Experimental investigation and theoretical modelling of the nonlinear acoustical behaviour of a liver tissue and comparison with a tissue mimicking hydrogel

Native harmonics generated by nonlinear distortion of ultrasound during propagation in a medium may cause misinterpretations in spectral analysis when studying contrast agents. The aim of this paper is to quantitatively evaluate nonlinear propagation effects of diagnostic ultrasound pulses in biological tissues and to assess whether a cellulose-based hydrogel can be a suitable material for tissue mimicking purposes. Hydrogel and pig liver tissue samples of various thicknesses were insonified in a through-transmission set-up, employing 2.25-MHz pulses with different mechanical index (MI) values (range 0.06–0.60). Second harmonic and first harmonic amplitudes were extracted from spectra of received signals and their ratio was then used to compare hydrogel and liver behaviours. Resulting trends are very similar for sample thicknesses up to 8 cm and highlight a significant increase in nonlinearity for MI > 0.3, for both liver and hydrogel. A numerical procedure was also employed to calculate pressure distribution along the beam axis: these theoretical results showed a very good agreement with experimental data in the low pressure range, though failed in predicting the MI threshold. In conclusion, the hydrogel resulted to be a suitable material for manufacturing tissue mimicking phantoms, in particular to study contrast agent behaviour with a “low power approach”.     

Development and characterization of a vitreous mimicking material for radiation force imaging

In many medical ultrasound applications tissue-mimicking phantoms are of fundamental importance for the performance of controlled experiments. Traditionally, such phantoms have been constructed using gelatin and agar gels. Although the use of these materials has become standard, few alternative materials have not been fully explored. In this paper, we present a protocol developed in our laboratory that reliably produces very soft, acrylamide-based phantoms that can mimic both acoustical and mechanical characteristics of the vitreous body of the eye. Following the described protocol, a series of phantoms were constructed ranging in acrylamide concentration from 1.60% to 1.70%. Measurements across the series yielded attenuation coefficients of 0.067-0.140 dB/cm/MHz, depending on acrylamide concentration. Speed of sound ranged between 1499 and 1510 m/s, also depending on acrylamide concentration. Published values for the vitreous gel indicate an attenuation of 0.10 dB/cm/MHz and a speed of sound of 1510 m/s, making our phantoms an excellent analog of this tissue. One application of these acrylamide phantoms is to test the efficacy of the Kinetic Acoustic Vitreoretinal Examination (KAVE), a tool developed in our laboratory with the potential to aid in the diagnosis of vitreoretinal disorders. KAVE utilizes acoustic radiation force to generate small, localized displacements within the vitreous-mimicking gel. These localized displacements are quantified to yield maximum displacement, relative elasticity, and relative viscosity images. We present KAVE images of a set of four phantoms with different gel concentrations. Although B-mode and relative viscosity images exhibit no significant differences, maximum displacement, and relative elasticity images clearly differentiate gels of different concentrations. Maximum displacements ranged between 30 and 5 microns, depending on acrylamide concentration. The results presented in this paper show that soft gel phantoms can be produced in a range of elasticities not previously reported, and that these phantoms are useful for testing ultrasound instruments designed for evaluation of the vitreous gel. Furthermore, the use of acrylamide-based gels may also offer a valuable and attractive alternative for many other ultrasound applications.     

Frequency-locked pulse sequencer for high-frame-rate monochromatic tissue motion imaging

To overcome the inherent low frame rate of conventional ultrasound, we have previously presented a system that can be implemented on conventional ultrasound scanners for high-frame-rate imaging of monochromatic tissue motion. The system employs a sector subdivision technique in the sequencer to increase the acquisition rate. To eliminate the delays introduced during data acquisition, a motion phase correction algorithm has also been introduced to create in-phase displacement images. Previous experimental results from tissue- mimicking phantoms showed that the system can achieve effective frame rates of up to a few kilohertz on conventional ultrasound systems. In this short communication, we present a new pulse sequencing strategy that facilitates high-frame-rate imaging of monochromatic motion such that the acquired echo signals are inherently in-phase. The sequencer uses the knowledge of the excitation frequency to synchronize the acquisition of the entire imaging plane to that of an external exciter. This sequencing approach eliminates any need for synchronization or phase correction and has applications in tissue elastography, which we demonstrate with tissue-mimicking phantoms.     

Improving thermal ablation delineation with electrode vibration elastography using a bidirectional wave propagation assumption

Thermal ablation procedures are commonly used to treat hepatic cancers and accurate ablation representation on shear wave velocity images is crucial to ensure complete treatment of the malignant target. Electrode vibration elastography is a shear wave imaging technique recently developed to monitor thermal ablation extent during treatment procedures. Previous work has shown good lateral boundary delineation of ablated volumes, but axial delineation was more ambiguous, which may have resulted from the assumption of lateral shear wave propagation. In this work, we assume both lateral and axial wave propagation and compare wave velocity images to those assuming only lateral shear wave propagation in finite element simulations, tissue-mimicking phantoms, and bovine liver tissue. Our results show that assuming bidirectional wave propagation minimizes artifacts above and below ablated volumes, yielding a more accurate representation of the ablated region on shear wave velocity images. Area overestimation was reduced from 13.4% to 3.6% in a stiff-inclusion tissue-mimicking phantom and from 9.1% to 0.8% in a radio-frequency ablation in bovine liver tissue. More accurate ablation representation during ablation procedures increases the likelihood of complete treatment of the malignant target, decreasing tumor recurrence.     

Adaptive imaging on a diagnostic ultrasound scanner at quasi real-time rates

Constructing an ultrasonic imaging system capable of compensating for phase errors in real-time is a significant challenge in adaptive imaging. We present a versatile adaptive imaging system capable of updating arrival time profiles at frame rates of approximately 2 frames per second (fps) with 1-D arrays and up to 0.81 fps for 1.75-D arrays, depending on the desired near-field phase correction algorithm. A novel feature included in this system is the ability to update the aberration profile at multiple beam locations for 1-D arrays. The features of this real-time adaptive imaging system are illustrated in tissue-mimicking phantoms with physical near-field phase screens and evaluated in clinical breast tissue with a 1.75-D array. The contrast-to-noise ratio (CNR) of anechoic cysts was shown to improve dramatically in the tissue-mimicking phantoms. In breast tissue, the width of point-like targets showed significant improvement: a reduction of 26.2% on average. Brightness of these targets, however, marginally decreased by 3.9%. For larger structures such as cysts, little improvement in features and CNR were observed, which is likely a result of the system assuming an infinite isoplanatic patch size for the 1.75-D arrays. The necessary requirements for constructing a real-time adaptive imaging system are also discussed.     

Real-time rectilinear volumetric imaging

Current real-time volumetric scanners use a 2-D array to scan a pyramidal volume consisting of many sector scans stacked in the elevation direction. This scan format is primarily useful for cardiac imaging to avoid interference from the ribs. However, a real-time rectilinear volumetric scan with a wider field of view close to the transducer could prove more useful for abdominal, breast, or vascular imaging. In previous work, computer simulations of very sparse array transducer designs in a rectilinear volumetric scanner demonstrated that a Mills cross array showed the best overall performance given current system constraints. Consequently, a 94×94 Mills cross array including 372 active channels operating at 5 MHz has been developed on a flexible circuit interconnect. In addition, the beam former delay software and scan converter display software of the Duke volumetric scanner were modified to achieve real-time rectilinear volumetric scanning consisting of a 30-mm×8-mm×60-mm scan at a rate of 47 volumes/s. Real-time rectilinear volumetric images were obtained of tissue-mimicking phantoms, showing a spatial resolution of 1 to 2 mm. Images of carotid arteries in normal subjects demonstrated tissue penetration to 6 cm     

A Review on the 3D Printing of Functional Structures for Medical Phantoms and Regenerated Tissue and Organ Applications

Medical models, or “phantoms,” have been widely used for medical training and for doctor-patient interactions. They are increasingly used for surgical planning, medical computational models, algorithm verification and validation, and medical devices development. Such new applications demand high-fidelity, patient-specific, tissue-mimicking medical phantoms that can not only closely emulate the geometric structures of human organs, but also possess the properties and functions of the organ structure. With the rapid advancement of three-dimensional (3D) printing and 3D bioprinting technologies, many researchers have explored the use of these additive manufacturing techniques to fabricate functional medical phantoms for various applications. This paper reviews the applications of these 3D printing and 3D bioprinting technologies for the fabrication of functional medical phantoms and bio-structures. This review specifically discusses the state of the art along with new developments and trends in 3D printed functional medical phantoms (i.e., tissue-mimicking medical phantoms, radiologically relevant medical phantoms, and physiological medical phantoms) and 3D bio-printed structures (i.e., hybrid scaffolding materials, convertible scaffolds, and integrated sensors) for regenerated tissues and organs.     

Skin color correction for tissue spectroscopy: demonstration of a novel approach with tissue-mimicking phantoms

The application of partial least squares (PLS) regression to visible-near-infrared (VIS-NIR) spectroscopy for modeling important blood and tissue parameters is generally complicated by the variation in skin pigmentation (melanin) across the human population. An orthogonal correction method for removing the influence of skin pigmentation has been demonstrated in diffuse reflectance spectra from two-layer tissue-mimicking phantoms. The absorption properties of the phantoms were defined by lyophilized human hemoglobin (bottom layer) and synthetic melanin (top layer). Tissue-like scattering was simulated in both layers with intralipid. The approach uses principal components analysis (PCA) loading vectors from a separate set of phantom spectra that encode the unwanted melanin variation to remove the effect of melanin from the test phantoms. The preprocessing of phantom spectra using this orthogonal correction method resulted in PLS models with reduced complexity and enhanced prediction performance. Preliminary results from a separate study that evaluates the feasibility of defining skin color variation in an experiment with a single human subject are also presented.     

Carboxymethyl derivative of scleroglucan: a novel thermosensitive hydrogel forming polysaccharide for drug delivery applications

A carboxymethyl derivative of scleroglucan (Scl-CM) with a derivatization degree of 65 ± 5% was synthesized. The rheological behaviour of this novel polymer was studied and compared with that of the starting polymer. We observed that the charged moieties carried on the chains could prevent the triple helix formation of Scl. Scl-CM aqueous solutions behave like true polymer solutions up to 1% w/v, whereas above this concentration a weak gel behaviour was observed. CaCl₂ addition to aqueous Scl-CM solutions led to a physical gel formation; the hydrogel strength was related to polymer and CaCl₂ concentrations. Temperature sweeps, registered at 1 Hz on hydrogels differing in CaCl₂ concentration, evidenced a gel → sol transition in the range of 30–40°C, depending on the molar ratio between carboxylic groups and Ca⁺². In order to verify a possible use of these hydrogels as drug delivery systems, acyclovir was loaded into the network. Rheological analysis evidenced that the loaded drug can affect the hydrogel elastic modulus. The release of acyclovir in phosphate buffer was evaluated at different temperatures in order to assess the suitability of this novel drug delivery system in topical applications.     

Spatial and temporal aberrator stability for real-time adaptive imaging

Reported real-time adaptive imaging systems use near-field phase correction techniques, which are desired because of their simple implementation and their compatibility with current system architectures. Aberrator stability is important to adaptive imaging because it defines the spatial and temporal limits for which the near-field phase estimates are valid. Spatial aberrator stability determines the required spatial sampling of the aberrator, and temporal aberrator stability determines the length of time for which the aberration profile can be used. In this study, the spatial and temporal stability of clinically measured aberrations is reported for breast, liver, and thyroid tissue. Cross correlations between aberration estimates revealed aberrators to have azimuthal isoplanatic patch sizes of 0.44, 0.28, and 0.20 mm for breast, liver, and thyroid tissue, respectively, at 80% correlation. Axial isoplanatic patch sizes were 1.26, 0.76, and 1.80 mm for the same tissue, respectively, at 80% correlation. Temporal stability at 80% correlation was determined to be greater than 1.5 seconds for breast and thyroid tissue, and 0.65 seconds for the liver. The effects of noise, motion, and target nonuniformity on aberrator stability are characterized by simulations and experiments in tissue mimicking phantoms.     

Lower-limb vascular imaging with acoustic radiation force elastography: Demonstration of in vivo feasibility

Acoustic radiation force impulse (ARFI) imaging characterizes the mechanical properties of tissue by measuring displacement resulting from applied ultrasonic radiation force. In this paper, we describe the current status of ARFI imaging for lower-limb vascular applications and present results from both tissue-mimicking phantoms and in vivo experiments. Initial experiments were performed on vascular phantoms constructed with polyvinyl alcohol for basic evaluation of the modality. Multilayer vessels and vessels with compliant occlusions of varying plaque load were evaluated with ARFI imaging techniques. Phantom layers and plaque are well resolved in the ARFI images, with higher contrast than B-mode, demonstrating the ability of ARFI imaging to identify regions of different mechanical properties. Healthy human subjects and those with diagnosed lower-limb peripheral arterial disease were imaged. Proximal and distal vascular walls are well visualized in ARFI images, with higher mean contrast than corresponding B-mode images. ARFI images reveal information not observed by conventional ultrasound and lend confidence to the feasibility of using ARFI imaging during lower-limb vascular workup.     

Graphite/poly (vinyl alcohol) hydrogel composite as porous ringy skirt for artificial cornea

Graphite/poly (vinyl alcohol) (PVA) hydrogel composites, which were designed as the porous ringy skirt surrounding the transparent core of a novel artificial cornea, were prepared by using the freeze/thawing process and the particle-leaching technique. The properties of the composites, including the water content, the mechanical strength, the porous architecture and the interactions between the graphite and PVA, were investigated. The tissue responses to the composite and pure PVA hydrogel were studied by in vivo implantation in the dorsal muscles of mice. The results showed that chemical interactions were present between the graphite and PVA in the composite, which benefited the combination of the two phases and enhanced the uniform distribution of graphite particles in the PVA matrix. However, the present of graphites in the PVA hydrogels reduced the tensile strength, elongation at break and water content of the composite. Moreover, the porous graphite/PVA hydrogel composite had interconnective pore structure with high porosity and enough mechanical strength. According to the histological analysis of 1 week and 12 weeks post-implantation, the graphite/PVA hydrogel composites showed less inflammatory reactions than the PVA hydrogels at the 1 week post-implantation. Moreover, compared to pure PVA hydrogel, the graphite/PVA hydrogel composite exhibited enhanced migration and infiltration of cells, and more neovascularization and tissue ingrowth. These in vivo characteristics will be beneficial for the long-term biofixation of artificial cornea. Therefore, the porous graphite/PVA hydrogel composite has a potential to be used as novel artificial cornea skirt.     

A new mammography dosimetric phantom

Breast phantoms produced with tissue-equivalent materials are used in an attempt to simulate glandular and adipose tissues, in terms of X-ray attenuation and density. In this work, a set of breast tissue-equivalent phantoms (BTE phantoms) with semicircular shapes of different thicknesses and compositions were produced. Such phantoms may be used in the measurement of the incident air kerma (K(i)) and the mean glandular dose (D(G)) delivered to patients undergoing mammography. To characterise the materials used to produce the phantoms, a series of 17-keV X-ray attenuation coefficient measurements were performed. The carbon-nitrogen-hydrogen elemental composition and the densities of the tissue-equivalent materials were also determined and compared with values available in the literature. Linear attenuation coefficients of 0.724 and 0.923 cm(-1) were determined, respectively, for adipose and glandular tissues. Such values agree with data available in the literature. On the basis of the results obtained in this work, it is suggested that BTE phantoms are used instead of polymethyl methacrylate phantoms to select exposure parameters (kV, mAs and target/filter combination) specific for breast glandularities from 0 to 50 % in the optimisation of doses in mammography.     

Microfluidic based platform for characterization of protein interactions in hydrogel nanoenvironments

Hydrogel posts in microfluidic devices were investigated as reaction environments for characterizing protein interactions with the goal of mimicking the complexity of a biological environment. The hydrogel environment can be easily tuned to study specific properties of the biological environment. In this study, the hydrogel pore size was tuned to mimic the effect of confinement/crowding on protein interactions. Arrays of polyacrylamide posts of different cross-link ratios (4 and 10%) were fabricated inside microfluidic channels via photopolymerization. Fluorescence-labeled proteins (protein A (PA) and immunoglobulins (IgG)) were transported into the posts via diffusion, and their interaction was studied using FRET. As the pore size of the hydrogel decreased, the binding between the proteins was enhanced. The degree to which crowding enhances a binding interaction depends on the intrinsic properties of the proteins; we observed that, inside the hydrogel post, the PA-goat IgG affinity was increased more than PA-rabbit IgG affinity. The integration of controlled nanoenvironments (hydrogels) with controlled microenvironments (microchannels) provides enhanced parametric control for studying protein interactions, which would be beneficial in developing sensors, in diagnostics, and for mimicking the biological environment at both the cell and the tissue level.     

Experimental and theoretical evaluation of a fiber-optic approach for optical property measurement in layered epithelial tissue

Improvements in measurement of epithelial tissue optical properties (OPs) in the ultraviolet and visible (UV-Vis) may lead to enhanced understanding of optical techniques for neoplasia detection. In this study, we investigated an approach based on fiber-optic measurement of reflectance to determine absorption and reduced scattering coefficients (μ(a) and μ(s)') in two-layer turbid media. Neural network inverse models were trained on simulation data for a wide variety of OP combinations (μ(a) = 1-22.5, μ(s)' = 5-42.5 cm(-1)). Experimental measurements of phantoms with top-layer thicknesses (D) ranging from 0.22 to 0.66 mm were performed at three UV-Vis wavelengths. OP estimation accuracy was calculated and compared to theoretical results. Mean prediction errors were strongly correlated with D and ranged widely, from 1.5 to 12.1 cm(-1). Theoretical analyses indicated the potential for improving accuracy with alternate probe geometries. Although numerous challenges remain, this initial experimental study of an unconstrained approach for fiber-optic-based OP determination in two-layer epithelial tissue indicates the potential to provide useful measurements.     

Coating process and early stage adhesion evaluation of poly(2-hydroxy-ethyl-methacrylate) hydrogel coating of 316L steel surface for stent applications

In this study, a spray-coating method has been set up with the aim to control the coating of poly(2-hydroxy-ethyl-methacrylate) (pHEMA), an hydrophilic polymeric hydrogel, onto the complex surface of a 316L steel stent for percutaneous coronary intervention (PCI). By varying process parameters, tuneable thicknesses, from 5 to 20 μm, have been obtained with uniform and homogeneous surface without crack or bridges. Surface characteristics of pHEMA coating onto metal surface have been investigated through FTIR-ATR, contact angle measurement, SEM, EDS and AFM. Moreover, results from Single-Lap-Joint and Pull-Off adhesion tests as well as calorimetric analysis of glass transition temperature suggested that pHEMA deposition is firmly adhered on metallic surface. The pHEMA coating evaluation of roughness, wettability together with its morphological and chemical stability after three cycles of expansion-crimping along with preliminary results after 6 months demonstrates the suitability of the coating for surgical implantation of stent.     

Adaptive clutter rejection filtering in ultrasonic strain-flow imaging

This paper introduces strain-flow imaging as a potential new technique for investigating vascular dynamics and tumor biology. The deformation of tissues surrounding pulsatile vessels and the velocity of fluid in the vessel are estimated from the same data set. The success of the approach depends on the performance of a digital filter that must separate echo signal components caused by flow from tissue motion components that vary spatially and temporally. Eigenfilters, which are an important tool for naturally separating signal components adaptively throughout the image, perform very well for this task. The method is examined using two tissue-mimicking flow phantoms that provide stationary and moving clutter associated with pulsatile flow.     

Effects of gold nanorod concentration on the depth-related temperature increase during hyperthermic ablation

The photothermal properties of gold nanorods (GNRs) provide an opportunity for the clinical application of highly efficient and tumor-specific photothermal therapy. For the effective hyperthermic ablation of tumor tissue using GNRs, it is essential to maintain a homogeneous therapeutic temperature in the target tissue during treatment. This study investigates whether the concentration of GNRs affects the distribution of the temperature increase during hyperthermal therapy. The investigation is conducted using polyacrylamide phantoms containing varying amounts of GNRs. In 0.1, 0.25, and 0.5 nM GNR-suspended phantoms, the change in temperature is relatively uniform along the depth of each phantom during laser irradiation at 2 W cm(-2) . In 1.0, 2.0, and 5.0 nM GNR-suspended phantoms, the rates of temperature increase in the deep regions of the phantoms decrease with increasing GNR concentration. At a laser irradiation of 5 W cm(-2) , the temperature of the GNR-suspended phantoms increases at a faster rate, whereas the range of GNR concentrations for maintaining the homogeneity of the temperature increase is not affected. This suggests that the concentration of GNRs is the major determinant of the depth-related temperature increase during hyperthermic ablation. Therefore, prior to the clinical application of hyperthermic ablation using GNRs, the concentration of GNRs has to be optimized to ensure a homogeneous distribution of therapeutic temperature in the targeted tissue.     

Use of frequency diversity and Nakagami statistics in ultrasonic tissue characterization

The Nakagami distribution was recently proposed as a generalized model for the envelope of the backscattered ultrasonic echo from tissue. The parameters of the Nakagami model were also shown to be useful in tissue characterization. This paper explores the possibility of enhancing the ability of these parameters for tissue characterization through the techniques of diversity and compounding. Frequency diversity has been used to create multiple versions of the envelope, which are then combined. This compounded envelope has been modeled, and its parameters have been analyzed. The ability of these new parameters to enhance tissue characterization is studied using computer simulation and experiments on tissue-mimicking phantoms. Results indicate that the use of frequency diversity and compounding may indeed improve the ability of the parameters of the Nakagami model to separate different number densities of scatterers. Therefore, it is suggested that such an approach may lead to better techniques in ultrasonic tissue characterization.     

The osteogenic differentiation of dog bone marrow mesenchymal stem cells in a thermo-sensitive injectable chitosan/collagen/β-glycerophosphate hydrogel: in vitro and in vivo

Type I collagen was added to the composite chitosan solution in a ratio of 1:2 to build a physical cross-linked self-forming chitosan/collagen/β-GP hydrogel. Osteogenic properties of this novel injectable hydrogel were evaluated. Gelation time was about 8 min which offered enough time for handling a mixture containing cells and the subsequent injection. Scanning electronic microscopy (SEM) observations indicated good spreading of bone marrow mesenchymal stem cells (BMSCs) in this hydrogel scaffold. Mineral nodules were found in the dog-BMSCs inoculated hydrogel by SEM after 28 days. After subcutaneous injection into nude mouse dorsum for 4 weeks, partial bone formation was observed in the chitosan/collagen/β-GP hydrogel loaded with pre-osteodifferentiated dog-BMSCs, which indicated that chitosan/collagen/β-GP hydrogel composite could induce osteodifferentiation in BMSCs without exposure to a continual supply of external osteogenic factors. In conclusion, the novel chitosan/collagen/β-GP hydrogel composite should prove useful as a bone regeneration scaffold.