京都大学 / 0048 / 新制・課程博士 / 博士(人間健康科学) / 甲第21460号 / 人健博第67号 / 新制||人健||5(附属図書館) / 京都大学大学院医学研究科人間健康科学系専攻 / (主査)教授 杉本 直三, 教授 藤井 康友, 教授 松田 秀一 / 学位規則第4条第1項該当 / Doctor of Human Health Sciences / Kyoto University / DFAM

No description available.

Patellar Tendinopathy (PT), commonly called "Jumper's Knee", is a condition resulting from repetitive loading of the patellar tendon that presents as anterior knee pain, which is commonly seen in basketball players due to the maneuvers in the sport. Diagnosis of PT often involves a clinical exam followed by ultrasound images for confirmation of the diagnosis to look for key factors of PT. Clinical assessment of ultrasound images of tendons is subjective and requires a high level of experience for reliable interpretation. Thus, there is a need for objective, quantitative methods to assess tendon abnormalities associated with pathology. Ultrasound image texture analysis has emerged as a reliable technique to augment the utility of conventional US imaging, and has recently been shown to distinguish healthy from abnormal tendon and myofascial tissues. The objective of the present study was to conduct image texture analysis to evaluate patellar tendons of collegiate basketball athletes over two seasons. Under an IRB-approved protocol with informed consent, a total of 33 Division 1 collegiate basketball athletes (16 male, 17 female, age 19.9 +/- 1.4 years) underwent clinical evaluation and ultrasound imaging. Four imaging sessions were collected over the course of two years (pre- and post-season). Participants were imaged using a GE LOGIQ S8 (General Electric, USA) ultrasound machine equipped with ML6-15 linear probe. At each imaging session, power Doppler images were collected in the longitudinal and transverse axis, at the proximal, central, and distal regions of the patellar tendon of both knees. Image texture analysis was performed using a custom MATLAB (Mathworks, USA) program to obtain first order (mean, median, variance, skewness, kurtosis, entropy), second order (contrast, energy, and homogeneity), and blob analysis (blob count, BC, and blob area, BA, for 5%, 25%, 50%, 75%, and 95% thresholding values) texture parameters in each image, based upon borders manually drawn by a single researcher. Statistical analysis was conducted to compare imaging sessions (JMP Pro 16, SAS). P-values <0.05 were considered statistically significant. Quantitative texture parameters are able to distinguish characteristics in patellar tendon ultrasound images to distinguish between anatomic region, gender, dominance and pre- to post- season. The 25% and 75% thresholding percentiles effectively showed characteristics of collagen fibers in the patellar tendon. The abnormal diagnosis does not greatly effect texture parameters, which needs to be investigated with more incorporation of grading criteria distinctions and a larger sample size. / Master of Science / Patellar Tendinopathy (PT) is a knee injury that commonly occurs in basketball players. The recovery for PT is often long and the player can still have knee pain when returning to the sport. Diagnosis of PT requires a high level of expertise to consider the patients history, conduct a physical exam and take ultrasound images to look for factors that indicate patellar tendon is damaged. The difficulty of diagnosing PT calls for an objective method to allow for accuracy in assessing patellar tendons. In order to create a more objective measure of ultrasound images, quantitative texture parameters are explored to understand what the brightness values of each pixel and the proximity of pixels together can convey about the image. The objective of this study is to understand what characteristics of the subject (anatomic region, knee dominance, gender, and time point) texture parameters are able to distinguish in patellar tendon ultrasound images.

Traumatic injury is a major burden in global healthcare systems, ranking among the leading causes of morbidity and mortality worldwide.1 Patients are first encountered at the pre-hospital scene by providers of varying levels of expertise, such as emergency medical technicians (EMTs) and paramedics, who provide temporizing measures while patients are transported to receiving hospitals to receive definitive care.2 Ultrasound is an ever-improving medical imaging modality which is increasingly portable, low cost, and provides diagnostic imaging rapidly without the harmful effects of radiation. The objective of this study is to determine whether introduction of prehospital ultrasound (PHUS) for use on ground ambulances by prehospital providers in order to improve choice of destination hospital and aid in needle thoracostomy for tension pneumothorax will have a positive impact on mortality rate of trauma patients in an urban EMS environment. In the proposed study, trauma patients in the city of Boston, Massachusetts receiving care from Boston Emergency Medical Services (EMS) prehospital providers will be recruited over a 12-month period with a minimum goal of 2,500 patients in total. Emergency responses coded as trauma by EMS dispatch will be randomized at a 1:1 ratio to either utilize PHUS or to refrain from utilizing PHUS. A z-test will be used to analyze primary outcome of 30-day mortality rate in patients who receive PHUS care as needed compared with patients who do not receive PHUS care. Study data will be collected directly from Boston EMS Electronic Medical Record (EMR). This study will be the first of its kind to randomize at the patient level, and the first to investigate a major clinical outcome of ultrasound in prehospital medical care: 30-day mortality. Point-of-care Ultrasound is an intriguing diagnostic modality that is becoming increasingly feasible in the prehospital environment, and may improve outcomes in trauma patients. Current studies provide convincing evidence for the diagnostic accuracy of these devices, especially in evaluating hemoperitoneum and pneumothorax. If an improvement in mortality rate of patients treated with prehospital ultrasound (PHUS) is demonstrated, this will be convincing evidence for the implementation of PHUS in ground ambulances and air medical services across the United States and worldwide.

No description available.

Fibers in biological scaffolds like fibronectin stiffen when they experience forces between cells. It will expose binding sites under contractile forces and then form disulfide bonds. This on-demand strain-stiffening is a desirable property in synthetic materials. Tran et.al. (2017) mimicked the “cryptic” design of fibronectin by copolymerizing thiol crosslinking sites with monomers containing poly (ethylene glycol) chains. When the PEG chain increased from 350 to 950 g/mol, the strains-stiffening became on-demand while the curing process extended from 3 hours to 15 hours. Extra steric hindrance brought by longer PEG chains caused decreasing mass transfer rates of cryptic sites while the same level of strain rate was introduced. I proposed to use stronger ultrasound mechanical perturbation so that higher strain rate can be induced, and the shielding effect brought by the PEG chain can be overcome more easily. Utilizing ultrasound as a stimuli has the potential to improve the gelation speed or achieve high mechanical performance while retain the long shelf life of the “cryptic” materials. To test this hypothesis, I synthesized “cryptic” polymer with aceto-acetoxy and primary amine as crosslinking sites such that, the only time limiting step is brought by the long PEG chain. This is because the bond formation reaction between these two reactive groups is rapid and spontaneous. When switching from weak to strong mechanical perturbation, the change in gelation speed owing to accelerated mass transfer between crosslinking sites can be easily compared. When the PEG chain is 300 Mw and 30 mol % crosslinking sites density, this “cryptic” polymer only showed strain-stiffening under ultrasound while strain under a rheometer was not able to overcome steric hindrance. Signs of chain scission appeared when the ultrasound amplitude was set at 75 %, but was counteracted by reducing amplitude mode over the time. The crosslinking was optimized by varying the ultrasound amplitude and intensity and a final mode of 1 hour 75 % amplitude, 0.5 hour 50 % amplitude and 3.5 hours 25% amplitude provided greatly improved on demand crosslinking. The estimated kinetic constant using this mode was two times higher than that of under simple shear strain. Through this study, I found that ultrasound can improve the curing time of this “cryptic” polymer system since it induces higher strain rate and expedite the mass transfer rate between crosslinking sites and optimizing the ultrasonic amplitude profile to limit chain scission provides improved crosslinking performance.

The purpose of this thesis was to understand the implications of ultrasound in the assessment of flexor tendon shear to establish its role in carpal tunnel syndrome. An in vitro and in vivo approach was used to examine ultrasound “shear” between the tendon and adjacent tenosynovium. Ultrasound shear is defined by the relative displacement between the tendon and tenosynovium, and has been considered a surrogate measure of tendon shear. However, the mechanical implications of relative displacement are not well understood. In Chapters 2 and 3, an in vitro approach was used to compare ultrasound to direct measurements of tendon displacement and tendon shear. Chapter 2 demonstrated the validity of colour Doppler ultrasonography in the evaluation of tendon displacement. Chapter 3 assessed the relationship between ultrasound shear and mechanical tendon shear using frictional work. We dispelled the notion that ultrasound shear represents tendon shear by showing it only captures the viscoelastic stretch of the tenosynovium in tendon shear; missing surface friction from neighbouring anatomical structures in the carpal tunnel. However, measuring viscoelastic resistance in tendon motion is important for the development of pathological fibrosis and thickening of the tenosynovium, a characteristic finding in carpal tunnel syndrome. In Chapter 4 we further established the clinical utility of ultrasound in vivo by showing ultrasound shear discriminated carpal tunnel syndrome symptomatic individuals from the healthy population. Ultrasound measures progressed with symptoms suggesting an etiological progression of fibrosis and thickening with carpal tunnel syndrome. This thesis concluded that ultrasound only partially represents tendon shear with the viscoelastic component, but underscored the clinical implications. Ultrasound provides a non-invasive assessment of viscoelastic resistance that will be highly valuable for our understanding of the role of wrist and hand motion in the etiology of injury with potential applications in the diagnosis of carpal tunnel syndrome. / Thesis / Master of Science in Kinesiology

Histotripsy is a non-invasive and non-thermal focused ultrasound therapy that can be used to ablate tissue within the body while overcoming many of the limitations of thermal ablation. Histotripsy utilizes short-duration, high pressure ultrasound pulses to create a cavitation bubble cloud of numerous rapidly expanding and collapsing bubbles, which cause mechanical stress on the targeted region. Histotripsy contains multiple subtypes including intrinsic threshold, shock scattering, and boiling histotripsy, where intrinsic threshold histotripsy utilizes single cycle pulses focused to a single point to create a bubble cloud from the peak negative pressure (p- ≥ 25 MPa for water-based tissues). Nanoparticle-mediated histotripsy (NMH) uses perfluorocarbon-filled nanoparticles to create bubble clouds at lower pressures than that of the intrinsic threshold of histotripsy. Prior studies have shown that nanodroplets (NDs) and nanocone clusters (NCCs) both reduce the cavitation threshold, but further investigation on different parameters to optimize treatments have not fully been studied. Additional research is needed for the characterization of these nanoparticles with different pulsing parameters such as cycle number and frequency in order to better predict and understand the mechanisms underlying NMH. In this thesis, I investigate the ability of new nanodroplets and nanocone clusters to reduce histotripsy cavitation threshold with NMH. I also investigate the effect that multi-cycle pulsing parameters have on NMH and stable bubble formation from acoustic droplet vaporization (ADV) for nancone clusters. The culmination of this thesis will advance our understanding of the behavior of acoustically-active nanoparticles when exposed to varied pulsing schemes and frequencies. This knowledge will allow for the further investigation of more efficient, effective, and safe methods for clinical focused ultrasound therapies. / Master of Science / Histotripsy is a non-invasive and non-thermal focused ultrasound therapy that can be used to destroy targeted tissue within the body. Histotripsy is currently being developed for non-invasive and non-thermal cancerous tissue destruction with the first-in-man trial having been conducted within the last year for the treatment of liver tumors. Histotripsy utilizes high-pressure, short-duration pulses focused to a single region to create a cloud of bubbles that are rapidly expanding and collapsing which causes mechanical damage to the targeted cells. Nanoparticle-mediated histotripsy (NMH) has been developed to utilize nanoparticles to reduce the pressure needed to induce cavitation. Despite many studies and advances in histotripsy, there are many areas within the topic that need additional research to better understand the capabilities of the treatment method. This additional research is crucial in allowing for the development of new nanoparticles, faster treatment times, and new parameters that could allow for more precision near critical structures. In this thesis, I investigate the ability of new nanoparticles to reduce histotripsy cavitation threshold with NMH. I also investigate the effect that multi-cycle pulsing parameters have on NMH and stable bubble formation for nanoparticles. The culmination of this thesis will advance our understanding of the behavior of acoustically-active nanoparticles when exposed to varied pulsing schemes and frequencies. This knowledge will allow for the further investigation of more efficient, effective, and safe methods for clinical focused ultrasound therapies.

Formalin-fixed, paraffin embedded (FFPE) tissue is the most abundant, accessible, and versatile tissue sample type available for genetic research and clinical applications. However, FFPE DNA extraction presents unique challenges and requires lengthy incubation periods, which can be impractical for certain applications. Here, we propose the use of focused ultrasound extraction (FUSE) technology for improved DNA extraction from FFPE tissue. FUSE generates a dense bubble cloud of acoustic cavitation capable of ablating tissue into an acellular lysate. FUSE treatment was applied to de-paraffinized porcine pancreas FFPE scrolls, followed by heated incubation for formaldehyde-induced DNA-protein crosslink reversal. When applied for 30 minutes, FUSE was found to successfully extract DNA from FFPE tissue as defined by increased DNA yield and improved purity ratios compared to conventional methods. DNA extracted via FUSE showed comparable fragmentation to conventional methods, and three out of four samples successfully amplified via PCR, indicating suitability for downstream analysis. These findings suggest that FUSE has the potential to increase the efficiency and effectiveness of DNA extraction from FFPE tissue. Further development and optimization of this protocol could develop a streamlined, easy to use extraction method that would simplify FFPE DNA extraction methods and address the primary time constraints which currently make FFPE DNA extraction time-consuming and impracticable for high-throughput applications. / Master of Science / Formalin-fixed, paraffin embedding (FFPE) has historically been the most popular method of biological tissue preservation, as it allows tissue to remain shelf stable for decades. As such, FFPE tissue is the most abundant, accessible, and versatile tissue sample type available for genetic research applications. Here, we propose the use of focused ultrasound extraction (FUSE) technology for improved DNA extraction from FFPE tissue. FUSE treatment applies rapid, focused ultrasound waves to tissue, resulting in the mechanical breakdown of cells and subsequent release of DNA. FUSE treatment was applied to pig pancreatic FFPE samples. When applied for 30 minutes, FUSE was found to successfully extract DNA from FFPE tissue as defined by increased DNA yield and improved purity compared to conventional methods. Three out of four DNA samples extracted via FUSE were successfully amplified, and DNA fragment lengths were comparable between FUSE and conventional methods, showing that FUSE did not fragment DNA beyond useful fragment lengths. These findings suggest that FUSE has the potential to increase the efficiency and effectiveness of DNA extraction from FFPE tissue. Further development and optimization of this protocol could develop a streamlined, easy to use extraction method that would simplify FFPE DNA extraction methods and address the primary time constraints which currently make FFPE DNA extraction time-consuming and impracticable for high-throughput applications.

Histotripsy is a noninvasive mechanical ablation method that uses focused ultrasound to disintegrate target tissues into acellular homogenate through the generation of acoustic cavitation and is currently being developed for numerous clinical applications. Histotripsy uses high-pressure (>10 MPa), short-duration (<15 cycles) pulses to cause the rapid expansion and collapse of nuclei at the focus resulting in large applied stress and strain in the adjacent tissue. At a sufficiently high pressure above the target medium's intrinsic cavitation threshold and an adequate number of applied pulses, cavitation "bubble clouds" create precise lesions with high fidelity to the region of the focus. Despite advances in histotripsy, additional research is still needed to better understand the acoustic cavitation nucleation process and its effects on therapies using focused ultrasound. This understanding is critical to better predict and control pulse dose for more rapid and efficient ablation procedures, to reduce off-target cavitation events for safer focused ultrasound therapies, and to localize ablation for high-precision procedures near critical structures or treatments without active imaging guidance. In this dissertation, I investigate the nucleation and dynamics of ultrasonically generated acoustic cavitation for novel applications of focused ultrasound. My Ph.D. thesis focuses on (1) investigating the effect of histotripsy pulsing parameters on bubble cloud cavitation nucleation, bubble dynamics, and ablation efficiency, (2) investigating the effect of nuclei characteristics on the threshold for cavitation nucleation and resulting bubble dynamics for therapeutic applications, and (3) developing methods alter select characteristics and dynamics of acoustic cavitation by adjusting pulsing parameters to optimize ablation efficiency in conventional and nanoparticle-mediated histotripsy. The culmination of this thesis will advance our understanding of the nucleation and behavior of acoustic cavitation from pulsed focused ultrasound and develop innovative systems to improve the efficacy, efficiency, and safety of clinical focused ultrasound therapies. / Doctor of Philosophy / Histotripsy is a noninvasive focused ultrasound method that precisely destroys target tissues such as tumors through the acoustic generation of cavitation and is currently being developed for numerous clinical applications. Histotripsy uses high-pressure, short-duration pulsed soundwaves to cause the bubbles to rapidly expand and collapse within a precise region called the focus. This rapid cavitation results in large mechanical strain in the targeted tissue. With increasingly higher pressure, numerous bubbles form in the shape of cavitation "bubble clouds" that create lesions, closely matching their shape, in the target tissue after a sufficient number of pulses have been applied. Despite advances in histotripsy, additional research is still needed to better understand the initiation of the acoustic cavitation process in histotripsy and its effects on focused ultrasound therapies. This understanding is critical to better predict and control ablation procedures, improve procedure efficiency, reduce off-target cavitation events for safer focused ultrasound therapies, and further increase ablation precision for procedures near critical structures or treatments without active image guidance. In this dissertation, I investigate the initiation, growth, and collapse of ultrasonically generated acoustic cavitation for novel applications of focused ultrasound. My Ph.D. thesis focuses on (1) investigating the effect of histotripsy pulsing parameters on bubble cloud cavitation initiation, bubble growth and collapse, and treatment efficiency, (2) investigating the effect of particle characteristics on the threshold for cavitation initiation and resulting bubble behavior for therapeutic applications, and (3) adjusting pulsing parameters to optimize ablation efficiency in conventional and particle mediated histotripsy. The culmination of this thesis will advance our understanding of the initiation and behavior of acoustic cavitation from pulsed focused ultrasound and develop innovative systems to improve the efficacy, efficiency, and safety of clinically focused ultrasound therapies.