Francisco J. Rojo

Decellularization of pericardial tissue and its impact on tensile viscoelasticity and glycosaminoglycan content

Bovine pericardium is a collagenous tissue commonly used as a natural biomaterial in the fabrication of cardiovascular devices. For tissue engineering purposes, this xenogeneic biomaterial must be decellularized to remove cellular antigens. With this in mind, three decellularization protocols were compared in terms of their effectiveness to extract cellular materials, their effect on glycosaminoglycan (GAG) content and, finally, their effect on tensile biomechanical behavior. The tissue decellularization was achieved by treatment with t-octyl phenoxy polyethoxy ethanol (Triton X-100), tridecyl polyethoxy ethanol (ATE) and alkaline treatment and subsequent treatment with nucleases (DNase/RNase). The quantified residual DNA content (3.0±0.4%, 4.4±0.6% and 5.6±0.7% for Triton X-100, ATE and alkaline treatment, respectively) and the absence of nuclear structures (hematoxylin and eosin staining) were indicators of effective cell removal. In the same way, it was found that the native tissue GAG content decreased to 61.6±0.6%, 62.7±1.1% and 88.6±0.2% for Triton X-100, ATE and alkaline treatment, respectively. In addition, an alteration in the tissue stress relaxation characteristics was observed after alkaline treatment. We can conclude that the three decellularization agents preserved the collagen structural network, anisotropy and the tensile modulus, tensile strength and maximum strain at failure of native tissue.

Fractional-order viscoelasticity applied to describe uniaxial stress relaxation of human arteries

Viscoelastic models can be used to better understand arterial wall mechanics in physiological and pathological conditions. The arterial wall reveals very slow time-dependent decays in uniaxial stress-relaxation experiments, coherent with weak power-law functions. Quasi-linear viscoelastic (QLV) theory was successfully applied to modeling such responses, but an accurate estimation of the reduced relaxation function parameters can be very difficult. In this work, an alternative relaxation function based on fractional calculus theory is proposed to describe stress relaxation experiments in strips cut from healthy human aortas. Stress relaxation (1 h) was registered at three incremental stress levels. The novel relaxation function with three parameters was integrated into the QLV theory to fit experimental data. It was based in a modified Voigt model, including a fractional element of order alpha, called spring-pot. The stress-relaxation prediction was accurate and fast. Sensitivity plots for each parameter presented a minimum near their optimal values. Least-squares errors remained below 2%. Values of order alpha = 0.1-0.3 confirmed a predominant elastic behavior. The other two parameters of the model can be associated to elastic and viscous constants that explain the time course of the observed relaxation function. The fractional-order model integrated into the QLV theory proved to capture the essential features of the arterial wall mechanical response.

Mechanical behaviour and rupture of normal and pathological human ascending aortic wall

The mechanical properties of aortic wall, both healthy and pathological, are needed in order to develop and improve diagnostic and interventional criteria, and for the development of mechanical models to assess arterial integrity. This study focuses on the mechanical behaviour and rupture conditions of the human ascending aorta and its relationship with age and pathologies. Fresh ascending aortic specimens harvested from 23 healthy donors, 12 patients with bicuspid aortic valve (BAV) and 14 with aneurysm were tensile-tested in vitro under physiological conditions. Tensile strength, stretch at failure and elbow stress were measured. The obtained results showed that age causes a major reduction in the mechanical parameters of healthy ascending aortic tissue, and that no significant differences are found between the mechanical strength of aneurysmal or BAV aortic specimens and the corresponding age-matched control group. The physiological level of the stress in the circumferential direction was also computed to assess the physiological operation range of healthy and diseased ascending aortas. The mean physiological wall stress acting on pathologic aortas was found to be far from rupture, with factors of safety (defined as the ratio of tensile strength to the mean wall stress) larger than six. In contrast, the physiological operation of pathologic vessels lays in the stiff part of the response curve, losing part of its function of damping the pressure waves from the heart.

Mechanical characterisation of the human thoracic descending aorta: experiments and modelling

This work presents experiments and modelling aimed at characterising the passive mechanical behaviour of the human thoracic descending aorta. To this end, uniaxial tension and pressurisation tests on healthy samples corresponding to newborn, young and adult arteries are performed. Then, the tensile measurements are used to calibrate the material parameters of the Holzapfel constitutive model. This model is found to adequately adjust the material behaviour in a wide deformation range; in particular, it captures the progressive stiffness increase and the anisotropy due to the stretching of the collagen fibres. Finally, the assessment of these material parameters in the modelling of the pressurisation test is addressed. The implication of this study is the possibility to predict the mechanical response of the human thoracic descending aorta under generalised loading states like those that can occur in physiological conditions and/or in medical device applications.

Mechanical properties of human coronary arteries

The lack of reliable mechanical data on coronary arteries and, more specifically, on their wall strength hampers the application of numerical models and simulations to vascular problems, and precludes physicians from knowing in advance the response of coronary arteries to the different interventions. Studies of the mechanical properties of coronary arteries have been carried out almost exclusively on animals. Only a few studies have tried to characterize the in vivo behavior of human coronaries through tests under physiological conditions. In this work, the mechanical properties of human coronary arteries have been characterized. Whole samples from human right (RC) and left anterior descending (LAD) coronary arteries aged between 23 and 83 years have been studied by means of in-vitro tensile testing up to failure.

Mechanical behavior of bilayered small-diameter nanofibrous structures as biomimetic vascular grafts

To these days, the production of a small diameter vascular graft (<6mm) with an appropriate and permanent response is still challenging. The mismatch in the grafts mechanical properties is one of the principal causes of failure, therefore their complete mechanical characterization is fundamental. In this work the mechanical response of electrospun bilayered small-diameter vascular grafts made of two different bioresorbable synthetic polymers, segmented poly(ester urethane) and poly(L-lactic acid), that mimic the biomechanical characteristics of elastin and collagen is investigated. A J-shaped response when subjected to internal pressure was observed as a cause of the nanofibrous layered structure, and the materials used. Compliance values were in the order of natural coronary arteries and very close to the bypass gold standard-saphenous vein. The suture retention strength and burst pressure values were also in the range of natural vessels. Therefore, the bilayered vascular grafts presented here are very promising for future application as small-diameter vessel replacements.

Increases of Corporal Temperature as a Risk Factor of Atherosclerotic Plaque Instability

Background This work explores for the first time the effects of temperature increments on the development of high shear stresses between plaque and arterial wall due to their different dilatational properties. Data from the literature report febrile reactions prior to myocardial infarction in patients with normal coronary arteries and that coronary syndromes seem to be triggered by bacterial and viral infections, being fever the common symptom. Methods The thermo-mechanical behavior of thoracic aortas of New Zealand White rabbits with different degrees of atherosclerosis was measured by means of pressure–diameter tests at different temperatures. In addition, specific measurements of the thermal dilatation coefficient of atheroma plaques and of healthy arterial walls were performed by means of tensile tests at different temperatures. Results Results show a different thermo-mechanical behavior, the dilatation coefficient of atheroma plaque being at least twice that of the arterial wall. The calculation of temperature-induced mechanical stress at the plaque–vessel interface yielded shear stress levels enough to promote plaque rupture. Conclusions Increases of corporal temperature either local—produced by the inflammatory processes associated with atherosclerosis—or systemic—by febrile reactions—can play a role in increasing the risk of acute coronary syndromes, and they deserve a more comprehensive study.

Stability and mechanical evaluation of bovine pericardium cross-linked with polyurethane prepolymer in aqueous medium

The present study investigates the potential use of non-catalyzed water-soluble blocked polyurethane prepolymer (PUP) as a bifunctional cross-linker for collagenous scaffolds. The effect of concentration (5, 10, 15 and 20%), time (4, 6, 12 and 24 h), medium volume (50, 100, 200 and 300%) and pH (7.4, 8.2, 9 and 10) over stability, microstructure and tensile mechanical behavior of acellular pericardial matrix was studied. The cross-linking index increased up to 81% while the denaturation temperature increased up to 12 °C after PUP crosslinking. PUP-treated scaffold resisted the collagenase degradation (0.167±0.14 mmol/g of liberated amine groups vs. 598±60 mmol/g for non-cross-linked matrix). The collagen fiber network was coated with PUP while viscoelastic properties were altered after cross-linking. The treatment of the pericardial scaffold with PUP allows (i) different densities of cross-linking depending of the process parameters and (ii) tensile properties similar to glutaraldehyde method.

Factors influencing the mechanical behaviour of healthy human descending thoracic aorta

In recent times, significant effort has been made to understand the mechanical behaviour of the arterial wall and how it is affected by the different vascular pathologies. However, to be able to interpret the results correctly, it is essential that the influence of other factors, such as aging or anisotropy, be understood. Knowledge of mechanical behaviour of the aorta has been customarily constrained by lack of data on fresh aortic tissue, especially from healthy young individuals. In addition, information regarding the point of rupture is also very limited. In this study, the mechanical behaviour of the descending thoracic aorta of 28 organ donors with no apparent disease, whose ages vary from 17 to 60 years, is evaluated. Tensile tests up to rupture are carried out to evaluate the influence of age and wall anisotropy. Results reveal that the tensile strength and stretch at failure of healthy descending aortas show a significant reduction with age, falling abruptly beyond the age of 30. This fact places age as a key factor when mechanical properties of descending aorta are considered.

Influencia de la presión y la temperatura en el comportamiento de la aorta y las carótidas humanas

Introduccion y objetivos La respuesta termomecanica de las arterias humanas es poco conocida a pesar de su importancia para la comprension de la fisiologia arterial, y para la evaluacion y mejora de los procedimientos quirurgicos. El objetivo de este trabajo es aportar por vez primera datos experimentales que muestren como se ve afectada la respuesta mecanica de dos tipos de arterias humanas –aorta y carotida– por los cambios de temperatura. Metodos La respuesta mecanica de las arterias se ha obtenido in vitro a traves de la medicion de las curves presion interior-diametro exterior para 4 temperaturas (17, 27, 37 y 42 °C). Se ha realizado un analisis termomecanico para obtener los coeficientes de dilatacion y la rigidez del material. El estado de la pared arterial se ha evaluado mediante analisis histologico. Resultados Las arterias aorta y carotida aumentan ligeramente su flexibilidad con la temperatura. El coeficiente de dilatacion de ambos vasos depende criticamente de la presion interior aplicada. A bajas presiones, el coeficiente de dilatacion es negativo (el vaso se contrae cuando se calienta), mientras que por encima de cierta presion umbral –distinta para cada tipo de arteria– el coeficiente de dilatacion se hace positivo. Conclusiones El efecto combinado de la presion interior y la temperatura afecta al comportamiento de las arterias y, por ello, debe ser tenido en cuenta al abordar situaciones clinicas que impliquen cambios de temperatura. La intensidad de este efecto depende del tipo de arteria estudiada, lo que requiere la obtencion de datos mas detallados, centrados en los vasos de interes clinico.