Active and passive mechanical characterization of a human descending thoracic aorta with Klippel-Trenaunay syndrome.

A human aorta from a female donor affected by Klippel-Trenaunay syndrome was retrieved during a surgery for organ donation for transplant. The aorta was preserved in refrigerated Belzer UW organ preservation solution and tested within a few hours for mechanical characterization with and without vascular smooth muscle activation. KCl and Noradrenaline were used as vasoactive agents in bubbled Krebs-Henseleit buffer solution at 37 °C. A quasi-static and a dynamic mechanical characterization of the full wall and the three individual layers were carried out for strips taken in longitudinal and circumferential directions. The full wall in the descending portion of the aorta underwent mechanical tests with and without smooth muscle activation. Results were compared to data obtained from healthy aortas and show a reduced stiffness of the full wall in circumferential direction. Also, a significant reduction of the response to vasoactive agents in circumferential direction was observed, while the longitudinal response was similar to healthy cases.

Development and mechanical characterization of decellularized scaffolds for an active aortic graft.

Decellularized porcine aortas are proposed as scaffolds for revolutionary active aortic grafts. A change in the static and dynamic mechanical properties, associated with the microstructure of elastin and collagen fibers, corresponds to alteration in the cyclic expansion and perfusion, in addition to possible graft damage. Therefore, the present study thoroughly investigates the mechanical response of the decellularized scaffolds of human and porcine origin to static and dynamic mechanical loads. The responses of the native human and porcine aortas are also compared; this is unavailable in the literature. Because the aorta is subjected to pulsatile blood pressure, dynamical responses to cyclic loads and their associated viscoelastic properties are particularly relevant for advanced graft design. In parallel, this study examines the microstructure of the decellularized aorta. The resulting data are compared to the analogous data obtained for the native human and porcine tissues. The results indicate that by using an optimized decellularization protocol - based on sodium dodecyl sulfate (SDS) and DNase - that minimizes mechanical and structural changes of the tissue, layered scaffolds with static and dynamic properties very similar to natural human aortas are obtained. In particular, a decellularized porcine aorta is non-inferior to a decellularized human aorta. STATEMENT OF SIGNIFICANCE: About 55,000 patients undergo abdominal aortic aneurysm repair annually in the USA. The currently implanted grafts present a large mechanical mismatch with the native tissue. This increases the pulsatile nature of the blood flow with negative consequences to the organ perfusion. For this reason, biomimetic and mechanically compatible grafts for aortic repair are urgently needed and they can be obtained through tissue engineering. In this study, scaffolds from porcine and human aortas are obtained from an optimized decellularization protocol. They are accurately compared to the native tissue and present the ideal static and dynamic mechanical properties for developing innovative aortic grafts.

Viscoelasticity of human descending thoracic aorta in a mock circulatory loop.

Healthy human descending thoracic aortas, obtained during organ donation for transplant and research, were tested in a mock circulatory loop to measure the mechanical response to physiological pulsatile pressure and flow. The viscoelastic properties of the aortic segments were investigated at three different pulse rates. The same aortic segments were also subjected to quasi-static pressure tests in order to identify the aortic dynamic stiffness ratio, which is defined as the ratio between the stiffness in case of pulsatile pressure and the stiffness measured for static pressurization, both at the same value of pressure. The loss factor was also identified. The shape of the deformed aorta under static and dynamic pressure was measured by image processing to verify the compatibility of the end supports with the natural deformation of the aorta in the human body. In addition, layer-specific experiments on 10 human descending thoracic aortas allowed to precisely identify the mass density of the aortic tissue, which is an important parameter in cardiovascular dynamic models.

Role of smooth muscle activation in the static and dynamic mechanical characterization of human aortas.

Experimental data and a suitable material model for human aortas with smooth muscle activation are not available in the literature despite the need for developing advanced grafts; the present study closes this gap. Mechanical characterization of human descending thoracic aortas was performed with and without vascular smooth muscle (VSM) activation. Specimens were taken from 13 heart-beating donors. The aortic segments were cooled in Belzer UW solution during transport and tested within a few hours after explantation. VSM activation was achieved through the use of potassium depolarization and noradrenaline as vasoactive agents. In addition to isometric activation experiments, the quasistatic passive and active stress-strain curves were obtained for circumferential and longitudinal strips of the aortic material. This characterization made it possible to create an original mechanical model of the active aortic material that accurately fits the experimental data. The dynamic mechanical characterization was executed using cyclic strain at different frequencies of physiological interest. An initial prestretch, which corresponded to the physiological conditions, was applied before cyclic loading. Dynamic tests made it possible to identify the differences in the viscoelastic behavior of the passive and active tissue. This work illustrates the importance of VSM activation for the static and dynamic mechanical response of human aortas. Most importantly, this study provides material data and a material model for the development of a future generation of active aortic grafts that mimic natural behavior and help regulate blood pressure.

Microstructural and mechanical characterization of the layers of human descending thoracic aortas.

The mechanical properties of human aortas are linked to the layered tissue and its microstructure at different length scales. Each layer has specific mechanical and structural properties. While the ground substance and the elastin play an important role in tissue stiffness at small strain, collagen fibers carry most of the load at larger strains, which corresponds to the physiological conditions of the aorta at maximum pulsatile blood pressure. In fact, collagen fibers are crimped in the unloaded state. Collagen fibers show different orientation distributions when they are observed in a plane that is tangent to the aortic wall (in-plane section) or along a direction orthogonal to it (out-of-plane section). This was systematically investigated using large images (2500 × 2500 µm) with high resolution obtained by second harmonic generation (SHG) in order to homogenize tissue heterogeneity after a convergence analysis, which is a main goal of the study. In addition, collagen fibers show lateral interactions due to entanglements and the presence of transverse elastin fibers, observed on varying length scales using atomic force microscopy and a three-dimensional rendering obtained by stacking a sequence of SHG and two-photon fluorescence images; this is another important contribution. Human descending thoracic aortas from 13 heartbeat donors aged 28 to 66 years were examined. Uniaxial tensile tests were carried out on the longitudinal and circumferential strips of the aortic wall and the three separated layers (intima, media and adventitia). A structurally-motivated material model with (i) a term to describe the combined response of ground substance and elastin and (ii) terms to consider four families of collagen fibers with different directions was applied. The exclusion of compressed fibers was implemented in the fitting process of the experimental data, which was optimized by a genetic algorithm. The results show that a single fiber family with directional and dispersion parameters measured from SHG images can describe the mechanical response of all 39 layers (3 layers for each of the 13 aortas) with very good accuracy when a second (auxiliary) family of aligned fibers is introduced in the orthogonal direction to account for lateral fiber interaction. Indeed, all observed distributions of collagen directions can be accurately fitted by a single bivariate von Mises distribution. Statistical analysis of in-plane and out-of-plane dispersion of fiber orientations reveals structural differences between the three layers and a change of collagen dispersion parameters with age. STATEMENT OF SIGNIFICANCE: The stiffness of healthy young aortas is adjusted so that a diameter expansion of about 10 % is possible during the heartbeat. This creates the Windkessel effect, which smooths out the pulsating nature of blood flow and benefits organ perfusion. The specific elastic properties of the aorta that are required to achieve this effect are related to the microstructure of the aortic tissue at different length scales. An increase in the aortic stiffness, in addition to reducing cyclic expansion and worsening perfusion, is a risk factor for clinical hypertension. The present study relates the microstructure of healthy human aortas to the mechanical response and examines the changes in microstructural parameters with age, which is a key factor in increasing stiffness.

Identification of viscoelastic properties of Dacron aortic grafts subjected to physiological pulsatile flow.

In vascular surgery, most synthetic vascular grafts currently used for large vessels replacements are made of Dacron (polyethylene terephthalate; PET). In this study, the dynamic response of these synthetic arterial substitutes to physiological pulsatile conditions is investigated in depth. Experiments were performed on a mock circulatory loop developed to replicate physiological pulsatile pressure and flow. Two different models of Dacron grafts (branched and straight) were tested at various heart rate conditions. Results are presented in terms of cyclic axisymmetric diameter changes, hysteretic loops of the pressure-diameter change, and viscoelastic parameters, such as loss factor and storage modulus that are identified from the hysteresis loop. The amplitude of cyclic diameter change of the Dacron graft was found to be always below 0.2% for all the heart rates considered (from 57 to 187 bpm). The loss factor of the Dacron graft slightly increased with the heart rate; almost no effect of the pulse rate was observed on the storage modulus, which was identified to be around 100 MPa. Both glycerol-water mixture (i.e. the blood analogue fluid) and saline solution were used in the circulatory loop and results did not present significant differences between the two cases. This shows that the effect of the shear load on the dynamic response of Dacron grafts is negligible. A comparison between Dacron vascular implants and human thoracic aortas shows a large mismatch in their viscoelastic mechanical properties.

Experiments on dynamic behaviour of a Dacron aortic graft in a mock circulatory loop.

Woven Dacron grafts are currently used for the surgical treatment of aortic aneurysm and acute dissection, two otherwise fatal pathologies when aortic wall rupture occurs. While Dacron is chosen for aortic grafts because of characteristics such as biocompatibility and durability, few data are available about the dynamic response of Dacron prosthetic devices and about their side effects on the cardiovascular system. In this study, a Dacron graft was subjected to physiological flow conditions in a specifically-developed mock circulatory loop. Experiments were conducted at different physiological pulsation-per-minute rates. Results show that, in comparison to an aortic segment of the same length, the prosthesis is extremely stiffer circumferentially, thus limiting the dynamical radial expansion responsible for the Windkessel effect in human arteries. The prosthesis is instead excessively compliant in the axial direction and develops preferentially bending oscillations. This very different dynamic behaviour with respect to the human aorta can alter cardiovascular pressure and flow dynamics resulting in long-term implant complications.