Testing the Efficacy of Contrast-Enhanced Ultrasound in Detecting Transplant Rejection Using a Murine Model of Heart Transplantation.

One of the key unmet needs to improve long-term outcomes of heart transplantation is to develop accurate, noninvasive, and practical diagnostic tools to detect transplant rejection. Early intragraft inflammation and endothelial cell injuries occur prior to advanced transplant rejection. We developed a novel diagnostic imaging platform to detect early declines in microvascular perfusion (MP) of cardiac transplants using contrast-enhanced ultrasonography (CEUS). The efficacy of CEUS in detecting transplant rejection was tested in a murine model of heart transplants, a standard preclinical model of solid organ transplant. As compared to the syngeneic groups, a progressive decline in MP was demonstrated in the allografts undergoing acute transplant rejection (40%, 64%, and 92% on days 4, 6, and 8 posttransplantation, respectively) and chronic rejection (33%, 33%, and 92% on days 5, 14, and 30 posttransplantation, respectively). Our perfusion studies showed restoration of MP following antirejection therapy, highlighting its potential to help monitor efficacy of antirejection therapy. Our data suggest that early endothelial cell injury and platelet aggregation contributed to the early MP decline observed in the allografts. High-resolution MP mapping may allow for noninvasive detection of heart transplant rejection. The data presented have the potential to help in the development of next-generation imaging approaches to diagnose transplant rejection.

Targeted disruption of the blood-brain barrier with focused ultrasound: association with cavitation activity.

Acoustic emission was monitored during focused ultrasound exposures in conjunction with an ultrasound contrast agent (Optison) in order to determine if cavitation activity is associated with the induction of blood-brain barrier disruption (BBBD). Thirty-four locations were sonicated (frequency: 260 kHz) at targets 10 mm deep in rabbit brain (N = 9). The sonications were applied at peak pressure amplitudes ranging from 0.11 to 0.57 MPa (burst length: 10 ms; repetition frequency of 1 Hz; duration: 20 s). Acoustic emission was recorded with a focused passive cavitation detector. This emission was recorded at each location during sonications with and without Optison. Detectable wideband acoustic emission was observed only at 0.40 and 0.57 MPa. BBBD was observed in contrast MRI after sonication at 0.29-0.57 MPa. The appearance of small regions of extravasated erythrocytes appeared to be associated with this wideband emission signal. The results thus suggest that BBBD resulting from focused ultrasound pulses in the presence of Optison can occur without indicators for inertial cavitation in vivo, wideband emission and extravasation. If inertial cavitation is not responsible for the BBBD, other ultrasound/microbubble interactions are likely the source. A significant increase in the emission signal due to Optison at the second and third harmonics of the ultrasound driving frequency was found to correlate with BBBD and might be useful as an online method to indicate when the disruption occurs.

Quantitative MRI-based temperature mapping based on the proton resonant frequency shift: review of validation studies.

MRI-based temperature imaging that exploits the temperature-sensitive water proton resonant frequency shift is currently the only available method for reliable quantification of temperature changes in vivo. Extensive pre-clinical work has been performed to validate this method for guiding thermal therapies. That work has shown the method to be useful for all stages of the thermal therapy, from resolving heating below the threshold for damage to ensuring that the thermal exposure is sufficient within the target volume and protecting surrounding critical structures and to accurately predicting the extent of the ablated volume. In this paper, these validation studies will be reviewed. In addition, clinical studies that have shown this method feasible in human treatments will be overviewed.

MRI guided and monitored focused ultrasound thermal ablation methods: a review of progress.

This paper reviews the current status in using magnetic resonance imaging (MRI) to guide and monitor thermal coagulation of tumours using focused ultrasound. The patient treatment procedure with a second generation phased array system will be described. Several clinical trials have found that patient treatments are feasible and that MRI thermometry allows noninvasive monitoring of clinical treatments. Overall, this emerging modality holds significant potential for non-invasive tumour treatment of both benign and malignant tumours.

Non-invasive opening of BBB by focused ultrasound.

Blood brain barrier (BBB) is a major barrier for delivering therapeutic agents in the brain. In this study we investigated the feasibility of open the BBB by using focused ultrasound. Rabbit brains were exposed to pulsed focused ultrasound while injecting ultrasound contrast agent containg microbubbles intravenously. The BBB opening was measured after the sonications by injecting MRI contrast agent i.v. and evaluating the local enhancement in the brain. Low ultrasound powers and pressure amplitudes were found to cause focal enhancement. Before sacrificing the animals trypan blue was also injected i.v.. After the sacrifice of the animals blue spots were found in the brain in the sonicated locations. This method may have potential for targeted delivery of macromolecules in the brain.

Noninvasive MR imaging-guided focal opening of the blood-brain barrier in rabbits.

PURPOSE: To determine if focused ultrasound beams can be used to locally open the blood-brain barrier without damage to surrounding brain tissue and if magnetic resonance (MR) imaging can be used to monitor this procedure. MATERIALS AND METHODS: The brains of 18 rabbits were sonicated (pulsed sonication) in four to six locations, with temporal peak acoustic power ranging from 0.2 to 11.5 W. Prior to each sonication, a bolus of ultrasonographic (US) contrast agent was injected into the ear vein of the rabbit. A series of fast or spoiled gradient-echo MR images were obtained during the sonications to monitor the temperature elevation and potential tissue changes. Contrast material-enhanced MR images obtained minutes after sonications and repeated 1-48 hours later were used to depict blood-brain barrier opening. Whole brain histologic evaluation was performed. RESULTS: Opening of the blood-brain barrier was confirmed with detection of MR imaging contrast agent at the targeted locations. The lowest power levels used produced blood-brain barrier opening without damage to the surrounding neurons. Contrast enhancement correlated with the focal signal intensity changes in the magnitude fast spoiled gradient-echo MR images. CONCLUSION: The blood-brain barrier can be consistently opened with focused ultrasound exposures in the presence of a US contrast agent. MR imaging signal intensity changes may be useful in the detection of blood-brain barrier opening during sonication.

Comparison of modelled and observed in vivo temperature elevations induced by focused ultrasound: implications for treatment planning.

Two numerical models for predicting the temperature elevations resulting from focused ultrasound heating of muscle tissue were tested against experimental data. Both models use the Rayleigh-Sommerfeld integral to calculate the pressure field from a source distribution. The first method assumes a source distribution derived from a uniformly radiating transducer whereas the second uses a source distribution obtained by numerically projecting pressure field measurements from an area near the focus backward toward the transducer surface. Both of these calculated ultrasound fields were used as heat sources in the bioheat equation to calculate the temperature elevation in vivo. Experimental results were obtained from in vivo rabbit experiments using eight-element sector-vortex transducers at 1.61 and 1.7 MHz and noninvasive temperature mapping with MRI. Results showed that the uniformly radiating transducer model over-predicted the peak temperature by a factor ranging from 1.4 to 2.8, depending on the operating mode. Simulations run using the back-projected sources were much closer to experimental values, ranging from 1.0 to 1.7 times the experimental results, again varying with mode. Thus, a significant improvement in the treatment planning can be obtained by using actual measured ultrasound field distributions in combination with backward projection.

Temperature monitoring with line scan echo planar spectroscopic imaging.

UNLABELLED: A new magnetic resonance imaging method, line scan echo planar spectroscopic imaging (LSEPSI), is shown capable of providing rapid, internally referenced temperature monitoring from water and fat chemical shifts. METHODS: Orthogonal 90 degrees and 180 degrees slice selective RF pulses inclined by 45 degrees from the image plane solicit a spin echo from a tissue column. The echo is read by asymmetric sampling of 32 gradient echoes spaced 1.4-1.8 ms apart. Sixty-four adjacent columns are sequentially sampled in 4.2-6.4 s with 4,096 voxels sampled with voxel volumes of 0.08-0.13 cm3. Mixed mayonnaise/water phantoms were used to correlate LSEPSI-derived chemical shifts and thermocouple-based temperature measurements from 23 to 60 degrees C with a 1.5 T scanner. Measurement artifacts unrelated to temperature were investigated with the phantom, as was the feasibility of applying the sequence in human breast in vivo. RESULTS: The correlation between LSEPSI and thermocouple-based temperature measurements in the phantom was excellent (r2>0.99). Field drifts affecting the temperature measurements using the water peak alone were corrected by using the water/lipid peak difference. The sequence had an average temperature resolution of 1.4 degrees C in the phantom. The frequency difference measurement reduced the sensitivity to artifacts related to temperature. Both water and lipid peaks were detectable throughout many locations in the breast, suggesting the applicability of LSEPSI in this organ. DISCUSSION: T1-saturation losses occur in conventional and echo-planar based 2D CSI sequences using phase encoding methods with short TR periods. These losses are eliminated when individual columns are sampled in snapshot fashion with LSEPSI since the effective TR becomes the time between scans rather than excitations. T1 saturation can make small spectral peaks difficult to detect at high temperatures and generally lowers the signal-to-noise ratio of the spectra. The rapid acquisition and insensitivity to T1 saturation effects make LSEPSI an attractive technique for monitoring thermal therapies in breast using the internally referenced fat/water frequency separation.

Apoptosis in ultrasound-produced threshold lesions in the rabbit brain.

Focused ultrasound (US) surgery has been used to induce high temperature elevations in tissue to coagulate the proteins and kill the tissue. The introduction of noninvasive online temperature monitoring has made it possible to induce well-controlled thermal exposures. In this study, we used magnetic resonance imaging (MRI) thermometry to monitor thermal exposures near the threshold of tissue damage, and then investigated if apoptosis was induced. Rabbit brains were sonicated with an eight-sector phased array to create a large region of uniform temperature elevation at the end of a 30-s sonication. Histological examination demonstrated that apoptosis was induced in some cells. At 4 h after the sonications, the apoptotic cells constituted 9 +/- 7% of identifiable cells. By 48 h after the sonications, the number of apoptotic cells had increased up to 17 +/- 9%. The impact of this finding for therapy needs to be explored further.

MR imaging-guided focused ultrasound surgery of fibroadenomas in the breast: a feasibility study.

PURPOSE: To test the feasibility of noninvasive magnetic resonance (MR) imaging-guided focused ultrasound surgery (FUS) of benign fibroadenomas in the breast. MATERIALS AND METHODS: Eleven fibroadenomas in nine patients under local anesthesia were treated with MR imaging-guided FUS. Based on a T2-weighted definition of target volumes, sequential sonications were delivered to treat the entire target. Temperature-sensitive phase-difference-based MR imaging was performed during each sonication to monitor focus localization and tissue temperature changes. After the procedure, T2-weighted and contrast material-enhanced T1-weighted MR imaging were performed to evaluate immediate and long-term effects. RESULTS: Thermal imaging sequences were improved over the treatment period, with 82% (279 of 342) of the hot spots visible in the last seven treatments. The MR imager was used to measure temperature elevation (12.8 degrees -49.9 degrees C) from these treatments. Eight of the 11 lesions treated demonstrated complete or partial lack of contrast material uptake on posttherapy T1-weighted images. Three lesions showed no marked decrease of contrast material uptake. This lack of effective treatment was most likely due to a lower acoustic power and/or patient movement that caused misregistration. No adverse effects were detected, except for one case of transient edema in the pectoralis muscle 2 days after therapy. CONCLUSION: MR imaging-guided FUS can be performed to noninvasively coagulate benign breast fibroadenomas.

MRI monitoring of the thermal ablation of tissue: effects of long exposure times.

MRI-derived thermometry based on the temperature-dependence of the proton resonant frequency (PRF) is extremely sensitive to changes in tissue unrelated to temperature changes, including tissue swelling. This study investigated the maximum amount of time that this phase-subtraction-based method can be used to accurately monitor temperature changes in vivo. Long-duration focused ultrasound sonications were delivered in rabbit thigh muscle with a phased-array transducer, and the time that tissue swelling began was monitored. Tissue swelling began to occur at about one minute. The temperature correlated well with an implanted thermocouple up to this time. After this time, severe artifacts in the phase-difference maps were observed. The thermal dose model predicted the extent of tissue damage well for subsequent one minute sonications. These results will have implications for MRI guidance of thermal therapies with long exposure times.

Usefulness of MR imaging-derived thermometry and dosimetry in determining the threshold for tissue damage induced by thermal surgery in rabbits.

PURPOSE: To investigate in vivo the feasibility of using magnetic resonance (MR) imaging-derived temperature and thermal dose measurements to find the threshold of thermal tissue damage. MATERIALS AND METHODS: Sonications were delivered in rabbit thigh muscles at varying powers. Temperature-sensitive MR images obtained during the sonications were used to estimate the temperature and thermal dose. The temperature, thermal dose, and applied power were then correlated to the occurrence of tissue damage observed on postsonication images. An eight-element phased-array transducer was used to produce spatially flat temperature profiles that allowed for averaging to reduce the effects of noise and the voxel size. RESULTS: The occurrence of tissue damage correlated well with the MR imaging-derived temperature and thermal dose measurements but not with the applied power. Tissue damage occurred at all locations with temperatures greater than 50.4 degrees C and thermal doses greater than 31.2 equivalent minutes at 43.0 degrees C. No tissue damage occurred when these values were less than 47.2 degrees C and 4.3 equivalent minutes. CONCLUSION: MR imaging thermometry and dosimetry provide an index to predict the threshold for tissue damage in vivo. This index offers improved online control over minimally invasive thermal treatments and should allow for more accurate target volume coagulation.

Temperature monitoring in fat with MRI.

The aim of the study was to test the hypothesis that fast spin echo T(1)-weighted images can be used to quantify the temperature in fat during thermal therapy in vivo. An MR compatible positioning device was used to manipulate focused ultrasound transducers in an MRI scanner. This system was used to sonicate fat tissue around the kidneys of 12 rabbits at various power levels for 10 to 20 sec. The scan parameters of T(1)-weighted fast spin echo (FSE) sequence were varied to optimize signal intensity characteristics while maintaining short scan times. An invasive optical probe was used to calibrate the temperature related signal intensity changes. For the T(1)-weighted FSE sequence, the signal intensity decreased with the temperature elevation at the rate of 0.97+/-0.02%/ degrees C. The single focused transducer produced a contrast-to-noise ratio more than 10 at power levels below the tissue damage threshold. The signal intensity was linearly dependent on the power, despite the measured temperatures being well above the coagulation threshold. This study demonstrates that T(1)-weighted FSE MRI sequences can be used to quantify the temperature elevation in fat in vivo during short focused ultrasound exposures. This can be very important for breast tumor surgery, fat ablation, and for treating deep seated tumors through superficial fat layers. Magn Reson Med 43:901-904, 2000.

Magnetic resonance image-guided thermal ablations.

Magnetic resonance imaging (MRI)-based monitoring has been shown in recent years to enhance the effectiveness of minimally or noninvasive thermal therapy techniques, such as laser, radiofrequency, microwave, ultrasound, and cryosurgery. MRI's unique soft-tissue contrast and ability to image in three dimensions and in any orientation make it extremely useful for treatment planning and probe localization. The temperature sensitivity of several intrinsic parameters enables MRI to visualize and quantify the progress of ongoing thermal treatment. MRI is sensitive to thermally induced tissue changes resulting from the therapies, giving the physician a method to determine the success or failure of the treatment. These methods of using MRI for planning, guiding, and monitoring thermal therapies are reviewed.

Magnetic resonance imaging-guided focused ultrasound synovectomy.

OBJECTIVE: To investigate the feasibility of magnetic resonance imaging (MRI)-guided high power focused ultrasound (FUS) to perform synovectomy noninvasively. METHODS: Five New Zealand white male rabbit knees with experimentally induced arthritis underwent MRI-guided thermal surgery by high power (60 W/10 s) sonication. Evidence of tissue coagulation was monitored during the procedure and confirmed by gross and microscopic evaluation and MRI. RESULTS: Partial synovectomy was performed in five animals. Necrotized synovial tissue was observed on gross and microscopic evaluation. Visible signal intensity alterations including high signal intensity on T2-weighted (T2W) images and lack of contrast-enhancement on T1-weighted (T1W) post-contrast, post-sonication images were characteristic and reproducible. CONCLUSION: Our results demonstrate the ability of high power sonication to destroy synovial tissue in vivo.

Determination of the optimal delay between sonications during focused ultrasound surgery in rabbits by using MR imaging to monitor thermal buildup in vivo.

PURPOSE: To use magnetic resonance (MR) imaging to monitor thermal buildup and its effects in treated tissues during sequentially delivered sonications in vivo to optimize the intersonication delay for any set of ultrasound and tissue parameters. MATERIALS AND METHODS: Sequential sonications were delivered next to each other in both thighs in 10 male New Zealand white rabbits. The time between sonications was 11-60 seconds. Phase-difference MR imaging was used to monitor temperature rise, which was used to estimate the thermal dose delivered to the tissue. T2-weighted and contrast agent-enhanced T1-weighted imaging were used to gauge the extent of tissue coagulation. RESULTS: With a short intersonication delay (11-40 seconds), the estimated temperature rise and the extent of tissue coagulation increased dramatically in subsequent sonications. However, when the delay was long (50-60 seconds), the size and shape of the destroyed tissue with subsequent sonications was uniform, and the temperature buildup was substantially lower. CONCLUSION: MR imaging can be used to monitor thermal buildup and its effects due to sequential, neighboring sonications in vivo to produce evenly shaped regions of tissue coagulation. The temperature information obtained from the monitoring can be used to optimize the intersonication delay for any set of ultrasound and tissue parameters.

Tissue temperature monitoring with multiple gradient-echo imaging sequences.

The inherent sensitivity of multiple gradient-echo sequences to the chemical shift is exploited to rapidly map muscle water frequency shifts caused by ultrasonic heating. The use of multiple echoes is shown to offer several advantages over single gradient-echo approaches previously proposed for temperature measurement. An increase in the effective bandwidth significantly reduces aliasing problems observed with single gradient-echo methods in high temperature applications. Of greater significance is the improved immunity to intrascan motion found for multi-echo versus single echo gradient methods, making the former more attractive for clinical applications. Finally, a sensitivity to the presence of multiple spectral components unavailable with single gradient-echo methods is obtained.