Non-invasive focused ultrasound nerve modulation for treating physiological conditions

Abstract

The subject matter of the present disclosure generally relates to techniques for applying focused ultrasound energy to a region of interest in a subject to induce neuromodulation of the celiac plexus for treating inflammatory bowel disease. The region of interest may include at least a portion of the peripheral ganglia of the celiac plexus.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the priority and benefit of U.S. Provisional Application No. 63 / 007,766, filed on April 9, 2020, and U.S. Provisional Application No. 63 / 027,724, filed on May 20, 2020.

Background Art

[0002] The subject matter disclosed herein relates to neuromodulation, and more specifically, to techniques for regulating physiological responses using energy applied from an energy source.

[0003] Neuromodulation has been used to treat a variety of clinical conditions. For example, electrical stimulation at various locations along the spinal cord has been used to treat chronic back pain. Implantable devices can periodically generate electrical energy that is applied to tissue to activate specific nerve fibers, thereby reducing the sensation of pain. With regard to spinal cord stimulation, the stimulating electrodes are generally placed in the epidural space, although the pulse generator may be placed some distance from the electrodes, for example in the abdomen or buttocks, but may be connected to the electrodes via a wire. In other embodiments, deep brain stimulation can be used to stimulate specific areas of the brain to treat motor disorders, and the stimulation location can be guided by neuroimaging. Such central nervous system stimulation generally targets the function of a local nerve or brain cell, delivering electrical pulses mediated by electrodes placed in or near the target nerve. However, placing electrodes in or near the target nerve is difficult. For example, such techniques may involve the surgical placement of electrodes to deliver energy. Furthermore, targeting specific tissues via neuromodulation is difficult. Electrodes placed on or near specific target nerves mediate neuromodulation by inducing action potentials in nerve fibers, resulting in sequential neurotransmitter release at nerve synapses and synaptic transmission with subsequent nerves. Such propagation can produce a relatively larger or more diffuse physiological effect than desired, as the implementation of current from the implanted electrodes stimulates many nerves or axons at once. Because neural pathways are complex and interconnected, more selective and targeted modulation may be more clinically useful. [Overview of the Initiative]

[0004] Specific embodiments are summarized below. These embodiments are not intended to limit the scope of the claimed subject matter, but rather to provide a brief overview of possible embodiments. In fact, this disclosure may encompass a variety of forms that may be similar to or different from the embodiments described below.

[0005] In one embodiment, a method is provided which involves applying focused ultrasound energy to a region of interest, wherein the region of interest includes at least a portion of the celiac plexus, in order to induce neural modulation of one or more nerve pathways.

[0006] In one embodiment, a system is provided comprising an ultrasound probe configured to apply focused ultrasound energy to a region of interest, including at least a portion of the celiac plexus, in order to neuralgeomodulate the peripheral ganglia of the celiac plexus of interest. The system also comprises a controller configured to acquire image data of a target from the ultrasound probe operating in imaging mode, select a region of interest based on the image data, and control the ultrasound probe to apply focused ultrasound energy to the region of interest as part of a therapeutic protocol for treating the target inflammatory bowel disease.

[0007] In one embodiment, a method is provided comprising acquiring image data of a subject from an ultrasound probe operating in imaging mode, wherein the subject is diagnosed with inflammatory bowel disease; selecting a region of interest, which includes at least a portion of the celiac plexus, based on the image data; and controlling the ultrasound probe to apply focused ultrasound energy to the region of interest as part of a treatment protocol for treating inflammatory bowel disease, wherein the region of interest includes at least a portion of the peripheral ganglia of the celiac plexus. [Brief explanation of the drawing]

[0008] These and other features, aspects, and advantages of this disclosure will be better understood by reading the following detailed description with reference to the accompanying drawings. In the accompanying drawings, similar reference numerals throughout the drawings represent similar parts.

[0009] [Figure 1] This is a schematic diagram of the experimental setup for applying ultrasonic energy according to the embodiments of this disclosure.

[0010] [Figure 2]This shows ultrasound imaging used to spatially select a region of interest for application of ultrasound energy.

[0011] [Figure 3] This shows the changes in disease activity index (DAI) over the course of repeated focused ultrasound (FUS) treatment in animal models ingesting sodium dextran sulfate (DSS) compared to a control.

[0012] [Figure 4] This shows the changes in the DAI subcategory of stool consistency over the course of daily focused ultrasound treatment in DSS-ingested animal models compared to a control.

[0013] [Figure 5] This shows the changes in the DAI subcategory of macroscopic bleeding over the course of daily focused ultrasound treatment in DSS-ingested animal models compared to controls.

[0014] [Figure 6] This shows the changes in the DAI subcategory of body weight over the course of daily focused ultrasound treatment in DSS-ingested animal models compared to controls.

[0015] [Figure 7] This is a schematic diagram of the postmortem colon of test animals, illustrating the effects of DSS and focused ultrasound on stool consistency.

[0016] [Figure 8] This shows the relationship between different concentrations of DSS on DAI scores in untreated animals.

[0017] [Figure 9] This shows the changes in DAI over time in an animal model of DSS ingestion treated with focused ultrasound twice daily, compared to a control.

[0018] [Figure 10]Shows the changes in the DAI subcategories of stool hardness over the course of twice-daily focused ultrasound treatment in a DSS-fed animal model compared to the control.

[0019] [Figure 11A] Shows the changes in the DAI subcategories of gross bleeding over the course of twice-daily focused ultrasound treatment in a DSS-fed animal model compared to the control.

[0020] [Figure 11B] Shows representative photographs of the changes in gross bleeding from rats on days 7 and 8 from various treatment groups and the control group.

[0021] [Figure 12] Shows the weight change as a percentage from baseline over the course of twice-daily focused ultrasound treatment in a DSS-fed animal model compared to the control.

[0022] [Figure 13] Shows the stool hardness scores of individual animals over the course of twice-daily focused ultrasound treatment in a DSS-fed animal model compared to the control.

[0023] [Figure 14] Shows the gross bleeding scores of individual animals over the course of twice-daily focused ultrasound treatment in a DSS-fed animal model compared to the control.

[0024] [Figure 15A] Shows the colon lengths of various groups in the study.

[0025] [Figure 15B] Shows representative photographs of colon lengths sized relative to each other.

[0026] [Figure 16A]The images show colonic hematoxylin and eosin Y stained sections from an animal model that received focused ultrasound treatment (DSS) twice daily, compared to a control.

[0027] [Figure 16B] The images show colonic hematoxylin and eosin Y stained sections from animal models that received daily focused ultrasound (DSS) compared to a control.

[0028] [Figure 16C] The histopathological scores of various groups in the study are shown.

[0029] [Figure 17] The array coordinates and array data for cytokine activity in rats, evaluated from the colon of rats in the "naive," "water + 2 × FUS," "DSS + 2 × pseudo-FUS," and "DSS + 2 × FUS" groups, are shown.

[0030] [Figure 18] Figure 17 shows the changes in the activity of individual cytokines in the animal population based on the data.

[0031] [Figure 19] Figure 17 shows the changes in the activity of individual cytokines in the animal population based on the data.

[0032] [Figure 20] Figure 17 shows the changes in the activity of individual cytokines in the animal population based on the data.

[0033] [Figure 21] This is a schematic diagram of a neuromodulatory system according to an embodiment of the present disclosure.

[0034] [Figure 22] This is a block diagram of a neural control system according to an embodiment of the present disclosure.

[0035] [Figure 23]This is a flowchart of a method for neural control according to an embodiment of the present disclosure. [Modes for carrying out the invention]

[0036] One or more specific embodiments are described below. In order to provide a concise description of these embodiments, not all features of the actual embodiments are described herein. It should be understood that in the development of any such actual embodiments, such as in an engineering or design project, numerous decisions specific to the embodiment must be made to achieve the developer's specific goals, such as addressing system-related or business-related constraints, and these constraints may differ from embodiment to embodiment. Furthermore, it should be understood that while such development efforts may be complex and time-consuming, they are nevertheless routine design, fabrication, and manufacturing tasks for those skilled in the art who are interested in this disclosure.

[0037] Any examples or illustrations provided herein should never be considered to represent any restriction, limitation, or definition of any one or more terms in which they are used. Rather, these examples or illustrations should be considered to be describing various particular embodiments and should be considered as illustrative only. Those skilled in the art will understand that any one or more terms in which these examples or illustrations are used encompass other embodiments, which may or may not be given with them or elsewhere herein, and that all such embodiments are intended to fall within the scope of that one or more terms. Wording referring to such non-limiting examples and illustrations includes, but is not limited to, “for example,” “for instance,” “such as,” “eg,” “including,” “in certain embodiments,” “in some embodiments,” and “in one (an) embodiment.”

[0038] When introducing elements of the various embodiments of this disclosure, the articles “a,” “an,” “the,” and “said” are intended to indicate that there is one or more of the elements. The terms “comprising,” “including,” and “having” are intended to be comprehensive and mean that additional elements other than those listed may exist. Furthermore, any numerical examples in the following description are intended to be non-limiting, and additional numbers, ranges, and percentages are within the scope of the disclosed embodiments.

[0039] Inflammatory bowel disease (IBD) encompasses a range of chronic and recurrent gastrointestinal (GI) disorders, with Crohn's disease (CD) and ulcerative colitis (UC) being two major notable mentions. IBD causes chronic inflammation of the GI ducts, accompanied by symptoms including persistent diarrhea, abdominal pain, bleeding, and / or rectal bleeding. Many IBD symptoms also manifest outside the GI ducts, including arthritis, episcleritis, and fatigue. As of 2015, approximately 3 million adults in the United States had been diagnosed with IBD, representing about 1% of the US population. The recent increase in diagnoses has positioned IBD as a global health issue, with an estimated 20% increase in new diagnoses occurring every decade, which is also posing a significant economic burden.

[0040] Despite efforts, treatment options for IBD are severely lacking in availability and / or effectiveness. Many patients become refractory to first-line drug therapy or develop further complications such as stenosis, perforation, or fistula, which may then necessitate surgical recommendations. The recent introduction of antitumor necrosis factor (TNF) agents for IBD has been a major advance in the treatment of the disease, but the non-responder rate can reach as high as 60%. Furthermore, 30% of individuals become refractory to treatment, relapse, and experience a recurrence of symptoms within 12 months of administration. Given that hospitalization and surgery rates for IBD have not decreased, and that data suggest a moderate increase in hospitalization and surgery rates for CD and UC, this only further highlights the unmet need and clinical utility for developing new therapeutic interventions to treat IBD.

[0041] There are various etiological components contributing to the development of IBD, including complex interactions between genetic, immunological, and environmental conditions. There is no single underlying pathophysiological mechanism for IBD, and the consensus in the field is that in individuals with inherent susceptibility to chronic disruption of the immune system and / or impaired interaction with the gut microbiota, certain environmental triggers disrupt gut homeostasis, leading to inflammation-mediated damage.

[0042] The direct link between the central nervous system (CNS) and the immune system may involve the spleen for its regulatory role in feedback control of cytokine release from the spleen and circulating macrophages. This is particularly relevant to IBD due to the role of dysregulated cytokine levels in regulating intestinal inflammation and colonic injury. Sensory nerve feedback from circulating cytokines / endotoxins may induce vagus-mediated signaling to splenic macrophages, thereby attenuating the inflammatory response. This may occur indirectly via adrenergic neurons in the celiac plexus within the cholinergic anti-inflammatory pathway (CAP). In light of this, peripheral pathways may serve as targets for novel bioelectrotherapy, although current electrical stimulation devices cannot stimulate the intestines.

[0043] A technique using focused ultrasound to neuromodulate the celiac plexus (or at least a portion thereof) for the purpose of improving IBD-like symptoms is provided herein. Results of a study in a rat model of colitis, including daily administration of dextran sulfate sodium (DSS) via a drinking bottle, are disclosed herein. This study evaluated endpoint metrics routinely used in rodent models of IBD, with and without administration of focused ultrasound. The focused ultrasound provided herein was shown to improve IBD-like symptoms, such as stool consistency, gross bleeding / diarrhea, and colon tissue integrity, in DSS rat models of colitis. Efficacy was revealed by using different formulations of DSS and was observed in both "mild" and "severe" versions of IBD. These results demonstrate the clinical utility of non-invasive focused ultrasound neuromodulation in IBD. Focused ultrasound can be used to target specific plexuses or ganglia of the peripheral nervous system for diseases such as IBD.

[0044] Figure 1 shows the experimental setup used to perform specific neuromodulatory experiments focused on the target provided herein (e.g., the celiac plexus). An energy application device, operating according to parameters set by a controller, applies focused ultrasound to the region of interest targeting the celiac plexus. In rats, this region is located directly below the diaphragm. The placement of the ultrasound transducer in rats was determined by initial ultrasound imaging, shown in Figure 2, which highlighted two major arteries, the hepatic artery and the splenic artery, as landmarks used to locate the celiac plexus. Subsequently, the transducer was positioned just lateral to the xiphoid process, approximately 25 mm above the celiac plexus, to activate the intestinal CAP pathway at the level of the superior mesenteric ganglion.

[0045] All experiments, including those involving animals, followed the guidelines of the National Institutes of Health and Albany Medical College (AMC) Institutional Animal Care and Use Committee (IACUC). Animals were purchased from Taconic Biosciences (Germantown, New York, USA) through procedures conducted during the light phase of the light-dark cycle (7:00 AM to 7:00 PM). Animals had free access to food and water.

[0046] Male Sprague Dawley rats with an initial body weight of 200–350 g were anesthetized by placing them in an inhalation chamber filled with 4% isoflurane (Harvard Apparatus, Massachusetts, USA) set to 2–3 L / min, allowing for easy handling of the animal's anus and subsequent collection of fecal samples for testing viscosity and blood analysis. Immediately afterward, the rats were placed supine with their heads in a sealed nose cone, while anesthesia was maintained at 2–3 L / min with 1.5% isoflurane via a tabletop vaporizer (Harvard Apparatus, Massachusetts, USA). The abdomen, specifically the area above the xiphoid process, was shaved and marked with a black pen to target the transducer for focused ultrasound administration. This location targets the anterior vagal trunk proximal to the superior mesenteric and both left and right celiac ganglia, as shown using an ultrasound imaging applicator (Figure 2). After applying focused ultrasound, the rats were removed from isoflurane anesthesia, weighed, and returned to their individual cages.

[0047] We used a dextran sulfate sodium (DSS) model to induce IBD-like pathophysiology in rats. Dextran sulfate sodium (DSS) is a water-soluble sulfated polysaccharide that induces intestinal inflammation in rats when administered daily over time in water and solution. This is because DSS is an epithelial cell irritant that concentrates in the large intestine, leading to ulcerative lesions throughout the lamina propria of the colonic mucosa. This induces IBD-like pathophysiology and symptoms by damaging the epithelial barrier and allowing luminal bacteria / antigens to enter the mucosa. Furthermore, the mucosal lesions also increase in number and size in the distal colon, inducing epithelial damage similar to that seen in human ulcerative colitis. DSS is a model of acute, chronic, and / or recurrent intestinal inflammation.

[0048] On the first day of the experiment, animals were given drinking water, including regular tap water, to which DSS was added. The concentration ranged from 4% to 9% depending on the experiment. The control group of animals consumed only tap water. For all groups, the drinking bottles were replaced every 3 to 5 days to ensure that the DSS in the water bottles had not decomposed and that there was no bacterial contamination that could affect DSS-induced symptoms.

[0049] The endpoint metric for evaluating symptoms in rodent models of IBD is the Disease Activity Index (DAI), which can be used as a metric for evaluating the effectiveness and / or success of neuromodulation, as provided herein. DAI scores were assessed using weight loss, stool consistency, and gross bleeding. Each criterion may range from 0 to 4 to reflect normal to the most severe symptoms. Weight loss was scored compared to baseline weight. This semi-quantitative analytical tool consists of daily measurements of animal weight, stool consistency, and the presence of blood in the stool and rectal bleeding according to the following criteria (Table 1). JPEG0007833404000001.jpg44170

[0050] Weight loss is assessed from 0 to 4 points, ranging from no weight loss (score 0) to weight loss of more than 15% (score 4); stool consistency is assessed from 0 to 4 points, ranging from normal texture and appearance (score 0) to watery diarrhea (score 4); and rectal bleeding is assessed from 0 to 4 points, ranging from no bleeding (score 0) to slight bleeding (score 2) to macroscopic bleeding (score 4). Combined, a maximum overall DAI score of 12 points is possible from these three subcategories.

[0051] The animals' weight was monitored daily to assess weight loss and its magnitude. Fecal consistency and macroscopic bleeding were visually evaluated using photographs taken as digital records. Fecal samples were further tested for blood using the Sure-Vue Fecal Occult Blood Slide Test System (Fisher Scientific, Hampton, NH). If the occult blood test was positive even though no bleeding was observed, the bleeding score was increased by 1 point. In addition to the DAI measurement criteria, beverage consumption over 12 days was also recorded.

[0052] Rats were given DSS orally for two weeks and then sacrificed with or without focused ultrasound treatment. Colons were collected for postmortem analysis, including comparison of colon length, cytokines, and histological damage, including epithelial damage and inflammatory cell infiltration.

[0053] To spatially restrict the size / scale of rat anatomical structures using focused ultrasound, the intestinal CAP pathway was targeted by modulating the celiac plexus (left and right celiac ganglia and / or superior mesenteric ganglia) with a focused ultrasound transducer probe positioned in the left ventral region between the xiphoid process and the inferior thoracic cage under isoflurane-mediated anesthesia (Figure 1).

[0054] The focused ultrasound system consisted of a function generator (Agilent 33120A), a radio frequency (RF) power amplifier (ENI 350L), and a custom-made 2.5 MHz focused ultrasound (FUS) transducer. The FUS transducer was 0.75 inches high and 0.75 inches in diameter, with a curved surface and a depth of focus of 25.4 mm. The FUS transducer was acoustically coupled to the animal through an ultrasound gel placed on a shaved abdomen. The function generator, amplified by the RF power amplifier, produced a pulsed sinusoidal waveform transmitted to the FUS transducer. FUS stimulation swept the transducer across the organ or 5 cm. 2 Unlike previous ultrasound techniques, which required leaving a footprint as unfocused ultrasound waves in place, this technology enables targeting of specific locations.

[0055] Focused ultrasound was applied to the target region for 3 minutes daily (carrier frequency: 2.5 MHz, amplitude: 300 mV, burst length: 300 carrier cycles, burst repetition period: 200 milliseconds). At a carrier frequency of 2.5 MHz, each carrier cycle is 0.4 microseconds, and 300 carrier cycles have a duration of 120 microseconds. Therefore, during each 200 millisecond burst repetition period, the transducer is activated and pulsed for 120 microseconds, and remains inactive for the remaining 199.88 milliseconds before repeating the next burst. The animals were divided into different groups described by treatment and control variables (Table 2). The animal groups were assigned according to their disease status and treatment. The left column is the animal group name, and the middle column is the treatment. A total of 60 rats were divided into the following groups: Animals that ingested dextran sulfate sodium, labeled "DSS"; Animals that received non-invasive focused ultrasound, labeled "FUS"; As a sham control group for focused ultrasound, labeled "Sham FUS," rats received focused ultrasound on a shaved abdomen, etc., under the same procedure as those anesthetized with isoflurane, marked with a pen, and the transducer probe was placed at the same spot between the xiphoid process and the lower rib, however, the RF power amplifier, function generator, and transducer were turned off. Animals that did not receive DSS or focused ultrasound and were removed from their cages only once a day to monitor weight changes and assess stool consistency and gross bleeding scores were labeled "Naive." Animals that received non-invasive focused ultrasound twice a day were labeled "2×" in the procedure naming. Specifically in the "2×FUS" group, rats received two focused ultrasound treatments daily: a first focused ultrasound session in the morning, approximately 6 hours after the first session, and a second focused ultrasound session in the afternoon. The settings for the second focused ultrasound were identical to those for the first focused ultrasound performed in the morning, but no endpoint metrics were collected during the second focused ultrasound treatment. For some animal groups, labeling was combined to reflect receiving multiple treatments. For example, rats ingesting DSS but receiving two sham focused ultrasound treatments daily were labeled "DSS+2×ShamFUS".

[0056] After the experiment was completed, the animals were anesthetized by IP injection of urethane in physiological saline (1.2–1.5 g / kg), followed by thoracotomy and incision to remove the colon from the cecum to the anus. Photographs were taken before and after flushing the colon with 1× phosphate buffer (PBS). The colon was divided into three sections approximately 20 mm long: the proximal section near the cecum, the distal section near the anus, and the central section between the two ends. Each section was weighed, rapidly frozen in liquid nitrogen, and stored at -80°C until the tissue was homogenized using a lysis buffer (tissue extraction reagent I, Invitrogen, Vienna, Austria) containing an inhibitor cocktail and ethylenediaminetetraacetic acid (Halt® protease inhibitor cocktail, Thermo Scientific, Rockford, Illinois, USA) for cytokine analysis. The samples were then centrifuged at 10,000 g for 30 minutes at 4°C, the supernatant was collected, and stored at -80°C. Protein concentrations were analyzed using a bicinconic acid assay (BCA), and cytokine activity was evaluated using the Proteome Profiler Rat Cytokine Array Kit (R&D Systems, Minneapolis, Minnesota). This rat cytokine array kit contained two sets of control antibodies on a nitrocellulose membrane. Blocking buffer was added to a 4-well plate, and the membrane was incubated for 1 hour. Solutions containing 26 different biotinylation detection antibodies (400 μg / μl) were incubated at room temperature for 1 hour and replaced with the blocking buffer removed from the wells. The membrane was placed in a shaker (Microjive shaker, Boekel Scientific, Feasterville, Pennsylvania) and incubated overnight at -4°C. The following day, the membrane was washed three times with washing buffer to remove unbound material. Subsequently, streptavidin-HRP and chemiluminescence detection reagent were added to the membrane, resulting in the appearance of dots. These positions, sizes, and colors correspond to the amount of bound selected cytokines (see Figures 17-18).The membrane was scanned using a digital block scanner (ChemiDoc Western Blot Digital imaging system, Bio-Rad, Hercules, California) and analyzed using HLImage++ (Western Vision Software, Salt Lake City, Utah).

[0057] Sections of the rectum and an adjacent 1.5 cm of colonic tissue were also obtained, preserved in 4% paraformaldehyde (PFA), transferred to 70% ethanol for hematoxylin and eosin (H&E) staining, and embedded in paraffin. This tissue was sectioned longitudinally to a thickness of 5 μm and placed on charged microscope slides (Globe Scientific; Mahway, New Jersey).

[0058] The slides were dewaxed with xylene and washed three times, followed by a series of alcohol washes with 100% ethanol (2x) and 95% ethanol (2x) to hydrate the tissue. The slides were placed in hematoxylin 7211 and immersed in a bluing reagent using Eosin-Y, followed by differentiation. Prior to the Eosin-Y wash, the slides were washed once with 95% alcohol. The slides were then dehydrated again by being immersed three times in 100% ethanol washing solution, washed with a xylene substitute washing solution, and covered with Cytoseal 60 (Fisher Scientific, Hampton, NH).

[0059] Hematoxylin-eosin Y stained sections were used to assess inflammatory cell infiltration, crypt distortion, and erosion using semi-quantitative scoring friction (Table 3). Histological disease severity was assessed using a scoring friction of 0–4 for epithelial damage and inflammatory cell infiltration. The sum of both scores was combined to obtain a total histological damage score of 8. The center column describes the magnitude of epithelial damage based on morphology, goblet cell loss, crypt loss, and crypt loss over a wide area. The right column describes the severity of inflammatory cell infiltration based on the depth of penetration into various colonic layers. JPEG0007833404000003.jpg61170

[0060] Specifically, epithelial damage and invasion were independently scored on a scale of 0 to 4: 0 = normal morphology; 1 = loss of goblet cells; 2 = loss of goblet cells over a wide area; 3 = loss of crypts; 4 = loss of crypts over a wide area. Invasion was scored as follows: 0 = no infiltration; 1 = infiltration near the base of the crypts; 2 = infiltration reaching the muscularis mucosa; 3 = extensive infiltration reaching the muscularis mucosa and mucosal thickening with abundant edema; 4 = submucosal infiltration. The sum of both epithelial damage and invasion results in a total maximum histopathological score of 8.

[0061] DSS chemicals consistent with symptoms of IBD considered "mild" were purchased from Sigma-Aldrich (MW: >500kDA, lot number BCBZ5763). DSS chemicals that induce symptoms of "severe" IBD were purchased from Fisher Scientific (MW: 35-50kDa, catalog number AAJ6360622). Unless otherwise specified, all other chemicals were purchased from Sigma-Aldrich, and any other chemicals and drugs purchased elsewhere are listed in this document.

[0062] DAI, stool consistency, and gross bleeding were assessed by at least two unblinded laboratory staff, and histopathological tissue was scored by two blinded laboratory staff. Data were analyzed using two-way analysis of variance (ANOVA) with Graphpad Prism software (v8.3.0; San Diego, California), with repeated measures where possible, and post-hoc analysis using Fisher's LSD for inter-treatment comparisons per day. Some datasets had missing values ​​from animals excluded from the study due to weight loss, mortality, and / or other exclusion criteria. For these data, a mixed-effects model (REML) two-way ANOVA was used instead of repeated measures. The decrease in animal numbers is expressed as sample size, with a range of values ​​where larger numbers represent the initial group size and smaller values ​​represent the lowest sample size after animals were excluded from the study. Sample size ranges are established for each animal group for analyses performed throughout the entire experimental schedule. In addition, n values ​​for multiple mentioned groups are separated by forward slashes representing the first and second listed groups, respectively. For example, comparing "DSS" and "DSS+FUS" (n=6-1 / 7-2) shows that the "DSS" group had a maximum of 6 animal data points and a minimum of 1 animal data point across all days, while the "DSS+FUS" group had a maximum of 7 animal data points and a minimum of 2 animal data points across all days. For daily analysis, comparing "DSS" and "DSS+FUS" on day 5 (n=4 / 5) shows that the "DSS" group had 4 data points and the "DSS+FUS" group had 5 data points on day 5. Colon lengths were compared using one-way ANOVA with multiple comparisons using the Holm-Sidak post-hoc test. Data are presented as mean ± standard error of the mean (SEM), and statistical significance was judged at p<0.05.

[0063] Figure 3 shows the DAI scores of DSS animals, regardless of whether or not focused ultrasound treatment was performed. The animals ingested either water or DSS. These two groups included cohorts of animals that received focused ultrasound (FUS) ("DSS+FUS" and "Water+FUS") and animals that did not ("DSS" and "Water+ShamFUS," the latter receiving sham ultrasound sessions). Rats in the "DSS" group (n=7-1) showed higher DAI scores immediately one day after drinking DSS compared to animals that drank tap water (n=11-2). The "DSS+FUS" group (n=8-4) had lower DAI scores and showed less severe IBD symptoms. Thus, as provided herein, focused ultrasound targeting the celiac plexus may result in improvement of IBD-related symptoms. The difference in DAI scores between the "DSS" group and the "DSS+FUS" group was first significant on day 5 (n=5 / 4), and remained significant on day 7 (n=2 / 4), day 9 (n=6 / 8), day 11 (n=6 / 8), and day 12 (n=4 / 4). An asterisk (*) indicates p≦0.05 for "DSS" vs. "DSS+FUS," and a pound (#) indicates p≦0.05 for "DSS+FUS" vs. "Water+FUS."

[0064] The cohort in Figure 3 represents 5 percent (5%) of the DSS group, which induced a significant increase in DAI scores in the DSS group compared to the water + sham FUS control group (n=7-1 / 11-2; p<0.001, Figure 3). The difference in DAI between these two groups was significant from 1 day after DSS intake (n=4 / 11; p=0.004) to the end of the experiment on day 12 (n=4 / 2; p<0.001). Comparing the DSS groups, those receiving focused ultrasound ("DSS+FUS" group) had significantly healthier DAI scores than those not receiving focused ultrasound ("DSS" group) on days 5 (n=5 / 4, p=0.014), 7 (n=2 / 4, p=0.047), 9 (n=6 / 8, p=0.007), 11 (n=6 / 8, p=0.022), and 12 (n=4 / 4, p=0.022) (n=8-4 / 7-2; p=0.045). On day 7 (n=2 / 4), when the greatest efficacy was observed between the "DSS" group and the "DSS+FUS" group, the former had a DAI score of 6±1.4, while the latter had a DAI score of 2.5±1.7. No difference in DAI scores was reported between the "water + FUS" and "water + sham FUS" groups (n=11-2 / 5-1; p=0.222). Interestingly, focused ultrasound was effective in measuring the overall DAI score of rats that ingested DSS, but no weight loss was observed in the DSS group (Figure 6), one of the subcategories that includes DAI score, which has also been frequently reported in other studies using 5% DSS by the end of the experimental period.

[0065] Each subcategory of the DAI was examined independently, rather than as an overall indicator, as shown in Figures 4-6. Subcategories of the DAI score included stool consistency (Figure 4), gross bleeding (Figure 5), and weight loss (Figure 6). In Figure 4, animals in the "DSS+FUS" group (n=7-2) showed lower stool consistency scores than animals in the "DSS" group (n=8-4). This effectiveness was observed as early as day 2, and improvement continued on days 3, 4, 5, and 11 compared to those without FUS. In Figure 5, gross bleeding symptoms were observable in the "DSS" group by day 5, but FUS delayed the onset of gross bleeding symptoms by 3 days. Specifically, the "DSS+FUS" group showed no difference in gross bleeding compared to the "Water+FUS" group until day 8. In Figure 6, no difference in weight loss was observed between the "DSS" group and the "DSS+FUS" group, or between any other group. An asterisk (*) indicates a p ≤ 0.05 comparison between "DSS" and "DSS+FUS," and a pound (#) indicates a p ≤ 0.05 comparison between "DSS+FUS" and "Water+FUS." DSS intake resulted in scores for loose stools and overly soft stools compared to rats in the "Water+SpuriousFUS" and "Water+FUS" groups (e.g., "DSS" vs. "Water+SpuriousFUS": n=6-1 / 5-1, p<0.001; "DSS" vs. "Water+FUS": n=6-1 / 11-2, p<0.001). Specifically, stool hardness scores worsened significantly by day 2 and remained higher than those of rats that drank water, with or without focused ultrasound, until the end of the experiment (p<0.001 for comparisons of both water groups over all days, Figure 4). Rats in the water group had similar stool hardness scores, regardless of whether they received focused ultrasound or not ("water + pseudo-FUS" vs. "water + FUS"; p=0.113). Stool hardness was significantly worse in the "DSS + FUS" group than in the "water + FUS" group, based on stools from days 4-5 and 7-12.However, rats in the "DSS+FUS" group showed better stool consistency than the "DSS" group (n=7-2 / 8-4; p=0.008), specifically on day 2 (n=8 / 7; p<0.001), day 3 (n=4 / 5; p=0.006), day 4 (n=8 / 7; p=0.006), day 5 (n=4 / 5; p=0.010), and day 11 (n=8 / 6; p=0.036) (Figure 4). The greatest improvement with focused ultrasound was observed on day 2 in the "DSS" group, where the stool consistency severity score was 2.2±1.0, while the "DSS+focused ultrasound" group had a score of 0.8±0.3.

[0066] The effects of DSS and FUS on fecal hardness can be observed in representative images (Figure 7) derived from postmortem photographic images of the rat colon. Animals receiving DSS, in contrast to the "water + 1 × FUS" group, had colons containing unformed pellets or no feces at all, particularly in the distal portion of the colon (upper arrow). In contrast, animals receiving both DSS and FUS ("DSS + 1 × FUS") showed intact "normal" pellet shape after two weeks of FUS treatment (lower arrow), similar to that observed in the "water + 1 × FUS" group. Fecal hardness scores were obtained from fresh fecal samples excreted from the animals daily in the morning. However, hardness was also visually verifiable when the rat colon was examined at the end of the experiment after the animals were sacrificed. Next, the "DSS+1×FUS" group (bottom of Figure 7) had healthier-looking tissues and harder fecal pellets that more closely resembled those of animals that drink only tap water (top of Figure 7, photograph), compared to the "DSS" group rats, which had little to no fecal shape or fecal pellets (center of Figure 7).

[0067] Animals in the "DSS" group had generally worse macroscopic hemorrhage scores than animals in the "water + 1 × pseudo-FUS" group (n=7-1 / 5-1; p=0.002), starting from day 5 (n=5 / 3; p=0.05) and up to day 12 (n=4 / 3; p=0.002), with the exception of day 6 (n=1 / 1; p=0.181) (Figure 5). Macroscopic hemorrhage was similar between the "water + FUS" group and the "water + pseudo-FUS" group throughout the 12 days (p=1.000). In particular, animals in the "DSS+FUS" group showed a significantly improved macroscopic bleeding score compared to animals in the "DSS" group (n=7-2 / 8-4; p=0.015), which was first observed on day 5 (n=4 / 5, p=0.048), followed by day 7 (n=4 / 2, p=0.047), day 9 (n=8 / 6, p=0.001), and day 11 (n=8 / 6, p=0.020) (Figure 5). The greatest improvement in macroscopic bleeding from the "DSS" group was seen on day 9, with the "DSS" group having a score of 3.0±0.7 and the "DSS+FUS" group having a score of 1.1±0.4. Furthermore, the appearance of macroscopic bleeding symptoms was delayed by approximately 3 days on focused ultrasound. Specifically, rats in the "DSS" group showed a macroscopic bleeding score of more than 1 by day 5. In contrast, the "DSS + focused ultrasound" group first showed a score of 1 or higher on day 8.

[0068] Rats that ingested a 5% DSS solution for 12 days exhibited IBD-like symptoms such as bloody stools with loose and watery stools, consistent with the DSS rodent model, but did not show significant weight loss. In fact, animals ingesting DSS gained weight despite ingesting the solution daily for two weeks, and this trend was observed in all groups ("DSS" vs. "DSS+FUS": n=7-1 / 8-4, p=0.534; "DSS+FUS" vs. "Water+FUS": n=8-4 / 5-1, p=0.577; "Water+FUS" vs. "Water+SpuriousFUS": n=5-1 / 5-1, p=0.15) Figure 6). Interestingly, only transient weight loss occurred in the "DSS+FUS" group on days 2 and 3.

[0069] Considering the absence of significant and long-term weight loss, the rats in the "DSS" group showed a maximum DAI score of only 6.7 out of 12, which is lower than the values ​​typically reported in high-concentration rodent DSS models. To induce more severe IBD symptoms, a 40kD formulation of DSS was used instead of the initial >500kD formulation used to induce the mild gastrointestinal injury described above. A second set of experiments was then conducted with a more potent DSS formulation to better reproduce severe IBD symptoms, allowing for an evaluation of the effectiveness of FUS for this severity of IBD symptoms.

[0070] We investigated smaller-sized DSS chemicals to determine whether they induced IBD symptoms that better replicated severe IBD models. Different groups of rats given 4%, 5%, 7%, or 9% DSS were monitored for two weeks (Figure 8). Based on the results, 7% DSS induced the most severe IBD-like symptoms with a maximum DAI score of 12 at day 10. In contrast, 9% DSS caused high mortality and / or led to the exclusion of rats from the study due to >20% weight loss by days 4–6, while 5% DSS induced only a maximum DAI score of 10. Considering the optimization data, we investigated the effectiveness of non-invasive focused ultrasound as a treatment for severe IBD symptoms using a 7% DSS concentration of a newer DSS formulation.

[0071] In the “severe” DSS model with a DSS concentration of 7%, the same focused ultrasound parameters as in the “mild” model were used to improve IBD symptoms. However, preliminary findings revealed that the DAI score did not improve in the “DSS+FUS” group animals, which had similar shortened colon length as seen in the “DSS” group (Figure 15A) (data not shown). From the initial experiments, it was determined that further administration of focused ultrasound was necessary to achieve effective intestinal CAP stimulation to mitigate the increasing severity of IBD. Therefore, the “dosage” of focused ultrasound treatment was increased in parallel by applying non-invasive focused ultrasound twice a day (“2×FUS”) instead of once a day.

[0072] Figure 9 shows the effect of 2× focused ultrasound on the DAI score in this group. When rats that ingested DSS were administered FUS twice daily, DAI symptoms improved compared to animals that ingested DSS and received sham 2×FUS. Improvement in DAI scores was again observed on day 5 (n=6 / 6) and day 7 (n=6 / 5). An asterisk (*) indicates a p<0.05 difference between the "DSS+2×FUS" group and the "DSS+sham 2×FUS" group. The "DSS+2×FUS" group had significantly higher (worse) DAI scores than the "water+2×FUS" group (n=6-1 / 6-1; p<0.001), but lower than the "DSS+sham 2×FUS" group (n=6-1 / 6-1; p=0.046). Specifically, rats that ingested DSS without focused ultrasound ("fake 2×FUS") had significantly higher scores than animals that received 2× focused ultrasound by day 5 (n=6 / 6; p=0.037) and day 7 (7.5±1.3 vs. 4.8±0.6, n=6 / 5; p=0.002). Animals on day 12 may also have shown FUS efficacy, but the sample size excluded any conclusions (10.0±0.0 vs. 6.0±0.0, n=1 / 1) (Figure 9). Maximum efficacy occurred on day 7. Interestingly, no changes were observed between the "DSS + sham 2×FUS" animals and the "DSS + 2×FUS" animals on days 8, 9, 10, and 11 (p = 0.11, 0.62, 0.80, and 0.29, respectively). This may be due to the three animals with the highest DAI scores in the "DSS + sham 2×FUS" group being excluded from the data due to mortality on days 7 (DAI: 11; 12) and 8 (DAI: 10). No animals in the "DSS + 2×FUS" group died during this period.

[0073] Figures 10-12 show each subcategory of DAI in detail. Figure 10 shows the effect of FUS on stool consistency scores. Animals that drank DSS and received 2×FUS (n=6-1) showed improved stool consistency compared to animals in the "DSS + 2×Spurious FUS" group (n=6-1). Significant improvement was observed from day 4 (n=6 / 6) to day 7 (n=6 / 5). An asterisk (*) indicates p<0.05 between the "DSS + 2×FUS" group and the "DSS + Spurious 2×FUS" group. A pound (#) indicates "DSS + 2×FUS" vs. "Water + 2×FUS" at p≦0.05. There was no difference between drinking animals that received 2× focused ultrasound or 2x sham-focused ultrasound ("Water + 2×FUS" vs. "Water + Sham 2×FUS": n=6-1 / 6-2; p=0.068) (Figure 10). In contrast, animals in the "DSS + Sham 2×FUS" group had worse stool hardness scores than animals in the "Water + Sham 2×FUS" group (n=6-1 / 6-2; p<0.001) from day 2 (n=6 / 6; p<0.001) to day 12 (significance not indicated). Ultrasound administered twice daily improved stool hardness in rats that drank DSS ("DSS + Sham 2×FUS" vs. "DSS + 2×FUS": n=6-1 / 6-1; p<0.001) (Figure 10), which was first observed by day 4. The latter group showed a stool consistency score of 0.5±0.34, while the former group had a score of 1.5±0.22 (n=6 / 6; p=0.003). The largest difference between the groups was on day 5, with the "DSS + sham 2× focused ultrasound" group (n=6) having a score of 2.2±0.40 and the "DSS + 2× focused ultrasound" group (n=6) having a score of 1.0±0.0. The improvement in stool consistency persisted until day 7, with the "DSS + 2× FUS" group (n=6) having a stool consistency of 2.4±0.24. In contrast, animals that ingested DSS with 2× sham FUS had a score of 3.3±0.21 (n=5; p=0.008). In particular, two animals in the "DSS+2 × pseudo-FUS" group, which had the highest stool hardness score of 4, died on day 7-8.

[0074] Figure 11A shows the effect of FUS on macroscopic bleeding scores. Animals receiving DSS but also FUS ("DSS + 2 × FUS"; n=6-1) had lower macroscopic bleeding scores than animals receiving sham FUS ("DSS + sham 2 × FUS"; n=6-1) from day 5 (n=6 / 6) to day 8 (n=4 / 5). Asterisks (*) indicate p<0.05 between the "DSS + 2 × FUS" group and the "DSS + sham 2 × FUS" group. Pounds (#) indicate "DSS + 0.05 2 × FUS" vs "Water + 2 × FUS" at p≦0.05. Figure 11B shows representative photographs of rats on day 7 and day 8 from various treatment and control groups. Animals receiving DSS + 1 × FUS, as well as DSS + 2 × sham FUS, had darker blood due to colonic blood stasis from a severe disease model. In contrast, animals treated with "DSS + 2×FUS" showed a visual improvement in macroscopic bleeding compared to animals treated with "DSS + pseudo 2×FUS". The "DSS + pseudo 2×FUS" group had macroscopic bleeding in the anus and feces, while animals in the "water + 2×FUS" group did not (n=6-1 / 6-1; p<0.001). Symptoms first appeared on day 5 (n=3 / 6; p=0.003), continued until day 9 (n=3 / 6; p<0.001), and possibly persisted until day 12, which was consistent with the limited sample size (n=3 / 1) (Figure 11A). Similar to findings regarding fecal consistency, animals in the "DSS + 2×FUS" group had reduced macroscopic bleeding compared to the "DSS + 2× pseudo FUS" group (n=6-1 / 6-1; p=0.012). Efficacy was first observed by day 5 (n=6 / 6; p=.006) in the group that received 2× focused ultrasound with a score of 0.0±0.0 and in the group that ingested 7% DSS without focused ultrasound with a score of 0.83±0.31. Improved macroscopic bleeding was extended to day 8 in the "DSS+2×fakeFUS" group with a score of 3.75±0.25 and the "DSS+2×FUS" group with a score of 3.0±0.0 (n=4 / 5; p=0.028). The largest difference in macroscopic bleeding scores between the two groups occurred on day 6. The "DSS+fake2×FUS" group showed a score of 2.0±0.26, while the "DSS+2×FUS" group showed a score of 0.83±0.30.In the "DSS+2 × pseudo-FUS" group, two animals with the most severe macroscopic hemorrhage score of 4 on day 7 died before day 8 (Figure 11A).

[0075] Figure 12 shows the change in body weight as a percentage from baseline, with negative values ​​representing weight loss. Both the DSS group with and without FUS experienced weight loss by the end of the experiment, while the water group experienced weight gain. No difference was observed between the DSS groups receiving 2×FUS (n=6-1) or sham 2×FUS (n=6-1). A cross symbol (†) indicates "water + sham 2×FUS" vs. "water + 2×FUS" for p≦0.05. Rats in the "water + sham 2×FUS" group (n=6-2) gained weight over 12 days, while rats that drank water and received 2× focused ultrasound (n=6-3) did not gain weight at the same rate (p<0.001). In fact, the "water + 2×FUS" group experienced weight loss during the first four days of the experiment. On day 5, the "water + sham 2×FUS" rats had gained 6.1 ± 0.8% of their body weight from day 1, while the "water + 2×FUS" group had lost 1.4 ± 0.5% of their body weight from day 1 (n=6 / 6; p<0.001). Weight loss was also observed in the "DSS + 2×FUS" group. However, considering that the animals had harder stools, less significant pain and bleeding, were more active, and exhibited exploratory behavior in their home cages, this may largely reflect the effect of 2×FUS rather than the severity of IBD symptoms. Furthermore, a similar weight loss trend was observed in the "water + 2×FUS" group. Conversely, the "DSS + sham 2×FUS" animals also lost weight (n=6-1), but despite having similar weight loss to the "DSS + 2×FUS" group, these animals showed significant impairments of motility and lethargy when handled (p=0.690), Figure 12.

[0076] Figures 13 and 14 show the stool consistency (Figure 13) and gross hemorrhage (Figure 14) scores of individual animals in the "DSS + 2 × FUS" group (n=6-1) and the "DSS + pseudo 2 × FUS" group (n=6-1). Each line on the graph represents an individual animal, and each arrow next to these lines indicates a "decrease" in the score. This highlights examples of improved symptoms. In the stool consistency score (Figure 13), as indicated by the number of arrows, there was a decrease of 14 in the "DSS + 2 × FUS" (n=6-1) group. In contrast, the "DSS + pseudo 2 × FUS" group (n=6-1) showed only two decreases. Figure 14 also shows that animals in the "DSS + 2 × FUS" group showed a greater number of decreases in gross hemorrhage score than animals in the "DSS + pseudo 2 × FUS" group. The scores were plotted to track changes over 12 days. In general, the animals in the "DSS+2×FUS" group (n=6-1) showed a greater daily decrease in DAI subcategory scores than the "DSS+fake 2×FUS" control animals (n=6-1). Specifically, rats in the "DSS+fake 2×FUS" group showed a sustained deterioration in the stool hardness severity score and only two defecations were observed. One occurred between days 2 and 3, and the other between days 4 and 5 (red downward arrows, Figure 13). In contrast, the ultrasound-treated rats ("DSS+2×FUS") showed more defecations and greater improvement in daily stool hardness severity, with 14 defecations observed throughout the 12-day experiment (downward arrows) (Figure 13). Similar findings were observed for the macroscopic hemorrhage scores of individual animals (Figure 14), with rats in the "DSS + pseudo-2×FUS" group showing a sustained increase (worsening) of macroscopic hemorrhage symptoms, except for two cases with fluctuations observed between days 8 and 9, and between days 10 and 11. However, rats in the "DSS + 2×FUS" group had many instances of downward fluctuations in the score, particularly in the first half of the experiment between days 3 and 5, with three fluctuations. Three more fluctuations also occurred between days 10 and 12 (Figure 14). When analyzed across fecal consistency and macroscopic hemorrhage endpoints, rats ingesting DSS with 2× focused ultrasound showed significantly higher daily improvement in the number of "downward" fluctuations or severity scores than rats ingesting DSS with 2× pseudo-focused ultrasound (p=0.012).

[0077] Figure 15A shows group data for colon length. As shown on the x-axis, certain groups ingested water, while others received DSS. Figure 15B shows representative photographs of the group data from Figure 15A, with a ruler along the colon for reference. The photographs are not scaled and are sized relative to each other. Compared to the DSS + sham 2×FUS control group, the pellet shape remained intact in the animals of the DSS + 2×FUS group. An asterisk (*) indicates p<0.05, and ns indicates no significant difference. Reduction in colon length is another important endpoint metric for DSS-induced IBD. The "DSS + 2×FUS" group had significantly shorter colon lengths than the "Water + 2×FUS" group (p=0.016 vs; n=5 / 6), but were even more intact and longer than the "DSS + Sham 2×FUS" group (18.5±1.8cm) (13.8±0.3cm, n=5 / 4; p=0.013) (Figure 15A~B). Rats that received focused ultrasound once daily ("DSS + 1×FUS") did not show improvement in colon length compared to the "DSS + Sham 1×FUS" control group, where the former was 14.1±0.50cm (n=4) and the latter was 13.0±0.3cm (n=3; p=0.958). No difference in colon length was observed between the "water + 2 × FUS" group (n=6) and the "water + 2 × pseudo-FUS" group (n=5), with lengths being 22.7 ± 0.5 cm and 22.6 ± 0.4 cm, respectively (p=0.958).

[0078] Figures 16A-C show histopathological data. A 2 cm section of the distal colon was preserved in paraformaldehyde immediately after sacrifice. Sections were treated with hematoxylin and eosin Y staining. Figure 16A shows mucosal damage in the distal colon of animals receiving DSS using pseudofocused ultrasound (left image) compared with DSS animals using 2× focused ultrasound (right image). Lesions and infiltrations in the crypt base and deep submucosa are increased. Figure 16B shows a magnified view (upper right image) of inflammatory factors in animals receiving DSS that are not observed in animals receiving DSS + 2× FUS (lower right image). Figure 16C shows grouped data derived from the scoring criteria for semi-quantitative analysis. Colons were collected from animals in the following groups: "water + pseudo 2×FUS" (n=2), "water + 2×FUS" (n=3), "DSS + pseudo 2×FUS" (n=6), and "DSS + 2×FUS" (n=5). The "DSS + pseudo 2×FUS" group had the highest histopathological score of 8 at the end of the experiment, indicating the greatest epithelial damage and inflammatory cell infiltration (Figure 16B: upper right image vs. lower right image). Rats that ingested DSS and received 2× focused ultrasound had higher histopathological scores (mean score: 6.3, SEM: 0.8) than the "water + 2×FUS" group (mean score: 0.4, SEM: 0.4) and the "water + pseudo 2×FUS" group (mean score: 1.8, SEM: 0.5) (p<0.001). However, receiving 2× focused ultrasound while ingesting 7% DSS was still associated with a healthier colon than the "DSS + 2× sham FUS" animals (mean score: 8.0, SEM: 0.0) (p=0.022) (Figure 13c). There was no difference between the "water + 2× FUS" group and the "water + sham 2× FUS" group (p=0.173) (Figure 16C).

[0079] Cytokine activity was evaluated from the colons of rats in the "naive," "water + 2x FUS," "DSS + 2x pseudo-FUS," and "DSS + 2x FUS" groups (n = 4 rats / group). Of the 29 cytokines detected using a rat cytokine array (Figure 17), seven cytokines were identified, and consistent results were obtained that led to significant differences between "naive" vs. "DSS + 2x pseudo-FUS," "water + 2x FUS" vs. "DSS + 2x pseudo-FUS," and "DSS + 2x pseudo-FUS" vs. "DSS + 2x FUS" (Figure 18). Specifically, ciliary neurotrophic factor (CNTF), fractalkines, granulocyte-macrophage colony-stimulating factor (GM-CSF), monokines induced and regulated by gamma interferon (MIG), normal T cell expression secretion (RANTES), tumor necrosis factor α (TNF-α), and thymic chemokines were all increased in rats ingesting pseudo-2×FUS DSS derived from naive rats or in rats ingesting 2×FUS water. Furthermore, all of these levels decreased to "naive" and "water + 2×FUS" levels in animals ingesting DSS but also receiving 2×FUS (Figure 18, bottom).

[0080] Similar response patterns were observed for eight out of ten interleukin cytokines, including IL-1β, IL-2, IL-3, IL-4, IL-6, IL-10, IL-13, and IL-17 (Figure 20), all of which were elevated in animals ingesting DSS with 2× pseudo-FUS from the “naive” and “water + 2× FUS” animal groups. These levels decreased to “naive” and “water + 2× FUS” levels in animals ingesting DSS but also receiving 2× FUS (Figure 19, bottom). Finally, eight other cytokines were identified that showed a more variable response to DSS and / or FUS (Figure 20). In particular, IL-1α and IL-1ra are two other interleukins in the assay kit. IL-1ra levels did not increase in animals ingesting DSS without 2×FUS from the "naive" or "water + 2×FUS" groups, while IL-1α levels were higher in "DSS + 2×FUS" animals than in "water + 2×FUS" animals, but not as high as in "naive" rats (Figure 21, bottom). Nevertheless, interleukins decreased in both animals ingesting DSS with 2×FUS and those ingesting DSS without 2×FUS. Interferon-gamma (IFN-gamma) levels increased in animals ingesting DSS without 2×FUS from the "naive" and "water + 2×FUS" groups, but decreased with 2×FUS (Figure 20, bottom). This decrease was still higher than the levels seen in "naive" rats, but no different from rats ingesting water with 2×FUS. Unsurprisingly, a similar pattern of change was observed for interferon-gamma-inducible protein 10 (IP-10, also known as CXCL10). The only cytokine that decreased in animals ingesting DSS without 2×FUS from "naive" rats was LIX (also known as CXCL5). Its levels were unaffected by 2×FUS.

[0081] Finally, seven cytokines remained invariant across all four animal groups, regardless of whether they were in drinking water or DSS with and without 2×FUS. These included metalloproteinase tissue inhibitor (TIMP-1) (Figure 20, bottom), along with cytokine-induced neutrophil chemotactic agent-1 (CINC-1), CINIC2α / β, CINC-3, macrophage inflammatory protein-1α (MIP-1α), soluble intercellular adhesion molecule-1 (sICAM-1), and L-selectin (data not shown).

[0082] The disclosed neuromodulation techniques may be used in conjunction with neuromodulation systems. Figure 21 schematically represents a system 10 for neuromodulation such as the release of neurotransmitters and / or activation of synaptic components (e.g., presynaptic cells, postsynaptic cells) within a ganglion of the celiac plexus in response to the application of energy. For example, one or more synapses within a ganglion of the celiac plexus can be modulated using focused ultrasound. The illustrated system includes a pulse generator 14 coupled to an energy application device 12 (e.g., an ultrasound transducer). The energy application device 12 is configured to receive energy pulses, for example via leads or wireless connections, directed during use to a region of interest in the internal tissue or organ of a target, resulting in a targeted physiological outcome. In certain embodiments, the pulse generator 14 and / or the energy application device 12 may be implanted in a biocompatible site (e.g., the abdomen), and one or more lead wires may internally couple the energy application device 12 and the pulse generator 14. For example, the energy application device 12 may be a MEMS transducer such as a capacitive micromachine ultrasonic transducer.

[0083] In certain embodiments, the energy application device 12 and / or pulse generator 14 can communicate wirelessly with, for example, a controller 16, which can then issue commands to the pulse generator 14. In other embodiments, the pulse generator 14 may be an extracorporeal device, for example, operated to apply energy percutaneously or non-invasively from an external location on the subject, and in certain embodiments may be integrated with the controller 16. In embodiments where the pulse generator 14 is extracorporeal, the energy application device 12 may be operated by a caregiver and positioned on or above the skin of the subject so that energy pulses are delivered percutaneously to the desired internal tissue. Once positioned to apply energy pulses to the desired site, the system 10 can initiate neuromodulation to achieve a targeted physiological outcome or clinical effect.

[0084] In certain embodiments, the system 10 may include an evaluation device 20 coupled to the controller 16, which evaluates characteristics indicating whether the targeted physiological outcome of the regulation has been achieved. In one embodiment, the targeted physiological outcome may be local. For example, the regulation may result in local tissue or functional changes such as changes in tissue structure, local changes in the concentration of a particular molecule, tissue migration, or increased fluid movement.

[0085] Regulation can result in systemic or non-local changes, and targeted physiological outcomes may relate to changes in the concentration of circulating molecules or changes in tissue properties that do not involve the region of interest to which energy is directly applied. In one example, migration may be a surrogate measure of desired regulation, and if the measured migration value falls below the expected migration value, the regulation parameter may be modified until the expected migration value is induced. Thus, the evaluation device 20 may be configured in some embodiments to evaluate changes in concentration. In some embodiments, the evaluation device 20 may be an imaging device configured to evaluate changes in organ size and / or location. Although the depicted elements of system 10 are shown separately, it should be understood that some or all of the elements can be combined with one another. Furthermore, some or all of the elements can communicate with one another in a wired or wireless manner.

[0086] Based on the evaluation, the adjustment parameters of the controller 16 may be changed. For example, if the desired adjustment relates to a change in concentration (circulating or tissue concentration of one or more molecules) within a defined time frame (e.g., 5 minutes, 30 minutes after the start of the energy application procedure) or relative to the baseline at the start of the procedure, then changes in adjustment parameters such as pulse frequency or other parameters may be desired, which can then be given to the controller 16 by the operator or via an automated feedback loop to determine or adjust the energy application parameters or adjustment parameters of the pulse generator 14.

[0087] The system 10 provided herein can deliver energy pulses as part of a treatment protocol according to various modulatory parameters. For example, the modulatory parameters may include various stimulation time patterns ranging from continuous to intermittent. In intermittent stimulation, energy is supplied for a certain period at a specific frequency during the signal-on time. Following the signal-on time, there is a period during which no energy is supplied, referred to as the signal-off time. The modulatory parameters may also include the frequency and duration of stimulation application. The application frequency may be continuous or delivered over various periods, e.g., within a day or a week. The treatment protocol duration is not limited but can last for various periods, including from a few minutes to several hours. In a particular embodiment, the treatment duration with a given stimulation pattern may last for one hour, repeated, for example, at 72-hour intervals. In a particular embodiment, the treatment may be delivered for a shorter duration, e.g., 30 minutes, and at a higher frequency, e.g., every three hours. According to modulatory parameters such as treatment duration and frequency, the application of energy can be controlled to be adjustable to achieve the desired result.

[0088] The focused ultrasound energy can be focused on a region of interest 22, which may be an internal structure, tissue, or organ including at least a portion of the celiac plexus. For example, the region of interest may include a ganglion or ganglion of the celiac plexus. Synapses within the region of interest 22 of the target tissue can be stimulated by directly applying energy to synapses within the focal area of ​​an energy application device 12 focused on the region of interest 22 of the target tissue, which may cause the release of molecules into the synaptic cleft and / or changes in ion channel activity, which may then cause downstream effects such as activation of intestinal CAP. The region of interest 22 can be selected to include a specific ganglion of the celiac plexus, while excluding different ganglia of the celiac plexus (and ganglia that are not part of the celiac plexus). Thus, the region of interest 22 can be selected to correspond to a desired ganglion or a portion of target tissue that has or is adjacent to a ganglion. The application of energy may be selected to preferentially induce the release of one or more molecules, such as neurotransmitters, from nerves within the synapse.

[0089] In one embodiment, energy may be applied to two or more regions of interest 22. In some embodiments, the energy application parameters may be selected to induce preferential activation of either neuronal or non-neuronal components in the tissue directly receiving the energy, thereby inducing a desired combined physiological effect. In certain embodiments, the energy is approximately 25 mm 3 It can be focused or concentrated within a volume of less than 0.5 mm. In certain embodiments, the energy is approximately 0.5 mm. 3 ~50mm 3 The energy can be focused or concentrated within a certain volume. The focusing volume and focusing depth for focusing or concentrating energy within the region of interest 22 may be influenced by the size / configuration of the energy application device 12. The focusing volume of the energy application can be determined by the focal region of the energy application device 12.

[0090] As provided herein, energy is applied substantially to only one or more regions of interest 22 to preferentially activate targeted synapses in order to achieve the targeted physiological outcome, and is not applied substantially in a general or nonspecific manner throughout the tissue. Thus, only subsets of multiple different types of ganglia are exposed to the direct application of energy. For example, a region of interest within an organ containing any of the blood vessels, nerves, or other anatomical landmarks can be spatially selected to identify regions having specific axon terminals and synapses. In one embodiment, the region of interest is selected by identifying the splenic artery or hepatic artery and spatially selecting a region close to or parallel to the splenic artery or hepatic artery. Organ structures may be segmented based on suborgan function, vascularity, and innervation, and subsets of axon terminals may be selected so as to be included in the region of interest to which energy is directly applied. Other ganglia or nerve structures may be outside the region of interest 22 and may not be exposed to the directly applied energy. The region of interest 22 can be selected based on factors including, but not limited to, historical or experimental data (e.g., data showing the association between a specific location and a desired or targeted physiological outcome). Alternatively or additionally, system 10 may apply energy to individual ganglia of the celiac plexus until the desired targeted physiological effect is achieved.

[0091] Figure 22 is a block diagram of a specific component of system 10. Where provided herein, system 10 for neuromodulation may include a pulse generator 14 adapted to produce multiple energy pulses for application to a target tissue. The pulse generator 14 may be separate or integrated into an external device such as a controller 16. The controller 16 includes a processor 30 for controlling the device. Software code or instructions are stored in the memory 32 of the controller 16 to be executed by the processor 30 to control various components of the device. The controller 16 and / or the pulse generator 14 may be connected to an energy application device 12 via one or more leads 33 or wirelessly.

[0092] The controller 16 also includes a user interface with input / output circuits 34 and a display 36 adapted to allow a clinician to present select inputs or adjustment parameters to an adjustment program. Each adjustment program may include one or more sets of adjustment parameters, including pulse amplitude, pulse width, pulse frequency, etc. The pulse generator 14 modifies its internal parameters in response to a control signal from the controller device 16 to alter the stimulating characteristics of the energy pulses transmitted to the target to which the energy application device 12 is applied via the leads 33. Any suitable type of pulse generation circuit can be used, including, but is not limited to, constant current, constant voltage, or multiple independent current or voltage sources. The applied energy is a function of the current amplitude and pulse width duration. The controller 16 allows for tunable control of the energy by changing the adjustment parameters and / or by initiating energy application at a particular time or canceling / suppressing energy application at a particular time. In one embodiment, the tunable control of the energy application device is based on information regarding the concentration of one or more molecules in the target (e.g., circulating molecules, e.g., biomarkers associated with IBD). If the information is from the evaluation device 20, the feedback loop can drive adjustable control. For example, if the circulating IBD biomarker concentration exceeds a predetermined threshold or range when measured by the evaluation device 20, the controller 16 can initiate energy application to the region of interest with an adjustment parameter related to the reduction of the biomarker. The initiation of energy application may be triggered by the IBD biomarker drifting above a predetermined (e.g., desired) threshold or outside a predetermined range. In another embodiment, the adjustable control may take the form of changing the adjustment parameter if the initial application of energy does not result in the expected change to the target physiological outcome (e.g., concentration of the molecule of interest) within a predetermined time frame (e.g., 1 hour, 2 hours, 4 hours, 1 day).

[0093] In one embodiment, memory 32 stores different operating modes that can be selected by the operator. For example, a stored operating mode may include instructions for executing a set of modulatory parameters associated with a particular treatment site. Different sites may have different associated modulatory parameters. Rather than requiring the operator to manually input a mode, the controller 16 may be configured to execute the appropriate instructions based on the selection. In another embodiment, memory 32 stores operating modes for different types of treatment. For example, activation may be associated with a range of stimulation pressure or frequency that is different from those associated with suppressing or blocking tissue function. In a particular example, if the energy application device is an ultrasonic transducer, the time-averaged power (time-averaged intensity) is 1 mW / cm². 2 ~30,000 mW / cm 2 (Time-averaged intensity) and peak positive pressure in the range of 0.1 MPa to 7 MPa (peak pressure) may be included. In one example, to avoid levels associated with thermal damage and ablation / cavitation, the time-averaged intensity in the region of interest is set to 35 W / cm². 2 It is less than [a certain value]. In another specific example, if the energy application device is a mechanical actuator, the amplitude of the vibration is in the range of 0.1 to 10 mm. The selected frequency may depend on the mode of energy application, e.g., ultrasonic or mechanical actuator.

[0094] In another embodiment, memory 32 stores calibration or setting modes that allow adjustment or modification of the control parameters to achieve a desired result. In one example, the stimulus starts with a lower energy parameter and increases incrementally, either automatically or upon receiving operator input. In this way, the operator can achieve adjustment of the inductive effect when the control parameters are changed.

[0095] The system may also include an imaging device, such as an imaging transducer 40, which facilitates focusing of the energy application device 12 by acquiring imaging data when the energy application device 12 is operating in imaging mode under command from the controller 16. The energy application device 12 may also include an ultrasound therapeutic transducer 42 that can apply focused ultrasound energy to a target within the region of interest 22 when operating in therapeutic mode under command from the controller 16. The energy application device 12 may include a control circuit for controlling the imaging transducer 40 and / or the ultrasound therapeutic transducer 42. The control circuit of the processor 30 may be integrated with the energy application device 12 (e.g., via the integrated controller 16) or may be a separate component. The imaging transducer 40 may also be configured to spatially select a desired or targeted region of interest and acquire image data to help focus the applied energy onto the region of interest of the target tissue or structure.

[0096] In one embodiment, the imaging device (transducer 40) may be integrated with or the same as the energy application device 12, thereby applying different ultrasound parameters (frequency, aperture, or energy) to select (e.g., spatially select) a region of interest for targeting and subsequent neural modulation, and to focus energy onto the selected region of interest. In another embodiment, memory 32 stores one or more targeting or focusing modes used to spatially select a region of interest within an organ or tissue structure. Spatial selection may include selecting a sub-region of an organ to identify the volume of the organ corresponding to the region of interest. Spatial selection may depend on the image data obtained herein. In one embodiment, the controller 16 can be programmed to automatically identify or select a region of interest based on the image data. In an embodiment, the image data can be displayed on a display 36, and the operator can specify a portion of the image corresponding to the region of interest. Based on user input, the controller 16 can select a region of interest. Based on the spatial selection, the energy application device 12 can focus on the selected volume corresponding to the region of interest. For example, the energy application device 12 may initially operate in imaging mode to apply imaging mode energy used to capture image data used to identify a region of interest. The imaging mode energy is not at a level of the modulatory parameters suitable for preferential activation, and / or is not applied at that level. However, once the region of interest is identified, the controller 16 can operate in therapeutic mode according to the modulatory parameters related to neuromodulation provided herein.

[0097] The controller 16 may also be configured to receive inputs related to the targeted physiological outcome as input to the selection of modulatory parameters. For example, if the imaging modality is used to evaluate tissue properties, the controller 16 may be configured to receive a calculated index or parameter of the property. The modulatory parameters can be modified based on whether the index or parameter is above or below a predetermined threshold. In one embodiment, the parameter may be a measure of tissue displacement of the affected tissue or a measure of the depth of the affected tissue. Other parameters may include evaluating the concentration of one or more molecules of interest (e.g., evaluating one or more of the change in concentration relative to a threshold or baseline / control, the rate of change, or determining whether the concentration is within a desired range). Furthermore, the energy application device 12 may operate under the control of the controller 16 to a) acquire tissue image data that can be used to spatially select a region of interest within the target tissue, b) apply modulatory energy to the region of interest, and c) acquire image data to determine that the targeted physiological outcome has occurred (e.g., via a motion measurement). In such embodiments, the imaging device, evaluation device 20, and energy application device 12 may be the same device.

[0098] In another embodiment, a desired set of adjustment parameters can also be stored by the controller 16. In this way, the target-specific parameters can be determined. Furthermore, the effectiveness of such parameters can be evaluated over time. If a particular set of parameters is less effective over time, the target may be developing a state of insensitivity to the activated pathway. If the system 10 includes an evaluation device 20, the evaluation device 20 can provide feedback to the controller 16. In certain embodiments, the feedback can be received from a user or the evaluation device 20, indicating the characteristics of the target's physiological outcome. The controller 16 can be configured to cause an energy application device to apply energy according to the adjustment parameters and to dynamically adjust the adjustment parameters based on the feedback. For example, based on the feedback, the processor 16 can automatically change the adjustment parameters (e.g., frequency, amplitude, or pulse width of an ultrasonic beam or mechanical vibration) in real time and in response to feedback from the evaluation device 20.

[0099] The disclosed techniques can be used to evaluate neuromodulatory effects, which can be used as input or feedback for sequentially selecting or modifying neuromodulatory parameters. The disclosed techniques can be used for direct assessment of tissue state or function as a target physiological outcome. Assessments may be performed before (i.e., baseline assessment), during, and / or after neuromodulation.

[0100] The evaluation technique may include at least one of functional magnetic resonance imaging, diffusion tensor magnetic resonance imaging, positive emission tomography, or acoustic monitoring or thermal monitoring. The evaluation technique may also include the evaluation of protein and / or marker concentrations. Images from the evaluation technique can be received by the system for automated or manual evaluation. Modification parameters may also be modified based on the image data. For example, changes in organ size or displacement may be used as markers of local neurotransmitter concentrations and can be effectively used as surrogate markers of local cell exposure to phenotypic modulatory neurotransmitters and as markers of predictive effects on the IBD pathway. Local concentration may refer to the concentration within the focal region of energy application.

[0101] Additionally or alternatively, the system can assess the presence or concentration of one or more molecules circulating in tissues or blood. Concentrations in tissues may be referred to as local concentrations or basal concentrations. Tissues may be obtained by fine-needle aspiration, and assessment of the presence or level of molecules of interest (e.g., metabolites, metabolic pathway markers, peptide mediators, catecholamines) may be performed by any suitable technique known to those skilled in the art. Where provided herein, molecules of interest may be one or more IBD biomarkers, which may be cytokines (TNF-α, IL-1β), perinuclear anti-neutrophil antibodies, anti-Saccharomyces cerevisiae antibodies, calprotectin, C-reactive protein, or anti-flagellin antibodies.

[0102] In other embodiments, the target physiological outcomes may include, but are not limited to, tissue displacement, changes in tissue size, changes in the concentration of one or more molecules (local, nonlocal, or circulating), changes in gene or marker expression, afferent activity, and cell migration. For example, tissue displacement (e.g., vascular displacement of adjacent arteries) may occur as a result of energy application to tissue. Other effects can be estimated by evaluating tissue displacement (e.g., via imaging). For example, a particular displacement may be characteristic of a particular change in molecular concentration.

[0103] Figure 23 is a flowchart of method 50 for neuromodulation of the celiac plexus. In method 50, image data of the subject is acquired in step 52 to identify a region of the subject that is likely to contain the desired peripheral ganglion to be modified. For example, individual ganglia may be difficult to visualize using ultrasound images, but the region of interest can be identified using the identification of one or more visualized anatomical landmarks. Such visual landmarks may include the identification of the abdominal aorta and one or more junctions with arteries adjacent to the peripheral ganglia of the celiac plexus. In one example, the region of interest may be positioned to include the junction of the abdominal aorta with the hepatic or splenic artery. In another example, the region of interest may be selected based on its relative position to such junctions (e.g., 1 to 10 mm away from the junctions). The energy application device is positioned in step 54 so that energy pulses are focused on a desired region of interest, and the pulse generator applies multiple energy pulses to the region of interest of the target tissue in step 56 to preferentially activate at least a portion of the peripheral ganglia located in the celiac plexus that are situated in the region of interest, for example, as provided herein, to stimulate the ganglia to release neurotransmitters and / or induce altered neurotransmitter release and / or altered activity. In embodiments, the region of interest includes the celiac plexus or at least a portion of the celiac plexus. In embodiments, the region of interest includes the peripheral ganglia of the celiac plexus. In certain embodiments, the method may include a step of evaluating the effect of the stimulation. For example, one or more direct or indirect assessments of the function or state of the tissue can be used. Based on the function of the evaluated tissue, the modulating parameters of one or more energy pulses can be modified (e.g., dynamically or tunably controlled) to achieve the target physiological outcome.

[0104] In one embodiment, evaluations can be performed before and after applying an energy pulse to assess changes in the concentration of the molecule of interest and characteristics of the stool (stool consistency, presence of blood in the stool). In one embodiment, the evaluation may include DAI determination. If the evaluation marker is above or below a threshold, appropriate modifications to the modulating parameters can be made. For example, if the marker is within or related to the desired physiological outcome, the energy applied during neuromodulation may be reduced to the minimum level supporting the desired outcome. If the change in characteristics relative to the threshold is related to an insufficient change in the marker, certain modulating parameters, including but not limited to modulating amplitude or frequency, pulse shape, stimulation pattern, and / or stimulation location, may be modified.

[0105] Furthermore, the characteristics or states being evaluated may be values ​​or indicators (e.g., DAI), such as flow rate, concentration, cell population (e.g., changes in the location or characteristics of leukocytes), or any combination thereof, which may be analyzed sequentially by appropriate techniques. For example, relative changes above a threshold can be used to determine whether a regulatory parameter has been modified. Desired regulation may be evaluated by measured clinical outcomes, such as the presence or absence of an increase in tissue structure size (e.g., characteristics of colon tissue) or a change in the concentration of one or more released molecules (e.g., relative to baseline concentration before neuromodulation). In one embodiment, desired regulation may include a concentration increase above a threshold, such as an increase in concentration of about 50%, 100%, 200%, 400%, or 1000% compared to baseline. In the case of blocking treatment, evaluation may include tracking a decrease in the concentration of a molecule over time, such as a decrease of at least 10%, 20%, 30%, 50%, or 75% of the molecule of interest. Furthermore, for a particular target, the desired blocking treatment may include maintaining a relatively stable concentration of a particular molecule in the context of other clinical events that may tend to increase the concentration of the molecule. That is, the desired blocking can prevent an increase in potential. Increases or decreases or other induced and measurable effects may be measured within a certain time frame from the start of treatment, e.g., within about 5 minutes, or within about 30 minutes. In certain embodiments, if neuromodulation is determined to be desirable, the change in neuromodulation is a command to stop the application of the energy pulse. In another embodiment, if neuromodulation is undesirable, one or more parameters of neuromodulation are changed. For example, a change in the modulation parameter may be an increase in the pulse repetition frequency, e.g., a stepwise increase in frequency from 10 to 100 Hz, and evaluation of the desired characteristics until the desired neuromodulation is achieved. In another embodiment, the pulse width may be changed. In yet another embodiment, two or more parameters may be changed together, in parallel, or sequentially. If neuromodulation is undesirable after changing multiple parameters, the focus (i.e., site) of energy application may be changed.

[0106] For example, this technique may be used to treat subjects with improved IBD-like symptoms to improve stool consistency, macroscopic bleeding / diarrhea, and colonic tissue integrity. In IBD, various pathogenic factors disrupt the delicate homeostasis of immune cells in the gut, and for example, T1, T2, T17, and Treg cells can generate a cascade of inflammatory cytokines via activated macrophages and dendritic cells in the self-maintenance cycle. Some notable candidates include tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin-(IL-)17, IL-22, IL-1, IL-6, IL-8, IL-12, and IL-18. While these inflammatory cytokines can induce benign local inflammatory responses, uncontrolled overproduction can lead to tissue damage, diffusion ultrasound coagulation, death, and / or hypotension. With this in mind, mitigating dysregulated cytokine levels in intestinal inflammation and tissue damage may be achieved via the cholinergic anti-inflammatory pathway (CAP). By targeting the celiac plexus, selective differentiation of the splenic ("systemic") versus intestinal ("local") cholinergic anti-inflammatory pathway and activation of intestinal CAP were facilitated at different locations within the mesentery, including the superior mesenteric ganglion, celiac ganglion, inferior mesenteric ganglion, dorsal root ganglion, or myelomyelitis plexus. In certain embodiments, the disclosed technology enables activation of intestinal CAP without activating splenic CAP, or with relatively low activation of splenic CAP. This technology offers advantages over electrical vagus stimulation devices that have been shown to require the splenic / splenic ganglion as a pathway for intestinal neuron activation. This technology precisely modulates the intestinal pathway rather than the splenic pathway. In certain embodiments, activation may be systemic / splenic, local / intestinal, or mixed / complex stimulation. Selection of CAP stimulation sites targeting specific peripheral ganglia versus direct activation of the myolacial plexus, and selection of specific ultrasound stimulation parameters, including stimulation power, duration, and timing of administration, can be further described for the optimization of IBD treatment.

[0107] This technique demonstrated the reduction of IBD symptoms in a given mild (5%) and more severe (7%) DSS by applying focused ultrasound once or twice daily. Targeting the celiac plexus demonstrated the effectiveness of focused ultrasound for IBD symptoms. The celiac plexus is a group of nerves involved in transmitting messages from the pancreas, liver, kidneys, gallbladder, spleen, and intestines. There are four major ganglia associated with the celiac plexus, and the disclosed example targeted the upper part of the celiac plexus, impairing the celiac ganglia and superior mesenteric ganglia. The superior mesenteric ganglia is responsible for innervating the intestines, making it an ideal target for focused ultrasound to treat IBD. Both the left and right celiac ganglia also play a role in enteric innervation, but these ganglia are also associated with hepatic activity. In rats, the superior mesenteric ganglia was measured to be approximately 3 mm from the celiac ganglia. Given that the transducer diameter is approximately 20 mm and the focal point is 1 mm, precisely positioning the transducer on this nerve plexus was a key factor in the success. Off-target focused ultrasound may explain why some animals showed much better stool consistency, bloody stools, and colon length with focused ultrasound than others. In line with similar considerations, animals breathing under isoflurane anesthesia may induce chest movement and changes in transducer focus, subsequently leading to mistargeting of the celiac plexus. Furthermore, this may also explain weight loss observed with focused ultrasound, as neural modulation at specific abdominal locations may result in direct activation of incretin pathways within the gastrointestinal system. Thus, targeting can be improved by considering respiratory-induced movement. Moreover, such variability can be directed towards the advantages of using a more targeted (focused), relatively small transducer with less diffusion effect.

[0108] The technical effects of the disclosed embodiments include techniques for non-invasive focused ultrasound over the celiac plexus to reduce the effects of IBD, such as bloody stools, improve stool consistency, and protect tissue length and integrity. These techniques may be used to treat patients who do not respond to pharmaceutical interventions, or to enhance pharmaceutical or surgical interventions.

[0109] This specification, as described herein, uses examples including the best mode and enables a person skilled in the art to carry out the disclosed embodiments, including constructing and using any device or system and performing any incorporated method. The patentable scope is defined by the claims and may include other examples that a person skilled in the art may conceive. Such other examples are intended to be within the claims if they have structural elements that are not different from the language of the claims, or if they include equivalent structural elements that are substantially different from the language of the claims.

Claims

1. It is a system, An ultrasound probe configured to apply focused ultrasound energy to a region of interest, including at least the upper part of the celiac plexus, in order to neurally modulate the peripheral ganglia of the celiac plexus of a target, and It is a controller, Image data of the target is acquired from the ultrasound probe operating in imaging mode. For the selection of the region of interest, the segmentation of the abdominal aorta and hepatic artery and / or the abdominal aorta and splenic artery in the image data is performed based on one or more of the suborgan tissue function, blood vessels, and nerve innervation. Based on the image data, the region of interest is selected so as to be adjacent to the junction of the abdominal aorta and the hepatic artery and / or the splenic artery. A system including a controller configured to control the ultrasound probe to apply the ultrasound energy focused on the region of interest as part of a treatment protocol for treating the aforementioned inflammatory bowel disease.

2. The system according to claim 1, wherein the ultrasound probe includes a therapeutic transducer configured to apply focused ultrasound energy and an imaging probe configured to acquire image data.

3. The system according to claim 1, wherein the region of interest includes at least a portion of the peripheral ganglia of the celiac plexus.

4. Includes an evaluation device for evaluating the response to the ultrasonic energy focused over a certain period of time, Based on the evaluation of the response, the controller modifies the treatment protocol to change the administration frequency. The system according to claim 1, wherein the treatment protocol includes applying the ultrasonic energy to the region of interest at least daily over a period of several days.

5. The aforementioned controller, Display the aforementioned image data, The system receives user input indicating the region of interest within the displayed image data, The system according to claim 1, configured to select the region of interest based on the user input.

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