Cardiac conduction abnormalities and arrhythmias are associated with stroke, heart failure, and sudden cardiac death, and remain a major cause of death and disability. However, the imaging tools currently available to the physician to guide these treatments by mapping the activation sequence of the heart are invasive, ionizing, time-consuming, and costly. In this dissertation, Electromechanical Wave Imaging (EWI) is described with an aim to characterize normal and abnormal rhythms noninvasively, transmurally, at the point of care, and in real time. More specifically, the methods to map the electromechanical wave (EW), i.e., the transient deformations occurring in response to the electrical activation of the heart, are developed and optimized. The correlation between EW and the electrical activation sequence during both normal and abnormal rhythms is demonstrated in canines in vivo and in silico. Finally, EWI is shown to noninvasively detect and characterize arrhythmias and conduction disorders in humans. Novel ultrasound imaging methodologies were developed to track the EW. Radio-frequency (RF) frames acquired at high frame rates were used in conjunction with cross-correlation algorithms to map the onset of the small, localized, transient deformations resulting from the electrical activation and forming the EW. To validate the capability of the EW to characterize cardiac rhythm, it was compared against the electrical activation in vivo and in silico. A high correlation between the electrical and electromechanical activations was obtained in normal canines in vivo during various pacing schemes and sinus rhythm. An in vivo-in silico framework was also developed to demonstrate that this correlation is maintained transmurally and independently of the imaging angle. EWI was also validated in abnormal canine hearts in vivo during ischemia, left bundle branch block, or atrio-ventricular dissociation. In a clinical feasibility study, we demonstrated that EWI was capable of noninvasively mapping normal and abnormal activation patterns in all four cardiac chambers of human subjects using a readily available clinical ultrasound scanner. Specifically, EWI maps were generated for three heart failure patients with cardiac resynchronization therapy (CRT) devices and for three patients with atrial flutter who subsequently underwent catheter mapping and radiofrequency ablation. Preliminary validation of EWI maps against invasive transcutaneous electroanatomical cardiac mapping was also demonstrated. EWI has the potential of becoming a noninvasive and highly translational technology that can serve as a unique imaging tool for the early detection, diagnosis and treatment monitoring and follow-up of arrhythmias and conduction disorders through ultrasound-based mapping of the transmural electromechanical activation sequence reliably, at the point of care, and in real time.

The overall goal of this dissertation is to describe the development of a dynamic diffuse optical tomographic (DDOT) imaging system for the diagnosis and monitoring of peripheral arterial disease (PAD) within the lower extremities. PAD affects 8-12 million individuals in the United States and is associated with significant morbidity and mortality. Early detection and monitoring of disease progression is crucial, but remains difficult. This is especially true for diabetic patients, as roughly 30 percent of all diabetic patients over the age of 50 are diagnosed with PAD. Diabetic patients have calcified arteries, which renders them incompressible. This falsely elevates blood pressure readings and causes false negative readings using traditional diagnostic techniques. DDOT offers an attractive opportunity to overcome current shortcomings in assessing PAD. This technology uses harmless near-infrared light to create three-dimensional, time-dependent images of biological tissues. Using DDOT to measure blood-perfusion in the foot should help diagnose and monitor the PAD. To test this hypothesis, I adapted an existing optical tomographic imaging system for the particular application of vascular imaging in the foot. In particular I design and tested various measuring probes that can accommodate different foot sizes and shapes. The result was a patient friendly interface that can be employed in a clinical setting. Using this modified DDOT imager, which we called vascular optical tomographic imaging (VOTI) system, I conducted a 40-subject pilot study to quantify its ability to diagnose PAD. The subjects were recruited into three cohorts, non-diabetic PAD patients (N=10), PAD Patients (N=10) and healthy volunteers (N=20). With this data in hand, I performed a comprehensive data analysis, in which I found imaging features that led to a good separation between the healthy and affected cohorts. In particular I demonstrated that statistically significant difference exist between the amount of blood pooling in the leg during a 1-minute, 60mmHg thigh cuff occlusion within healthy subjects and both affected cohorts (P=0.006, P=0.006). In addition, using receiver operating characteristic (ROC) curve analysis, I identified that the new VOTI system could diagnose PAD with a sensitivity and specificity of over 80%, even within the diabetic patients. This imaging modality was also capable of identifying the severity of the disease with similar accuracy to the existing diagnostic methods while not being inhibited by arterial calcifications. Furthermore, the VOTI system provided spatial information, helping identify which regions of the foot suffered from mal-perfusion. When combined with angiosome theory, the spatial information could help physicians in deciding how to intervene in PAD patients. After completing this first clinical study, I developed a dedicated VOTI system by entirely redesigning the hard and software. This new system has many novelties over its predecessor. First it employs a contact-free patient interface that allows to imaging patients with ulcerations. The illumination fibers used do not need to make physical contact with the patient. Second, instead of using individual silicon photodiodes as detectors, a highly sensitive CCD camera is use to detect transmitted light intensity. The system has two wavelengths of light (660 and 860 nm), which can be illuminated at up to 20 different positions along the surface of the foot. The system is built for dynamic imaging and is capable of imaging at a multispectral-volumetric frame rate speeds of 1 Hz. This set-up allows us to create three-dimensional images of large portions of the foot. This imaging system was tested on phantom studies and healthy volunteers and was shown to be able to image blood flow dynamics within a three-dimensional volume of the foot.

Adipose tissue is traditionally regarded as a source of energy storage and cushion for skeletal functions. Soft tissue injuries take place in war and peace time, as a result of tumor resection such as breast cancer, and in rare disorders such as lipoatrophy. Adipose tissue reconstruction is one of the key challenges in medicine. Here we report the robust differentiation of human adipose derived stem cells using a small molecule into adipocytes. This thesis describes the biological effects and actions of novel unknown small-molecule inhibitor of BMP signaling--Pyrintegrin, which was previously found to promote human embryonic stem cells survival. The overall objective of this thesis is to test the hypothesis that this novel drug, Pyrintegrin treatment will induce, promote and accelerate adipogenic differentiation of stem cells in vitro and in vivo. We found that Pyrintegrin promotes the adipogenesis-dependent transcriptional changes of multiple gene products involved in the adipogenic process, including peroxisome proliferator-activated receptor (PPARgamma), CCAAT/enhancer-binding protein alpha; (C/EBPalpha), adiponectin and leptin secretion, and total triglyceride secretion. When transplanted into mice, Pyrintegrin treated adipose cells/progenitors gave rise to ectopic fat pads with the morphological and functional characteristics of white adipose tissue. This was further confirmed by higher expression of human PPARgamma gene in Pyrintegrin treated cells group than any other group. We further tested the presence of human nuclear staining that confirmed the presence of human cells in all the transplanted groups. However, the number of positive human cells was substantially low in all the groups, which was likely due to transplanted cell death because of lack of vascularization. Hence, we further tested the Pyrintegrin adsorbed scaffolds capacity to regenerate better soft tissue compared to Ptn-free scaffold. We found that Ptn adsorbed scaffolds was positive for adipocytes as evident by positive Oil Red O staining. We further investigated the signaling mechanism of Pyrintegrin. Using a human PPARgamma reporter assay system, based on non-human mammalian cells engineered to express human PPARgamma protein, we found that Pyrintegrin is not a PPARgamma agonist as witnessed by lack of any luciferase activity. In contrast, Rosiglitazone, a known PPARgamma agonist demonstrated significant amount of luciferase activity in these reporter cells. We also found that Pyrintegrin selectively inhibits the BMP pathway and thus blocks BMP-mediated SMAD1/5 phosphorylation, target gene transcription and osteogenic differentiation. In vitro studies showed that Pyrintegrin inhibited the differentiation of stem cells into putative osteoblasts, as evident by decreased alizarin red staining. A striking finding was that Pyrintegrin up regulated markers of adipogenesis and stimulated lipid droplets accumulation in stem cells undergoing osteoblastic differentiation in vitro. This came at the expense of down regulating markers of osteogenesis and osteoblastic differentiation of stem cells, as compared to the cells undergoing osteogenic differentiation in the absence of the drug treatment. These findings show that the novel small-molecule Pyrintegrin is a potent promoter of adipogenesis and thus may have therapeutic potential for soft tissue reconstruction.

In multicellular organisms, one-cell-thick monolayer sheets are the simplest tissues, yet they play crucial roles in physiology and tissue engineering. Cells within these sheets are tightly connected to each other through specialized cell-adhesion molecules that typically cluster into in discrete patches called cell-cell junctions. Working together, these junctional organelles glue cells to their neighbors, integrate the cytoskeletons into a mechanical syncytium and transduce a variety of mechanical signals. Human bodies offer many vivid illustration of how a cell sheet physiology changes considerably during development and diseases, as shown in epidermal blistering and certain cardiomyopathy. Despite the extensive molecular and clinical work on cell junctions, relevant in vitro experimental data are often masked by cell-substrate interactions due to a lack of suitable experimental methods. It is therefore important to develop novel in vitro methods for characterizing how junctional proteins, as well as tightly associated cytoskeletal proteins, may modulate various cellular behaviors, such as viability and apoptosis, cell-cell adhesiveness and tissue integrity. Control over cell viability is a fundamental property underlying numerous physiological processes. Cell-cell contact is likely to play a significant role in regulating cell vitality, but its function is easily masked by cell-substrate interactions, thus remains incompletely characterized. In the first part of this thesis, we developed an enzyme-based whole cell sheet lifting method and generated substrate- and scaffold-free keratinocyte (N/TERT-1) cell sheets. Cells within the suspended cell sheets have persisting intercellular contacts and remain viable, in contrast to trypsinized cells suspended without either cell-cell or cell-substrate contact, which underwent apoptosis at high rates. Suppression of junctional protein plakoglobin weakened cell-cell adhesion in cell sheets and suppressed apoptosis in suspended, trypsinized cells. These results demonstrate that cell-cell contact may be a fundamental control mechanism governing cell viability and that the plakoglobin is a key regulator of this process. The study also laid groundwork for subsequent characterization and manipulation of viable cell sheets for cell sheet engineering purpose. Cell sheet engineering, characterized by harvest of cultured cell monolayer as a scaffold-free sheet, was recently developed. Particularly, cell sheet engineering based cardiac tissue engineering has emerged as an alternative method for the repair of damaged heart tissue. Such an engineered cell sheet offers a new way to study cell junctions when substrate interactions are no longer dominant. While this method is promising, it is limited by the fragility and shrinkage of the sheets as well as the lack of information regarding the characteristics of such sheets. In next part of the thesis we pursued two related research projects by developing a novel partial-lift method to generate strong, unshrunk substrate-free and scaffold-free cell sheets, first using skin cells and then refined and expanded to cardiac cells. The rationales for this approach are the ease with which skin cells can be manipulated, the similarities in cell junctions between skin and cardiac cells, and their potential clinical applications. These partially-lifted cell sheets engage primarily in cell-cell interactions, yet are amenable to biological and chemical perturbations and, importantly, mechanical conditioning. This simple yet powerful method was then deployed to test the hypothesis that the lifted cells would exhibit substantial reinforcement of key cytoskeletal and junctional components at cell-cell contacts, and that such reinforcement would be enhanced by mechanical conditioning. Results further demonstrate that the mechanical strength and cohesion of the substrate-free cell sheets strongly depend on the integrity of the actomyosin cytoskeleton and expression of the junctional protein plakoglobin. Moreover, our results showed that dissociating cell-substrate interactions and implementing mechanical conditioning enhances contraction, calcium signaling, alters viscoelastic property, and thus improves the functional cell-cell coupling in the cardiac sheets. In sum, this thesis represents a first systematic examination of junctional regulation of cell viability and mechanical conditioning on cells with primarily cell-cell interactions. The information gained from this study will help advance our understanding of cell-cell interactions and improve cell sheets biomechanical properties. For tissue engineering purposes, our dispase-based partial-lift cell sheet harvesting method has the advantage of being biocompatible, easily applicable, rapidly collectable and stretchable, compared to currently available techniques. This simple yet powerful partial lift technique has enormous potential for fabricating clinically applicable skin and cardiac tissues.

Magnetic resonance imaging (MRI) is a medical imaging technique which can provide fine tissue contrast with relatively high image resolution in human. Besides the image quality, imaging speed is the other major concern in modern MRI, especially in human experiments where sufficient volumetric coverage is necessary. One approach to increase imaging speed is increasing image acquisition speed so that the same amount of volumetric coverage can be achieved within shorter time under conventional experiment paradigms. In this dissertation, the application of pseudo-random amplitude modulation (PRAM) in MRI was explored to increase imaging speed by designing more efficient experiment paradigms for the human brain. Two relatively slow MRI studies were investigated. The first study was measuring longitudinal relaxation time. A novel method "Relaxation by Amplitude Modulation" (RLXAM) was invented. The RLXAM modulation code can be chosen from a large family of binary sequences. PRAM is a specific implementation using the maximum length sequence, also known as pseudo-random sequence. The other study was measuring transit time distribution in arterial spin labeling. The application of PRAM in transit time measurement was reported before on a 3T Philips Acheiva scanner using a single-slice protocol with standard gradient echo acquisition. The original theory was extended and multi-slice sequences with two different acquisition strategies were developed on a 3T Siemens Trio scanner. Both methods were applied to both phantom and human to demonstrate the theories and evaluate their performance.

Osteoarthritis (OA), the most prevalent form of joint disease, afflicts nine percent of the US population over the age of thirty and costs the economy nearly $100 billion annually in healthcare and socioeconomic costs. It is characterized by joint pain and dysfunction, though the pathophysiology remains largely unknown. The progressive loss of cartilage followed by inadequate repair and remodeling of subchondral bone are common hallmarks of this degenerative disease. Due to its avascular nature and limited cellularity, articular cartilage exhibits a poor intrinsic healing response following injury. As such, significant research efforts are aimed at producing engineered cartilage as a cell-based approach for articular cartilage repair. However, the knee joint is mechanically demanding, and during injury, also a milieu of harsh inflammatory agents. The unforgiving mechanochemical environment requires constructs that are capable of bearing such burdens. To this end, previous work in our laboratory has explored the application of stimuli inspired by the native joint environment in attempts to create tissue with functional properties similar to native cartilage so that it may restore loading to the joint. While we have had success at producing these replacement tissues, there is little evidence in the literature that the biological functionality (i.e. response to in vivo-like conditions) of engineered cartilage matches native cartilage. Therefore, in an effort to provide a more complete characterization of the functional nature of developing tissues and facilitate their use clinically, the overarching motivation of the work described in this dissertation is two-fold: 1) characterize the response of engineered cartilage to chemical and mechanical injury; and 2) develop strategies for enhancing the performance and protection of engineered cartilage for in vivo success. Studies in the literature have extensively characterized the effects of wounding to native articular cartilage as well as the effects of an inflammatory environment. For mechanical injuries, cell death is immediate and progressive, ultimately leading to failure of the tissue. Chemical insult has been shown to promote degradation of the matrix components, also leading to failure of the tissue. Under a controlled application of injury (mechanical and chemical), it was found that engineered cartilage, in contrast to native cartilage, has the potential to repair itself following an injury event, as long as there is no catastrophic damage to the matrix. Additionally, when this matrix is intact and well-developed, engineered cartilage constructs exhibit a resistance to degradation, highlighting the potential utility of engineered cartilage as replacement tissues. Enhancing functionality in engineered cartilage was also explored, with the aim of developing strategies to improve, repair, and protect engineered cartilage constructs for their use in vivo. For these purposes, the studies in this dissertation spanned both 2D migration studies to influence the limited wound repair potential of cells as well as 3D culture studies to explore the possibility of protection effects at a tissue level. Together, these models allowed us to capture the complexity needed to fully develop approaches for cartilage repair. Though it has previously been found that applied DC electric fields modulate cell migration, we have developed a novel strategy of employing this technique to screen for desirable populations of cells (those with the greatest capacity for directed migration) to use in cartilage repair. We also found that the AQP1 water channel plays a key role in mechanosensing the extracellular environment, highlighting the potential for its use in therapeutic strategies. For tissue engineering efforts at creating functional cartilage replacement, we uncovered novel strategies to foster better tissue development via co-culture systems and promote the resistance of engineered cartilage to catabolic factors. These findings motivate their potential use in therapeutics and in tissue engineering efforts at creating clinically relevant tissue-engineered constructs for the treatment of OA or following injury. The research described in this dissertation has characterized the biological functionality of engineered tissues and identified strategies for enhancing their use in vivo by modulating the subsequent response to injury, laying the foundation for their use in clinical applications.

Mathematical formulation and physical models are the foundation of scientific understanding and technological advancement. Our ability to design experiments effectively is heavily dependent on our physical understanding of the system under investigation, and careful mathematical analysis is required in order to effectively progress from scientific concepts towards viable technologies. With increasing system complexity, the focus of mathematical formulation has shifted from simple, elegant models which rely on basic physical concepts to tailored, increasingly complex solutions using high-powered simulations and numerical solutions. While these methods may provide insights into specific systems, adapting these models to different systems is generally difficult, even when the systems under question operate according to the same physical laws. This is especially evident in nanobiotechnology, where the complexity of the systems studied has given rise to experiment-driven focus. Our aim is to focus on the mathematical modeling of transport processes in nanoscale systems, and to construct generalized, conceptual models for three model systems, which in turn could be applied to many biological and engineered systems. The three model systems we use - enzyme cascades, coupled molecular motors and self-assembling molecular shuttles provide a broad basis for generalized transport systems in nanoscale systems. These systems combine diffusive and active transport, as well as diverse assembly conditions and multi-scale systems with size scales spanning nano- to millimeter sizes and system complexity ranging from isolated two-component systems to multimolecular, highly-coupled systems. By applying and adapting these basic models to increasingly complex systems, we can both understand the physics behind nanoscale systems, as well as design these systems with increased robustness, scalability and repeatability.

Osteoarthritis (OA) is the most common joint disease and the leading cause of disability among Americans. OA afflicts 20 million Americans and costs $128 billion in direct medical and work-related losses each year. Nearly 1/3 of OA patients in the United States are over 65 years of age and given the aging population of the "baby boomer" generation, the prevalence of this disease is predicted to increase dramatically in the coming decades. The disease is characterized by the degeneration of cartilage and progressive loss of normal structure and function. However, the harsh loading environment and the avascular nature of mature cartilage lead to a poor intrinsic healing capacity after injury. As a result, cell-based therapies, including tissue engineering strategies for growing clinically relevant grafts, are being intensively researched. An autologous cell source would be ideal for growing clinically relevant engineered cartilage; however, using cells from an osteoarthritic or injured tissue to grow engineered cartilage with mechanical and biochemical properties similar to healthy native tissue poses several challenges, including lack of healthy donor tissues and donor site morbidity. As a result, the clinical potential of mesenchymal stem cells (MSCs) has driven forward efforts toward their optimization for tissue engineering applications. Of these MSCs, synovium-derived stem cells (SDSCs) are being intensively researched due to their proximity to the defect site and high chondrogenic potential. To address the need for cell-based therapies, functional tissue engineering aims to restore cartilage function by culturing grafts in vitro that recapitulate the mechanical, biochemical, and structural framework of the tissue in order to have an increased chance of integration and survival upon in vivo implantation. While previous work in the lab has explored the utility of physiologically relevant stimuli for creating tissue grafts with chondrocytes, it has not yet been investigated for SDSCs. Therefore, in order to determine the potential of SDSCs as a tissue engineering strategy for growing clinically relevant cartilage grafts, this dissertation had four primary aims: (1) to initially produce tissue growth utilizing synovium-derived stem cells, (2) to utilize additional chemical, physical, and physico-chemical factors to further optimize growth of tissue engineered cartilage using SDSCs, (3) to characterize the response of SDSCs to the factors applied, and (4) to utilize the optimized culture techniques to translate the findings to clinically-relevant human cells. Our initial studies investigated the potential of using physiologically relevant growth factors during both 2D expansion and 3D culture conditions, from which a baseline culture protocol was established. We then sought to explore additional strategies to further optimize tissue growth. Motivated by the discrepancy in osmolarities between native and in vitro culture conditions, we first assessed the influence of adjusting the osmolarity of the baseline culture media. We found that culturing constructs under a more physiologic osmolarity (400 mOsM) was beneficial for tissue growth. Based on these findings implicating osmolarity as a key influencer of growth potential, we sought to determine and potentially manipulate some of the pathways involved in the osmotic response in an effort to further optimize and characterize our tissue-engineered cartilage constructs. Our results supported the role of the TRPV4 ion channel in our SDSC-seeded constructs as a key mechano-osmosensing mechanism. Through the culturing techniques evaluated, we were able to achieve native mechanical and biochemical measures of juvenile bovine cartilage using SDSCs. As has been shown in the literature, observed results in other species (bovine or canine) may not always correlate to findings using human cell sources, thereby prompting the emphasis for more relevant pre-clinical models. Therefore, our final studies sought to translate our treatment strategies to clinically relevant human cells from normal (non-diseased) and diseased (OA) SDSCs and chondrocytes in order to determine their utility. We were able to create a complete set of micropellet data for both SDSCs and chondrocytes to allow for comparisons. Overall, our micropellet results indicate that tissue condition (non-diseased vs OA) is the primary determinant of matrix synthesis. The research described in this dissertation has demonstrated the utility of SDSCs for strategies aimed at cartilage regeneration. We present the first studies to grow SDSC-seeded constructs to native properties of juvenile bovine chondrocytes. Therefore, utilization of the culture techniques presented here and other optimization strategies may hold key insights to developing a tissue using autologous/allogeneic SDSCs that can fully recreate native cartilage. In addition, the findings support the clinical potential of human SDSCs as a cell source for cartilage repair strategies.

The overall goal of this thesis is to develop a new non-contact, whole-body, fluorescence molecular tomography system for small animal imaging. Over the past decade, small animal in vivo imaging has led to a better understanding of many human diseases and improved our ability to develop and test new drugs and medical compounds. Among various imaging modalities, optical imaging techniques have emerged as important tools. In particular, fluorescence and bioluminescence imaging systems have opened new ways for visualizing many molecular pathways inside living animals including gene expression and protein functions. While substantial progress has been made in available prototype and commercial optical imaging systems, there still exist areas for further improvement in the outcome of existing instrumentations. Currently, most small animal optical imaging systems rely on 2D planar imaging that provides limited ability to accurately locate lesions deep inside an animal. Furthermore, most existing tomographic imaging systems use a diffusion model of light propagation, which is of limited accuracy. While more accurate models using the equation of radiative transfer have become available, they have not been widely applied to small animal imaging yet. To overcome the limitations of existing optical small animal imaging systems, a novel imaging system that makes use of the latest hardware and software advances in the field was developed. At the heart of the system is a new double-conical-mirror-based imaging head that enables a single fixed position camera to capture multi-directional views simultaneously. Therefore, the imaging head provides 360-degree measurement data from an entire animal surface in one step. Another benefit provided by this design is the substantial reduction of multiple back-reflections between the animal and mirror surfaces. These back reflections are common in existing mirror-based imaging heads and tend to degrade the quality of raw measurement data. Furthermore, the conical-mirror design offers the capability to measure angular-resolved data from the animal surface. To make full use of this capability, a novel equation of radiative transfer-based ray-transfer operator was introduced to map the spatial and angular information of emitted light on the animal surface to the captured image data. As a result, more data points are involved into the image reconstructions, which leads to a higher image resolution. The performance of the imaging system was evaluated through numerical simulations, experiments using a well-defined tissue phantom, and live-animal studies. Finally, the double reflection mirror scheme presented in this dissertation can be cost-effectively employed with all camera-based imaging systems. The shapes and sizes of mirrors can be varied to accommodate imaging of other objects such as larger animals or human body parts, such as the breast, head, or feet.

The rate limiting factor for the treatment of neurodegenerative diseases is the blood-brain barrier (BBB), which protects the brain microenvironment from the efflux of large molecules, and thus it constitutes a major obstacle in therapeutic drug delivery. All state-of-the-art strategies to circumvent the BBB are invasive or non-localized, include side-effects and limited distribution of the molecule of interest to the brain. Focused Ultrasound (FUS) in conjunction with microbubbles has been shown to open the BBB non-invasively, locally and transiently to allow large molecules diffusion in rodents and non-human primates. This thesis entails a quantitative analysis of the FUS-induced BBB opening in vivo for drug delivery in neurodegenerative diseases. First, quantitative analysis and modeling of the physiologic changes of the BBB opening, such as permeability changes, volume of opening, and reversibility timeline, were studied in wild-type mice, in brain areas related to Alzheimer's and Parkinson's disease. This study provided in vivo tools for BBB opening analysis, as well as the design of a FUS method with optimized parameters for efficient and safe drug delivery. Second, the neurotrophic factor Neurturin, which has been shown to have neuroregenerative and neuroprotective effects in dopaminergic neurons was successfully delivered in wild-type mice and MPTP-lesion parkinsonism model mice. It was shown that FUS enhanced the delivery of Neurturin to the entire regions of interest associated with the disease, downstream signaling for neuronal proliferation was also detected, and finally neuroregeneration was observed in the FUS-treated side compared to the contralateral side. In the third part of this thesis, a pre-clinical translation of the pharmacodynamic analysis was designed and analyzed in non-human primates. The permeability changes, the volume of opening separately in grey and white matter, as well as the concentration of an MR-contrast agent were measured in vivo for the first time. The interaction of FUS with the inhomogeneous primate brain was investigated and the drug delivery efficiency of the FUS technique for BBB opening was measured non-invasively; rather critical findings for safe and optimal drug delivery using FUS in a pre-clinical setting.