Compromised homeostasis in aged carotid arteries revealed by microstructural studies of elastic lamellae.

Healthy arteries are continuously subjected to diverse mechanical stimuli and adapt in order to maintain a mechanical homeostasis which is characterized by a uniform distribution of wall stresses. However, aging may compromise the homeostatic microenvironment within arteries. Structural heterogeneity has been suggested as a potential microstructural mechanism that could lead to homogeneous stress distribution across the arterial wall. Our previous study on the unfolding and stretching of the elastic lamellae revealed the underlying microstructural mechanism for equalizing the circumferential stresses through wall; inner elastic layers are wavier and unfold more than the outer layers which helps to evenly distribute lamellar stretching (Yu et al., 2018). In this study, we investigated the effect of aging on lamellar deformation and its implications for tissue homeostasis. Common carotid arteries from aged mice were imaged under a multi-photon microscope while subjected to biaxial extension and inflation at five different pressures ranging from 0 up to 120 mmHg. Lamellar unfolding during pressurization was then determined from the reconstructed cross-sectional images of elastic lamellae. Tissue-level circumferential stretch was combined with the lamellar unfolding to calculate lamellar stretching. Our results revealed that the straightness gradient of aged elastic lamellae is similar to the young ones. However, during pressurization, the inner elastic lamella of the aged mice unfolded significantly more than the inner layer in young arteries. An important finding of our study is the uneven increase in inter-lamellar space which contributed to a nonuniform stretching of the elastic lamellae of aged mice arteries, elevated stress gradient, and a shifting of the load-bearing component to adventitia. Our results shed light into the complex microstructural mechanisms that take place in aging and adversely affect arterial mechanical behavior and homeostasis.

A novel constitutive model considering the role of elastic lamellae' structural heterogeneity in homogenizing transmural stress distribution in arteries.

Understanding how the homeostatic stress state can be reached in arterial tissues can provide new insights into vascular physiology. Even though the function of maintaining homeostasis is often linked to the concentric layers of medial elastic lamellae, how the lamellae are capable of evenly distributing the stress transmurally remains to be understood. The recent microstructural study by Yu et al. (2018 J. R. Soc. Interface 15, 20180492) revealed that, circumferentially, lamellar layers closer to the lumen are wavier than the ones further away from it and, thus, experience more unfolding when subjected to blood pressure. Motivated by this peculiar finding, the current study, for the first time, proposes a novel approach to model elastic lamellae and such structural heterogeneity using the extensible worm-like chain model. When implemented into the material description of the conventional two-layer artery model, in which adventitial collagen is modelled using the inextensible worm-like chain model, it is demonstrated that structural heterogeneity in elastic lamellae plays an important role in dictating transmural stress distribution and, therefore, the homeostasis of the arterial wall.

A classification approach to improve out of sample predictability of structure-based constitutive models for ascending thoracic aortic tissue.

In this research, a pipeline was developed to assess the out-of-sample predictive capability of structure-based constitutive models of ascending aortic aneurysmal tissue. The hypothesis being tested is that a biomarker can help establish similarities among tissues sharing the same level of a quantifiable property, thus enabling the development of biomarker-specific constitutive models. Biomarker-specific averaged material models were constructed from biaxial mechanical tests of specimens that shared similar biomarker properties such as level of blood-wall shear stress or microfiber (elastin or collagen) degradation in the extracellular matrix. Using a cross-validation strategy commonly used in classification algorithms, biomarker-specific averaged material models were assessed in contrast to individual tissue mechanics of out of sample specimens that fell under the same category but did not contribute to the averaged model's generation. The normalized root means square errors (NRMSE) calculated on out-of-sample data were compared with average models when no categorization was performed versus biomarker-specific models and among different level of a biomarker. Different biomarker levels exhibited statistically different NRMSE when compared among each other, indicating more common features shared by the specimens belonging to the lower error groups. However, no specific biomarkers reached a significant difference when compared to the average model created when No Categorization was performed, possibly on account of unbalanced number of specimens. The method developed could allow for the screening of different biomarkers or combinations/interactions in a systematic manner leading the way to larger datasets and to more individualized constitutive approaches.

Magnetic resonance imaging-based hemodynamic wall shear stress alters aortic wall tissue biomechanics in bicuspid aortic valve patients.

OBJECTIVE: In this study we aimed to conclusively determine whether altered aortic biomechanics are associated with wall shear stress (WSS) independent of region of tissue collection. Elevated WSS in the ascending aorta of patients with bicuspid aortic valve has been shown to contribute to local maladaptive aortic remodeling and might alter biomechanics. METHODS: Preoperative 4-dimensional flow magnetic resonance imaging was performed on 22 patients who underwent prophylactic aortic root and/or ascending aorta replacement. Localized elevated WSS was identified in patients using age-matched healthy atlases (n = 60 controls). Tissue samples (n = 78) were collected and categorized according to WSS (elevated vs normal) and region. Samples were subjected to planar biaxial testing. To fully quantify the nonlinear biomechanical response, the tangential modulus (local stiffness) at a low-stretch (LTM) and high-stretch (HTM) linear region and the onset (TZo) and end stress of the nonlinear transition zone were measured. A linear mixed effect models was implemented to determine statistical relationships. RESULTS: A higher LTM in the circumferential and axial direction was associated with elevated WSS (P = .007 and P = .018 respectively) independent of collection region. Circumferential TZo and HTM were higher with elevated WSS (P = .024 and P = .003); whereas the collection region was associated with variations in axial TZo (P = .013), circumferential HTM (P = .015), and axial HTM (P = .001). CONCLUSIONS: This study shows strong evidence that biomechanical changes in the aorta are strongly associated with hemodynamics, and not region of tissue collection for bicuspid valve aortopathy patients. Elevated WSS is associated with tissue behavior at low stretch ranges (ie, LTM and TZo).

Biomechanics in ascending aortic aneurysms correlate with tissue composition and strength.

Objective: This study correlates low strain tangential modulus (LTM) and transition zone onset (TZo) stress, biomechanical parameters that occur within the physiological range of stress seen in vivo, with tissue strength and histopathologic changes in aneurysmal ascending aortic tissue. Method: Ascending aortic aneurysm tissue samples were collected from 41 patients undergoing elective resection. Samples were subjected to planar biaxial testing to quantify LTM and TZo. These were then correlated with strength assessed from uniaxial testing and with histopathologic quantification of pathologic derangements in elastin, collagen, and proteoglycan (PG). Results: Decreased LTM and TZo were correlated with reduced strength (P < .05), PG content (P < .05), and elastin content (P < .05). Reduced TZo also was correlated with increased elastin fragmentation (P < .05). Conclusions: LTM and TZo are correlated with common biomechanical and histopathologic alterations in ascending aortic aneurysm tissue that are thought to relate to the risk of acute aortic syndromes. LTM and TZo are measured under conditions approximating in vivo physiology and have the potential to be obtained noninvasively using medical imaging techniques. Therefore, they represent parameters that warrant future study as potential contributors to our growing knowledge of pathophysiology, disease progression, and risk stratification of aortic disease.

Biomechanical properties of ascending aortic aneurysms: Quantification of inter- and intra-patient variability.

This study investigates the biomechanical properties of ascending aortic aneurysms focusing on the inter-patient differences vs. the heterogeneity within a patient's aneurysm. Each specimen was tested on a biaxial testing device and the resulting stress-strain response was fitted to a four-parameter Fung constitutive model. We postulate that the inter-patient variability (differences between patients) blurs possible intra-patient variability (regional heterogeneity) and, thus, that both effects must be considered to shed light on the role of heterogeneity in aneurysm progression. We propose, demonstrate, and discuss two techniques to assess differences by, first, comparing conventional biomechanical properties and, second, the overall constitutive response. Results show that both inter- and intra-patient variability contribute to errors when using population averaged models to fit individual tissue behaviour. When inter-patient variability was accounted for and its effects excluded, intra-patient heterogeneity could be assessed, showing a wide degree of heterogeneity at the individual patient level. Furthermore, the right lateral region (from the patient's perspective) appeared different (stiffer) than the other regions. We posit that this heterogeneity could be a consequence of maladaptive remodelling due to altered loading conditions that hastens microstructural changes naturally occurring with age. Further validation of these results should be sought from a larger cohort study.

Biomechanics of Wound Healing in an Equine Limb Model: Effect of Location and Treatment with a Peptide-Modified Collagen-Chitosan Hydrogel.

The equine distal limb wound healing model, characterized by delayed re-epithelialization and a fibroproliferative response to wounding similar to that observed in humans, is a valuable tool for the study of biomaterials poised for translation into both the veterinary and human medical markets. In the current study, we developed a novel method of biaxial biomechanical testing to assess the functional outcomes of healed wounds in a modified equine model and discovered significant functional and structural differences in both unwounded and injured skin at different locations on the distal limb that must be considered when using this model in future work. Namely, the medial skin was thicker and displayed earlier collagen engagement, medial wounds experienced a greater proportion of wound contraction during closure, and proximal wounds produced significantly more exuberant granulation tissue. Using this new knowledge of the equine model of aberrant wound healing, we then investigated the effect of a peptide-modified collagen-chitosan hydrogel on wound healing. Here, we found that a single treatment with the QHREDGS (glutamine-histidine-arginine-glutamic acid-aspartic acid-glycine-serine) peptide-modified hydrogel (Q-peptide hydrogel) resulted in a higher rate of wound closure and was able to modulate the biomechanical function toward a more compliant healed tissue without observable negative effects. Thus, we conclude that the use of a Q-peptide hydrogel provides a safe and effective means of improving the rate and quality of wound healing in a large animal model.

Effect of testing conditions on the mechanical response of aortic tissues from planar biaxial experiments: Loading protocol and specimen side.

This study concerns procedural aspects of planar biaxial experiments on aortic tissues that have not been exhaustively addressed in the literature. The following questions are explored. First, is there a difference in the apparent mechanical properties if the experiments are conducted in a force-controlled regime or a displacement-controlled regime. Second, does it matter whether the deformations of the surface are tracked from one side of the tissue or the other (luminal vs. abluminal surface). The study provides answers to these questions with the help of a series of experiments on porcine aortic tissue, constitutive modelling and statistical analysis. It was found that the loading protocol does not substantially affect the constitutive response, while the surface orientation does.

"Hands-On, Minds-On, and Science-Up": A Concept-Based Learning Laboratory With a Taste of Research Experience for an Undergraduate Biomedical Engineering Course.

In this paper, we bridged faculty research expertise with concept-based learning pedagogy to design and implement a unique laboratory experience for biomedical engineering undergraduate students enrolled in the biomechanics of tissues course at the University of Calgary. This laboratory aimed to increase student engagement, facilitate deeper understanding of course content, and provide an opportunity for accelerated undergraduate research through "hands-on," "minds-on," and "science-up" learning components, respectively. The laboratory exercise involves testing aortic tissues using a novel miniaturized planar biaxial machine. This type of machine is normally reserved for use in the context of research. The relevance of the proposed laboratory as a teaching tool was assessed using student feedback. Results indicate an overall valuable and positive learning experience for students.

Multi-sector approximation method for arteries: the residual stresses of circumferential rings with non-trivial openings.

The opening angle method is a popular choice in biomechanics to estimate residual stresses in arteries. Experimentally, it means that an artery is cut into rings; then the rings are cut axially or radially allowing them to open into sectors; finally, the corresponding opening angles are measured to give residual stress levels by solving an inverse problem. However, for many tissues, for example in pathological tissues, the ring does not open according to the theory into a neat single circular sector, but rather creates an asymmetric geometry, often with abruptly changing curvature(s). This phenomenon may be due to a number of reasons including variation in thickness, microstructure, mechanical properties, etc. As a result, these samples are often eliminated from studies relying on the opening angle method, which limits progress in understanding and evaluating residual stresses in real arteries. With this work, we propose an effective approach to deal with these non-trivial openings of rings. First, we digitize pictures of opened rings to split them into multiple, connected circular sectors. Then we measure the corresponding opening angles for each sub-sector. Subsequently, we can determine the residual stresses for individual sectors in a closed-ring configuration and, thus, approximate the circumferential residual bending effects.

Anisotropic residual stresses in arteries.

The paper provides a deepened insight into the role of anisotropy in the analysis of residual stresses in arteries. Residual deformations are modelled following Holzapfel and Ogden (Holzapfel and Ogden 2010, J. R. Soc. Interface 7, 787-799. ( doi:10.1098/rsif.2009.0357 )), which is based on extensive experimental data on human abdominal aortas (Holzapfel et al. 2007, Ann. Biomed. Eng. 35, 530-545. ( doi:10.1007/s10439-006-9252-z )) and accounts for both circumferential and axial residual deformations of the individual layers of arteries-intima, media and adventitia. Each layer exhibits distinctive nonlinear and anisotropic mechanical behaviour originating from its unique microstructure; therefore, we use the most general form of strain-energy function (Holzapfel et al. 2015, J. R. Soc. Interface 12, 20150188. ( doi:10.1098/rsif.2015.0188 )) to derive residual stresses for each layer individually. Finally, the systematic experimental data (Niestrawska et al. 2016, J. R. Soc. Interface 13, 20160620. ( doi:10.1098/rsif.2016.0620 )) on both mechanical and structural properties of the different layers of the human abdominal aorta facilitate our discussion on (i) the importance of anisotropy in modelling residual stresses; (ii) the variability of residual stresses within the same class of tissue, the abdominal aorta; (iii) the limitations of conventional opening angle method to account for complex residual deformations; and (iv) the effect of residual stresses on the loaded configuration of the aorta mimicking in vivo conditions.

Wrinkles and creases in the bending, unbending and eversion of soft sectors.

We study what is clearly one of the most common modes of deformation found in nature, science and engineering, namely the large elastic bending of curved structures, as well as its inverse, unbending, which can be brought beyond complete straightening to turn into eversion. We find that the suggested mathematical solution to these problems always exists and is unique when the solid is modelled as a homogeneous, isotropic, incompressible hyperelastic material with a strain-energy satisfying the strong ellipticity condition. We also provide explicit asymptotic solutions for thin sectors. When the deformations are severe enough, the compressed side of the elastic material may buckle and wrinkles could then develop. We analyse, in detail, the onset of this instability for the Mooney-Rivlin strain energy, which covers the cases of the neo-Hookean model in exact nonlinear elasticity and of third-order elastic materials in weakly nonlinear elasticity. In particular, the associated theoretical and numerical treatment allows us to predict the number and wavelength of the wrinkles. Guided by experimental observations, we finally look at the development of creases, which we simulate through advanced finite-element computations. In some cases, the linearized analysis allows us to predict correctly the number and the wavelength of the creases, which turn out to occur only a few per cent of strain earlier than the wrinkles.