Unravelling anisotropic nonlinear shear elasticity in muscles: Towards a non-invasive assessment of stress in living organisms.

Acoustoelasticity theory describes propagation of shear waves in uniaxially stressed medium and allows the retrieval of nonlinear elastic coefficients of tissues. In transverse isotropic medium such as muscles the theory leads to 9 different configurations of propagating shear waves (stress axis vs. fibers axis vs. shear wave polarization axis vs. shear wave propagation axis). In this work we propose to use 4 configurations to quantify these nonlinear parameters ex vivo and in vivo. Ex vivo experiments combining ultrasound shear wave elastography and mechanical testing were conducted on iliopsoas pig muscles to quantify three third-order nonlinear coefficients A, H and K that are possibly linked to the architectural structure of muscles. In vivo experiments were performed with human volunteers on biceps brachii during a stretching exercise on an ergometer. A combination of the third order nonlinear elastic parameters was assessed. The knowledge of this nonlinear elastic parameters paves the way to quantify in vivo the local forces produced by muscle during exercise, contraction or movements.

Tumor Solid Stress: Assessment with MR Elastography under Compression of Patient-Derived Hepatocellular Carcinomas and Cholangiocarcinomas Xenografted in Mice.

Malignant tumors have abnormal biomechanical characteristics, including high viscoelasticity, solid stress, and interstitial fluid pressure. Magnetic resonance (MR) elastography is increasingly used to non-invasively assess tissue viscoelasticity. However, solid stress and interstitial fluid pressure measurements are performed with invasive methods. We studied the feasibility and potential role of MR elastography at basal state and under controlled compression in assessing altered biomechanical features of malignant liver tumors. MR elastography was performed in mice with patient-derived, subcutaneously xenografted hepatocellular carcinomas or cholangiocarcinomas to measure the basal viscoelasticity and the compression stiffening rate, which corresponds to the slope of elasticity versus applied compression. MR elastography measurements were correlated with invasive pressure measurements and digital histological readings. Significant differences in MR elastography parameters, pressure, and histological measurements were observed between tumor models. In multivariate analysis, collagen content and interstitial fluid pressure were determinants of basal viscoelasticity, whereas solid stress, in addition to collagen content, cellularity, and tumor type, was an independent determinant of compression stiffening rate. Compression stiffening rate had high AUC (0.87 ± 0.08) for determining elevated solid stress, whereas basal elasticity had high AUC for tumor collagen content (AUC: 0.86 ± 0.08). Our results suggest that MR elastography compression stiffening rate, in contrast to basal viscoelasticity, is a potential marker of solid stress in malignant liver tumors.

Ultrafast ultrasound coupled with cervical magnetic stimulation for non-invasive and non-volitional assessment of diaphragm contractility.

KEY POINTS: Twitch transdiaphragmatic pressure elicited by cervical magnetic stimulation of the phrenic nerves is a fully non-volitional method for assessing diaphragm contractility in humans, yet it requires invasive procedures such as oesophageal and gastric catheter balloons.  Ultrafast ultrasound enables a very high frame rate allowing the capture of transient events, such as muscle contraction elicited by nerve stimulation (twitch). Whether indices derived from ultrafast ultrasound can be used as an alternative to the invasive measurement of twitch transdiaphragmatic pressure is unknown.  Our findings demonstrate that maximal diaphragm tissue velocity assessed using ultrafast ultrasound following cervical magnetic stimulation is reliable, sensitive to change in cervical magnetic stimulation intensity, and correlates to twitch transdiaphragmatic pressure.  This approach provides a novel fully non-invasive and non-volitional tool for the assessment of diaphragm contractility in humans. ABSTRACT: Measuring twitch transdiaphragmatic pressure (Pdi,tw ) elicited by cervical magnetic stimulation (CMS) is considered as a reference method for the standardized evaluation of diaphragm function. Yet, the measurement of Pdi requires invasive oesophageal and gastric catheter-balloons. Ultrafast ultrasound is a non-invasive imaging technique enabling frame rates high enough to capture transient events such as evoked muscle contractions. This study investigated relationships between indices derived from ultrafast ultrasound and Pdi,tw , and how these indices might be used to estimate Pdi,tw . CMS was performed in 13 healthy volunteers from 30% to 100% of maximal stimulator intensity in units of 10% in a randomized order. Pdi,tw was measured and the right hemidiaphragm was imaged using a custom ultrafast ultrasound sequence with 1 kHz framerate. Maximal diaphragm axial velocity (Vdi ,max ) and diaphragm thickening fraction (TFdi,tw ) were computed. Intra-session reliability was assessed. Repeated-measures correlation (R) and Spearman correlation coefficients (ρ) were used to assess relationships between variables. Intra-session reliability was strong for Pdi,tw and Vdi,max and moderate for TFdi,tw . Vdi,max correlated with Pdi,tw in all subjects (0.64 < ρ < 1.00, R = 0.75; all P < 0.05). TFdi,tw correlated with Pdi,tw in eight subjects only (0.85 < ρ < 0.93, R = 0.69; all P < 0.05). Coupling ultrafast ultrasound and CMS shows promise for the non-invasive and fully non-volitional assessment of diaphragm contractility. This approach opens up the prospect of both diagnosis and follow-up of diaphragm contractility in clinical populations.

Robust sound speed estimation for ultrasound-based hepatic steatosis assessment.

Hepatic steatosis is a common condition, the prevalence of which is increasing along with non-alcoholic fatty liver disease (NAFLD). Currently, the most accurate noninvasive imaging method for diagnosing and quantifying hepatic steatosis is MRI, which estimates the proton-density fat fraction (PDFF) as a measure of fractional fat content. However, MRI suffers several limitations including cost, contra-indications and poor availability. Although conventional ultrasound is widely used by radiologists for hepatic steatosis assessment, it remains qualitative and operator dependent. Interestingly, the speed of sound within soft tissues is known to vary slightly from muscle (1.575 mm · µs-1) to fat (1.450 mm · µs-1). Building upon this fact, steatosis could affect liver sound speed when the fat content increases. The main objectives of this study are to propose a robust method for sound speed estimation (SSE) locally in the liver and to assess its accuracy for steatosis detection and staging. This technique was first validated on two phantoms and SSE was assessed with a precision of 0.006 and 0.003 mm · µs-1 respectively for the two phantoms. Then a preliminary clinical trial (N = 17 patients) was performed. SSE results was found to be highly correlated with MRI proton density fat fraction (R 2 = 0.69) and biopsy (AUROC = 0.952) results. This new method based on the assessment of spatio-temporal properties of the local speckle noise for SSE provides an efficient way to diagnose and stage hepatic steatosis.

Feasibility and Diagnostic Accuracy of Supersonic Shear-Wave Elastography for the Assessment of Liver Stiffness and Liver Fibrosis in Children: A Pilot Study of 96 Patients.

PURPOSE: To evaluate the feasibility of using supersonic shear-wave elastography (SSWE) in children and normal values of liver stiffness with the use of control patients of different ages (from neonates to teenagers) and the diagnostic accuracy of supersonic shear wave elastography for assessing liver fibrosis by using the histologic scoring system as the reference method in patients with liver disease, with a special concern for early stages of fibrosis. MATERIALS AND METHODS: The institutional review board approved this prospective study. Informed consent was obtained from parents and children older than 7 years. First, 51 healthy children (from neonate to 15 years) were analyzed as the control group, and univariate and multivariate comparisons were performed to study the effect of age, transducer, breathing condition, probe, and position on elasticity values. Next, 45 children (from 1 month to 17.2 years old) who underwent liver biopsy were analyzed. SSWE measurements were obtained in the same region of the liver as the biopsy specimens. Biopsy specimens were reviewed in a blinded manner by a pathologist with the use of METAVIR criteria. The areas under the receiver operating characteristics curve (AUCs) were calculated for patients with fibrosis stage F0 versus those with stage F1-F2, F2 or higher, F3 or higher, and F4 or higher. RESULTS: A successful rate of SSWE measurement was 100% in 96 patients, including neonates. Liver stiffness values were significantly higher when an SC6-1 probe (Aixplorer; SuperSonic Imagine SA, Aix-enProvence, France) was used than when an SL15-4 probe (Aixplorer) was used (mean ± standard deviation, 6.94 kPa ± 1.42 vs 5.96 kPa ± 1.31; P = .006). There was no influence of sex, the location of measurement, or respiratory status on liver elasticity values (P = .41-.93), although the power to detect such a difference was low. According to the degree of liver fibrosis at liver biopsy, 88.5%-96.8% of patients were correctly classified, with AUCs of 0.90-0.98 (95% confidence interval [CI]: 0.8, 1.0). The AUC for patients with stage F0 versus stage F1-F2 was 0.93 (95% CI: 0.87, 0.99). CONCLUSION: SSWE allows accurate assessment of liver fibrosis, even in children with early stage (F1-F2) disease, and the choice of transducer influences liver stiffness values.

Assessment of the Cervix in Pregnant Women Using Shear Wave Elastography: A Feasibility Study.

The quantitative assessment of the cervix is crucial for the estimation of pre-term delivery risk and the prediction of the success of labor induction. We conducted a cross-sectional study using shear wave elastography based on the supersonic shear imaging technique. The shear wave speed (SWS) of the lower anterior part of the cervix was quantified over an 8-mm region of interest in 157 pregnant women. Cervical SWS is slightly but significantly reduced in patients diagnosed with pre-term labor and in patients who actually delivered pre-term.

Evaluation of Nonradiative Clinical Imaging Techniques for the Longitudinal Assessment of Tumour Growth in Murine CT26 Colon Carcinoma.

Background and Objectives. To determine the most appropriate technique for tumour followup in experimental therapeutics, we compared ultrasound (US) and magnetic resonance imaging (MRI) to characterize ectopic and orthotopic colon carcinoma models. Methods. CT26 tumours were implanted subcutaneously (s.c.) in Balb/c mice for the ectopic model or into the caecum for the orthotopic model. Tumours were evaluated by histology, spectrofluorescence, MRI, and US. Results. Histology of CT26 tumour showed homogeneously dispersed cancer cells and blood vessels. The visualization of the vascular network using labelled albumin showed that CT26 tumours were highly vascularized and disorganized. MRI allowed high-resolution and accurate 3D tumour measurements and provided additional anatomical and functional information. Noninvasive US imaging allowed good delineation of tumours despite an hypoechogenic signal. Monitoring of tumour growth with US could be accomplished as early as 5 days after implantation with a shorter acquisition time (<5 min) compared to MRI. Conclusion. MRI and US afforded excellent noninvasive imaging techniques to accurately follow tumour growth of ectopic and orthotopic CT26 tumours. These two techniques can be appropriately used for tumour treatment followup, with a preference for US imaging, due to its short acquisition time and simplicity of use.

Application of 1-D transient elastography for the shear modulus assessment of thin-layered soft tissue: comparison with supersonic shear imaging technique.

Elasticity estimation of thin-layered soft tissues has gained increasing interest propelled by medical applications like skin, corneal, or arterial wall shear modulus assessment. In this work, the authors propose one-dimensional transient elastography (1DTE) for the shear modulus assessment of thin-layered soft tissue. Experiments on three phantoms with different elasticities and plate thicknesses were performed. First, using 1DTE, the shear wave speed dispersion curve inside the plate was obtained and validated with finite difference simulation. No dispersive effects were observed and the shear wave speed was directly retrieved from time-of-flight measurements. Second, the supersonic shear imaging (SSI) technique (considered to be a gold standard) was performed. For the SSI technique, the propagating wave inside the plate is guided as a Lamb wave. Experimental SSI dispersion curves were compared with finite difference simulation and fitted using a generalized Lamb model to retrieve the plate bulk shear wave speed. Although they are based on totally different mechanical sources and induce completely different diffraction patterns for the shear wave propagation, the 1DTE and SSI techniques resulted in similar shear wave speed estimations. The main advantage of the 1DTE technique is that bulk shear wave speed can be directly retrieved without requiring a dispersion model.

Assessment of the mechanical properties of the musculoskeletal system using 2-D and 3-D very high frame rate ultrasound.

One of the great challenges for understanding muscular diseases is to assess noninvasively the active and passive mechanical properties of the musculoskeletal system. In this paper we report the use of ultrafast ultrasound imaging to explore with a submillimeter resolution the behavior of the contracting tissues in vivo (biceps brachii). To image the contraction, which is a very brief phenomenon (100 ms), a recently designed ultrasound scanner prototype able to take up to 6000 frames/s was used. A very high frame rate from 1000 to 2500 frames/s was used to image the cross section plane of the muscle (transverse to fibers) enabling us to catch in real time the muscle contraction during a transient electrostimulation. Tissue velocities were obtained from radiofrequency based speckle tracking techniques and their profiles are discussed with respect to electrostimulation intensities and pulse repetition frequencies for different volunteers. Three-dimensional (3-D) very high frame rate movies were also acquired by repeating the experiment for different acquisition planes while triggering the imaging system with the electrostimulation device. The reconstructed 3-D velocity field allows the full localization of the contracting fibers bundle. This ultrasound technique, referred to as echo mechanomyography, offers new perspectives for in vivo and in situ noninvasive muscle diagnosis of an active contractile tissue.

Quantitative assessment of breast lesion viscoelasticity: initial clinical results using supersonic shear imaging.

This paper presents an initial clinical evaluation of in vivo elastography for breast lesion imaging using the concept of supersonic shear imaging. This technique is based on the combination of a radiation force induced in tissue by an ultrasonic beam and an ultrafast imaging sequence capable of catching in real time the propagation of the resulting shear waves. The local shear wave velocity is recovered using a time-offlight technique and enables the 2-D mapping of shear elasticity. This imaging modality is implemented on a conventional linear probe driven by a dedicated ultrafast echographic device. Consequently, it can be performed during a standard echographic examination. The clinical investigation was performed on 15 patients, which corresponded to 15 lesions (4 cases BI-RADS 3, 7 cases BI-RADS 4 and 4 cases BI-RADS 5). The ability of the supersonic shear imaging technique to provide a quantitative and local estimation of the shear modulus of abnormalities with a millimetric resolution is illustrated on several malignant (invasive ductal and lobular carcinoma) and benign cases (fibrocystic changes and viscous cysts). In the investigated cases, malignant lesions were found to be significantly different from benign solid lesions with respect to their elasticity values. Cystic lesions have shown no shear wave propagate at all in the lesion (because shear waves do not propage in liquid). These preliminary clinical results directly demonstrate the clinical feasibility of this new elastography technique in providing quantitative assessment of relative stiffness of breast tissues. This technique of evaluating tissue elasticity gives valuable information that is complementary to the B-mode morphologic information. More extensive studies are necessary to validate the assumption that this new mode potentially helps the physician in both false-positive and false-negative rejection.

Assessment by transient elastography of the viscoelastic properties of blood during clotting.

Blood clotting is a natural process that can be both beneficial and life-threatening for the human body. It allows the maintenance of hemostasis after vascular injury, but it can also cause deep vein thrombosis and heart stroke. This study aimed better to understand the clotting process from a biomechanical point of view by using an acoustic method. The long-term objective is the staging of the age of clots in deep veins for therapy planning. The transient elastography method using a shear elasticity probe served to evaluate the shear wave velocity (V(S)) and shear wave attenuation (alpha(S)) of porcine whole blood during in vitro clot formation. By solving an inverse problem, it was then possible to provide images of the elasticity (mu(B)) and of the viscosity (eta(B)) from clotting blood. The time-varying elasticity and viscosity were very similar to what has been observed for the sol-gel transition of polymers. The mechanical properties of blood clot, which were modified by varying the hematocrit and by adding heparin or fibrinogen, were clearly assessed by the transient elastography technique. It is concluded that the shear elasticity probe is an appropriate tool to quantify and follow the sol-gel transition of blood during clotting.

Human muscle hardness assessment during incremental isometric contraction using transient elastography.

The aim of this study was to investigate the relationship between biceps brachii hardness using the transient elastography technique, and its activity level by quantifying the surface electromyographic signal (sEMG). Ten healthy subjects volunteered for this protocol. To assess the maximal biceps brachii myoelectric activity (sEMG-RMSm), subjects had to achieve their maximal voluntary contraction trial during an elbow flexion effort. They were then asked to perform an isometric biceps sEMG-RMS ramp trial in elbow flexion from 0% to 50% of their sEMG-RMSm in 120 s. A low-frequency pulse was sent every 5 s during all trials by an innovative shear elasticity probe previously placed over the belly of the biceps brachii allowing the calculation of a transverse shear modulus. The main results of this study were (i) the finding of a systematic linear relationship between the biceps brachii transverse shear moduli and the corresponding sEMG-RMS values. This was not the case when plotting transverse shear modulus versus the elbow flexion torque production. Therefore, the computation of a hardness index from the slope of individual transverse shear modulus-sEMG-RMS linear relationship was enabled; (ii) It was also found that the higher is the rest shear modulus, the lower is the hardness index, indicating that the transverse shear modulus change during contraction depends on its level at rest. Therefore, this non-invasive technique could be useful in the medical field to explore deep muscles which are unreachable by classical testing methods. It could also be applied for the follow-up of neuromuscular diseases inducing stiffness changes such as in Duchenne muscular dystrophy.

Assessment of elastic parameters of human skin using dynamic elastography.

Sonoelastography and transient elastography are two ultrasound-based techniques that facilitate noninvasive characterization of the viscoelastic properties of soft tissues by investigating their response to shear mechanical excitation. Young's modulus is the principle assessment parameter. Because it defines local tissue stiffness, it is of major interest for the medical imaging and cosmetic industries as it could replace subjective palpation by yielding local, quantitative information. In this paper, we describe a new high-resolution device capable of measuring local Young's modulus in very thin layers (1-5 mm) and devoted to the in vivo evaluation of the elastic properties of human skin. It uses an ultrasonic probe (50 MHz) for tracking the displacements induced by a 300 Hz shear wave generated by a ring surrounding the transducer. The displacements are measured using a conventional cross-correlation technique between successive ultrasonic back-scattered echoes. First, this noninvasive technique has been experimentally proven to be accurate for investigating elasticity in different skin-mimicking phantoms. Second, data were acquired in vivo on human forearms. As expected, Young's modulus was found to be higher in the dermis than in the hypodermis and other soft tissues.