.011 and, in the direction perpendicular to sectioning, of 0.023+/-0.005 (n=5). These strains are assumed uniform and form the basis for correcting section geometry. Reconstructions using corrections for sectioning and shrinkage-related distortions had errors of 1.6+/-0.5% (n=5) and 4.0+/-1.7% (n=5), respectively.

Characterizing intramural stress and inflammation in hypertensive arterial bifurcations.

A histology-based methodology was developed and used to determine whether intramural stress and combined monocyte/macrophage density positively correlate within hypertensive bifurcations. Hypertension was induced in Sprague-Dawley rats using Angiotensin II pumps. Analysis focused on mesenteric bifurcations harvested 7 days (n = 4) post implant, but also included normotensive (n = 2) and 21-day hypertensive (n = 1) samples. Mesentery was processed in a manner that preserves morphology, corrects for histology-related distortions and results in reconstructions suitable for finite element analysis. Peaks in intramural stress and monocyte/macrophage density occurred near bifurcations after the onset of hypertension. Cell density peaks occurred in regions where surface curvature is complex and tends to heighten intramural stress. Also, a strong positive correlation between mean stress and mean cell density suggests that they are related phenomena. A point-by-point comparison of stress and cell density throughout each bifurcation did not exhibit a consistent pattern. We offer reasons why this most stringent test did not corroborate our other findings that high intramural stress is correlated with increased inflammation near the center of the bifurcation.

Effect of stenosis asymmetry on blood flow and artery compression: a three-dimensional fluid-structure interaction model.

A nonlinear three-dimensional thick-wall model with fluid-structure interactions is introduced to simulate blood flow in carotid arteries with an asymmetric stenosis to quantify the effects of stenosis severity, eccentricity, and pressure conditions on blood flow and artery compression (compressive stress in the wall). Mechanical properties of the tube wall are measured using a thick-wall stenosis model made of polyvinyl alcohal hydrogel whose mechanical properties are close to that of carotid arteries. A hyperelastic Mooney-Rivlin model is used to implement the experimentally measured nonlinear elastic properties of the tube wall. A 36.5% pre-axial stretch is applied to make the simulation physiological. The Navier-Stokes equations in curvilinear form are used for the fluid model. Our results indicate that severe stenosis causes critical flow conditions, high tensile stress, and considerable compressive stress in the stenosis plaque which may be related to artery compression and plaque cap rupture. Stenosis asymmetry leads to higher artery compression, higher shear stress and a larger flow separation region. Computational results are verified by available experimental data.

Constitutive modeling of porcine coronary arteries using designed experiments.

The development of new coronary artery constitutive models is of critical importance in the design and analysis of coronary replacement grafts. In this study, a two-parameter logarithmic complementary energy function, with normalized measured force and internal pressure as the independent variables and strains as the dependent variables, was developed for healthy porcine coronary arteries. Data was collected according to an experimental design with measured force ranging from 9.8 to 201 mN and internal pressure ranging from 0.1 to 16.1 kPa (1 to 121 mmHg). Comparisons of the estimated constitutive parameters showed statistically significant differences between the left anterior descending [LAD] and right coronary artery [RCA], but no differences between the LAD and left circumflex [LCX] or between the LCX and RCA. Point-by-point strain comparisons confirm the findings of the model parameter study and isolate the difference to the axial strain response. Average axial strains for the LAD, LCX, and RCA are 0.026 +/- 0.009, 0.015 +/- 0.005, and 0.011 +/- 0.009, respectively, at all physiologic loads, suggesting that the axial strains in the LAD are significantly higher than in the other regions.

Blood vessel constitutive models-1995-2002.

Knowledge of blood vessel mechanical properties is fundamental to the understanding of vascular function in health and disease. Analytic results can help physicians in the clinic, both in designing and in choosing appropriate therapies. Understanding the mechanical response of blood vessels to physiologic loads is necessary before ideal therapeutic solutions can be realized. For this reason, blood vessel constitutive models are needed. This article provides a critical review of recent blood vessel constitutive models, starting with a brief overview of the structure and function of arteries and veins, followed by a discussion of experimental techniques used in the characterization of material properties. Current models are classified by type, including pseudoelastic, randomly elastic, poroelastic, and viscoelastic. Comparisons are presented between the various models and existing experimental data. Applications of blood vessel constitutive models are also briefly presented, followed by the identification of future directions in research.

Arterial wall adaptation under elevated longitudinal stretch in organ culture.

Arteries in vivo are subjected to large longitudinal stretch which may change significantly due to vascular disease and surgery. However, little is known about the effect of longitudinal stretch on vascular function and wall remodeling, although the effects of tensile and shear stress from blood pressure and flow have been well documented. To study the effect of longitudinal stretch on vascular function and wall remodeling, porcine carotid arteries were longitudinally stretched 20% more than in vivo for 5 days while being maintained in an ex vivo organ culture system under conditions of pulsatile flow at physiologic pressure. Vessel viability was demonstrated by strong vasomotor responses to norepinephrine (NE, 10(-6) M), carbachol (10(-6) M), and sodium nitroprusside (10(-5) M), as well as by dense staining for mitochondrial activity and a low occurrence of cell necrosis. Cell proliferation was examined by incorporation of bromodeoxyuridine (BrdU). Results showed that arteries maintain normal structure and viability after 5 days in organ culture. Both the stretched and control arteries demonstrated significant contractile responses. For example, both stretched and control arteries showed approximately 10% diameter contraction in response to NE. Stretched arteries contained 8% BrdU-positive cells compared to 5% in controls (p<0.05). These results indicate that longitudinal stretch promotes cell proliferation in arteries while maintaining arterial function.