Inflation-extension Tests Research Articles

False aneurysms of the thoracic aorta: anastomosis investigation using the inflation-extension test.

False aneurysms in the thoracic aorta are dangerous complications that can occur after cardiac surgery. They often result in high mortality rates. These aneurysms are caused by damage to all layers of the aortic wall. This study aimed to pinpoint the area of the experimental specimen (native vessel, anastomosis, or prosthetic graft) with the greatest deformation, to determine whether a false aneurysm is likely to develop in the anastomotic portion. We conducted the inflation-extension test by performing eight cycles ranging from 0 to 20. The pressure sampling frequency was 100 Hz, and each cycle lasted approximately 34 seconds, resulting in a loading frequency of 0.03 Hz. During the experiment, each camera captured 3,000 frames. Based on the data collected, we evaluated and compared the loading stages of cycle 1 and cycle 8. During loading, the native vessel experienced a dominant deformation of approximately 7% in the circumferential direction. The prosthetic graft, which had a longitudinal construction, deformed by approximately 8% in the axial direction. The prosthetic graft, on the other hand, only experienced a deformation of up to 1.5% in the circumferential direction, which was about 5 times smaller than the deformation of the native vessel. The anastomosis area was very stiff and showed minimal deformation. Additionally, there was little difference in the mechanical response between the first C1 and the eighth C8 cycle. Based on the available evidence, it can be inferred that aortic false aneurysms are more likely to form just behind the suture lines in the native aorta, which is more elastic compared to stiff sections of anastomosis and prosthetic graft. Numerous pulsations of the native vessel will likely cause the impairment of the aorta at the margin of the anastomosis. This will lead to disruption of the aortic wall and false aneurysm formation in the native vessel near the area of anastomosis.

Development and Initial Characterisation of a Localised Elastin Degradation Ex Vivo Porcine Aortic Aneurysm Model

Aortic aneurysms (AA) occur in 4.8% of people causing 150,000 deaths annually. While endovascular aneurysm repairs reduce surgical morbidity, device-related failures (leak/displacement) are frequent highlighting the need for test models that better represent the mural geometry and compliance changes in human AAs. We aimed to develop and characterise an ex vivo porcine aortic model of AA. The optimal duration of tissue elastase exposure to emulate AA changes in elastin microstructure and content was determined using porcine aortic rings. Elastase-induced changes were quantified morphologically, and mechanical properties assessed via ring tensile testing. Subsequent experiments tested the potential for localised elastase treatment in a 1 cm segment of porcine aorta using a specially designed 3D printed rig. The effect on pressure-diameter behaviour was investigated via inflation-extension testing. Elastase treatment produced time dependent decreases in elastin, resulting in an increased tensile modulus and circumferential length in the ring samples in the final phase of the J-shaped tissue stress-strain curves. In whole aortic segments, localised elastase-induced luminal degradation was successfully limited to a central region. The degree of elastin degradation achieved was sufficient to cause localised dilation with respect to controls under physiological pressures. Localised elastin degradation in porcine aortic segments is feasible and emulates the changes seen clinically in aortic aneurysms.

Experimental surgery as part of the development of degradable biomaterials for cardiovascular surgery

Introduction: Cardiovascular diseases are responsible for significant morbidity and mortality in the population. Artificial vascular grafts are often essential for surgical procedures in radical or palliative treatment. Many new biodegradable materials are currently under development. Preclinical testing of each new material is imperative, both in vitro and in vivo, and therefore animal experiments are still a necessary part of the testing process before any clinical use. The aim of this paper is to present the options of using various experimental animal models in the field of cardiovascular surgery including their extrapolation to clinical medicine. Methods: The authors present their general experience in the field of experimental surgery. They discuss the selection process of an optimal experimental animal model to test foreign materials for cardiovascular surgery and of an optimal region for implantation. Results: The authors present rat, rabbit and porcine models as optimal experimental animals for material hemocompatibility and degradability testing. Intraperitoneal implantation in the rat is a simple and feasible procedure, as well as aortic banding in the rabbit or pig. The carotid arteries can also be used, as well. Porcine pulmonary artery banding is slightly more difficult with potential complications. The banded vessels, explanted after a defined time period, are suitable for further mechanical testing using biomechanical analyses, for example, the inflation-extension test. Conclusion: An in vivo experiment cannot be avoided in the last phases of preclinical research of new materials. However, we try to strictly observe the 3R concept – Replacement, Reduction and Refinement; in line with this concept, the potential of each animal should be used as much as possible to reduce the number of animals.

Cell contractility and Focal Adhesion Kinase control circumferential arterial stiffness.

Arterial stiffening is a hallmark of aging and cardiovascular disease. While it is well established that vascular smooth muscle cells (SMCs) contribute to arterial stiffness by synthesizing and remodeling the arterial extracellular matrix, the direct contributions of SMC contractility and mechanosensors to arterial stiffness, and particularly the arterial response to pressure, remain less well understood despite being a long-standing question of biomedical importance. Here, we have examined this issue by combining use of pressure myography of intact carotid arteries, pharmacologic inhibition of contractility, and genetic deletion of SMC focal adhesion kinase (FAK). Biaxial inflation-extension tests performed at physiological pressures showed that acute inhibition of cell contractility with blebbistatin or EGTA altered vessel geometry and preferentially reduced circumferential, as opposed to axial, arterial stiffness in wild-type mice. Similarly, genetic deletion of SMC FAK, which attenuated arterial contraction to KCl, reduced vessel wall thickness and circumferential arterial stiffness in response to pressure while having minimal effect on axial mechanics. Moreover, these effects of FAK deletion were lost by treating arteries with blebbistatin or by inhibiting myosln light chain kinase. The expression of arterial fibrillar collagens, the integrity of arterial elastin, or markers of SMC differentiation were not affected by deletion of SMC FAK. Our results connect cell contractility and SMC FAK to the regulation of arterial wall thickness and directionally-specific arterial stiffening.

Biomechanical Properties of Mouse Carotid Arteries With Diet-Induced Metabolic Syndrome and Aging.

Metabolic syndrome increases the risk of cardiovascular diseases. Arteries gradually stiffen with aging; however, it can be worsened by the presence of conditions associated with metabolic syndrome. In this study, we investigated the combined effects of diet-induced metabolic syndrome and aging on the biomechanical properties of mouse common carotid arteries (CCA). Male mice at 2 months of age were fed a normal or a high fat and high sucrose (HFHS) diet for 2 (young group), 8 (adult group) and 18–20 (old group) months. CCAs were excised and subjected to in vitro biaxial inflation-extension tests and the Cauchy stress-stretch relationships were determined in both the circumferential and longitudinal directions. The elastic energy storage of CCAs was obtained using a four-fiber family constitutive model, while the material stiffness in the circumferential and longitudinal directions was computed. Our study showed that aging is a dominant factor affecting arterial remodeling in the adult and old mice, to a similar extent, with stiffening manifested with a significantly reduced capability of energy storage by ∼50% (p < 0.05) and decreases in material stiffness and stress (p < 0.05), regardless of diet. On the other hand, high fat high sucrose diet resulted in an accelerated arterial remodeling in the young group at pre-diabetic stage by affecting the circumferential material stiffness and stress (p < 0.05), which was eventually overshadowed by aging progression. These findings have important implications on the effects of metabolic syndrome on elastic arteries in the younger populations.

Histological and biomechanical properties of systemic arteries in young and old Warmblood horses.

Arterial rupture is a well-recognized cause of sudden death in horses, which mainly affects older horses. The arterial wall is known to stiffen with age, although the underlying age-related histological and biomechanical changes remain unclear. The purpose of this study was to investigate the effect of aging by histological analysis of the arterial wall and examination of the arterial wall biomechanical properties using an inflation-extension test. Entire circular samples of the proximal and distal aorta, cranial and caudal common carotid, external iliac, femoral and median artery were collected from 6 young (6 years) and 14 old horses (≥15 years). Samples of all arteries were histologically examined and intima media thickness as well as area % of elastin, smooth muscle actin and collagen type I and III were determined. Older horses had a significantly larger intima media thickness and a significantly higher area % of smooth muscle actin compared to young horses. Samples of the proximal and distal aorta, the caudal common carotid and the external iliac artery were mechanically assessed using an in-house developed inflation-extension device with ultrasound analysis. Rupture occurred in a minority of arteries (8/78) at high pressures (between 250–300 mmHg), and mostly occurred in older horses (7/8). Pressure-area, pressure-compliance and pressure-distensibility curves were constructed. A significant difference in the pressure-area curves of the distal aorta, common carotid artery and external iliac artery, the pressure-compliance curves of the proximal aorta and carotid artery and the pressure-distensibility curve of the proximal aorta was observed between young and old horses. Results demonstrate an effect of age on the histological and biomechanical properties of the arterial wall, which might explain why arterial rupture occurs more often in older horses.

The Role of Biaxial Loading on Smooth Muscle Contractility in the Nulliparous Murine Cervix.

Throughout the estrus cycle, the extracellular matrix (ECM) and cervical smooth muscle cells (cSMC) coordinate to accomplish normal physiologic function in the non-pregnant cervix. While previous uniaxial experiments provide fundamental knowledge about cervical contractility and biomechanics, the specimen preparation is disruptive to native organ geometry and does not permit simultaneous assessment of circumferential and axial properties. Thus, a need remains to investigate cervical contractility and passive biomechanics within physiologic multiaxial loading. Biaxial inflation-extension experiments overcome these limitations by preserving geometry, ECM-cell interactions, and multiaxially loading the cervix. Utilizing in vivo pressure measurements and inflation-extension testing, this study presented methodology and examined maximum biaxial contractility and biomechanics in the nulliparous murine cervix. The study showed that increased pressure resulted in decreased contractile potential in the circumferential direction, however, axial contractility remained unaffected. Additionally, total change in axial stress ([Formula: see text]) increased significantly (p < 0.05) compared to circumferential stress ([Formula: see text]) with maximum contraction. However, passive stiffness was significantly greater (p < 0.01) in the circumferential direction. Overall, axial cSMC may have a critical function in maintaining cervical homeostasis during normal function. Potentially, a loss of axial contractility in the cervix during pregnancy may result in maladaptive remodeling such as cervical insufficiency.

Longitudinal histomechanical heterogeneity of the internal thoracic artery

The internal thoracic artery (ITA) is the principal choice for coronary artery bypass grafting (CABG) due to its mechanical compatibility, histological composition, anti-thrombogenic lumen, and single anastomotic junction. Originating at the subclavian artery, traversing the thoracic cavity, and terminating at the superior epigastric and musculophrenic bifurcation, bilateral ITAs follow a protracted circuitous pathway. The physiological hemodynamics, anatomical configuration, and perivascular changes that occur throughout this length influence the tissue's microstructure and gross mechanical properties. Since histomechanics play a major role in premature graft failure we used inflation-extension testing to quantify the regional material and biaxial mechanical properties at four distinct locations along the left (L) and right (R) ITA and fit the results to a structurally-motivated constitutive model. Our comparative analysis of 44 vessel segments revealed a significant increase in the amount of collagen but not smooth muscle and a significant decrease in elastin and elastic lamellae present with distance from the heart. A subsequent decrease in the total deformation energy and isotropic contribution to the strain energy was present in the LITA but not RITA. Circumferential stress and compliance generally decreased along the length of the LITA while axial stress increased in the RITA. When comparing RITAs to LITAs, some morphological and histological differences were found in proximal sections while distal sections revealed differences predominantly in compliance and axial stress. Overall, this information can be used to better guide graft selection, graft preparation, and xenograft-based tissue-engineering strategies for CABG.

Null strain analysis of submerged aneurysm analogues using a novel 3D stereomicroscopy device

To measure the inhomogeneous 3D-strain fields present during inflation-extension testing of physiologically submerged micro-aneurysms, a Stereo Digital Image Correlation (StereoDIC) microscopy system is developed that revolves 15° stereo-angle cameras around a centrally-mounted target. Calibration is performed using submerged dot patterns and system accuracy verified using strain and deformation analyses for rigid body motions of speckle-patterned, micro-aneurysmal surrogates. In terms of the Green-Lagrange strain tensor and the 3D displacement fields, the results are stable even after 120 minutes, with maxima in both strain bias and strain standard deviation less than 2E-03 for all components, and micron-level displacement standard deviation.

Iliac Veins Are More Compressible Than Iliac Arteries: A New Method of Testing.

Incompressibility implies that a tissue preserves its volume regardless of the loading conditions. Although this assumption is well-established in arterial wall mechanics, it is assumed to apply for the venous wall without validation. The objective of this study is to test whether the incompressibility assumption holds for the venous wall. To investigate the vascular wall volume under different loading conditions, inflation-extension testing protocol was used in conjunction with intravascular ultrasound (IVUS) in both common iliac arteries (n = 6 swine) and common iliac veins (n = 9 dogs). Use of IVUS allows direct visualizations of lumen dimensions simultaneous with direct measurements of outer dimensions during loading. The arterial tissue was confirmed to preserve volume during various load conditions (p = 0.11) consistent with the literature, while the venous tissue was found to lose volume (about 35%) under loaded conditions (p < 0.05). Using a novel methodology, this study shows the incompressibility assumption does not hold for the venous wall especially at higher pressures, which suggests that there may be fluid loss through the vein wall during loading. This has important implications for coupling of fluid transport across the wall and biomechanics of the wall in healthy and diseased conditions.

Biaxial biomechanical properties of the nonpregnant murine cervix and uterus

From a biomechanical perspective, female reproductive health is an understudied area of research. There is an incomplete understanding of the complex function and interaction between the cervix and uterus. This, in part, is due to the limited research into multiaxial biomechanical functions and geometry of these organs. Knowledge of the biomechanical function and interaction between these organs may elucidate etiologies of conditions such as preterm birth. Therefore, the objective of this study was to quantify the multiaxial biomechanical properties of the murine cervix and uterus using a biaxial testing set-up. To accomplish this, an inflation-extension testing protocol (n = 15) was leveraged to quantify biaxial biomechanical properties while preserving native matrix interactions and geometry. Ultrasound imaging and histology (n = 10) were performed to evaluate regional geometry and microstructure, respectively. Histological analysis identified a statistically significant greater collagen content and significantly smaller smooth muscle content in the cervix as compared to the uterus. No statistically significant differences in elastic fibers were identified. Analysis of bilinear fits revealed a significantly stiffer response from the circumferentially orientated ECM fibers compared to axially orientated fibers in both organs. Bilinear fits and a two-fiber family constitutive model showed that the cervix was significantly less distensible than the uterus. We submit that the regional biaxial information reported in this study aids in establishing an appropriate reference configuration for mathematical models of the uterine-cervical complex. Thus, may aid future work to elucidate the biomechanical mechanisms leading to cervical or uterine conditions.

Development of vascular substitutes for low-flow peripheral bypass grafting – a review

The development of alow-flow vascular prosthesis is avery topical issue. The authors present apathway for the development of aprosthesis with optimal properties based on the idea of mimicking the characteristics of abiological model (saphenous vein graft) and programming these properties in the model of the prosthetic substitute. The vascular prosthesis presented consists of three layers - anon-absorbable scaffold representing vascular &#8220;media&#8221;, and two absorbable collagen layers - pseudointima and pseudoadventitia. The basic methods of physical testing are presented - the single axis stretch test and inflation-extension test, as well as other procedures that affect the final properties. These include collagen curing, antithrombotic treatment of the inner layer and the use of sterilization methods. The designed new graft was successfully implanted in an ovine model.

Constitutive modeling of the passive inflation-extension behavior of the swine colon.

In the present work, we propose the first structural constitutive model of the passive mechanical behavior of the swine colon that is validated against physiological inflation-extension tests, and accounts for residual strains. Sections from the spiral colon and the descending colon were considered to investigate potential regional variability. We found that the proposed constitutive model accurately captures the passive inflation-extension behavior of both regions of the swine colon (coefficient of determination R2=0.94±0.02). The model revealed that the circumferential muscle layer does not provide significant mechanical support under passive conditions and the circumferential load is actually carried by the submucosa layer. The stress analysis permitted by the model showed that the colon tissue can distend up to 30% radially without significant increase in the wall stresses suggesting a highly compliant behavior of the tissue. This is in-line with the requirement for the tissue to easily accommodate variable quantities of fecal matter. The analysis also showed that the descending colon is significantly more compliant than the spiral colon, which is relevant to the storage function of the descending colon. Histological analysis showed that the swine colon possesses a four-layer structure similar to the human colon, where the longitudinal muscle layer is organized into bands called taeniae, a typical feature of the human colon. The model and the estimated parameters can be used in a Finite Element framework to conduct simulations with realistic geometry of the swine colon. The resulting computational model will provide a foundation for virtual assessment of safe and effective devices for the treatment of colonic diseases.

Inflation Test of Developed Biological Composite Reinforced by Polyester Mesh

The inflation-extension tests were performed with developed hybrid composite manufactured from biological collagenous matrix and reinforced with polyester mesh. One sample was made without the mesh. This control specimen showed compliant and almost linear behavior. Tests of samples with integrated mesh in the matrix demonstrated that mechanical properties of final composite could be modulated by placing of its reinforced component in the matrix. It was shown that we can obtain response from stiff and linear to nonlinear and very compliant depending on the position of the mesh within biological matter.

Identifying Hyper-Viscoelastic Model Parameters from an Inflation-Extension Test and Ultrasound Images

The aim of the present study was to develop and validate a method based on medical imaging to identify the parameters of a hyper-viscoelastic model suitable for describing the mechanical behavior of vascular tissues, focusing on the aorta. The method uses an inflation-extension test, and the model comprises one hyperelastic element in parallel with one or more Maxwell elements. Cylindrical samples of elastomeric silicone materials with a mechanical behavior similar to vascular tissues were placed in a circuit simulating hemodynamic flow through adequate controlled-pressure variation. Ultrasound B-mode image sequences were analyzed to measure the cyclic circumferential and longitudinal elongations. Precautions were taken a posteriori to resynchronize pressure and deformation signals, and thus minimize errors in the viscosity parameters estimated. The hyper-viscoelastic parameters of the samples were identified with reasonable accuracy as compared with the values obtained via standard measurements, namely tensile tests and dynamic mechanical analysis. However, the estimates of the viscosity parameters can be hampered in the case of stiffer samples. This limitation is bound to a restricted range of frequencies analyzed by the test, which mainly depends on the image acquisition rate. The use of the present method in the clinical environment for in vivo experiments can be foreseen provided that the local pressure measurements are available.

Biomechanical evaluation of ascending aortic aneurysms.

The biomechanical properties of ascending aortic aneurysms were investigated only in the last decade in a limited number of studies. Indeed, in recent years, there has been a growing interest in this field in order to identify new predictive parameters of risk of dissection, which may have clinical relevance. The researches performed so far have been conducted according to the methods used in the study of abdominal aortic aneurysms. In most cases, uniaxial or biaxial tensile tests were used, while in a smaller number of studies other methods, such as opening angle, bulge inflation, and inflation-extension tests, were used. However, parameters and protocols of these tests are at present very heterogeneous in the studies reported in the literature, and, therefore, the results are not comparable and are sometimes conflicting. The purpose of this review then thence to provide a comprehensive analysis of the experimental methodology for determination of biomechanical properties in the specific field of aneurysms of the ascending aorta to allow for better comparison and understanding of the results.

Constitutive Modelling and Histology of &lt;i&gt;Vena saphena&lt;/i&gt;

The inflation-extension test was performed in order to obtain the mechanical response (stress-strain curves) of the human vein - vena saphena magna (usually used for coronary artery bypass graft surgery). Tubular samples of the vein were inflated four times up to the pressure approx. 4 kPa (vein pressure) and then four times up to approx. 16 kPa (systolic pressure). The experiments were recorded by the CCD camera. The longitudinal and circumferential deformations of the tube were evaluated using the edge detection method. The experimental data were fitted by anisotropic, nonlinear, constitutive model in order to obtain model parameters, especially the parameter which can be explained as collagen fibres orientation approximation. This parameter was then compared with the findings from histology. The histology analyses based on label-free imaging were performed additionally to the mechanical testing. Collagen (most important load-bearing component of the vein wall) was visualized using second harmonic generation imaging (SHG, excitation at 860 nm by a tunable IR pulse laser, detection at 430±10 nm). This method enabled us to observe collagen through the vein wall. It was found that the collagen fibres are helically aligned within the vein at an angle 37±6° measured from circumferential axis. The results of collagen orientation angle show a good agreement of findings obtained from histology and from constitutive model.

A Preliminary Analysis of the Data From an in Vitro Inflation-Extension Test Can Validate the Assumption of Arterial Tissue Elasticity

The objective of this study is to propose a method for preliminary processing of the experimental data from an inflation-extension test on tubular arterial specimens. The method is based on the condition for existence of a strain energy function (SEF) and can be used to verify whether the data from a certain experiment validate the assumption that the tissue can be considered as an elastic solid. As an illustrative example of the proposed method, experimental data for a porcine renal artery are used and the sources of the error in satisfying the condition of elasticity are analyzed. The results lead to the conclusion that the experimental data for a renal artery validate that the artery exhibits an elastic mechanical response and a constitutive formulation based on the existence of the SEF is justified. A modification of the proposed method for the case of an in-plane biaxial stretching test of mechanically isotropic and orthotropic tissues is considered.

A structural constitutive model considering angular dispersion and waviness of collagen fibres of rabbit facial veins

BackgroundStructural constitutive models of vascular wall integrate information on composition and structural arrangements of tissue. In blood vessels, collagen fibres are arranged in coiled and wavy bundles and the individual collagen fibres have a deviation from their mean orientation. A complete structural constitutive model for vascular wall should incorporate both waviness and orientational distribution of fibres. We have previously developed a model, for passive properties of vascular wall, which considers the waviness of collagen fibres. However, to our knowledge there is no structural model of vascular wall which integrates both these features.MethodsIn this study, we have suggested a structural strain energy function that incorporates not only the waviness but also the angular dispersion of fibres. We studied the effect of parameters related to the orientational distribution on macro-mechanical behaviour of tissue during inflation-extension tests. The model was further applied on experimental data from rabbit facial veins.ResultsOur parametric study showed that the model is less sensitive to the orientational dispersion when fibres are mainly oriented circumferentially. The macro-mechanical response is less sensitive to changes in the mean orientation when fibres are more dispersed. The model accurately fitted the experimental data of veins, while not improving the quality of the fit compared to the model without dispersion. Our results showed that the orientational dispersion of collagen fibres could be compensated by a less abrupt and shifted to higher strain collagen engagement pattern. This should be considered when the model is fitted to experimental data and model parameters are used to study structural modifications of collagen fibre network in physiology and disease.ConclusionsThe presented model incorporates structural features related to waviness and orientational distribution of collagen fibres and thus offers possibilities to better understand the relation between structure and function in the vascular wall. Also, the model can be used to further study mechanically induced collagen remodelling in vascular tissue in health and disease.

Role of elastin anisotropy in structural strain energy functions of arterial tissue

The vascular wall exhibits nonlinear anisotropic mechanical properties. The identification of a strain energy function (SEF) is the preferred method to describe its complex nonlinear elastic properties. Earlier constituent-based SEF models, where elastin is modeled as an isotropic material, failed in describing accurately the tissue response to inflation-extension loading. We hypothesized that these shortcomings are partly due to unaccounted anisotropic properties of elastin. We performed inflation-extension tests on common carotid of rabbits before and after enzymatic degradation of elastin and applied constituent-based SEFs, with both an isotropic and an anisotropic elastin part, on the experimental data. We used transmission electron microscopy (TEM) and serial block-face scanning electron microscopy (SBFSEM) to provide direct structural evidence of the assumed anisotropy. In intact arteries, the SEF including anisotropic elastin with one family of fibers in the circumferential direction fitted better the inflation-extension data than the isotropic SEF. This was supported by TEM and SBFSEM imaging, which showed interlamellar elastin fibers in the circumferential direction. In elastin-degraded arteries, both SEFs succeeded equally well in predicting anisotropic wall behavior. In elastase-treated arteries fitted with the anisotropic SEF for elastin, collagen engaged later than in intact arteries. We conclude that constituent-based models with an anisotropic elastin part characterize more accurately the mechanical properties of the arterial wall when compared to models with simply an isotropic elastin. Microstructural imaging based on electron microscopy techniques provided evidence for elastin anisotropy. Finally, the model suggests a later and less abrupt collagen engagement after elastase treatment.