Ultrasound shear wave elastography methods are commonly used for estimation of mechanical properties of soft biological tissues in diagnostic medicine. A limitation of most currently used elastography methods is that they yield only the shear storage modulus ( G ) but not the loss modulus ( G ). Therefore, no information on viscosity or loss tangent (tan δ) is provided. In this paper, an ultrasound shear wave viscoelastography method is developed for model-independent quantification of frequency-dependent viscoelastic complex shear modulus of macroscopically homogeneous tissues. Three in vitro tissue-mimicking phantoms and two ex vivo porcine liver samples were evaluated. Shear waves were remotely induced within the samples using several acoustic radiation force pushes to generate a semicylindrical wave field similar to those generated by most clinically used elastography systems. The complex shear modulus was estimated over a broad frequency range (up to 1000 Hz) through the analytical solution of the developed inverse wave propagation problem using the measured shear wave speed and amplitude decay versus propagation distance. The shear storage and loss moduli obtained for the in vitro phantoms were compared with those from a planar shear wave method and the average differences over the whole frequency range studied were smaller than 7% and 15%, respectively. The reliability of the proposed method highlights its potential for viscoelastic tissue characterization, which may improve noninvasive diagnosis.
In vivo quantification of shear-wave attenuation in soft tissues may help to better understand human tissue rheology and lead to new diagnostic strategies. Attenuation is difficult to measure in acoustic radiation force elastography because the shear-wave amplitude decreases due to a combination of diffraction and viscous attenuation. Diffraction correction requires assuming a cylindrical wavefront and an isotropic propagation medium, which may not be the case in some applications. In this paper, the frequency-shift method, used in ultrasound imaging and seismology, was adapted for shear-wave attenuation measurement in elastography. This method is not sensitive to diffraction effects. For a linear frequency dependence of the attenuation, a closed-form relation was obtained between the decrease in the peak frequency of the gamma-distributed wave amplitude spectrum and the attenuation coefficient of the propagation medium. The proposed method was tested against a plane-wave reference method in homogeneous agar-gelatin phantoms with 0%, 10%, and 20% oil concentrations, and hence different attenuations of 0.117, 0.202, and 0.292 [Formula: see text]/Hz, respectively. Applicability to biological tissues was demonstrated with two ex vivo porcine liver samples (0.79 and 1.35 [Formula: see text]/Hz) and an in vivo human muscle, measured along (0.43 [Formula: see text]/Hz) and across (1.77 [Formula: see text]/Hz) the tissue fibers. In all cases, the data supported the assumptions of a gamma-distributed spectrum for the source and linear frequency attenuation for the tissue. This method provides tissue attenuation, which is relevant diagnostic information to model viscosity, in addition to shear-wave velocity used to assess elasticity. Data processing is simple and could be performed automatically in real time for clinical applications.
A characterization method based on Rayleigh wave propagation was developed for the quantification of the frequency-dependent viscoelastic properties of soft materials at high frequencies; i.e., up to 4 kHz. Planar harmonic surface waves were produced on the surface of silicone rubber samples. The phase and amplitude of the propagating waves were measured at different locations along the propagation direction, which allowed the calculation of the complex Rayleigh wavenumbers at each excitation frequency using a transfer function method. An inverse wave propagation problem was then solved to obtain the complex shear/elastic moduli from the measured wavenumbers. In a separate, related investigation, dynamic indentation tests using atomic force microscopy (AFM) were performed at frequencies up to 300 Hz. No systematic verification study is available for the AFM-based method, which can be used when the dimensions of the test samples are too small for other existing testing methods. The results obtained from the Rayleigh wave propagation and AFM-based indentation methods were compared with those from a well-established method, which involves the generation of standing longitudinal compression waves in rod-shaped test specimens. The results were cross validated and qualitatively confirmed theoretical expectations presented in the literature for the frequency-dependence of polymers.
The human vocal folds (VFs) undergo complex biomechanical stimulation during phonation. The aim of the present study was to develop and validate a phono-mimetic VF flow perfusion bioreactor, which mimics the mechanical microenvironment of the human VFs in vitro. The bioreactor uses airflow-induced self-oscillations, which have been shown to produce mechanical loading and contact forces that are representative of human phonation. The bioreactor consisted of two synthetic VF replicas within a silicone body. A cell-scaffold mixture (CSM) consisting of human VF fibroblasts, hyaluronic acid, gelatin, and a polyethylene glycol cross-linker was injected into cavities within the replicas. Cell culture medium (CCM) was perfused through the scaffold by using a customized secondary flow loop. After the injection, the bioreactor was operated with no stimulation over a 3-day period to allow for cell adaptation. Phonation was subsequently induced by using a variable speed centrifugal blower for 2 h each day over a period of 4 days. A similar bioreactor without biomechanical stimulation was used as the nonphonatory control. The CSM was harvested from both VF replicas 7 days after the injection. The results confirmed that the phono-mimetic bioreactor supports cell viability and extracellular matrix proteins synthesis, as expected. Many scaffold materials were found to degrade because of challenges from phonation-induced biomechanical stimulation as well as due to biochemical reactions with the CCM. The bioreactor concept enables future investigations of the effects of different phonatory characteristics, that is, voice regimes, on the behavior of the human VF cells. It will also help study the long-term functional outcomes of the VF-specific biomaterials before animal and clinical studies.
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