Quantifying Active and Passive Stiffness in Plantar Flexor Muscles Following Intermittent Maximal Isometric Contractions Using Shear Wave Elastography.

OBJECTIVE: This study aimed: (i) to investigate the impact of fatigue, triggered by maximal isometric contraction exercises, on the active and passive stiffness of plantar flexors (PF), and (ii) to examine the relationship between changes in mechanical parameters and neuromuscular alterations after fatigue. METHODS: A healthy cohort (n = 12; age = 27.3 ± 5.5 y; BMI = 24.4 ± 2.35 kg/m²) was instructed to perform 60 isometric contractions, each lasting 4 s with a 1-s rest interval, using an ergometer. Several measures were taken before and after the fatigue protocol. First, the stiffness of the PF-tendon complex (PFC) was quantified during passive ankle mobilization both during and after the fatigue protocol using the ergometer. Additionally, from shear wave elastography, the active and passive stiffness of the gastrocnemius medialis (GM) were measured during passive ankle mobilization and isometric maximal voluntary contraction (MVC), respectively. Finally, the peak torque and the rate of torque development (RFD) of PF were assessed during the MVC using the ergometer. Ankle muscle activities (surface electromyograph [SEMG]) were recorded during all evaluations using electromyography. RESULTS: After the fatigue protocol, the results revealed a decline in active stiffness, peak torque of PF, RFD and SEMG activity of the GM (p < 0.001). Furthermore, significant correlation was identified between the decrease of the peak torque of PF and the active stiffness of the GM (r = 0.6; p < 0.05). A decrease in the PFC stiffness (p < 0.001) and a decrease in the shear modulus of the GM at 20° (p < 0.001) were also observed. CONCLUSION: Isometric fatiguing exercises modify the mechanical properties of both the contractile and elastic components. Notably, decreases in both passive and active stiffness may be critical for athletes, as these changes could potentially increase the risk of injury.

Quantifying Plantar Flexor Muscles Stiffness During Passive and Active Force Generation Using Shear Wave Elastography in Individuals With Chronic Stroke.

OBJECTIVES: This study aims to investigate the mechanical properties of paretic and healthy plantar flexor muscles and assesses the spatial distribution of stiffness between the gastrocnemius medialis (GM) and lateralis (GL) during active force generation. METHODS: Shear wave elastography measurements were conducted on a control group (CNT, n=14; age=59.9±10.6 years; BMI=24.5±2.5 kg/m2) and a stroke survivor group (SSG, n=14; age=63.2±9.6 years; BMI=23.2±2.8 kg/m2). Shear modulus within the GM and GL was obtained during passive ankle mobilization at various angles of dorsiflexion (P0 =0°; P1=10°; P2=20°; P3=-20° and P4=-30°) and during different levels (30%, 50%, 70%, 100%) of maximal voluntary contraction (MVC). Muscle activations of GM, GL, soleus and tibialis anterior were also evaluated. RESULTS: The results revealed a significant increase in passive stiffness within the paretic plantar flexor muscles under high tension during passive mobilization (p<0.05). Yet, during submaximal and maximal MVC, the paretic plantar flexors exhibited decreased active stiffness levels (p<0.05). A notable discrepancy was found between the stiffness of the GM and GL, with the GM demonstrating greater stiffness from 0° of dorsiflexion in the SSG (p<0.05), and from 10° of dorsiflexion in the CNT (p<0.05). No significant difference in stiffness was observed between the GM and GL muscles during active condition. CONCLUSION: Stroke affects the mechanical properties differently depending on the state of muscle activation. Notably, the distribution of stiffness among synergistic plantar flexor muscles varied in passive condition, while remaining consistent in active condition.

Dataset of Inter and intramuscular variability of stiffness in paretic individuals during prone and standing positions.

Several studies have investigated muscle rigidity using SWE. However, the assessments may not consider the most affected regions within the same muscle tissue nor the intramuscular variability of rigidity between muscles of the same muscle group, e.g., plantar flexors. The data presented in this article aimed to explore the inter-and intramuscular variability of plantar flexors stiffness during prone and standing positions at different muscle lengths in healthy and paretic individuals. Shear wave ultrasound images were acquired for the three plantar flexor muscles (gastrocnemius medialis [GM], gastrocnemius lateralis [GL], and soleus [SOL]) in two positions: prone and standing. The imaging was conducted at various dorsiflexion angles (0°, 10°, and 20°), and measurements were taken at different proximo-distal regions within each muscle. This data set allowed us to highlight the impact of stroke on mechanical properties that varies depending on whether ankle muscles are in an active or passive state during dorsiflexion. Additionally, the modification of the ankle muscle state influences the distribution of stiffness both within and between the plantar flexors.

Mechanical properties change of immobilized skeletal muscle in short position measured by shear wave elastography and pure shearing test.

The purpose of this study was to evaluate the effects of immobilization on mechanical properties of skeletal muscle over the time. An in vivo rat model was used to investigate the shear modulus change of the flexor carpi ulnaris (FCU) in a short position. Measurements were performed by shear wave elastography (SWE) to compare contralateral and immobilized cases. The results showed a significant increase of 18.1% (p = 3.86. 10-7) in the shear modulus of immobilized skeletal muscle after two weeks (D14) when compared with the contralateral case. For the purposes of comparison, in vitro mechanical pure shearing tests were performed on samples collected from the skeletal muscles of the same rats. Although the difference between contralateral and immobilized cases was 17.6% (p = 0.32) at D14, the shear modulus difference was 35.7% (p = 0.0126 and p = 1.57.10-5 for immobilization and contralateral respectively) between in vivo and in vitro approaches. The mechanical properties changes were then correlated with the density of collagen from histological analysis, and it was shown that the contralateral collagen surface density was 25.4% higher than the immobilized density at D14 (p = 0.001). Thus, the results showed the feasibility of the comparison between the two approaches, which can surely be improved by optimizing the experimental protocols.

Spatial distribution of stiffness between and within muscles in paretic and healthy individuals during prone and standing positions.

This study investigated the inter- and intramuscular variability of plantar flexors stiffness during prone and standing positions at different muscle lengths in healthy and paretic individuals. To access tissue stiffness, shear wave elastography (SWE) measurements were carried out on two groups: control group (CG; n=14; age 43.9±9.6 years; body mass index [BMI]=24.5±2.5 kg/m2) and stroke survivor group (SSG; n=14; age 43.9±9.6 years; BMI=24.5±2.5 kg/m2). Shear Modulus (µ, kPa) within three plantar flexors (the gastrocnemius medialis [GM], gastrocnemius lateralis [GL], and soleus [SOL]) was obtained during two conditions: prone and standing position, at different angles of dorsiflexion (0°, 10°, and 20°). Measurements were also performed in different proximo-distal regions of each muscle. Muscle activation of the GM, GL, SOL, and tibialis anterior were evaluated during the two conditions. Results showed a high spatial stiffness variability between and within plantar flexors during dorsiflexion. The highest stiffness was observed in the GM, especially in the distal region at 20° in healthy and paretic muscles. In the prone position, the paretic muscle exhibits greater stiffness compared to the healthy muscle (p < 0.05). In contrast, in the standing position, an increase of stiffness in the healthy muscle compared to the paretic muscle was observed (p < 0.05). Thus, mechanical properties are differently affected by stroke depending on active and passive states of ankle muscles during dorsiflexion. In addition, the modification of ankle muscle state change stiffness distribution between and within plantar flexors.

Regional variation in the mechanical properties of the skeletal muscle.

Regional mechanics of skeletal muscle were investigated from equibiaxial testing in vitro on tissue samples. Samples were collected in three excising zones in transversal direction to the myofibers. Thus, the transverse plane stiffness, likely to be dictated by extracellular matrix collagen (ECM), was studied. For that, distal, middle, and proximal samples of healthy brachial biceps of rats have been tested. Data was used to generate the material parameters of the first order Ogden constitutive model at these different zones of skeletal muscle. In addition to having a nonlinear mechanical behavior, the analysis of the material parameters of the model showed that the stiffness value of the skeletal muscle tissue may on average have doubled depending on the collected sample location (p < 0.001). Furthermore, it was also shown that during the tests, when the storage temperature of the samples increases from 22 °C to 37 °C, the stiffness of the muscle tissue becomes more important (p < 0.05), which may be due to the rigor mortis phenomenon. Thus, these results contribute to investigating the regional change of mechanical properties of skeletal muscle, particularly those of ECM that play a major role in stiffness tissue, which is essential for the development of accurate computational models.

Mechanical and microstructural changes of skeletal muscle following immobilization and/or stroke.

Patient management following a stroke currently represents a medical challenge. The presented study investigates the effect of immobilization on skeletal muscles in short positions after a stroke. A rat model was implemented in order to compare four situations within 14 days including control group, immobilization of one forelimb without stroke, stroke without immobilization and stroke with immobilization of the paretic forelimb. To analyze the changes of the mechanical properties of the passive skeletal muscle, the biological tissue is assumed to behave as a visco-hyperelastic and incompressible material characterized by the first-order Ogden's strain energy function coupled with second-order Maxwell's model. The material parameters were identified from inverse finite element method by using uniaxial relaxation tests data of skeletal muscle samples. Based on measurements of histological parameters, we observe that muscle immobilization led to microconstituents changes of skeletal muscles that were correlated with degradations of its mechanical properties. In the case of immobilization without stroke, the neurological behavior was also altered in the same manner as in the case of a stroke. We showed that immobilization of skeletal muscles in short positions produced contractile tissue atrophy, connective tissue thickening and alteration of passive mechanical behavior that were more damaging than the effects produced by a stroke. These results showed then that immobilization of skeletal muscles in short positions is highly deleterious with or without a stroke.

Influence of experimental conditions on visco-hyperelastic properties of skeletal muscle tissue using a Box-Behnken design.

The Mechanical characterization of skeletal muscles is strongly dependent on numerous experimental design factors. Nevertheless, significant knowledge gaps remain on the characterization of muscle mechanics and a large number of experiments should be implemented to test the influence of a large number of factors. In this study, we propose a design of experiment method (DOE) to study the parameter sensitivity while minimizing the number of tests. A Box-Behnken design was then implemented to study the influence of strain rate, preconditioning and preloading conditions on visco-hyperelastic mechanical parameters of two rat forearm muscles. The results show that the strain rate affects the visco-hyperelastic parameters for both muscles. These results are consistent with previous work demonstrating that stiffness and viscoelastic contributions increase with strain rate. Thus, DOE has been shown to be a valid method to determine the effect of the experimental conditions on the mechanical behaviour of biological tissues such as skeletal muscle. This method considerably reduces the number of experiments. Indeed, the presented study using 3 parameters at 3 levels would have required at least 54 tests per muscle against 14 for the proposed DOE method.

Arterial Stiffness Assessment by Shear Wave Elastography and Ultrafast Pulse Wave Imaging: Comparison with Reference Techniques in Normotensives and Hypertensives.

Shear wave elastography and ultrafast imaging of the carotid artery pulse wave were performed in 27 normotensive participants and 29 age- and sex-matched patients with essential hypertension, and compared with reference techniques: carotid-femoral pulse wave velocity (cfPWV) determined via arterial tonometry and carotid stiffness (carPWV) determined via echotracking. Shear wave speed in the carotid anterior (a-SWS) and posterior (p-SWS) walls were assessed throughout the cardiac cycle. Ultrafast PWV was measured in early systole (ufPWV-FW) and in end-systole (dicrotic notch, ufPWV-DN). Shear wave speed in the carotid anterior appeared to be the best candidate to evaluate arterial stiffness from ultrafast imaging. In univariate analysis, a-SWS was associated with carPWV (r = 0.56, p = 0.003) and carotid-to-femoral PWV (r = 0.66, p < 0.001). In multivariate analysis, a-SWS was independently associated with age (R²â€¯= 0.14, p = 0.02) and blood pressure (R²â€¯= 0.21, p = 0.004). Moreover, a-SWS increased with blood pressure throughout the cardiac cycle and did not differ between normotensive participants and patients with essential hypertension when compared at similar blood pressures.

Loss of anisotropic properties in abdominal aorta aneurysm obtained from the xenograft rat model.

BACKGROUND: Cellular treatments using mesenchymal stem cells (MSCs) cultured in 3D conditions constitute a solution to the classical surgery in treating abdominal aortic aneurysm (AAA). The recurrent question is: how this type of biotherapy changes the mechanical behavior of artery? METHODS: Experiments measurements based on xenograft rat model showed that the proposed cellular treatment leads to a decreasing radius and length of the AAA during its growth. An inverse finite element method was used to investigate the mechanical hyperelastic behavior of the AAA in the untreated case compared to the treated one. RESULTS: Although AAA leads a loss anisotropy while the cellular treatment does not restore it, it was shown that the stiffness of the arterial wall was improved. The numerical analysis of the stress distributions permitted to localize the stress concentration through the arterial wall and the probable zone of the rupture of the aneurysm developed from the xenograft rat model. CONCLUSIONS: The treatment of AAA with MSCs cultured in a 3D conditions constitutes a new challenge. Based on xenograft rat model, this study shows the potential of this cellular treatment to reduce the variation of the growth, the stiffness and the stress distributions.

Effect of cryopreservation at -80°C on visco-hyperelastic properties of skeletal muscle tissue.

This paper investigated the effect of cryopreservation at -80°C on mechanical visco-hyperelastic properties of skeletal muscle. For that, both tensile and compression relaxation tests were performed on porcine tissues samples in fibre and cross-fibre directions. Material parameters were identified by using first order Ogden's strain energy function coupled with second order Maxwell's model. The results revealed that the cryopreservation conditions at -80°C with conservative and cryoprotectant solution significantly affected the mechanical properties of the skeletal muscle. Thus, cryopreserved tissue showed a higher instantaneous initial Young's modulus than for the fresh tissue in both tensile and compression deformations, and in the two muscular fibre directions. Furthermore, in compression tests, cryopreserved tissue exhibited a smaller non-linearity and a higher total relaxation ratio in both muscle fibre directions than for the fresh tissue.

Mechanical behavior of the abdominal aortic aneurysm assessed by biaxial tests in the rat xenograft model.

This paper addresses the mechanical biaxial behavior of degraded arteries obtained by the rat xenograft model. For that, a pressure myograph was used to perform extension-inflation tests on abdominal aortic aneurysms (AAAs). Furthermore, residual stresses in the aneurismal wall were assessed by opening angle tests. Thus, the changes in mechanical behavior between native murine aortas, decellularized guinea pig aortas (the grafts) and degraded aortas (AAAs) were investigated. It was shown that decellularized and degraded aortas exhibited a different mechanical behavior than native murine aortas. Indeed, decellularized aortas presented a marked decrease in circumferential stretch and distensibility compared with native aortas. Moreover, we evidenced an exacerbation of these changes in mechanical behavior for AAAs, which showed the lowest distension and distensibility at 100mmHg. The opening angle test also revealed a complete loss of residual stresses in the degraded arterial wall given the non opening of rings extracted from AAAs.

Diameter and thickness-related variations in mechanical properties of degraded arterial wall in the rat xenograft model.

The purpose of this study was to evaluate the diameter and thickness-related variations in mechanical properties of degraded arterial wall. To this end, ring tests were performed on 31 samples from the rat xenograft model of abdominal aortic aneurysm (AAA) and failure properties were determined. An inverse finite element method was then employed to identify the material parameters of a hyperelastic and incompressible strain energy function. Correlations with outer diameter and wall thickness of the rings were examined. Furthermore, we investigated the changes in mechanical properties between the grafts, which consist in guinea pig decellularized aortas, native murine aortas and degraded aortas (AAAs). Decellularized aortas presented a significantly lower ultimate strain associated with a higher stiffening rate compared to native aortas. AAAs exhibited a significantly lower ultimate stress than other groups and an extensible-but-stiff behavior. The proposed approach revealed correlations of ultimate stress and material parameters of aneurysmal aortas with outer diameter and thickness. In particular, the negative correlations of the material parameter accounting for the response of the non-collagenous matrix with diameter and thickness (r=-0.67 and r=-0.73, p<0.001) captured the gradual loss of elastin with dilatation observed in histology (r=-0.97, p<0.001). Moreover, it exposed the progressive weakening of the wall with enlargement and thickening (r=-0.64 and r=-0.69, p<0.001), suggesting that wall thickness and diameter may be indicators of rupture risk in the rat xenograft model.

Influence of viscoelastic properties of an hyaluronic acid-based hydrogel on viability of mesenchymal stem cells.

BACKGROUND: The present research is involved in the framework of the biotherapy using mesenchymal stem cells (MSCs). Here, MSC encapsulation in a hydrogel based on hyaluronic acid (HA) is investigated to optimize the composition of the biomaterial. METHODS: Several formulations candidates of the hydrogel (9 in total) are postulated as a scaffold for the 3D MSC culture in order to investigate their potential to mimic the in vivo cellular environment. Rheological measurements in oscillation mode of complex modulus and complex viscosity are performed on the different hydrogels. Biological tests are carried out for the measurement of the cell viability of MSC encapsulated in the hydrogels. RESULTS: Rheological and biological findings are correlated together in order to establish relationships between the viscoelastic properties of the hydrogel and the cellular viability of MSC. CONCLUSIONS: In the light of the viability results, the composition of the hydrogel was related to the MSC proliferation. Thus, such relations are useful tools for scientists offering them more flexibility in the design of their hydrogels while ensuring an acceptable level of MSC viability.

Mechanical behavior of abdominal aorta aneurysm in rat model treated by cell therapy using mesenchymal stem cells.

Regenerative medicine to substitute conventional surgery or an endovascular stent constitutes currently a challenge to treat abdominal aneurysm artery (AAA). The present paper addresses the following question: Can a cellular therapy from mesenchymal stem cells reestablish the mechanical properties of damaged abdominal aorta? For that, the xenograft rat model that mimics arterial dilatation due to aneurysmal disease is used to study the effects of the proposed cellular therapy. To investigate the changes in the mechanical behavior of the arterial wall, the artery is assumed to be made of a hyperelastic and incompressible material characterized by a strain energy function fitted to the average data set of uniaxial tests of AAA tissue samples. In order to compute the stresses in the artery by using an analytical approach, the aneurysm is represented as a "parabolic-exponential" thin membrane. Thus, when compared to healthy, untreated and treated arteries, the obtained results demonstrate that the cellular therapy stabilizes the geometry of AAAs, improves the stiffness of the tissue and decreases stress variations in the arterial wall.

Optimized hyaluronic acid-hydrogel design and culture conditions for preservation of mesenchymal stem cell properties.

A novel approach that preserved most mesenchymal stem cell (MSC) characteristics was developed using MSC encapsulation in a hydrogel based on hyaluronic acid (HA). An optimized HA-hydrogel composition, whose characteristics were assessed by scanning electron microscopy and viscoelastic property analyses, as well as the more favorable MSC seeding density, was established. These optimal three-dimensional MSC culture conditions allowed morphological cell remodeling, maintained the expression of stem cell markers over 28 days of culture, and preserved MSC differentiation plasticity. In addition, MSCs in HA-hydrogel submitted for 7 days to mechanical constraint that aimed at mimicking in vivo cardiac beat displayed enhanced cell survival by more than 40% compared to static culture conditions. Thus, the optimized HA-based hydrogel provides a niche for MSCs, which preserves their properties and opens ways for cell therapy, in particular in aortic repair medicine.

Stochastic modelling of wall stresses in abdominal aortic aneurysms treated by a gene therapy.

A stochastic mechanical model using the membrane theory was used to simulate the in vivo mechanical behaviour of abdominal aortic aneurysms (AAAs) in order to compute the wall stresses after stabilisation by gene therapy. For that, both length and diameter of AAAs rats were measured during their expansion. Four groups of animals, control and treated by an endovascular gene therapy during 3 or 28 days were included. The mechanical problem was solved analytically using the geometric parameters and assuming the shape of aneurysms by a 'parabolic-exponential curve'. When compared to controls, stress variations in the wall of AAAs for treated arteries during 28 days decreased, while they were nearly constant at day 3. The measured geometric parameters of AAAs were then investigated using probability density functions (pdf) attributed to every random variable. Different trials were useful to define a reliable confidence region in which the probability to have a realisation is equal to 99%. The results demonstrated that the error in the estimation of the stresses can be greater than 28% when parameters uncertainties are not considered in the modelling. The relevance of the proposed approach for the study of AAA growth may be studied further and extended to other treatments aimed at stabilisation AAAs, using biotherapies and pharmacological approaches.

Carotid artery mechanical properties and stresses quantified using in vivo data from normotensive and hypertensive humans.

The goal of this study was to model the in vivo non-linear mechanical behavior of human common carotid arteries (CCAs) and then to compare wall stresses and associated contributions of micro-constituents in normotensive (NT) and treated hypertensive (HT) subjects. We used an established theoretical model of 3D arterial mechanics that assumes a hyperelastic, anisotropic, active-passive, and residually stressed wall. In vivo data were obtained non-invasively from CCAs in 16 NT (21-64 years old) and 25 treated HT (44-69 years old) subjects. The associated quasi-static boundary value problem was solved semi-analytically over a cardiac cycle while accounting for surrounding perivascular tissue. Best-fit values of model parameters, including those describing contributions by intramural elastin, fibrillar collagen, and vascular smooth muscle, were estimated by a non-linear least-squares method. The model (1) captured temporal changes in intraluminal pressure, (2) estimated wall stress fields that appeared to reflect the presence or absence of age and disease, and (3) suggested changes in mechanical characteristics of wall micro-constituents despite medical treatment of hypertension. For example, age was positively correlated with residual stresses and altered fibrillar collagen in NT subjects, which indirectly validated the modeling, and HT subjects had higher levels of stresses, increased smooth muscle tone, and a stiffer elastin-dominated matrix despite treatment. These results are consistent with prior reports on effects of age and hypertension, but provide increased insight into evolving contributions of cell and matrix mechanics to arterial behavior in vivo.

Mechanical properties of arteries cryopreserved at -80 degrees C and -150 degrees C.

A new protocol for cryopreservation of arteries frozen at -80 degrees C was compared to the reference protocol for cryopreservation at -150 degrees C and to freshly harvested arteries. The aim of the study is to evaluate both protocols as global procedures to freeze and thaw arteries commonly used in tissue banks. Changes in mechanical properties of rabbit common carotid arteries were studied. Vascular segments were tested in vitro under dynamics loading conditions. Pressure and diameter were recorded simultaneously by a high fidelity transducer and an echotracking device, respectively. The pressure-diameter relationship was fitted by the arctangent Langewouters' model and the arterial thickness was derived from histological measurements. Histological sections showed that the fresh and -80 degrees C groups were less damaged by hemodynamic load and histological preparation than the -150 degrees C group (p<0.05). No differences between fresh and cryopreserved arteries regarding the structural (diameter, intimal-media thickness) and mechanical parameters (distensibility, circumferential stress, elastic modulus) were found. The isobaric circumferential stress was reduced in frozen arteries. These results demonstrate that the cryopreservation at -80 degrees C preserves the histological structure and mechanical properties better than the cryopreservation at -150 degrees C, suggesting that the new cryopreservation protocol at -80 degrees C is a method of choice for treating vessel replacement in vascular surgery.

Bone remodeling regulation under unloading conditions: numerical investigations.

The present paper addresses the following question: can a simple regulatory bone remodeling model predict effects of unloading conditions on the trabecular bone morphology? In an attempt to answer this question, rat tail-suspension was chosen as a model that mimics the microgravity environment. Over 23 days, histomorphometric analysis was carried out on cross-sections of tibias of the suspended animals. The slices were digitalized and images discretized to obtain osteocyte distribution and apparent bone density. Based on these experimental data, finite element simulations were conducted to evaluate the bone loss and the change in trabecular architecture similar to those observed after a spaceflight. The numerical model is driven by a remodeling law that takes into account the nonuniform osteocyte distribution that may itself provide mechanoreception. We used the bone density rate of change from the remodeling theory and a time stepping algorithm witch are implemented in a finite element software. This approach takes into account the unloading effects on bone remodeling process and permits to confront experimental and numerical data. We showed that there is a good agreement between these data, particularly at the beginning of the simulated bone mass loss during the rat tail-suspension experiment. Indeed, we obtained a variation of 5.25% at day 7 (D7), 2.09% at day 13 (D13) and finally, 51.03% at day 23 (D23). Despite that last variation, the proposed theoretical model can be suitable to simulate the alteration of bone mineral density under the specific unloading conditions of the rat tail-suspension model.