Attenuation Compensation and Estimation.

Estimating the loss of ultrasound signal with propagation depth as a function of frequency is essential for quantifying tissue properties. Specifically, ultrasound attenuation is used to correct for spectral distortion prior to estimating quantitative ultrasound parameters to assess the tissue. Ultrasound attenuation can also be used independently to characterize the tissue. In this chapter, we review the primary algorithms for estimating both the local attenuation within a region of interest as well as the total attenuation between a region of interest and an ultrasound source. The strengths and weaknesses of each algorithm are also discussed.

Diagnostic Ultrasound Safety Review for Point-of-Care Ultrasound Practitioners.

Potential ultrasound exposure safety issues are reviewed, with guidance for prudent use of point-of-care ultrasound (POCUS). Safety assurance begins with the training of POCUS practitioners in the generation and interpretation of diagnostically valid and clinically relevant images. Sonographers themselves should minimize patient exposure in accordance with the as-low-as-reasonably-achievable principle, particularly for the safety of the eye, lung, and fetus. This practice entails the reduction of output indices or the exposure duration, consistent with the acquisition of diagnostically definitive images. Informed adoption of POCUS worldwide promises a reduction of ionizing radiation risks, enhanced cost-effectiveness, and prompt diagnoses for optimal patient care.

Scan Parameter Optimization for Histotripsy Treatment of S. Aureus Biofilms on Surgical Mesh.

There is a critical need to develop new noninvasive therapies to treat bacteria biofilms. Previous studies have demonstrated the effectiveness of cavitation-based ultrasound histotripsy to destroy these biofilms. In this study, the dependence of biofilm destruction on multiple scan parameters was assessed by conducting exposures at different scan speeds (0.3-1.4 beamwidths/s), step sizes (0.25-0.5 beamwidths), and the number of passes of the focus across the mesh (2-6). For each of the exposure conditions, the number of colony-forming units (CFUs) remaining on the mesh was quantified. A regression analysis was then conducted, revealing that the scan speed was the most critical parameter for biofilm destruction. Reducing the number of passes and the scan speed should allow for more efficient biofilm destruction in the future, reducing the treatment time.

Impact of High-Intensity Ultrasound on Strength of Surgical Mesh When Treating Biofilm Infections.

The use of cavitation-based ultrasound histotripsy to treat infections on surgical mesh has shown great potential. However, any impact of the therapy on the mesh must be assessed before the therapy can be applied in the clinic. The goal of this study was to determine if the cavitation-based therapy would reduce the strength of the mesh thus compromising the functionality of the mesh. First, Staphylococcus aureus biofilms were grown on the surgical mesh samples and exposed to high-intensity ultrasound pulses. For each exposure, the effectiveness of the therapy was confirmed by counting the number of colony forming units (CFUs) on the mesh. Most of the exposed meshes had no CFUs with an average reduction of 5.4-log10 relative to the sham exposures. To quantify the impact of the exposure on mesh strength, the force required to tear the mesh and the maximum mesh expansion before damage were quantified for control, sham, and exposed mesh samples. There was no statistical difference between the exposed and sham/control mesh samples in terms of ultimate tensile strength and corresponding mesh expansion. The only statistical difference was with respect to mesh orientation relative to the applied load. The tensile strength increased by 1.36 N while the expansion was reduced by 1.33 mm between different mesh orientations.

Histotripsy Treatment of S. Aureus Biofilms on Surgical Mesh Samples Under Varying Scan Parameters.

Cavitation-based ultrasound histotripsy has shown potential for treating infections on surgical mesh. The goal of this paper was to explore a new scan strategy while assessing the impact of scan speed, scan step size, and the number of cycles in the tone burst on the destruction of S. aureus biofilms grown on surgical mesh samples using ultrasound histotripsy pulses (150 MPa/-17 MPa). For each exposure, the number of colony forming units (CFUs) on the mesh and released onto the surrounding gel was quantified. Most of the exposed mesh samples had no CFUs, and there was a statistically significant reduction in CFUs on the mesh for each of the exposures, with an average reduction of 3.8 log10 relative to the sham. Compared with the sham, there was also a statistically significant reduction in CFUs on the gel with the highest exposures.

Histotripsy Treatment of S. Aureus Biofilms on Surgical Mesh Samples Under Varying Pulse Durations.

Prior studies demonstrated that histotripsy generated by high-intensity tone bursts to excite a bubble cloud adjacent to a medical implant can destroy the bacteria biofilm responsible for the infection. The goal of this paper was to treat Staphylococcus aureus (S. aureus) biofilms on surgical mesh samples while varying the number of cycles in the tone burst to minimize collateral tissue damage while maximizing therapy effectiveness. S. aureus biofilms were grown on 1-cm square surgical mesh samples. The biofilms were then treated in vitro using a spherically focused transducer (1.1 MHz, 12.9-cm focal length, 12.7-cm diameter) using either a sham exposure or histotripsy pulses with tone burst durations of 3, 5, or 10 cycles (pulse repetition frequency of 333 Hz, peak compressional pressure of 150 MPa, peak rarefactional pressure of 17 MPa). After treatment, the number of colony forming units (CFUs) on the mesh and the surrounding gel was independently determined. The number of CFUs remaining on the mesh for the sham exposure (4.8 ± 0.9-log10) (sample mean ± sample standard deviation-log10 from 15 observations) was statistically significantly different from the 3-cycle (1.9 ± 1.5-log10), 5-cycle (2.2 ± 1.1-log10), and 10-cycle exposures (1 ± 1.5-log10) with an average reduction in the number of CFUs of 3.1-log10. The numbers of CFUs released into the gel for both the sham and exposure groups were the same within a bound of 0.86-log10, but this interval was too large to deduce the fate of the bacteria in the biofilm following the treatment.

Beyond Cervical Length: A Pilot Study of Ultrasonic Attenuation for Early Detection of Preterm Birth Risk.

The purpose of this study was to determine whether cervical ultrasonic attenuation could identify women at risk of spontaneous preterm birth. During pregnancy, women (n = 67) underwent from one to five transvaginal ultrasonic examinations to estimate cervical ultrasonic attenuation and cervical length. Ultrasonic data were obtained with a Zonare ultrasound system with a 5- to 9-MHz endovaginal transducer and processed offline. Cervical ultrasonic attenuation was lower at 17-21 wk of gestation in the SPTB group (1.02 dB/cm-MHz) than in the full-term birth groups (1.34 dB/cm-MHz) (p = 0.04). Cervical length was shorter (3.16 cm) at 22-26 wk in the SPTB group than in the women delivering full term (3.68 cm) (p = 0.004); cervical attenuation was not significantly different at this time point. These findings suggest that low attenuation may be an additional early cervical marker to identify women at risk for SPTB.

Correlation of Hemorrhage Near Developing Opossum Skull With Pulsed Ultrasound Exposure Parameters.

OBJECTIVES: High-intensity focused ultrasound (HIFU) has been used noninvasively for therapeutic applications. Before HIFU can be used therapeutically on a human fetus, the bioeffects related to HIFU must be studied, and the mechanism causing the bioeffects should be understood. Previous studies have shown that HIFU, when targeted on fetal rat and mice bones. resulted in hemorrhage. However, the mechanism responsible has not been identified. In this study, we looked at ultrasound parameters related to hemorrhage in an effort to better understand the mechanism. METHODS: Brazilian opossum pups (7-8 postnatal days) were exposed to a 1.1-MHz f/1 spherically focused transducer (6.3 cm focal length). Four treatment groups of n = 14 and a control group of n = 14 were exposed to rarefactional pressures of 3.6 to 6 MPa with spatial-peak temporal average intensity values of 5.4 to 10.8 W/cm(2). The pulse repetition frequency was varied from 500 to 1000 Hz with exposure durations of 1 to 4 minutes. RESULTS: Four groups with sample sizes of 14 had hemorrhage percentages of 43%, 36%, 29%, and 36%, respectively. Hemorrhage occurrence and size were found to correlate strongly with the nonlinear product of energy density and number of pulses, with correlation values of 0.92 and 0.97, respectively. CONCLUSIONS: The dependence of hemorrhage on energy density and the number of pulses suggests that the hemorrhage may be due to high-stress, low-cycle mechanical fatigue damage. Hence, for therapeutic applications, the product of energy density and number of pulses should not exceed a certain predetermined limit.

Development of an ultrasonic method to detect cervical remodeling in vivo in full-term pregnant women.

The objective of this study was to determine whether estimates of ultrasonic attenuation could detect changes in the cervix associated with medically induced cervical remodeling. Thirty-six full-term pregnant women underwent two transvaginal ultrasonic examinations separated in time by 12 h to determine cervical attenuation, cervical length and changes thereof. Ultrasonic attenuation and cervical length data were acquired from a zone (Zonare Medical Systems, Mountain View, CA, USA) ultrasound system using a 5-9 MHz endovaginal probe. Cervical attenuation and cervical length significantly decreased in the 12 h between the pre-cervical ripening time point and 12 h later. The mean cervical attenuation was 1.1 ± 0.4 dB/cm-MHz before cervical ripening agents were used and 0.8 ± 0.4 dB/cm-MHz 12 h later (p < 0.0001). The mean cervical length also decreased from 3.1 ± 0.9 cm before the cervical ripening was administered to 2.0 ± 1.1 cm 12 h later (p < 0.0001). Cervical attenuation and cervical length detected changes in cervical remodeling 12 h after cervical ripening administration.

Dependence of ablative ability of high-intensity focused ultrasound cavitation-based histotripsy on mechanical properties of agar.

Cavitation-based histotripsy uses high-intensity focused ultrasound at low duty factor to create bubble clouds inside tissue to liquefy a region, and provides better fidelity to planned lesion coordinates and the ability to perform real-time monitoring. The goal of this study was to identify the most important mechanical properties for predicting lesion dimensions, among these three: Young's modulus, bending strength, and fracture toughness. Lesions were generated inside tissue-mimicking agar, and correlations were examined between the mechanical properties and the lesion dimensions, quantified by lesion volume and by the width and length of the equivalent bubble cluster. Histotripsy was applied to agar samples with varied properties. A cuboid of 4.5 mm width (lateral to focal plane) and 6 mm depth (along beam axis) was scanned in a raster pattern with respective step sizes of 0.75 and 3 mm. The exposure at each treatment location was either 15, 30, or 60 s. Results showed that only Young's modulus influenced histotripsy's ablative ability and was significantly correlated with lesion volume and bubble cluster dimensions. The other two properties had negligible effects on lesion formation. Also, exposure time differentially affected the width and depth of the bubble cluster volume.

Flow rate and duty cycle effects in lysis of Chlamydomonas reinhardtii using high-energy pulsed focused ultrasound.

To consider microalgae lipid biofuel as a viable energy source, it is a necessity to maximize algal cell lysis, lipid harvest, and thus biofuel production versus the energy used to lyse the cells. Previous techniques have been to use energy consumptive ultrasound waves in the 10-40 kHz range in a stationary exposure environment. This study evaluated the potential of using 1.1 MHz ultrasound pulses in a new flow through type chamber on Chlamydomonas reinhardtii as a model organism for cell breakage. The ultrasound was generated using a spherically focused transducer with a focal length of 6.34 cm and an active diameter of 6.36 cm driven by 20 cycle sine-wave tone bursts at varied pulse repetition frequencies. First, variations in flow rate were examined at a constant duty cycle of 3.6%. After assessing flow rates, the duty cycle was varied to further explore the dependence on the tone burst parameters. Cell lysis was assessed by quantifying protein and chlorophyll release into the supernatant as well as by lipid extractability. Appropriate flow rates with higher duty cycles led to statistically significant increases in cell lysis relative to controls and other exposure conditions.

Lysis of Chlamydomonas reinhardtii by high-intensity focused ultrasound as a function of exposure time.

Efficient lysis of microalgae for lipid extraction is an important concern when processing biofuels. Historically, ultrasound frequencies in the range of 10-40 kHz have been utilized for this task. However, greater efficiencies might be achievable if higher frequencies could be used. In our study, we evaluated the potential of using 1.1 MHz ultrasound to lyse microalgae for biofuel production while using Chlamydomonas reinhardtii as a model organism. The ultrasound was generated using a spherically focused transducer with a focal length of 6.34 cm and an active diameter of 6.36 cm driven by 20 cycle sine-wave tone bursts at a pulse repetition frequency of 2 kHz (3.6% duty cycle). The time-average acoustic power output was 26.2 W while the spatial-peak-pulse-average intensity (ISPPA) for each tone burst was 41 kW/cm(2). The peak compressional and rarefactional pressures at the focus were 102 and 17 MPa, respectively. The exposure time was varied for the different cases in the experiments from 5s to 9 min and cell lysis was assessed by quantifying the percentage of protein and chlorophyll release into the supernate as well as the lipid extractability. Free radical generation and lipid oxidation for the different ultrasound exposures were also determined. We found that there was a statistically significant increase in lipid extractability for all of the exposures compared to the control. The longer exposures also completely fragmented the cells releasing almost all of the protein and chlorophyll into the supernate. The cavitation activity did not significantly increase lipid oxidation while there was a minor trend of increased free radical production with increased ultrasound exposure.

Effect of pulse repetition frequency and scan step size on the dimensions of the lesions formed in agar by HIFU histotripsy.

Histotripsy uses high-intensity focused ultrasound pulses at low duty cycle to generate energetic bubble clouds inside tissue to fractionate a region. As a potential tumor treatment modality, this cavitation-based non-invasive technique has the advantages of easy monitoring and sharp borders. Aiming at therapy efficiency, we experimentally investigated the effects of pulse repetition frequency (PRF) and lateral scan step size on the dimensions of lesions formed through HIFU histotripsy in agar mimicking tissue in terms of mechanical (not acoustical) properties. The single-element spherically focused source (1.1 MHz, 6.34 cm focal length, f/1) was excited to reach the peak compressional and rarefactional pressures of ~102 and 17 MPa, respectively. A targeted rectangular block of 4.5 mm wide (lateral) and 6mm deep (axial) was scanned in a raster pattern with a constant axial step size of 3mm. The lateral step size was varied between 375, 750, 1500, 2250 and 4500 µm. Pulses at each treatment location consisted of 5000 20-cycle sine wave tone bursts with the PRF of 167, 333 or 1000 Hz. Results suggested that the bubble activity region could extend beyond the -3 dB region and that refining the lateral scan mesh and/or increasing PRF enlarged the lesion extent. The 1500 µm-333 Hz and the 1500 µm-1 kHz conditions were in a more favorable position to be viewed as optimal with regard to lesion volume generation rate, bubble activity region width, and the potential for thermal damage.

Techniques and evaluation from a cross-platform imaging comparison of quantitative ultrasound parameters in an in vivo rodent fibroadenoma model.

This contribution demonstrates that quantitative ultrasound (QUS) capabilities are platform independent, using an in vivo model. Frequency-dependent attenuation estimates, backscatter coefficient, and effective scatterer diameter estimates are shown to be comparable across four different ultrasound imaging systems with varied processing techniques. The backscatter coefficient (BSC) is a fundamental material property from which several QUS parameters are estimated; therefore, consistent BSC estimates among different systems must be demonstrated. This study is an intercomparison of BSC estimates acquired by three research groups (UIUC, UW, ISU) from four in vivo spontaneous rat mammary fibroadenomas using three clinical array systems and a single-element laboratory scanner system. Because of their highly variable backscatter properties, fibroadenomas provided an extreme test case for BSC analysis, and the comparison is across systems for each tumor, not across the highly heterogeneous tumors. RF echo data spanning the 1 to 12 MHz frequency range were acquired in three dimensions from all animals using each system. Each research group processed their RF data independently, and the resulting attenuation, BSC, and effective scatterer diameter (ESD) estimates were compared. The attenuation estimates across all systems showed the same trends and consistently fit the power-law dependence on frequency. BSCs varied among the multiple slices of data acquired by each transducer, with variations between transducers being of a similar magnitude as those from slice to slice. Variation between BSC estimates was assessed via functional signal-to-noise ratios derived from backscatter data. These functional signal-to-noise ratios indicated that BSC versus frequency variations between systems ranged from negligible compared with the noise level to roughly twice the noise level. The corresponding functional analysis of variance (fANOVA) indicated statistically significant differences between BSC curves from different systems. However, root mean squared difference errors of the BSC values (in decibels) between different transducers and imaging platforms were less than half of the BSC magnitudes in most cases. Statistical comparison of the effective scatterer diameter (ESD) estimates resulted in no significant differences in estimates from three of the four transducers used for those estimates, demonstrating agreement among estimates based on the BSC. This technical advance demonstrates that these in vivo measurements can be made in a system-independent manner; the necessary step toward clinical implementation of the technology.

Precision control of lesions by high-intensity focused ultrasound cavitation-based histotripsy through varying pulse duration.

The goal of this experimental study was to explore the feasibility of acquiring controllable precision through varying pulse duration for lesions generated by cavitation-based histotripsy. Histotripsy uses high-intensity focused ultrasound (HIFU) at low duty factor to create energetic bubble clouds inside tissue to liquefy a region. It uses cavitation-mediated mechanical effects while minimizing heating, and has the advantages of real-time monitoring and lesion fidelity to treatment planning. In our study, histotripsy was applied to three groups of tissue-mimicking agar samples of different stiffnesses (29.4 ± 5.3, 44.8 ± 5.9, and 66.4 ± 7.1 kPa). B-mode imaging was used first to quantify bubble cluster dimensions in both water and agar. Then, a 4.5-mm-wide square (lateral to the focal plane) was scanned in a raster pattern with a step size of 0.75 mm in agar histotripsy experiments to estimate equivalent bubble cluster dimensions based on the histotripsyinduced damage. The 15-s exposure at each treatment location comprised 5000 sine-wave tone bursts at a spatial-peak pulseaverage intensity of 41.1 kW/cm2, with peak compressional and rarefactional pressures of 102 and 17 MPa, respectively. The results showed that bubble cluster width and length increased with pulse duration and decreased with agar stiffness. Therefore, a significant improvement in histotripsy precision could be achieved by reducing the pulse duration.

Assessment of ultrasound histotripsy-induced damage to ex vivo porcine muscle.

OBJECTIVES: Cavitation-based histotripsy uses high-intensity focused ultrasound pulses at a low duty cycle to generate energetic bubble clouds inside tissue to fractionate cells and is a potential noninvasive tumor treatment modality. Aiming at determining therapy efficiency, we experimentally investigated the effects of pulse repetition frequency and lateral scan step size on the degree of damage of histotripsy-induced lesions in porcine muscle tissue. METHODS: A single-element spherically focused source (1.1 MHz, 6.34-cm focal length, f/1) was excited to reach the peak compressional and rarefactional pressures of approximately 102 and 17 MPa, respectively. A targeted square of 9 mm wide (lateral to focal plane) was scanned in a raster pattern with the step sizes of 375, 750, 1500, 2250, and 4500 µm. Pulses at each treatment location consisted of 5000 20-cycle sine wave tone bursts with a pulse repetition frequency of 167, 333, or 1000 Hz. Histopathologic examination and image processing were performed to evaluate the tissue damage for each experimental condition. RESULTS: Skeletal myofiber damage was successfully created with our 7 exposure conditions. Three scales for muscle damage were identified through performing quad-tree decomposition to photomicrograph images and then relating decomposition with lesion homogeneity. CONCLUSIONS: Using a finer scan step size promoted the lesion homogeneity. Selection of the optimal condition does not depend solely on the comparison of tissue damage. Given the uncertainty on which of the 3 scales for tissue damage allows muscle repair, 2 conditions were identified as optimal: the 1500 µm-333 Hz condition for scale 3 (related to mild damage) and the 750 µm-333 Hz condition for scale 1 (related to severe damage).

Cross-imaging system comparison of backscatter coefficient estimates from a tissue-mimicking material.

A key step toward implementing quantitative ultrasound techniques in a clinical setting is demonstrating that parameters such as the ultrasonic backscatter coefficient (BSC) can be accurately estimated independent of the clinical imaging system used. In previous studies, agreement in BSC estimates for well characterized phantoms was demonstrated across different laboratory systems. The goal of this study was to compare the BSC estimates of a tissue mimicking sample measured using four clinical scanners, each providing RF echo data in the 1-15 MHz frequency range. The sample was previously described and characterized with single-element transducer systems. Using a reference phantom for analysis, excellent quantitative agreement was observed across the four array-based imaging systems for BSC estimates. Additionally, the estimates from data acquired with the clinical systems agreed with theoretical predictions and with estimates from laboratory measurements using single-element transducers.

Impact of preconditioning pulse on lesion formation during high-intensity focused ultrasound histotripsy.

Therapeutic applications with high-intensity focused ultrasound (HIFU) fall into two classifications-one using thermal effect for coagulation or ablation while generally avoiding cavitation and the other using cavitation-mediated mechanical effects while suppressing heating. Representative of the latter, histotripsy uses HIFU at low duty factor to create energetic bubble clouds inside tissue to liquefy a region and has the advantages in real-time monitoring and lesion fidelity to treatment planning. We explored the impact of a preconditioning/heating pulse on histotripsy lesion formation in porcine muscle samples. During sonication, a targeted square region 9 mm wide (lateral to the focal plane) was scanned in a raster pattern with a step size of 0.75 mm. The 20-s exposure at each treatment location consisted of a 5-s duration preconditioning burst at spatial-peak intensities from 0-1386 W/cm² followed by 5000 tone bursts at high intensity (with spatial-peak pulse-average intensity of 47.34 kW/cm², spatial-peak temporal-average intensity of 284 W/cm², peak compressional pressure of 102 MPa and peak rarefactional pressure of 17 MPa). The temperature increase for all exposures was measured using a thermal imager immediately after each exposure. Lesion volume increased with increasing amplitude of the preconditioning pulse until coagulation was observed, but lesion width/area did not change significantly with the amplitude. In addition, the lesion dimensions became smaller when the global tissue temperature was raised before applying the histotripsy pulsing sequence. Therefore, the benefit of the preconditioning pulse was not caused by global heating.