Mechanical properties of elastin along the thoracic aorta in the pig

Understanding the mechanical environment of each component within the arterial wall is fundamental for understanding vascular growth and remodelling and for engineering artificial vascular conduits. We have investigated the mechanical status of arterial elastin by measuring the circumferential mechanical properties of purified elastin as function of position along the descending thoracic aorta of the pig. The tensile circumferential secant modulus, E(sec), measured in uniaxial mechanical tests, increased 30% (P<0.001), from a value of 0.88 MPa in the proximal tissue near the aortic arch to 1.14 MPa in the distal tissue near the diaphragm, indicating the stiffness of the elastin sample increased with position. Breaking stress was 54% higher in the distal tissue compared to the proximal (P<0.001), but the breaking stretch ratio did not change. E(sec) correlated with the ratio of radius to wall thickness measured in the no load state, r(nl)/h(nl), suggesting that the rise in stiffness was linked to ring morphology. The higher stiffness and strength of the distal tissue might be explained by a higher proportion of circumferentially oriented fibres in the distal tissue, which would indicate that the elastin meshwork in the thoracic aorta may become progressively anisotropic with distance from the heart. The ratio r(nl)/(h(nl)E (sec))rose only 7%, which suggests that the in vivo circumferential strain on the elastin may be constant along the pig thoracic aorta. The positional variation in elastin's properties should be taken into account in mechanical studies on purified elastin and in mathematical models of aorta mechanics.     

Circumferential variations of mechanical behavior of the porcine thoracic aorta during the inflation test

We developed an extension-inflation experimental apparatus with a stereo vision system and a stress-strain analysis method to determine the regional mechanical properties of a blood vessel. Seven proximal descending thoracic aortas were investigated during the inflation test at a fixed longitudinal stretch ratio of 1.35 over a transmural pressure range from 1.33 to 21.33 kPa. Four circumferential regions of each aorta were designated as the anterior (A), left lateral (L), posterior (P), and right lateral (R) regions, and the inflation test was repeated for each region of the aortas. We used continuous functions to approximate the surfaces of the regional aortic wall in the reference configuration and the deformed configuration. Circumferential stretch and stress at the four circumferential regions of the aorta were computed. Circumferential stiffness, defined as the tangent of the stress-stretch curve, and physiological aortic stiffness, named pressure-strain elastic modulus, were also computed for each region. In the low pressure range, the stress increased linearly with increased stretch, but the mechanical response became progressively stiffer in the high-pressure range above a transition point. At a transmural pressure of 12.00 kPa, mean values of stiffness were 416±104 kPa (A), 523±99 kPa (L), 634±91 kPa (P), and 489±82 kPa (R). The stiffness of the posterior region was significantly higher than that of the anterior region, but no significant difference was found in pressure-strain elastic modulus.     

Distribution of stress and strain along the porcine aorta and coronary arterial tree

The existence of a homeostatic state of stresses and strains has been axiomatic in the cardiovascular system. The objective of this study was to determine the distribution of circumferential stress and strain along the aorta and throughout the coronary arterial tree to test this hypothesis. Silicone elastomer was perfused through the porcine aorta and coronary arterial tree to cast the arteries at physiological pressure. The loaded and zero-stress dimensions of the vessels were measured. The aorta (1.8 cm) and its secondary branches were considered down to 1.5 mm diameter. The left anterior descending artery (4.5 mm) and its branches down to 10 microm were also measured. The Cauchy mean circumferential stress and midwall stretch ratio were calculated. Our results show that the stretch ratio and Cauchy stress were lower in the thoracic than in the abdominal aorta and its secondary branches. The opening angle (theta) and midwall stretch ratio (lambda) showed a linear variation with order number (n) as follows: theta = 10.2n + 63.4 (R(2) = 0.989) and lambda = 4.47 x 10(-2)n + 1.1 (R(2) = 0.995). Finally, the stretch ratio and stress varied between 1.2 and 1.6 and between 10 and 150 kPa, respectively, along the aorta and left anterior descending arterial tree. The relative uniformity of strain (50% variation) from the proximal aorta to a 10-microm arteriole implies that the vascular system closely regulates the degree of deformation. This suggests a homeostasis of strain in the cardiovascular system, which has important implications for mechanotransduction and for vascular growth and remodeling.     

Linked mechanical and biological aspects of remodeling in mouse pulmonary arteries with hypoxia-induced hypertension

Supravalvular aortic stenosis (SVAS) is associated with decreased elastin and altered arterial mechanics. Mice with a single deletion in the elastin gene (ELN(+/-)) are models for SVAS. Previous studies have shown that elastin haploinsufficiency in these mice causes hypertension, decreased arterial compliance, and changes in arterial wall structure. Despite these differences, ELN(+/-) mice have a normal life span, suggesting that the arteries remodel and adapt to the decreased amount of elastin. To test this hypothesis, we performed in vitro mechanical tests on abdominal aorta, ascending aorta, and left common carotid artery from ELN(+/-) and wild-type (C57BL/6J) mice. We compared the circumferential and longitudinal stress-stretch relationships and residual strains. The circumferential stress-stretch relationship is similar between genotypes and changes <3% with longitudinal stretch at lengths within 10% of the in vivo value. At mean arterial pressure, the circumferential stress in the ascending aorta is higher in ELN(+/-) than in wild type. Although arterial pressures are higher, the increased number of elastic lamellae in ELN(+/-) arteries results in similar tension/lamellae compared with wild type. The longitudinal stress-stretch relationship is similar between genotypes for most arteries. Compared with wild type, the in vivo longitudinal stretch is lower in ELN(+/-) abdominal and carotid arteries and the circumferential residual strain is higher in ELN(+/-) ascending aorta. The increased circumferential residual strain brings the transmural strain distribution in ELN(+/-) ascending aorta close to wild-type values. The mechanical behavior of ELN(+/-) arteries is likely due to the reduced elastin content combined with adaptive remodeling during vascular development.     

Mechanics of the human femoral adventitia including the high-pressure response

Adventitial mechanics were studied on the basis of adventitial tube tests and associated stress analyses utilizing a thin-walled model. Inflation tests of 11 nonstenotic human femoral arteries (79.3 +/- 8.2 yr, means +/- SD) were performed during autopsy. Adventitial tubes were separated anatomically and underwent cyclic, quasistatic extension-inflation tests using physiological pressures and high pressures up to 100 kPa. Associated circumferential and axial stretches were typically <20%, indicating "adventitiosclerosis." Adventitias behaved nearly elastically for both loading domains, demonstrating high tensile strengths (>1 MPa). The anisotropic and strongly nonlinear mechanical responses were represented appropriately by two-dimensional Fung-type stored-energy functions. At physiological pressure (13.3 kPa), adventitias carry ~25% of the pressure load in situ, whereas their circumferential and axial stresses were similar to the total wall stresses (~50 kPa in both directions), supporting a "uniform stress hypothesis." At higher pressures, they became the mechanically predominant layer, carrying >50% of the pressure load. These significant load-carrying capabilities depended strongly on circumferential and axial in-vessel prestretches (mean values: 0.95 and 1.08). On the basis of these results, the mechanical role of the adventitia at physiological and hypertensive states and during balloon angioplasty was characterized.     

Passive elastic properties of the rat aorta

The passive anisotropic elastic properties of rat's aorta were studied in vitro by subjecting cylindrical segments of thoracic and abdominal aorta to a wide range of deformations. Using data on pressure, axial stretch, outer diameter, axial force and wall thickness, incremental moduli of elasticity in the circumferential, axial and radial directions were computed. Results indicate that while the elastic behavior of the aortic wall is globally anisotropic, there exists a state of deformation at which the vessel displays incremental isotropy. This state of deformation corresponds approximately to the loading conditions to which the aorta is exposed in situ. Values of the moduli, analyzed as a function of transmural pressure, show that the stiffness of the aortic wall is fairly constant at low pressures but raises steeply for pressures higher than physiological. For axial stretches as occurring in situ, the magnitudes of the circumferential and radial moduli do not differ significantly for the thoracic aorta; hence this vessel can be regarded as transversely isotropic over a wide range of pressures. The same observation is valid also for the abdominal aorta when pressures equal or smaller than physiological are considered. For both the thoracic and abdominal segments of the aorta, the circumferential and radial moduli are smaller than the axial modulus at low pressures, while the reverse is true for large pressures.     

Differential effects of static and cyclic stretching during elastase digestion on the mechanical properties of extracellular matrices.

Enzyme activity plays an essential role in many physiological processes and diseases such as pulmonary emphysema. While the lung is constantly exposed to cyclic stretching, the effects of stretch on the mechanical properties of the extracellular matrix (ECM) during digestion have not been determined. We measured the mechanical and failure properties of elastin-rich ECM sheets loaded with static or cyclic uniaxial stretch (40% peak strain) during elastase digestion. Quasistatic stress-strain measurements were taken during 30 min of digestion. The incremental stiffness of the sheets decreased exponentially with time during digestion. However, digestion in the presence of static stretch resulted in an accelerated stiffness decrease, with a time constant that was nearly 3 x smaller (7.1 min) than during digestion alone (18.4 min). These results were supported by simulations that used a nonlinear spring network model. The reduction in stiffness was larger during static than cyclic stretch, and the latter also depended on the frequency. Stretching at 20 cycles/min decreased stiffness less than stretching at 5 cycles/min, suggesting a rate-dependent coupling between mechanical forces and enzyme activity. Furthermore, pure digestion reduced the failure stress of the sheets from 88 +/- 21 kPa in control to 29 +/- 15 kPa (P < 0.05), while static and cyclic stretch resulted in a failure stress of 7 +/- 5 kPa (P < 0.05). We conclude that not only the presence but the dynamic nature of mechanical forces have a significant impact on enzyme activity, hence the deterioration of the functional properties of the ECM during exposure to enzymes.     

A fiber-based constitutive model predicts changes in amount and organization of matrix proteins with development and disease in the mouse aorta

Decreased elastin in mice (Eln+/?) yields a functioning vascular system with elevated blood pressure and increased arterial stiffness that is morphologically distinct from wild-type mice (WT). Yet, function is retained enough that there is no appreciable effect on life span and some mechanical properties are maintained constant. It is not understood how the mouse modifies the normal developmental process to produce a functioning vascular system despite a deficiency in elastin. To quantify changes in mechanical properties, we have applied a fiber-based constitutive model to mechanical data from the ascending aorta during postnatal development of WT and Eln+/? mice. Results indicate that the fiber-based constitutive model is capable of distinguishing elastin amounts and identifying trends during development. We observe an increase in predicted circumferential stress contribution from elastin with age, which correlates with increased elastin amounts from protein quantification data. The model also predicts changes in the unloaded collagen fiber orientation with age, which must be verified in future work. In Eln+/? mice, elastin amounts are decreased at each age, along with the predicted circumferential stress contribution of elastin. Collagen amounts in Eln+/? aorta are comparable to WT, but the predicted circumferential stress contribution of collagen is increased. This may be due to altered organization or structure of the collagen fibers. Relating quantifiable changes in arterial mechanics with changes in extracellular matrix (ECM) protein amounts will help in understanding developmental remodeling and in producing treatments for human diseases affecting ECM proteins.     

In vivo estimation of the contribution of elastin and collagen to the mechanical properties in the human abdominal aorta: effect of age and sex

The mechanical properties of the aorta affect cardiac function and are related to cardiovascular morbidity/mortality. This study was designed to evaluate the isotropic (mainly elastin, elastin(iso)) and anisotropic (mainly collagen, collagen(ani)) material parameters within the human aorta in vivo. Thirty healthy men and women in three different age categories (23-30, 41-54, and 67-72 yr) were included. A novel mechanical model was used to identify the mechanical properties and the strain field with aid of simultaneously recorded pressure and radius in the abdominal aorta. The magnitudes of the material parameters relating to both the stiffness of elastin(iso) and collagen(ani) were in agreement with earlier in vitro studies. The load-bearing fraction attributed to collagen(ani) oscillated from 10 to 30% between diastolic and systolic pressures during the cardiac cycle. With age, stiffness of elastin(iso) increased in men, despite the decrease in elastin content that has been found due to elastolysis. Furthermore, an increase in stiffness of collagen(ani) at high physiological pressure was found. This might be due to increased glycation, as well as changed isoforms of collagen in the aortic wall with age. A marked sex difference was observed, with a much less age-related effect, both on elastin(iso) and collagen(ani) stiffness in women. Possible factors of importance could be the effect of sex hormones, as well as differing collagen isoforms, between the sexes.     

Mechanical anisotropy of inflated elastic tissue from the pig aorta

Uniaxial and biaxial mechanical properties of purified elastic tissue from the proximal thoracic aorta were studied to understand physiological load distributions within the arterial wall. Stress–strain behaviour was non-linear in uniaxial and inflation tests. Elastic tissue was 40% stiffer in the circumferential direction compared to axial in uniaxial tests and～100% stiffer in vessels at an axial stretch ratio of 1.2 or 1.3 and inflated to physiological pressure. Poisson’s ratio v_θz averaged 0.2 and v_zθ increased with circumferential stretch from ～0.2 to ～0.4. Axial stretch had little impact on circumferential behaviour. In intact (unpurified) vessels at constant length, axial forces decreased with pressure at low axial stretches but remained constant at higher stretches. Such a constant axial force is characteristic of incrementally isotropic arteries at their in vivo dimensions. In purified elastic tissue, force decreased with pressure at all axial strains, showing no trend towards isotropy. Analysis of the force–length–pressure data indicated a vessel with v_θz≈0.2 would stretch axially 2–4% with the cardiac pulse yet maintain constant axial force. We compared the ability of 4 mathematical models to predict the pressure-circumferential stretch behaviour of tethered, purified elastic tissue. Models that assumed isotropy could not predict the stretch at zero pressure. The neo-Hookean model overestimated the non-linearity of the response and two non-linear models underestimated it. A model incorporating contributions from orthogonal fibres captured the non-linearity but not the zero-pressure response. Models incorporating anisotropy and non-linearity should better predict the mechanical behaviour of elastic tissue of the proximal thoracic aorta.     

Effects of elastin degradation and surrounding matrix support on artery stability

Tortuous arteries are often associated with aging, hypertension, atherosclerosis, and degenerative vascular diseases, but the mechanisms are poorly understood. Our recent theoretical analysis suggested that mechanical instability (buckling) may lead to tortuous blood vessels. The objectives of this study were to determine the critical pressure of artery buckling and the effects of elastin degradation and surrounding matrix support on the mechanical stability of arteries. The mechanical properties and critical buckling pressures, at which arteries become unstable and deform into tortuous shapes, were determined for a group of five normal arteries using pressurized inflation and buckling tests. Another group of nine porcine arteries were treated with elastase (8 U/ml), and the mechanical stiffness and critical pressure were obtained before and after treatment. The effect of surrounding tissue support was simulated using a gelatin gel. The critical pressures of the five normal arteries were 9.52 kPa (SD 1.53) and 17.10 kPa (SD 5.11) at axial stretch ratios of 1.3 and 1.5, respectively, while model predicted critical pressures were 10.11 kPa (SD 3.12) and 17.86 kPa (SD 5.21), respectively. Elastase treatment significantly reduced the critical buckling pressure (P < 0.01). Arteries with surrounding matrix support buckled into multiple waves at a higher critical pressure. We concluded that artery buckling under luminal pressure can be predicted by a buckling equation. Elastin degradation weakens the arterial wall and reduces the critical pressure, which thus leads to tortuous vessels. These results shed light on the mechanisms of the development of tortuous vessels due to elastin deficiency.     

A theoretical and non-destructive experimental approach for direct inclusion of measured collagen orientation and recruitment into mechanical models of the artery wall

Gradual collagen recruitment has been hypothesized as the underlying mechanism for the mechanical stiffening with increasing stress in arteries. In this work, we investigated this hypothesis in eight rabbit carotid arteries by directly measuring the distribution of collagen recruitment stretch under increasing circumferential loading using a custom uniaxial (UA) extension device combined with a multi-photon microscope (MPM). This approach allowed simultaneous mechanical testing and imaging of collagen fibers without traditional destructive fixation methods. Fiber recruitment was quantified from 3D rendered MPM images, and fiber orientation was measured in projected stacks of images. Collagen recruitment was observed to initiate at a finite strain, corresponding to a sharp increase in the measured mechanical stiffness, confirming the previous hypothesis and motivating the development of a new constitutive model to capture this response. Previous constitutive equations for the arterial wall have modeled the collagen contribution with either abrupt recruitment at zero strain, abrupt recruitment at finite strain or as gradual recruitment beginning at infinitesimal strain. Based on our experimental data, a new combined constitutive model was presented in which fiber recruitment begins at a finite strain with activation stretch represented by a probability distribution function. By directly including this recruitment data, the collagen contribution was modeled using a simple Neo-Hookean equation. As a result, only two phenomenological material constants were required from the fit to the stress stretch data. Three other models for the arterial wall were then compared with these results. The approach taken here was successful in combining stress-strain analysis with simultaneous microstructural imaging of collagen recruitment and orientation, providing a new approach by which underlying fiber architecture may be quantified and included in constitutive equations.     

Flow-induced shear strain in intima of porcine coronary arteries.

The in vivo circumferential strain has a small variation throughout the vascular system (aorta to arterioles). The axial strain has also been shown to be nearly the same as the circumferential strain under physiological loading. Since the endothelium is mechanically much softer than the media-adventitia in healthy arteries, the porcine intima was considered as a mechanically distinct layer from the media-adventitia in a two-layer computational model. Based on the simulation result, we hypothesize that the flow-induced shear strain in intima can be of similar value as the pressure-induced circumferential strain in healthy coronary arteries, even though the shear stress is orders of magnitude smaller than the circumferential stress. The nearly isotropic deformation (circumferential, axial, and shear strains) may have important implications for mechanical homeostasis of endothelial cells, mechanotransduction, growth, and remodeling of blood vessels.     

Biomechanical and morphological properties in rat large intestine

Intestinal stress-strain distributions are important determinants of intestinal function and are determined by the mechanical properties of the intestinal wall, the physiological loading conditions and the zero-stress state of the intestine. In this study the distribution of morphometric measures, residual circumferential strains and stress-strain relationships along the rat large intestine were determined in vitro. Segments from four parts of the large intestine were excised, closed at both ends, and inflated with pressures up to 2kPa. The outer diameter and length were measured. The zero-stress state was obtained by cutting rings of large intestine radially. The geometric configuration at the zero-stress state is of fundamental importance because it is the basic state with respect to which the physical stresses and strains are defined. The outer and inner circumferences, wall thickness and opening angle were measured from digitised images. Subsequently, residual strain and stress-strain distributions were calculated. The wall thickness and wall thickness-to-circumference ratio increased in the distal direction. The opening angle varied between approximately 40 and approximately 125 degrees with the highest values in the beginning of proximal colon (F=1.739, P<0.05). The residual strain at the inner surface was negative indicating that the mucosa-submucosal layers of the large intestine in no-load state are in compression. The four segments showed stress-strain distributions that were exponential. All segments were stiffer in longitudinal direction than in the circumferential direction (P<0.05). The transverse colon seemed stiffest both in the circumferential and longitudinal directions. In conclusion, significant variations were found in morphometric and biomechanical properties along the large intestine. The circumferential residual strains and passive elastic properties must be taken into account in studies of physiological problems in which the stress and strain are important, e.g. large intestinal bolus transport function.     

Arterial Extracellular Matrix: A Mechanobiological Study of the Contributions and Interactions of Elastin and Collagen

The complex network structure of elastin and collagen extracellular matrix (ECM) forms the primary load bearing components in the arterial wall. The structural and mechanobiological interactions between elastin and collagen are important for properly functioning arteries. Here, we examined the elastin and collagen organization, realignment, and recruitment by coupling mechanical loading and multiphoton imaging. Two-photon excitation fluorescence and second harmonic generation methods were performed with a multiphoton video-rate microscope to capture real time changes to the elastin and collagen structure during biaxial deformation. Enzymatic removal of elastin was performed to assess the structural changes of the remaining collagen structure. Quantitative analysis of the structural changes to elastin and collagen was made using a combination of two-dimensional fast Fourier transform and fractal analysis, which allows for a more complete understanding of structural changes. Our study provides new quantitative evidence, to our knowledge on the sequential engagement of different arterial ECM components in response to mechanical loading. The adventitial collagen exists as large wavy bundles of fibers that exhibit fiber engagement after 20% strain. The medial collagen is engaged throughout the stretching process, and prominent elastic fiber engagement is observed up to 20% strain after which the engagement plateaus. The fiber orientation distribution functions show remarkably different changes in the ECM structure in response to mechanical loading. The medial collagen shows an evident preferred circumferential distribution, however the fiber families of adventitial collagen are obscured by their waviness at no or low mechanical strains. Collagen fibers in both layers exhibit significant realignment in response to unequal biaxial loading. The elastic fibers are much more uniformly distributed and remained relatively unchanged due to loading. Removal of elastin produces similar structural changes in collagen as mechanical loading. Our study suggests that the elastic fibers are under tension and impart an intrinsic compressive stress on the collagen.     

Arterial Extracellular Matrix: A Mechanobiological Study of the Contributions and Interactions of Elastin and Collagen

The complex network structure of elastin and collagen extracellular matrix (ECM) forms the primary load bearing components in the arterial wall. The structural and mechanobiological interactions between elastin and collagen are important for properly functioning arteries. Here, we examined the elastin and collagen organization, realignment, and recruitment by coupling mechanical loading and multiphoton imaging. Two-photon excitation fluorescence and second harmonic generation methods were performed with a multiphoton video-rate microscope to capture real time changes to the elastin and collagen structure during biaxial deformation. Enzymatic removal of elastin was performed to assess the structural changes of the remaining collagen structure. Quantitative analysis of the structural changes to elastin and collagen was made using a combination of two-dimensional fast Fourier transform and fractal analysis, which allows for a more complete understanding of structural changes. Our study provides new quantitative evidence, to our knowledge on the sequential engagement of different arterial ECM components in response to mechanical loading. The adventitial collagen exists as large wavy bundles of fibers that exhibit fiber engagement after 20% strain. The medial collagen is engaged throughout the stretching process, and prominent elastic fiber engagement is observed up to 20% strain after which the engagement plateaus. The fiber orientation distribution functions show remarkably different changes in the ECM structure in response to mechanical loading. The medial collagen shows an evident preferred circumferential distribution, however the fiber families of adventitial collagen are obscured by their waviness at no or low mechanical strains. Collagen fibers in both layers exhibit significant realignment in response to unequal biaxial loading. The elastic fibers are much more uniformly distributed and remained relatively unchanged due to loading. Removal of elastin produces similar structural changes in collagen as mechanical loading. Our study suggests that the elastic fibers are under tension and impart an intrinsic compressive stress on the collagen.     

Mechanical efficiency of the trout heart during volume and pressure-loading: metabolic implications of the stiffness of the ventricular tissue

In the mammalian heart the metabolic costs of pressure loading exceed those of volume loading. As evidence suggests that the opposite may be true in fish, we evaluated the metabolic costs of volume and pressure loading in the isolated trout heart and compared the results with the mammalian heart based on the biomechanical properties of cardiac muscle. The highest power output (2.33+/-0.32 mW g(-1), n=5) appeared at the highest preload pressure tested (0.3 kPa) and at an afterload of 5 kPa. At a higher afterload, power did not increase because stroke volume fell. The highest mechanical efficiency (20.7+/-2.0%, n=5) was obtained at a preload of 0.15 kPa and an afterload of 5 kPa. Further increases in preload or afterload did not increase mechanical efficiency, probably because of increases in ventricular wall stress which increased the oxygen consumed disproportionately more than the stroke work. Under pressure unloading (25% decrease in power output), mechanical efficiency was significantly higher in comparison with volume unloading. Given that stiffness of the ventricular tissue is larger in trout than in rat papillary muscles, it is suggested that the increased strain during volume loading is energetically disadvantageous for stiff muscles like those of trout, but it is advantageous when muscle stiffness is lower as it occurs in the rat papillary muscle.     

Changes in the structure-function relationship of elastin and its impact on the proximal pulmonary arterial mechanics of hypertensive calves

Extracellular matrix remodeling has been proposed as one mechanism by which proximal pulmonary arteries stiffen during pulmonary arterial hypertension (PAH). Although some attention has been paid to the role of collagen and metallomatrix proteins in affecting vascular stiffness, much less work has been performed on changes in elastin structure-function relationships in PAH. Such work is warranted, given the importance of elastin as the structural protein primarily responsible for the passive elastic behavior of these conduit arteries. Here, we study structure-function relationships of fresh arterial tissue and purified arterial elastin from the main, left, and right pulmonary artery branches of normotensive and hypoxia-induced pulmonary hypertensive neonatal calves. PAH resulted in an average 81 and 72% increase in stiffness of fresh and digested tissue, respectively. Increase in stiffness appears most attributable to elevated elastic modulus, which increased 46 and 65%, respectively, for fresh and digested tissue. Comparison between fresh and digested tissues shows that, at 35% strain, a minimum of 48% of the arterial load is carried by elastin, and a minimum of 43% of the change in stiffness of arterial tissue is due to the change in elastin stiffness. Analysis of the stress-strain behavior revealed that PAH causes an increase in the strains associated with the physiological pressure range but had no effect on the strain of transition from elastin-dominant to collagen-dominant behavior. These results indicate that mechanobiological adaptations of the continuum and geometric properties of elastin, in response to PAH, significantly elevate the circumferential stiffness of proximal pulmonary arterial tissue.     

The Effect of Static Stretch on Elastin Degradation in Arteries

Previously we have shown that gradual changes in the structure of elastin during an elastase treatment can lead to important transition stages in the mechanical behavior of arteries [1]. However, in vivo arteries are constantly being loaded due to systolic and diastolic pressures and so understanding the effects of loading on the enzymatic degradation of elastin in arteries is important. With biaxial tensile testing, we measured the mechanical behavior of porcine thoracic aortas digested with a mild solution of purified elastase (5 U/mL) in the presence of a static stretch. Arterial mechanical properties and biochemical composition were analyzed to assess the effects of mechanical stretch on elastin degradation. As elastin is being removed, the dimensions of the artery increase by more than 20% in both the longitude and circumference directions. Elastin assays indicate a faster rate of degradation when stretch was present during the digestion. A simple exponential decay fitting confirms the time constant for digestion with stretch (0.11±0.04 h⁻¹) is almost twice that of digestion without stretch (0.069±0.028 h⁻¹). The transition from J-shaped to S-shaped stress vs. strain behavior in the longitudinal direction generally occurs when elastin content is reduced by about 60%. Multiphoton image analysis confirms the removal/fragmentation of elastin and also shows that the collagen fibers are closely intertwined with the elastin lamellae in the medial layer. After removal of elastin, the collagen fibers are no longer constrained and become disordered. Release of amorphous elastin during the fragmentation of the lamellae layers is observed and provides insights into the process of elastin degradation. Overall this study reveals several interesting microstructural changes in the extracellular matrix that could explain the resulting mechanical behavior of arteries with elastin degradation.     

The effect of elastin damage on the mechanics of the aortic valve

Porcine bioprosthetic heart valves degenerate and fail mechanically through a mechanism that is currently not well understood. It has been suggested that damage to the elastin component of prosthetic valve cusps could be responsible for changes in the mechanical function of the valve that would predispose it to increased damage and ultimate failure. To determine whether damage to elastin can produce the structural and mechanical changes that could initiate the process of bioprosthetic valve degeneration, we developed an elastase treatment protocol that fragments elastin and negates its mechanical contribution to the valve tissue. Valve cusps were mechanically tested before and after digestion to measure the mechanical changes resulting from elastin damage. Elastin damage produced a decrease in radial and circumferential extensibility (from 43 to 18% strain radially and 12 to 7% strain circumferentially), with a slight increase in stiffness (1.3-2.6kN/m for radial and 10.6-11.9kN/m for circumferential directions). Digestions with trypsin, which does not cleave elastin, confirmed that the changes in mechanics of the circumferential samples were likely due to the nonspecific removal of proteoglycans by elastase, while the changes in the radial samples were indeed due to elastin damage. Removing the mechanical contribution of elastin alters the mechanical behavior of the aortic valve cusp, primarily in the radial direction. This finding implies that damage to elastin will distend the cusps, reduce their extensibility, and increase their stiffness. Damage to elastin may therefore contribute to the degeneration and failure of prosthetic valves.