Investigating the potential of catheter-assisted pulsed focused ultrasound ablation for atherosclerotic plaques.

BACKGROUND: Atherosclerosis is a condition in which an adhesive substance called plaque accumulates over time inside the arteries. Plaque buildup results in the constriction of arteries, causing a shortage of blood supply to tissues and organs. Removing atherosclerotic plaques controls the development of acute ischemic stroke and heart diseases. It remains imperative for positive patient outcomes. PURPOSE: This study sought to develop a minimally invasive technique for removing arterial plaques by applying focused ultrasound (FUS) energy on the metal surface of a nitinol catheter wire to induce inertial cavitation. The induced cavitation can deplete plaque mechanically inside the arteries, leading towards improved recanalization of blood vessels. METHODS: The enhanced cavitation effect induced by combining FUS with a metal catheter was first verified by exposing agar phantom gels with or without a 0.9-mm diameter nitinol wire to an acoustic field produced by a 0.5-MHz FUS transducer. The phenomenon was further confirmed in pork belly fat samples with or without a 3-mm diameter nitinol catheter wire. Cavitation was monitored by detecting the peaks of emitted ultrasound signals from the samples using a passive cavitation detector (PCD). Cavitation threshold values were determined by observing the jump in the peak amplitude of signals received by the PCD when the applied FUS peak negative pressure (PNP) increased. To simulate arterial plaque removal, FUS with or without a catheter was used to remove tissues from pork belly fat samples and the lipid cores of human atherosclerotic plaque samples using 2500-cycle FUS bursts at 10% duty cycle and a burst repetition rate of 20 Hz. Treatment outcomes were quantified by subtracting the weight of samples before treatment from the weight of samples after treatment. All measurements were repeated 5 times (n = 5) unless otherwise indicated, and paired t-tests were used to compare the means of two groups. A p-value of <0.05 will be considered significant. RESULTS: Our results showed that with a nitinol wire, the cavitation threshold in agar phantoms was reduced to 2.6 MPa from 4.3 MPa PNP when there was no nitinol wire in the focal region of FUS. For pork belly fat samples, cavitation threshold values were 1.0 and 2.0 MPa PNP, with and without a catheter wire, respectively. Pork belly fat tissues and lipid cores of atherosclerotic plaques were depleted at the interface between a catheter and the samples at 2 and 4 MPa FUS PNP, respectively. The results showed that with a catheter wire in the focal region of a 3-min FUS treatment session, 24.7 and 25.6 mg of lipid tissues were removed from pork belly fat and human atherosclerotic samples, respectively. In contrast, the FUS-only group showed no reduction in sample weight. The differences between FUS-only and FUS-plus-catheter groups were statistically significant (p < 0.001 for the treatment on pork belly samples, and p < 0.01 for the treatment on human atherosclerotic samples). CONCLUSION: This study demonstrated the feasibility of catheter-assisted FUS therapy for removing atherosclerotic plaques.

Real-time cavitation monitoring during optical coherence tomography guided photo-mediated ultrasound therapy of the retina.

Photo-mediated ultrasound therapy (PUT) is a novel antivascular therapeutic modality based on cavitation-induced bioeffects. During PUT, synergistic combinations of laser pulses and ultrasound bursts are used to remove the targeted microvessels selectively and precisely without harming nearby tissue. In the current study, an integrated system combining PUT and spectral domain optical coherence tomography (SD-OCT) was developed, where the SD-OCT system was used to guide PUT by detecting cavitation in real time in the retina of the eye. METHOD: We first examined the capability of SD-OCT in detecting cavitation on a vascular-mimicking phantom and compared the results with those from a passive cavitation detector. The performance of the integrated system in treatment of choroidal microvessels was then evaluated in rabbit eyes in vivo. RESULTS: During the in vivo PUT experiments, several biomarkers at the subretinal layer in the rabbit eye were identified on OCT images. The findings indicate that, by evaluating biomarkers of treatment effect, real-time SD-OCT monitoring could help to avoid micro-hemorrhage, which is a potential major side effect. CONCLUSION: Real-time OCT monitoring can thus improve the safety and efficiency of PUT in removing the retinal and choroidal microvasculature.

Photoacoustic imaging of squirrel monkey cortical responses induced by peripheral mechanical stimulation.

Non-human primates (NHPs) are crucial models for studies of neuronal activity. Emerging photoacoustic imaging modalities offer excellent tools for studying NHP brains with high sensitivity and high spatial resolution. In this research, a photoacoustic microscopy (PAM) device was used to provide a label-free quantitative characterization of cerebral hemodynamic changes due to peripheral mechanical stimulation. A 5 × 5 mm area within the somatosensory cortex region of an adult squirrel monkey was imaged. A deep, fully connected neural network was characterized and applied to the PAM images of the cortex to enhance the vessel structures after mechanical stimulation on the forelimb digits. The quality of the PAM images was improved significantly with a neural network while preserving the hemodynamic responses. The functional responses to the mechanical stimulation were characterized based on the improved PAM images. This study demonstrates capability of PAM combined with machine learning for functional imaging of the NHP brain.

Cutaneous Hypervascularization Treatment Using Photo-Mediated Ultrasound Therapy.

Photo-mediated ultrasound therapy (PUT) is a cavitation-based, highly selective antivascular technique. In this study, the effectiveness and safety of PUT on cutaneous vascular malformation was examined through in vivo experiments in a clinically relevant chicken wattle model, whose microanatomy is similar to that of port-wine stain and other hypervascular dermal diseases in humans. Assessed by optical coherence tomography angiography, the blood vessel density in the chicken wattle decreased by 73.23% after one session of PUT treatment in which 0.707 J/cm2 fluence laser pulses were applied concurrently with ultrasound bursts (n = 7, P < .01). The effectiveness of removing blood vessels in the skin at depth up to 1 mm was further assessed by H&E-stained histology at multiple time points, which included days 1, 3, 7, 14, and 21 after treatment. Additional immunohistochemical analyses with CD31, caspase-3, and Masson's trichrome stains were performed on day 3 after treatment. The results show that the PUT-induced therapeutic effect was confined and specific to blood vessels only, whereas unwanted collateral damage in other skin tissues such as collagen was avoided. The findings from this study demonstrate that PUT can efficiently and safely remove hypervascular dermal capillaries using laser fluence at a level that is orders of magnitude smaller than that used in conventional laser treatment of vascular lesions, thus offering a safer alternative technique for clinical management of cutaneous vascular malformations.

A review on photo-mediated ultrasound therapy.

Photo-mediated ultrasound therapy (PUT) is a novel therapeutic technique based on the combination of ultrasound and laser. The underlying mechanism of PUT is the enhanced cavitation effect inside blood vessels. The enhanced cavitation activity can result in bio-effects such as reduced perfusion in microvessels. The reduced perfusion effect in microvessels in the eye has the potential to control the progression of eye diseases such as diabetic retinopathy and age-related macular degeneration. Several in vivo studies have demonstrated the feasibility of PUT in removing microvasculature in the eye using rabbit eye model and vasculature in the skin using rabbit ear model. Numerical studies using a bubble dynamics model found that cavitation is enhanced during PUT due to the dramatic increase in size of air/vapor nuclei in blood. In addition, the study conducted to model cavitation dynamics inside a blood vessel during PUT found stresses induced on the vessel wall during PUT are higher than that at normal physiological levels, which may be responsible for bio-effects. The concentration of vasodilators such as nitric oxide and prostacyclin were also found to be affected during PUT in an in vitro study, which may limit blood perfusion in vessels. The main advantage of PUT over conventional techniques is non-invasive, precise, and selective removal of microvessels with high efficiency at relatively low energy levels of ultrasound and laser, without affecting the nearby structures. However, the main limitation of vessel rupture/hemorrhage needs to be overcome through the development of real-time monitoring of treatment effects during PUT. In addition to the application in removing microvessels, PUT-based techniques were also explored in treating other diseases. Studies have found a combination of ultrasound and laser to be effective in removing blood clots inside veins, which has the potential to treat deep-vein thrombosis. The disruption of atherosclerotic plaque using combined ultrasound and laser was also tested, and the feasibility was demonstrated.

Recanalize ureteral stents with focused ultrasound.

BACKGROUND: Maintaining ureteral patency is imperative to preventing renal injury and systemic infection. Ureteral stents are small conduits connecting the kidney and the bladder. They have been widely used to treat ureteral obstructions and ureteral leaks. The most problematic and frequent stent-associated complication is stent encrustation. This occurs when mineral crystals (e.g. calcium, oxalate, phosphorus, struvite) are deposited onto the surface and internal lumen of the stent. Encrustation can lead to the obstruction of a stent and increases risk of systemic infection. As a result, ureteral stents need to be replaced typically every 2-3 months. PURPOSE: In this study, we present a non-invasive, high-intensity focused ultrasound (HIFU)-based technique to recanalize obstructed stents. By taking advantage of the mechanical force produced by a HIFU beam, including acoustic radiation force, acoustic streaming, and cavitation, HIFU can break up encrustations, clearing the stent of obstruction. METHODS: The ureteral stents for this study were obtained from patients undergoing ureteral stent removal. Under the guidance of ultrasound imaging, the encrustation in the stents were located, and then targeted by HIFU at frequencies of 0.25 and 1 MHz. The duty cycle of HIFU was 10%, and the HIFU burst repetition rate was 1 Hz, while the HIFU amplitude was varied to find the threshold pressure that would displace encrustations. The treatment duration was limited at 2 min (or 120 shots from HIFU). The treatments were carried out in two different orientations (parallel and perpendicular) of the ureteral stent with respect to the HIFU beam. For each setting, five treatments were conducted for a maximum duration of 2 min. During the entire treatment, an ultrasound imaging system was used to monitor the movement of encrustations inside the stent. The peak negative HIFU pressures needed to move the encrustations inside the stent was recorded for quantitative analysis. RESULTS: Our results demonstrated that at both 0.25 and 1 MHz ultrasound frequencies, obstructed stents could be recanalized. At 0.25 MHz, the needed average peak negative pressure was 0.52 MPa in parallel orientation and 0.42 MPa in perpendicular orientation. At 1 MHz, the needed average peak negative pressure was 1.10 MPa in parallel orientation and 1.15 MPa in perpendicular orientation CONCLUSIONS: This first in-vitro study has demonstrated the feasibility of non-invasive HIFU to recanalize ureteral stents. This technology has a potential to reduce the need for ureteral stent exchange.

Choroidal neovascularization removal with photo-mediated ultrasound therapy.

BACKGROUND: Age-related macular degeneration (AMD) is a major cause of irreversible central vision loss. The main reason for lost vision due to AMD is choroidal neovascularization (CNV). In the clinic, current treatments for CNV include photodynamic therapy, laser photocoagulation, and anti-vascular endothelial growth factor (VEGF) therapy. PURPOSE: This study evaluates a novel treatment technique combining synchronized nanosecond laser pulses and ultrasound bursts, namely photo-mediated ultrasound therapy (PUT) as a potential treatment method for CNV, for its efficacy and safety in the treatment of CNV via the experiments in a clinically-relevant rabbit model in vivo. METHODS: CNV was created by subretinal injection of Matrigel and vascular endothelial growth factor (M&V) in 10 New Zealand white rabbits. Six rabbits were used in the PUT group. In the control groups, two rabbits were treated by laser-only, and two rabbits were treated by ultrasound-only. The treatment efficacy was evaluated through fundus photography and fluorescein angiography (FA) longitudinally for up to 4 weeks. Rabbits were sacrificed for histopathology 3 months after treatment to examine the safety of PUT. RESULTS: The fluorescein leakage on FA was quantified to longitudinally evaluate treatment outcome. Compared with baseline, the relative intensity index was reduced by 26.57% ± 8.66% at 3 days after treatment, 27.24% ± 6.21% at 1 week after treatment, 27.79% ± 2.61% at 2 weeks after treatment, and 32.12% ± 3.23% at 4 weeks after treatment, all with a statistically significant difference of p < 0.01. The comparison between the relative intensity indexes from the two control groups (laser-only treatment and ultrasound-only treatment) did not show any statistically significant difference at all time points. Safety evaluation at 3 months with histopathology demonstrated that the PUT did not result in morphologic changes to the neurosensory retina. CONCLUSIONS: This study introduces PUT for the first time for the treatment of CNV. The results demonstrated good efficacy and safety of PUT to treat CNV in a clinically-relevant rabbit model. With a single session of treatment, PUT can safely reduce the leakage of CNV for at least 1 month after treatment.

A feasibility study on removing lipid deposition in atherosclerotic plaques with ultrasound-assisted laser ablation.

Objective. Atherosclerosis is the buildup of fats, cholesterol, and other substances on the inner walls of arteries. It can affect arteries of heart, brain, arms, legs, pelvis and kidney, resulting in ischemic heart disease, carotid artery disease, peripheral artery disease and chronic kidney disease. Laser-based treatment techniques like laser atherectomy can be used to treat many common atherosclerostic diseases. However, the use of laser-based treatment remains limited due to the high risk of complications and low efficiency in removing atherosclerostic plaques as compared with other treatment methods. In this study, we developed a technology that used high intensity focused ultrasound to assist laser treatment in the removal of the lipid core of atherosclerotic plaques.Approach. The fundamental mechanism to disrupt atherosclerostic plaque was to enhance the mechanical effect of cavitation during laser/ultrasound therapy. To promote cavitation, spatiotemporally synchronized ultrasound bursts of 2% duty cycle at 0.5 MHz and nanosecond laser pulses at 532 nm wavelength were used. Experiments were first performed on pig belly fat samples to titrate ultrasound and laser parameters. Then, experiments were conducted on human plaque samples, where the lipid depositions of the plaques were targeted.Main results. Our results showed that fat tissue could be removed with an ultrasound peak negative pressure (PNP) of 2.45 MPa and a laser radiant exposure as low as 3.2 mJ mm-2. The lipid depositions on the atherosclerostic plaques were removed with laser radiant exposure of 16 mJ mm-2in synchronizing with an ultrasound PNP of 5.4 MPa. During all the experiments, laser-only and ultrasound-only control treatments at the same energy levels were not effective in removing the lipid.Significance. The results demonstrated that the addition of ultrasound could effectively reduce the needed laser power for atherosclerotic plaque removal, which will potentially improve treatment safety and efficiency of current laser therapies.

Effect of Photo-Mediated Ultrasound Therapy on Nitric Oxide and Prostacyclin from Endothelial Cells.

Several studies have investigated the effect of photo-mediated ultrasound therapy (PUT) on the treatment of neovascularization. This study explores the impact of PUT on the release of the vasoactive agents nitric oxide (NO) and prostacyclin (PGI2) from the endothelial cells in an in vitro blood vessel model. In this study, an in vitro vessel model containing RF/6A chorioretinal endothelial cells was used. The vessels were treated with ultrasound-only (0.5, 1.0, 1.5 and 2.0 MPa peak negative pressure at 0.5 MHz with 10% duty cycle), laser-only (5, 10, 15 and 20 mJ/cm2 at 532 nm with a pulse width of 5 ns), and synchronized laser and ultrasound (PUT) treatments. Passive cavitation detection was used to determine the cavitation activities during treatment. The levels of NO and PGI2 generally increased when the applied ultrasound pressure and laser fluence were low. The increases in NO and PGI2 levels were significantly reduced by 37.2% and 42.7%, respectively, from 0.5 to 1.5 MPa when only ultrasound was applied. The increase in NO was significantly reduced by 89.5% from 5 to 20 mJ/cm2, when only the laser was used. In the PUT group, for 10 mJ/cm2 laser fluence, the release of NO decreased by 76.8% from 0.1 to 1 MPa ultrasound pressure. For 0.5 MPa ultrasound pressure in the PUT group, the release of PGI2 started to decrease by 144% from 15 to 20 mJ/cm2 laser fluence. The decreases in NO and PGI2 levels coincided with the increased cavitation activities in each group. In conclusion, PUT can induce a significant reduction in the release of NO and PGI2 in comparison with ultrasound-only and laser-only treatments.

A 3D finite element model to study the cavitation induced stresses on blood-vessel wall during the ultrasound-only phase of photo-mediated ultrasound therapy.

Photo-mediated ultrasound therapy (PUT) is a novel technique utilizing synchronized ultrasound and laser to generate enhanced cavitation inside blood vessels. The enhanced cavitation inside blood vessels induces bio-effects, which can result in the removal of micro-vessels and the reduction in local blood perfusion. These bio-effects have the potential to treat neovascularization diseases in the eye, such as age-related macular degeneration and diabetic retinopathy. Currently, PUT is in the preclinical stage, and various PUT studies on in vivo rabbit eye models have shown successful removal of micro-vessels. PUT is completely non-invasive and particle-free as opposed to current clinical treatments such as anti-vascular endothelial growth factor therapy and photodynamic therapy, and it precisely removes micro-vessels without damaging the surrounding tissue, unlike laser photocoagulation therapy. The stresses produced by oscillating bubbles during PUT are responsible for the induced bio-effects in blood vessels. In our previous work, stresses induced during the first phase of PUT due to combined ultrasound and laser irradiation were studied using a 2D model. In this work, stresses induced during the third or last phase of PUT due to ultrasound alone were studied using a 3D finite element method-based numerical model. The results showed that the circumferential and shear stress increased as the bubble moves from the center of the vessel toward the vessel wall with more than a 16 times increase in shear stress from 1.848 to 31.060 kPa as compared to only a 4 times increase in circumferential stress from 211 to 906 kPa for a 2 µm bubble placed inside a 10 µm vessel on the application of 1 MHz ultrasound frequency and 130 kPa amplitude. In addition, the stresses decreased as the bubble was placed in smaller sized vessels with a larger decrease in circumferential stress. The changes in shear stress were found to be more dependent on the bubble-vessel wall distance, and the changes in circumferential stress were more dependent on the bubble oscillation amplitude. Moreover, the bubble shape changed to an ellipsoidal with a higher oscillation amplitude in the vessel's axial direction as it was moved closer to the vessel wall, and the bubble oscillation amplitude decreased drastically as it was placed in vessels of a smaller size.

Photo-mediated ultrasound therapy for the treatment of retinal neovascularization in rabbit eyes.

OBJECTIVES: Retinal neovascularization (RNV) is the growth of abnormal microvessels on the retinal surface and into the vitreous, which can lead to severe vision loss. By combining relatively low-intensity ultrasound and nanosecond-pulse-duration laser, we developed a novel treatment method, namely photo-mediated ultrasound therapy (PUT), which holds a potential to remove RNV with minimal or no damage to the adjacent tissues. METHODS: RNV was created in both albino and pigmented rabbits (n = 10) through a single intravitreal injection with DL-α-aminoadipic acid. RNV was treated with PUT 8 weeks postinjection. After PUT treatment, animals were evaluated longitudinally for up to 6 weeks. Treatment outcomes were evaluated through fundus photography, red-free fundus photography, fluorescein angiography (FA), and histopathology. RESULTS: In both albino and pigmented rabbits, there were no leakage in the treatment area immediately after PUT treatment as demonstrated by FA, indicating the cessation of blood perfusion of the RNV in the treated area. The fluorescence leakage did not recover in albino rabbits during the 6-week posttreatment monitoring period, and only 9.9 ± 9.8% of the neovascularization remained at the end of 6 weeks. In the pigmented rabbits, the fluorescence leakage partially returned, but the level of leakage decreased over time during the 6-week posttreatment monitoring period, and only 10.8 ± 9.8% of the neovascularization remained at the end of 6 weeks. Histology demonstrated removal of vasculature without damage to the surrounding neurosensory retina. CONCLUSIONS: These results demonstrate that PUT could precisely remove RNV without damage to the surrounding neurosensory retina in both rabbit strains.

Photoacoustic imaging of squirrel monkey cortical and subcortical brain regions during peripheral electrical stimulation.

The investigation of neuronal activity in non-human primate models is of critical importance due to their genetic similarity to human brains. In this study, we tested the feasibility of using photoacoustic imaging for the detection of cortical and subcortical responses due to peripheral electrical stimulation in a squirrel monkey model. Photoacoustic computed tomography and photoacoustic microscopy were applied on squirrel monkeys for real-time deep subcortical imaging and optical-resolution cortical imaging, respectively. The electrically evoked hemodynamic changes in primary somatosensory cortex, premotor cortices, primary motor cortex, and underlying subcortical areas were measured. Hemodynamic responses were observed in both cortical and subcortical brain areas at the cortices during external stimulation, demonstrating the feasibility of photoacoustic technique for functional imaging of non-human primate brain.

Hyaluronic acid carrier-based photodynamic therapy for head and neck squamous cell carcinoma.

PURPOSE: Conventional photosensitizers for photodynamic therapy (PDT) typically have wide tissue distribution and poor water solubility. A hyaluronic acid (HA) polymeric nanoparticle with specific lymphatic uptake and highly water solubility was developed to deliver pyropheophorbide-a (PPa) for locally advanced head and neck squamous cell carcinoma (HNSCC) treatment. METHODS AND RESULTS: PPa was chemically conjugated to the HA polymeric nanoparticle via an adipic acid dihydrazide (ADH) linker. The conjugates were injected subcutaneously in a region near the tumor. Near-infrared (NIR) imaging was used to monitor distribution, and diode laser was used to activate PPa. The singlet oxygen generation efficiency of PPa was not affected by conjugation to HA nanoparticles at a PPa loading degree of 1.89 w.t.%. HA-ADH-PPa inhibited human HNSCC MDA-1986 cell growth only when photo-irradiation was applied. After HA-ADH-PPa treatment and radiation, NU/NU mice bearing human HNSCC MDA-1986 tumors showed reduced tumor growth and significantly enhanced survival time compared with an untreated group (p < 0.05). CONCLUSIONS: These results demonstrate that HA-ADH-PPa could be useful for in vivo locoregional photodynamic therapy of HNSCC.

The feasibility of ultrasound-assisted endovascular laser thrombolysis in an acute rabbit thrombosis model.

PURPOSE: This study aimed to test the feasibility of combined ultrasound and laser technique, namely, ultrasound-assisted endovascular laser thrombolysis (USELT), for thrombolysis by conducting in vivo tests in a rabbit thrombosis model. MATERIALS AND METHODS: An acute thrombus was created in the right jugular vein of rabbit and then was treated with ultrasound only, laser only, and USELT to dissolve the blood clot. A total of 20 rabbits were used. Out of which, the first three rabbits were used to titrate the laser and ultrasound parameters. Then, five rabbits were treated with ultrasound only, five rabbits were treated with laser only, and seven rabbits were treated with USELT. During USELT, 532-nm laser pulses were delivered endovascularly directly to the clot through a fiber optic, and 0.5 MHz ultrasound pulses were applied noninvasively to the same region. A laser fluence of 4 to 12 mJ/cm2 and ultrasound amplitude of 1 to 2 MPa were used. Recanalization of the jugular vein was assessed by performing ultrasound Doppler imaging immediately after the treatment. The maximum blood flow speed after the treatment as compared to its value before the treatment was used to calculate the blood flow recovery in vessel. RESULTS: The blood flow was fully recovered (100%) in three rabbits, partially recovered in two rabbits (more than 50% and less than 100%) with mean percentage recovery of 69.73% and poorly recovered in two rabbits (<50%) with mean percentage recovery of 6.2% in the USELT group. In contrast, the treatment group with ultrasound or laser alone did not show recanalization of vein in any case, all the five rabbits were poorly/not recovered with a mean percentage recovery of 0%. CONCLUSIONS: The USELT technology was shown to effectively dissolve the blood clots in an acute rabbit jugular vein thrombosis model.

The effect of ultrasound cavitation on endothelial cells.

Acoustic cavitation has been widely explored for both diagnostic and therapeutic purposes. Ultrasound-induced cavitation, including inertial cavitation and non-inertial cavitation, can cause microstreaming, microjet, and free radical formation. The acoustic cavitation effects on endothelial cells have been studied for drug delivery, gene therapy, and cancer therapy. Studies have demonstrated that the ultrasound-induced cavitation effect can treat cancer, ischaemia, diabetes, and cardiovascular diseases. In this minireview, we will review the impact of ultrasound-induced cavitation on the endothelial cells such as cell permeability, cell proliferation, gene expression regulation, cell viability, hemostasis interaction, oxygenation, and variation in the level of calcium ions, ceramide, nitric oxide (NO) and nitric oxide synthase (NOS) activity. The applications of these effects and the cavitation mechanism involved will be summarized, demonstrating the important role of acoustic cavitation in non-invasive ultrasound treatment of various physiological conditions.

Ultrasound-assisted laser thrombolysis with endovascular laser and high-intensity focused ultrasound.

PURPOSE: The combination of laser and ultrasound can significantly improve the efficiency of thrombolysis through an enhanced cavitation effect. We developed a fiber optics-based laser-ultrasound thrombolysis device and tested the feasibility and efficiency of this technology for restoring blood flow in an in vitro blood clot model. METHODS: An in vitro blood flow-clot model was setup, and then an endovascular laser thrombolysis system was combined with high-intensity focused ultrasound to remove the clot. The laser and ultrasound pulses were synchronized and delivered to the blood clot concurrently. The laser pulses of 532 nm were delivered to the blood clot endovascularly through an optical fiber, whereas the ultrasound pulses of 0.5 MHz were applied noninvasively to the same region. Effectiveness of thrombolysis was evaluated by the ability to restore blood flow, which was monitored by ultrasound Doppler. RESULTS: As laser powers increased, the ultrasound threshold pressures for effective thrombolysis decreased. For laser fluence levels of 0, 2, and 4 mJ/cm2 , the average negative ultrasound threshold pressures were 1.26 ± 0.114, 1.05 ± 0.181, and 0.59 ± 0.074 MPa, respectively. The periods of time needed to achieve effective thrombolysis were measured at 0.8, 2, and 4 mJ/cm2 laser fluence levels and 0.42, 0.70, and 0.98 MPa negative ultrasound pressures. In general, thrombolysis could be achieved more rapidly with higher laser powers or ultrasound pressures. CONCLUSIONS: Effective thrombolysis can be achieved by combining endovascular laser with noninvasive ultrasound at relatively low power and pressure levels, which can potentially improve both the treatment efficiency and safety.

Photo-Mediated Ultrasound Therapy for the Treatment of Corneal Neovascularization in Rabbit Eyes.

Purpose: Corneal neovascularization (CNV) is the invasion of new blood vessels into the avascular cornea, leading to reduced corneal transparency and visual acuity, impaired vision, and even blindness. Current treatment options for CNV are limited. We developed a novel treatment method, termed photo-mediated ultrasound therapy (PUT), that combines laser and ultrasound, and we tested its feasibility for treating CNV in a rabbit model. Methods: A suture-induced CNV model was established in New Zealand White rabbits, which were randomly divided into two groups: PUT and control. For the PUT group, the applied light fluence at the corneal surface was estimated to be 27 mJ/cm2 at 1064-nm wavelength with a pulse duration of 5 ns, and the ultrasound pressure applied on the cornea was 0.43 MPa at 0.5 MHz. The control group received no treatment. Red-free photography and fluorescein angiography were utilized to evaluate the efficiency of PUT. Safety was evaluated by histology and immunohistochemistry. For comparison with the PUT safety results, conventional laser photocoagulation (LP) treatment was performed with standard clinical parameters: 532-nm continuous-wave (CW) laser with 0.1-second pulse duration, 450-mW power, and 75-µm spot size. Results: In the PUT group, only 1.8% ± 0.8% of the CNV remained 30 days after treatment. In contrast, 71.4% ± 7.2% of the CNV remained in the control group after 30 days. Safety evaluations showed that PUT did not cause any damage to the surrounding tissue. Conclusions: These results demonstrate that PUT is capable of removing CNV safely and effectively in this rabbit model. Translational Relevance: PUT can remove CNV safely and effectively.

Photo-mediated Ultrasound Therapy to Treat Retinal Neovascularization.

This report describes a novel therapeutic technique called photo-mediated ultrasound therapy (PUT). PUT applies synchronized short pulse duration (nanosecond) laser and ultrasound burst on targeted tissue, offering high-precision localized treatment. PUT is based on controlled induction and promotion of micro-cavitation activity in the target tissue. PUT is able to safely and effectively treat retinal neovascularization in rabbits with persistent nonperfusion up to 4 weeks after PUT in the choroidal vasculature.Clinical Relevance- PUT can selectively remove retinal angiogenesis in clinically-relevant disease models in humansized eyes (rabbit) without damaging surrounding tissue.

Removing Subcutaneous Microvessels Using Photo-Mediated Ultrasound Therapy.

BACKGROUND AND OBJECTIVES: We have developed a novel anti-vascular technique, termed photo-mediated ultrasound therapy (PUT), which utilizes nanosecond duration laser pulses synchronized with ultrasound bursts to remove the microvasculature through cavitation. The objective of the current study is to explore the potential of PUT in removing subcutaneous microvessels. STUDY DESIGN/MATERIALS AND METHODS: The auricular blood vessels of two New Zealand white rabbits were treated by PUT with a peak negative ultrasound pressure of 0.45 MPa at 0.5 MHz, and a laser fluence of 0.056 J/cm2 at 1064 nm for 10 minutes. Blood perfusion in the treated area was measured by a commercial laser speckle imaging (LSI) system before and immediately after treatment, as well as at 1 hour, 3 days, 2 weeks, and 4 weeks post-treatment. Perfusion rates of 38 individual vessels from four rabbit ears were tracked during this time period for longitudinal assessment. RESULTS: The measured perfusion rates of the vessels in the treated areas, as quantified by the relative change in perfusion rate, showed a statistically significant decrease for all time points post-treatment (P < 0.001). The mean decrease in perfusion is 50.79% immediately after treatment and is 32.14% at 4 weeks post-treatment. Immediately after treatment, the perfusion rate decreased rapidly. Following this, there was a partial recovery in perfusion rate up to 3 days post-treatment, followed by a plateau in the perfusion from 3 days to 4 weeks. CONCLUSIONS: This study demonstrated that a single PUT treatment could significantly reduce blood perfusion by 32.14% in the skin for up to 4 weeks. With unique advantages such as low laser fluence as compared with photothermolysis and agent-free treatment as compared with photodynamic therapy, PUT holds the potential to be developed into a new tool for the treatment of cutaneous vascular lesions. Lasers Surg. Med. © 2020 Wiley Periodicals, LLC.

The Effect of Laser and Ultrasound Synchronization in Photo-Mediated Ultrasound Therapy.

OBJECTIVE: Photo-mediated ultrasound therapy (PUT) is a novel, non-invasive, agent-free, highly selective, and precise anti-vascular technique. PUT removes microvessels through promoting cavitation activity precisely in targeted microvessels by applying synchronized nanosecond laser pulses and ultrasound bursts. The synchronization between laser and ultrasound is critical to the outcome of PUT. METHODS: Through theoretical simulation and experimental study, the effect of synchronization between laser pulses and ultrasound bursts on cavitation activity during PUT is evaluated. RESULTS: By using a theoretical model, we found that cavitation activity was enhanced when laser pulses and ultrasound bursts were synchronized such that the produced photoacoustic wave overlaid the rarefactional phase of the ultrasound wave. This finding was then verified through in vitro studies where cavitation was monitored by using a passive cavitation detector. Furthermore, we demonstrated that the in vivo treatment outcome of PUT in rabbits was directly related to the synchronization between laser and ultrasound. The anti-vascular effect could only be observed when laser and ultrasound were properly synchronized in vivo. CONCLUSION: PUT is more efficient when the laser-induced photoacoustic wave overlays the rarefactional phase of the ultrasonic wave. SIGNIFICANCE: This is a systematic study to investigate the synchronization effect of PUT, which would be significant for further understanding the mechanism and further improving the treatment efficiency of PUT.