Size and proximity of micro-scale hard-inclusions increase the risk of rupture in fibroatheroma-like laboratory models.

Increased mechanical stresses of the fibroatheroma cap tissue is a crucial risk factor on the pathogenesis of asymptomatic coronary artery disease events. Moreover, both numerical and analytical studies have shown that microcalcifications (µCalcs) located in the fibrous cap can multiply the cap tissue stress by a factor of 2-7. This stress amplification depends on the ratio of the gap between particles (h) and their diameter (D) when they are aligned along the tensile axis. However, the synergistic effect of cap stiffness and uCalcs on the ultimate stress and rupture risk of the atheroma cap has not been fully investigated. In this context, we studied the impact of micro-beads (µBeads) of varying diameters and concentration on the rupture of silicone-based laboratory models mimicking human fibroatheroma caps of different stiffness (shear moduli µsoft = 40 kPa, µstiff = 400 kPa) and thickness (650 µm and 100 µm). A total of 145 samples were tested under uniaxial tension up to failure and the true stress and strain response of each model was derived by means of Digital Image Correlation (DIC). Before testing, samples were scanned using high-resolution Micro-CT, to perform morphometry analyses of the embedded micro-beads and determine the number of closely spaced particles (h/D<0.5). The micro-beads structural and spatial features were then compared to the case of 29 non-ruptured human atheroma fibrous caps presenting µCalcs. Samples with and without µBeads exhibited a distinct hyperelastic behavior typical of arterial tissues. Regardless of the sample stiffness, large µBeads (>80 µm) significantly reduced the ultimate tensile stress (UTS) of the thick cap models with the effect being more pronounced as the particle diameter increases. Stiff models experienced early rupture in the presence of µBeads with 40 µm diameter. Smaller µBeads of 6 µm and 20 µm didn't affect the ultimate strength of the thick cap models. However, when 6 µm µBeads where introduced in thinner cap models, we observed more than 20% drop in UTS. Increasing the µBeads concentration was also positively correlated with lower stresses at rupture as more clusters formed resulting in lower values of h/D. Morphometry analyses of cap models and human atheroma show that the 6 µm µBeads groups present very similar size distributions to µCalcs and that human µCalcs occupy an average volume ratio of 0.79 ± 0.85%. Our results clearly capture the influence of µBeads on the rupture threshold of a vascular tissue mimicking material. This effect appears to be dependent on the µBeads-to-cap thickness size ratio as well as their proximity. These findings support previous numerical and analytical studies suggesting that µCalcs located within the fibroatheroma cap may be responsible for significantly increasing the risk of cap rupture that precedes myocardial infarction and sudden death.

The effect of plaque morphology, material composition and microcalcifications on the risk of cap rupture: A structural analysis of vulnerable atherosclerotic plaques.

Background: The mechanical rupture of an atheroma cap may initiate a thrombus formation, followed by an acute coronary event and death. Several morphology and tissue composition factors have been identified to play a role on the mechanical stability of an atheroma, including cap thickness, lipid core stiffness, remodeling index, and blood pressure. More recently, the presence of microcalcifications (µCalcs) in the atheroma cap has been demonstrated, but their combined effect with other vulnerability factors has not been fully investigated. Materials and methods: We performed numerical simulations on 3D idealized lesions and a microCT-derived human coronary atheroma, to quantitatively analyze the atheroma cap rupture. From the predicted cap stresses, we defined a biomechanics-based vulnerability index (VI) to classify the impact of each risk factor on plaque stability, and developed a predictive model based on their synergistic effect. Results: Plaques with low remodeling index and soft lipid cores exhibit higher VI and can shift the location of maximal wall stresses. The VI exponentially rises as the cap becomes thinner, while the presence of a µCalc causes an additional 2.5-fold increase in vulnerability for a spherical inclusion. The human coronary atheroma model had a stable phenotype, but it was transformed into a vulnerable plaque after introducing a single spherical µCalc in its cap. Overall, cap thickness and µCalcs are the two most influential factors of mechanical rupture risk. Conclusions: Our findings provide supporting evidence that high risk lesions are non-obstructive plaques with softer (lipid-rich) cores and a thin cap with µCalcs. However, stable plaques may still rupture in the presence of µCalcs.

Biaxial testing system for characterization of mechanical and rupture properties of small samples.

The study of damage and rupture of soft tissues using a tensile testing system is essential to understand the limits of mechanical behavior and loss of function in diseased tissues. However, commercial material testing systems are often expensive and may not be fully suitable for rupture tests of small samples. While several research laboratories have developed custom, less expensive, uniaxial or biaxial devices, there is a need for an open source, inexpensive, accurate and easy to customize biaxial material testing system to perform rupture tests in small soft samples. We designed a testing system (BiMaTS) that (a) was shown able to perform uniaxial and biaxial tests, (b) offers a large travel range for rupture tests of small samples, (c) maintains a centered field of view for effective strain mapping using digital image correlation, (d) provides a controlled temperature environment, (e) utilize many off-the-shelve components for easy manufacture and customization, and it is cost effective (∼$15 K). The instrument performance was characterized using 80%-scaled down, ASTM D412-C shaped PDMS samples. Our results demonstrate the ability of this open source, customizable, low-cost, biaxial materials testing system to successfully characterize the mechanical and rupture properties of small samples with high repeatability and accuracy.

Tunable elastomer materials with vascular tissue-like rupture mechanics behavior.

Purpose. Laboratory models of human arterial tissues are advantageous to examine the mechanical response of blood vessels in a simplified and controllable manner. In the present study, we investigated three silicone-based materials for replicating the mechanical properties of human arteries documented in the literature.Methods. We performed uniaxial tensile tests up to rupture on Sylgard184, Sylgard170 and DowsilEE-3200 under different curing conditions and obtained their True (Cauchy) stress-strain behavior and Poisson's ratios by means of digital image correlation (DIC). For each formulation, we derived the constitutive parameters of the 3-term Ogden model and designed numerical simulations of tubular models under a radial pressure of 250 mmHg.Results. Each material exhibits evident non-linear hyperelasticity and dependence on the curing condition. Sylgard184 is the stiffest formulation, with the highest shear moduli and ultimate stresses at relative low strains (µ184 = 0.52-0.88 MPa,σ184 = 15.90-16.54 MPa,ε184 = 0.72-0.96). Conversely, Sylgard170 and DowsilEE-3200 present significantly lower shear moduli and ultimate stresses that are closer to data reported for arterial tissues (µ170 = 0.33-0.7 MPaσ170 = 2.61-3.67 MPa,ε170 = 0.69-0.81;µdow= 0.02-0.09 MPaσdow= 0.83-2.05 MPa,εdow= 0.91-1.05). Under radial pressure, all formulations except DowsilEE-3200 at 1:1 curing ratio undergo circumferential stresses that remain in the elastic region with values ranging from 0.1 to 0.18 MPa.Conclusion. Sylgard170 and DowsilEE-3200 appear to better reproduce the rupture behavior of vascular tissues within their typical ultimate stress and strain range. Numerical models demonstrate that all three materials achieve circumferential stresses similar to human common carotid arteries (Sommeret al2010), making these formulations suited for cylindrical laboratory models under physiological and supraphysiological loading.

Increased cochlear otic capsule thickness and intracortical canal porosity in the oim mouse model of osteogenesis imperfecta.

Osteogenesis imperfecta (OI or brittle bone disease) is a group of genetic disorders of the connective tissues caused mainly by mutations in the genes encoding collagen type I. Clinical manifestations of OI include skeletal fragility, bone deformities, and severe functional disabilities, such as hearing loss. Progressive hearing loss, usually beginning in childhood, affects approximately 70% of people with OI with more than half of the cases involving the inner ear. There is no cure for OI nor a treatment to ameliorate its corresponding hearing loss, and very little is known about the properties of OI ears. In this study, we investigate the morphology of the otic capsule and the cochlea in the inner ear of the oim mouse model of OI. High-resolution 3D images of 8-week old oim and WT inner ears were acquired using synchrotron microtomography. Volumetric morphometric measurements were conducted for the otic capsule, its intracortical canal network and osteocyte lacunae, and for the cochlear spiral ducts. Our results show that the morphology of the cochlea is preserved in the oim ears at 8 weeks of age but the otic capsule has a greater cortical thickness and altered intracortical bone porosity, with a larger number and volume density of highly branched canals in the oim otic capsule. These results portray a state of compromised bone quality in the otic capsule of the oim mice that may contribute to their hearing loss.

Stenting-induced Vasa Vasorum compression and subsequent flow resistance: a finite element study.

Vascular stenting is a common intervention for the treatment for atherosclerotic plaques. However, stenting still has a significant rate of restenosis caused by intimal hyperplasia formation. In this study, we evaluate whether stent overexpansion leads to Vasa Vasorum (VV) compression, which may contribute to vascular wall hypoxia and restenosis. An idealized multilayered fibroatheroma model including Vasa Vasorum was expanded by three coronary stent designs up to a 1.3:1 stent/artery luminal diameter ratio (exp1.1, exp1.2, exp1.3) using a finite element analysis approach. Following Poiseuille's law for elliptical sections, the fold increase in flow resistance was calculated based on VV compression in the Intima (Int), Media (Med) and Adventitia (Adv). The VV beneath the plaque experiences the smallest degree of compression, while the opposite wall regions are highly affected by stent overexpansion. The highest compressions for Adv, Med and Int at exp1.1 are 60.7, 65.9, 72.3%, at exp1.2 are 62.1, 67.3, 73.5% and at expp1.3 are 63.2, 68.7, 74.8%. The consequent fold increase in resistance to flow for Adv, Med and Int at exp1.1 is 3.3, 4.4, 6.6, at exp1.2 is 3.5, 4.7, 7.2 and at exp1.3 is 3.8, 5.1, 7.9. Stent overexpansion induces significant VV compression, especially in the Intima and Media layers, in agreement with previously observed Media necrosis and loss in elasticity after stenting. The observed steep increase in flow resistance suggests the blood flow and associated oxygen delivery would drop up to five times in the Media and almost eight in the Intima, which may lead to intimal hyperplasia and restenosis.

Defining the relationship between maternal care behavior and sensory development in Wistar rats: Auditory periphery development, eye opening and brain gene expression.

Defining the relationship between maternal care, sensory development and brain gene expression in neonates is important to understand the impact of environmental challenges during sensitive periods in early life. In this study, we used a selection approach to test the hypothesis that variation in maternal licking and grooming (LG) during the first week of life influences sensory development in Wistar rat pups. We tracked the onset of the auditory brainstem response (ABR), the timing of eye opening (EO), middle ear development with micro-CT X-ray tomography, and used qRT-PCR to monitor changes in gene expression of the hypoxia-sensitive pathway and neurotrophin signaling in pups reared by low-LG or high-LG dams. The results show the first evidence that the transcription of genes involved in the hypoxia-sensitive pathway and neurotrophin signaling is regulated during separate sensitive periods that occur before and after hearing onset, respectively. Although the timing of ABR onset, EO, and the relative mRNA levels of genes involved in the hypoxia-sensitive pathway did not differ between pups from different LG groups, we found statistically significant increases in the relative mRNA levels of four genes involved in neurotrophin signaling in auditory brain regions from pups of different LG backgrounds. These results suggest that sensitivity to hypoxic challenge might be widespread in the auditory system of neonate rats before hearing onset, and that maternal LG may affect the transcription of genes involved in experience-dependent neuroplasticity.

Analytical and numerical modeling of the hearing system: Advances towards the assessment of hearing damage.

Hearing is an extremely complex phenomenon, involving a large number of interrelated variables that are difficult to measure in vivo. In order to investigate such process under simplified and well-controlled conditions, models of sound transmission have been developed through many decades of research. The value of modeling the hearing system is not only to explain the normal function of the hearing system and account for experimental and clinical observations, but to simulate a variety of pathological conditions that lead to hearing damage and hearing loss, as well as for development of auditory implants, effective ear protections and auditory hazard countermeasures. In this paper, we provide a review of the strategies used to model the auditory function of the external, middle, inner ear, and the micromechanics of the organ of Corti, along with some of the key results obtained from such modeling efforts. Recent analytical and numerical approaches have incorporated the nonlinear behavior of some parameters and structures into their models. Few models of the integrated hearing system exist; in particular, we describe the evolution of the Auditory Hazard Assessment Algorithm for Human (AHAAH) model, used for prediction of hearing damage due to high intensity sound pressure. Unlike the AHAAH model, 3D finite element models of the entire hearing system are not able yet to predict auditory risk and threshold shifts. It is expected that both AHAAH and FE models will evolve towards a more accurate assessment of threshold shifts and hearing loss under a variety of stimuli conditions and pathologies.

Human cochlear hydrodynamics: A high-resolution µCT-based finite element study.

Measurements of perilymph hydrodynamics in the human cochlea are scarce, being mostly limited to the fluid pressure at the basal or apical turn of the scalae vestibuli and tympani. Indeed, measurements of fluid pressure or volumetric flow rate have only been reported in animal models. In this study we imaged the human ear at 6.7 and 3-µm resolution using µCT scanning to produce highly accurate 3D models of the entire ear and particularly the cochlea scalae. We used a contrast agent to better distinguish soft from hard tissues, including the auditory canal, tympanic membrane, malleus, incus, stapes, ligaments, oval and round window, scalae vestibule and tympani. Using a Computational Fluid Dynamics (CFD) approach and this anatomically correct 3D model of the human cochlea, we examined the pressure and perilymph flow velocity as a function of location, time and frequency within the auditory range. Perimeter, surface, hydraulic diameter, Womersley and Reynolds numbers were computed every 45° of rotation around the central axis of the cochlear spiral. CFD results showed both spatial and temporal pressure gradients along the cochlea. Small Reynolds number and large Womersley values indicate that the perilymph fluid flow at auditory frequencies is laminar and its velocity profile is plug-like. The pressure was found 102-106° out of phase with the fluid flow velocity at the scalae vestibule and tympani, respectively. The average flow velocity was found in the sub-µm/s to nm/s range at 20-100Hz, and below the nm/s range at 1-20kHz.

Posttraumatic Malalignment of the Humeral Shaft: Challenging the Existing Paradigm.

OBJECTIVE: To investigate the impact of posttraumatic humeral shaft malalignment on the ability to position the hand in space. METHODS: Two unique models were created: a cadaveric model and a computerized 3-dimensional (3D) model. In the cadaveric model, a midshaft transverse osteotomy of the humerus was created to simulate fracture. The osteotomy was fixed in varying degrees of coronal and sagittal malalignment. The hand's ability to reach 6 different bony landmarks was assessed as a surrogate measure of function. Subsequently, a healthy male volunteer underwent full-body magnetic resonance imaging with subsequent 3D skeletal recreation. A "virtual" midshaft transverse osteotomy was created. The osteotomy was angulated in various degrees of coronal and sagittal malalignment, and the hand's ability to reach the same 6 bony landmarks was measured. RESULTS: In the cadaveric model, varus angulation was better tolerated than valgus and sagittal deformity. Varus deformity less than 25 degree did not have a negative influence. Valgus angulation of 20 degree resulted in a more severe deficit. Estimated function of the upper extremity was most sensitive to sagittal deformity. These trends were confirmed in the 3D model. CONCLUSIONS: The direction and magnitude of posttraumatic humeral shaft malalignment independently affect the ability to position the hand in space, a surrogate measure of function. Upper extremity function may be more sensitive to posttraumatic humeral shaft malalignment than previously reported. Clinical studies investigating the impact of humeral shaft malalignment on the functional use of the upper extremity are warranted to clinically confirm these findings. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.

Technique for bone volume measurement from human femur head samples by classification of micro-CT image histograms.

INTRODUCTION: Micro-CT analysis is a powerful technique for a non-invasive evaluation of the morphometric parameters of trabecular bone samples. This elaboration requires a previous binarization of the images. A problem which arises from the binarization process is the partial volume artifact. Voxels at the external surface of the sample can contain both bone and air so thresholding operates an incorrect estimation of volume occupied by the two materials. AIM: The aim of this study is the extraction of bone volumetric information directly from the image histograms, by fitting them with a suitable set of functions. METHODS: Nineteen trabecular bone samples were extracted from femoral heads of eight patients subject to a hip arthroplasty surgery. Trabecular bone samples were acquired using micro-CT Scanner. Hystograms of the acquired images were computed and fitted by Gaussian-like functions accounting for: a) gray levels produced by the bone x-ray absorption, b) the portions of the image occupied by air and c) voxels that contain a mixture of bone and air. This latter contribution can be considered such as an estimation of the partial volume effect. RESULTS: The comparison of the proposed technique to the bone volumes measured by a reference instrument such as by a helium pycnometer show the method as a good way for an accurate bone volume calculation of trabecular bone samples.