Effects of swelling and anatomical location on the viscoelastic behavior of the porcine urinary bladder wall.

The ability of the urinary bladder to perform its physiological function depends largely on its mechanical characteristics. Understanding the mechanics of this tissue is crucial to the development of accurate models of not just this specific organ, but of the pelvic floor overall. In this study, we tested porcine bladder to identify variations in the tissue's viscoelastic characteristics associated with anatomical locations and swelling. We investigated this relationship using a series of stress-relaxation experiments as well as a modified Maxwell-Wiechert model to aid in the interpretation of the experimental data. Our results highlight that tissue located near the neck of the bladder presents significantly different viscoelastic characteristics than the body of the organ. This supports what was previously observed and is a valuable contribution to the understanding of the location-specific properties of the bladder. We also tested the effect of swelling, revealing that the bladder's viscoelastic behavior is mostly independent of solution osmolarity in hypoosmotic solutions, but the use of a hyperosmotic solution can significantly affect its behavior. This is significant, since several urinary tract pathologies can lead to chronic inflammation and disrupt the urothelial barrier causing increased urothelial permeability, thus subjecting the bladder wall to non-physiologic osmotic challenge.

Quantifying whole bladder biomechanics using the novel pentaplanar reflected image macroscopy system.

Optimal bladder compliance is essential to urinary bladder storage and voiding functions. Calculated as the change in filling volume per change in pressure, bladder compliance is used clinically to characterize changes in bladder wall biomechanical properties that associate with lower urinary tract dysfunction. But because this method calculates compliance without regard to wall structure or wall volume, it gives little insight into the mechanical properties of the bladder wall during filling. Thus, we developed Pentaplanar Reflected Image Macroscopy (PRIM): a novel ex vivo imaging method to accurately calculate bladder wall stress and stretch in real time during bladder filling. The PRIM system simultaneously records intravesical pressure, infused volume, and an image of the bladder in five distinct visual planes. Wall thickness and volume were then measured and used to calculate stress and stretch during filling. As predicted, wall stress was nonlinear; only when intravesical pressure exceeded ~ 15 mmHg did bladder wall stress rapidly increase with respect to stretch. This method of calculating compliance as stress vs stretch also showed that the mechanical properties of the bladder wall remain similar in bladders of varying capacity. This study demonstrates how wall tension, stress and stretch can be measured, quantified, and used to accurately define bladder wall biomechanics in terms of actual material properties and not pressure/volume changes. This method is especially useful for determining how changes in bladder biomechanics are altered in pathologies where profound bladder wall remodeling occurs, such as diabetes and spinal cord injury.

Compound 48/80 increases murine bladder wall compliance independent of mast cells.

A balance between stiffness and compliance is essential to normal bladder function, and changes in the mechanical properties of the bladder wall occur in many bladder pathologies. These changes are often associated with the release of basic secretagogues that in turn drive the release of inflammatory mediators from mast cells. Mast cell degranulation by basic secretagogues is thought to occur by activating an orphan receptor, Mas-related G protein-coupled receptor B2 (Mrgprb2). We explored the effects of the putative mast cell degranulator and Mrgprb2 agonist Compound 48/80 on urinary bladder wall mechanical compliance, smooth muscle contractility, and urodynamics, and if these effects were mast cell dependent. In wild-type mice, Mrgprb2 receptor mRNA was expressed in both the urothelium and smooth muscle layers. Intravesical instillation of Compound 48/80 decreased intermicturition interval and void volume, indicative of bladder overactivity. Compound 48/80 also increased bladder compliance while simultaneously increasing the amplitude and leading slope of transient pressure events during ex vivo filling and these effects were inhibited by the Mrgprb2 antagonist QWF. Surprisingly, all effects of Compound 48/80 persisted in mast cell-deficient mice, suggesting these effects were independent of mast cells. These findings suggest that Compound 48/80 degrades extracellular matrix and increases urinary bladder smooth muscle excitability through activation of Mrgprb2 receptors located outside of mast cells. Thus, the pharmacology and physiology of Mrgprb2 in the urinary bladder is of potential interest and importance in terms of treating lower urinary tract dysfunction.

Store-operated calcium entry: From physiology to tubular aggregate myopathy.

Store-Operated Ca2+ entry (SOCE) is recognized as a key mechanism in muscle physiology necessary to refill intracellular Ca2+ stores during sustained muscle activity. For many years the cell structures expected to mediate SOCE in skeletal muscle fibres remained unknown. Recently, the identification of Ca2+ Entry Units (CEUs) in exercised muscle fibres opened new insights into the role of extracellular Ca2+ in muscle contraction and, more generally, in intracellular Ca2+ homeostasis. Accordingly, intracellular Ca2+ unbalance due to alterations in SOCE strictly correlates with muscle disfunction and disease. Mutations in proteins involved in SOCE (STIM1, ORAI1, and CASQ1) have been linked to tubular aggregate myopathy (TAM), a disease that causes muscle weakness and myalgia and is characterized by a typical accumulation of highly ordered and packed membrane tubules originated from the sarcoplasmic reticulum (SR). Achieving a full understanding of the molecular pathways activated by alterations in Ca2+ entry mechanisms is a necessary step to design effective therapies for human SOCE-related disorders.

Increased extracellular matrix stiffness accompanies compromised bladder function in a murine model of radiation cystitis.

Radiation cystitis, a long-term bladder defect due to pelvic radiation therapy, results in lower urinary tract symptoms, such as urinary frequency and nocturia, suggestive of compromised bladder compliance. The goal of this study was to identify alterations to the mechanical behavior of the urinary bladder extracellular matrix of a murine model of radiation cystitis, at 3 and 6 months after radiation exposure. The results of this study demonstrated that the extracellular matrix of irradiated bladders was significantly less distensible when compared to age matching controls. These findings coincided with functional bladder changes, including increased number of voids and decreased voided volume. Both mechanical and functional changes were apparent at 3 months post-irradiation and were statistically significant at 6 months, demonstrating the progressive nature of radiation cystitis. Overall, the results of this study indicate that irradiation exposure changes both the mechanical and physiological properties of the bladder. STATEMENT OF SIGNIFICANCE: In humans, radiation cystitis results in lower urinary tract symptoms, such as urinary frequency and nocturia, suggestive of compromised bladder compliance. This pathology can significantly affect recovery and quality of life for cancer survivors. Gaining knowledge about how alterations to the mechanical behavior of the urinary bladder extracellular matrix can affect urinary function will have a significant impact on this population. The results of this study demonstrated that the extracellular matrix of irradiated bladders was significantly less distensible when compared to age matching controls, in a mouse model of radiation cystitis. These findings were accompanied by functional voiding changes, including increased number of voids and decreased voided volume. The results of this study uncovered that irradiation exposure changes the mechanical and physiological properties of the bladder.

Aortic stiffness is lower when PVAT is included: a novel ex vivo mechanics study.

Perivascular adipose tissue (PVAT) is increasingly recognized as an essential layer of the functional vasculature, being responsible for producing vasoactive substances and assisting arterial stress relaxation. Here, we test the hypothesis that PVAT reduces aortic stiffness. Our model was the thoracic aorta of the male Sprague-Dawley rat. Uniaxial mechanical tests for three groups of tissue were performed: aorta with PVAT attached (+PVAT) or removed (-PVAT), and isolated PVAT (PVAT only). The output of the mechanical test is reported in the form of a Cauchy stress-stretch curve. This work presents a novel, physiologically relevant approach to measure mechanical stiffness ex vivo in isolated PVAT. Low-stress stiffness (E0), high-stress stiffness (E1), and the stress corresponding to a stretch of 1.2 (σ1.2) were measured as metrics of distensibility. The low-stress stiffness was largest in the -PVAT samples and smallest in PVAT only samples. Both the high-stress stiffness and the stress at 1.2 stretch were significantly higher in -PVAT samples when compared with +PVAT samples. Taken together, these results suggest that -PVAT samples are stiffer (less distensible) both at low stress (not significant) as well as at high stress (significant) when compared with +PVAT samples. These conclusions are supported by the results of the continuum mechanics material model that we also used to interpret the same experimental data. Thus, tissue stiffness is significantly lower when considering PVAT as part of the aortic wall. As such, PVAT should be considered as a target for improving vascular function in diseases with elevated aortic stiffness, including hypertension.NEW & NOTEWORTHY We introduce a novel and physiologically relevant way of measuring perivascular adipose tissue (PVAT) mechanical stiffness which shows that PVAT's low, yet measurable, stiffness is linearly correlated with the amount of collagen fibers present within the tissue. Including PVAT in the measurement of the aortic wall's mechanical behavior is important, and it significantly affects the resulting metrics by decreasing aortic stiffness.

Remodeling of extracellular matrix in the urinary bladder of paraplegic rats results in increased compliance and delayed fiber recruitment 16 weeks after spinal cord injury.

The ability of the urinary bladder to maintain low intravesical pressures while storing urine is key in ensuring proper organ function and highlights the key role that tissue mechanics plays in the lower urinary tract. Loss of supraspinal neuronal connections to the bladder after spinal cord injury can lead to remodeling of the structure of the bladder wall, which may alter its mechanical characteristics. In this study, we investigate if the morphology and mechanical properties of the bladder extracellular matrix are altered in rats 16 weeks after spinal cord injury as compared to animals who underwent sham surgery. We measured and quantified the changes in bladder geometry and mechanical behavior using histological analysis, tensile testing, and constitutive modeling. Our results suggest bladder compliance is increased in paraplegic animals 16 weeks post-injury. Furthermore, constitutive modeling showed that increased distensibility was driven by an increase in collagen fiber waviness, which altered the distribution of fiber recruitment during loading. STATEMENT OF SIGNIFICANCE: The ability of the urinary bladder to store urine under low pressure is key in ensuring proper organ function. This highlights the important role that mechanics plays in the lower urinary tract. Loss of control of neurologic connection to the bladder from spinal cord injury can lead to changes of the structure of the bladder wall, resulting in altered mechanical characteristics. We found that the bladder wall's microstructure in rats 16 weeks after spinal cord injury is more compliant than in healthy animals. This is significant since it is the longest time post-injury analyzed, to date. Understanding the extreme remodeling capabilities of the bladder in pathological conditions is key to inform new possible therapies.

Comparative Study of Human and Murine Aortic Biomechanics and Hemodynamics in Vascular Aging.

Introduction: Aging has many effects on the cardiovascular system, including changes in structure (aortic composition, and thus stiffening) and function (increased proximal blood pressure, and thus cardiac afterload). Mouse models are often used to gain insight into vascular aging and mechanisms of disease as they allow invasive assessments that are impractical in humans. Translation of results from murine models to humans can be limited, however, due to species-specific anatomical, biomechanical, and hemodynamic differences. In this study, we built fluid-solid-interaction (FSI) models of the aorta, informed by biomechanical and imaging data, to compare wall mechanics and hemodynamics in humans and mice at two equivalent ages: young and older adults. Methods: For the humans, 3-D computational models were created using wall property data from the literature as well as patient-specific magnetic resonance imaging (MRI) and non-invasive hemodynamic data; for the mice, comparable models were created using population-based properties and hemodynamics as well as subject-specific anatomies. Global aortic hemodynamics and wall stiffness were compared between humans and mice across age groups. Results: For young adult subjects, we found differences between species in pulse pressure amplification, compliance and resistance distribution, and aortic stiffness gradient. We also found differences in response to aging between species. Generally, the human spatial gradients of stiffness and pulse pressure across the aorta diminished with age, while they increased for the mice. Conclusion: These results highlight key differences in vascular aging between human and mice, and it is important to acknowledge these when using mouse models for cardiovascular research.

Investigation of Fiber-Driven Mechanical Behavior of Human and Porcine Bladder Tissue Tested Under Identical Conditions.

The urinary bladder is a highly dynamic organ that undergoes large deformations several times per day. Mechanical characteristics of the tissue are crucial in determining the function and dysfunction of the organ. Yet, literature reporting on the mechanical properties of human bladder tissue is scarce and, at times, contradictory. In this study, we focused on mechanically testing tissue from both human and pig bladders using identical protocols to validate the use of pigs as a model for the human bladder. Furthermore, we tested the effect of two treatments on tissue mechanical properties. Namely, elastase to digest elastin fibers, and oxybutynin to reduce smooth muscle cell spasticity. Additionally, mechanical properties based on the anatomical direction of testing were evaluated. We implemented two different material models to aid in the interpretation of the experimental results. We found that human tissue behaves similarly to pig tissue at high deformations (collagen-dominated behavior) while we detected differences between the species at low deformations (amorphous matrix-dominated behavior). Our results also suggest that elastin could play a role in determining the behavior of the fiber network. Finally, we confirmed the anisotropy of the tissue, which reached higher stresses in the transverse direction when compared to the longitudinal direction.

Potential damage in pulmonary arterial hypertension: An experimental study of pressure-induced damage of pulmonary artery.

Pulmonary arterial hypertension (PAH) is associated with elevated pulmonary arterial pressure. PAH prognosis remains poor with a 15% mortality rate within 1 year, even with modern clinical management. Previous clinical studies proposed wall shear stress (WSS) to be an important hemodynamic factor affecting cell mechanotransduction, growth and remodeling, and disease progress in PAH. However, WSS in vivo is typically at most 2.5 Pa and a doubt has been cast whether WSS alone can drive disease progress. Furthermore, our current understanding of PAH pathology largely comes from small animals' studies in which caliber enlargement, a hallmark of PAH in humans, is rarely reported. Therefore, a large-animal experiment on pulmonary arteries (PAs) is needed to validate whether increased pressure can induce enlargement of PAs caliber. In this study, we use an inflation testing device to characterize the mechanical behavior, both nonlinear elastic behavior and irreversible damage of porcine arteries. The parameters of elastic behavior are estimated from the inflation test at a low-pressure range before and after over-pressurization. Then, histological images are qualitatively examined for medial and adventitial layers. This study sheds light on the relevance of pressure-induced damage mechanism in human PAH.

The effects of body position on the material properties of soft tissue in the human thigh.

Forty percent of patients with a spinal cord injury acquire a pressure ulcer during rehabilitation, and sixty percent of individuals in elderly care facilities have at least one pressure ulcer upon admittance. A commonality between those populations is the increased amount of time they spend in the seated position. The loading on the buttocks and thighs while in the seated position has been cited as a risk factor for pressure ulcer formation, especially for wheelchair users. Finite element models provide a tool with which to evaluate the internal tissue stresses, but they are reliant upon accurate material properties for the soft tissue. Thus the goals of this research were to determine and compare the material properties of the soft tissue in the thigh and buttock regions in the seated, quadruped (a universally accessible position with the knee and hip articulations similar to the seated position), and prone positions. A custom indenter was designed to collect force and deflection data for the buttocks/proximal thigh, middle thigh, and distal thigh regions of twenty able-bodied individuals. The force and deflection data were converted into stress and stretch data, which were used to obtain parameters from an Ogden material model. Our results indicated that the prone position yielded significantly stiffer tissue properties than in the seated and quadruped positions for both males and females, meaning that position should be taken into account when obtaining material properties that are input into finite element models. Realistic material properties of the soft tissue will lead to better understanding of tissue injury risk.

A New Function for Perivascular Adipose Tissue (PVAT): Assistance of Arterial Stress Relaxation.

In health, PVAT secretes anti-contractile factors that relax the underlying artery. PVAT's contributions to vascular function include more than production of vasoactive substances. We hypothesized that PVAT benefits the artery by assisting the function of stress (-induced) relaxation. Thoracic aorta rings from Sprague Dawley rats were mounted in isolated tissue baths with (+) and without (-) PVAT. A cumulative length tension (0-6 grams) was generated. The tension to which the tissue stress relaxed over 30 minutes was recorded; the tension lost was stress relaxation. The presence of PVAT increased the amount of stress relaxation (final tension in mgs; aortic ring -PVAT = 4578 ± 190; aortic ring + PVAT = 2730 ± 274, p < 0.05). PVAT left attached but not encompassing the aorta provided no benefit in cumulative stress relaxation (aortic ring +/- PVAT = 4122 ± 176; p > 0.05 vs -PVAT). A PVAT ring separated from the aorta demonstrated more profound stress relaxation than did the aortic ring itself. Finally, PVAT-assisted stress relaxation was observed in an artery with white fat (superior mesenteric artery) and in aorta from both male and female of another rat strain, the Dahl S rat. Knowledge of this new PVAT function supports PVAT as an essential player in vascular health.

Assessment of time to completion, number of errors, and knot-holding capacity of square knots and Aberdeen knots tied by veterinary students and student perceptions of knot security and knot-tying difficulty.

OBJECTIVE: To assess the time to completion, number of errors, and knot-holding capacity (KHC) for starting and ending square knots (SSKs and ESKs) of a continuous pattern and Aberdeen knots tied by veterinary students and to investigate student perceptions of knot security and knot-tying difficulty for the 3 knot types. SAMPLE: 16 second-year veterinary students. PROCEDURES: Students created 3 (4-throw) SSKs, 3 (5-throw) ESKs, and 3 (3 + 1 configuration) Aberdeen knots with 2-0 polydioxanone on a custom test apparatus. Time to complete each knot, the number of errors in each knot, and student ratings of knot-tying difficulty and confidence in knot security were recorded. Each knot was tested to failure on a uniaxial tensiometer to determine KHC and mode of failure. Variables of interest were compared by repeated-measures ANOVA or the Friedman test with post hoc pairwise comparisons. RESULTS: Mean knot completion time for Aberdeen knots was significantly less than mean completion time for SSKs or ESKs. Mean KHC was significantly lower for ESKs than for SSKs; KHC for Aberdeen knots was not compared with these values because of methodological differences. Median error rate was higher for ESKs than for other knot types. Mean difficulty rating for Aberdeen knots was lower than that for ESKs. Most tested knots failed by breakage at the knot. CONCLUSIONS AND CLINICAL RELEVANCE: Aberdeen knots appeared to be easy for veterinary students to learn and were completed more rapidly and with fewer errors than ESKs. Including this type of knot in surgical skills curriculum for novices may be beneficial.

Mechanical behavior of SiNC layers on PDMS: effects of layer thickness, PDMS modulus, and SiNC surface functionality.

Thin layers of nanomaterials on stretchable substrates have the potential to enable stretchable, bendable optoelectronic devices, wearable diagnostics, and more. Recently, our group reported on a novel method for finding the neo-Hookean coefficient of thin layers of silicon nanocrystals (SiNCs) on polydimethylsiloxane (PDMS). Here we elaborate on that initial study by examining the effects of the SiNC layer thickness, PDMS neo-Hookean coefficient, and SiNC surface functionality on the neo-Hookean coefficient of the SiNC layers. We found that, while the layer thickness and PDMS neo-Hookean coefficient influence the behavior of the SiNC layers, layers of surface-functionalized SiNCs do not exhibit disparate behavior from layers of bare SiNCs.

Inverse finite element characterization of the human thigh soft tissue in the seated position.

Pressure ulcers are localized damage to the skin and underlying tissues caused by sitting or lying in one position for a long time. Stresses within the soft tissue of the thigh and buttocks area play a crucial role in the initiating mechanism of these wounds. Therefore, it is crucial to develop reliable finite element models to evaluate the stresses caused by physiological loadings. In this study, we compared how the choice of material model and modeling area dimension affect prediction accuracy of a model of the thigh. We showed that the first-order Ogden and Fung orthotropic material models could approximate the mechanical behavior of soft tissue significantly better than neo-Hookean and Mooney-Rivlin. We also showed that, significant error results from using a semi-3D model versus a 3D model. We then developed full 3D models for 20 participants employing Ogden and Fung material models and compared the estimated material parameters between different sexes and locations along the thigh. We showed that males tissues are less deformable overall when compared to females and the material parameters are highly dependent on location, with tissues getting softer moving distally for both men and women.

A microstructurally inspired constitutive model for skin mechanics.

This study investigates the link between the mechanical properties of skin and its microstructural characteristics. Rat back skin samples from different locations, orientations, and sexes were collected and subjected to quasi-static uniaxial tensile tests. Stress-stretch behavior at low stress ranges and rupture data at high stress ranges were collected. The influence of location, orientation, and sex on skin mechanical properties was examined by comparing the mechanical parameters (i.e., initial slope, maximum slope, ultimate tensile strength, rupture stretch, and toughness) evaluated from the tensile testing data. Location and orientation were both found to have a significant effect on the mechanical properties. Collagen structural data (i.e., fiber orientation distribution, relative content, and fiber straightness) were evaluated using histology images. It was found that the rat lower (caudal) back had higher relative collagen content when compared to the upper (cranial) back. A microstructurally based constitutive model was proposed to describe the mechanical behavior of preconditioned rat back skin. The constitutive model incorporated the distribution of collagen fiber bundle orientations and relative collagen content measured from histology, and showed good agreement with the tensile test data. The influence of location and orientation was also evident in the optimized constitutive parameters. This study was a comprehensive investigation that combines skin mechanical behavior, micro-structure, and constitutive modeling.

Influences of sodium and glycosaminoglycans on skin oedema and the potential for ulceration: a finite-element approach.

Venous ulcers are chronic transcutaneous wounds common in the lower legs. They are resistant to healing and have a 78% chance of recurrence within 2 years. It is commonly accepted that venous ulcers are caused by the insufficiency of the calf muscle pump, leading to blood pooling in the lower legs, resulting in inflammation, skin oedema, tissue necrosis and eventually skin ulceration. However, the detailed physiological events by which inflammation contributes to wound formation are poorly understood. We therefore sought to develop a model that simulated the inflammation, using it to determine the internal stresses and pressure on the skin that contribute to venous ulcer formation. A three-layer finite-element skin model (epidermis, dermis and hypodermis) was developed to explore the roles in wound formation of two inflammation identifiers: glycosaminoglycans (GAG) and sodium. A series of parametric studies showed that increased GAG and sodium content led to oedema and increased tissue stresses of 1.5 MPa, which was within the reported range of skin tissue ultimate tensile stress (0.1-40 MPa). These results suggested that both the oedema and increased fluid pressure could reach a threshold for tissue damage and eventual ulcer formation. The models presented here provide insights to the pathological events associated with venous insufficiency, including inflammation, oedema and skin ulceration.

A novel approach to finding mechanical properties of nanocrystal layers.

Flexible, bendable, stretchable devices represent the future of electronics for a wide range of real-world applications. Due to the fact that these technologies deviate significantly from traditional wafer technologies there is a need to understand and engineer material systems that allow large elastic deformations present in such devices, which requires knowledge about the mechanical properties of these material systems. Here we evaluate the mechanical properties of a bilayer polydimethylsiloxane (PDMS)/silicon nanocrystal system. By observing the formation of instabilities due to finite bending deformation and applying theoretical modeling, we estimated the neo-Hookean coefficient (analogous to shear modulus at low stress/strain) of the silicon nanocrystal film to be 345 ± 23 kPa. The method used here represents a novel approach to evaluating these properties and is widely applicable to many different combinations of systems of nanocrystals and elastomers.

Contribution of left ventricular residual stress by myocytes and collagen: existence of inter-constituent mechanical interaction.

We quantify the contribution of myocytes, collagen fibers and their interactions to the residual stress field found in the left ventricle (LV) using both experimental and theoretical methods. Ring tissue samples extracted from normal rat, male and female, LV were treated with collagenase and decellularization to isolate myocytes and collagen fibers, respectively. Opening angle tests were then performed on these samples as well as intact tissue samples containing both constituents that served as control. Our results show that the collagen fibers are the main contributor to the residual stress fields found in the LV. Specifically, opening angle measured in collagen-only samples (106.45[Formula: see text] ± 23.02[Formula: see text]) and myocytes-only samples (21.00[Formula: see text] ± 4.37[Formula: see text]) was significantly higher and lower than that of the control (57.88[Formula: see text] ± 12.29[Formula: see text]), respectively. A constrained mixture (CM) modeling framework was then used to infer these experimental results. We show that the framework cannot reproduce the opening angle found in the intact tissue with measurements made on the collagen-only and myocytes-only samples. Given that the CM framework assumes that each constituent contributes to the overall mechanics simply by their mere presence, this result suggests the existence of some myocyte-collagen mechanical interaction that cannot be ignored in the LV. We then propose an extended CM formulation that takes into account of the inter-constituent mechanical interaction in which constituents are deformed additionally when they are physically combined into a mixture. We show that the intact tissue opening angle can be recovered in this framework.