京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18864号 / 医博第3975号 / 新制||医||1008(附属図書館) / 31815 / 京都大学大学院医学研究科医学専攻 / (主査)教授 木村 剛, 教授 坂田 隆造, 教授 山下 潤 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

The project is an analysis of technology production of pressure pipes made of aluminum alloys. The basis is a literary study of TIG technology, aluminum heat-tretable and non-heat-treatable materials. The flange-material is EN AW 5083 and the pipe is made of EN AW 6005A. The design of the welding is compromise between the preparation, the cleaning of the welding edges and the weld metal backing strip. Weld was made in real production. Examined impacts are evaluated on the basis of destructive and non-destructive welding test methods. After heating process of weldment material exhibits better mechanical properties. Using the economical and technological evaluation were selected sutiable proces parameters. The result is a suitable weld of the pressure vessel. Further optimization is possible through automation and robotics.

Microglia, the resident macrophages of the central nervous system (CNS), are dynamic cells, constantly extending and retracting their processes as they contact and functionally regulate neurons and other glial cells. There is far less known about how microglia interact with the CNS vasculature, particularly under healthy steady-state conditions. Here, I provide the first extensive characterization of juxtavascular microglia in the healthy, postnatal brain and identify a molecular mechanism regulating the timing of these interactions during development. Using the mouse cerebral cortex, I show that microglia are intimately associated with the vasculature in the CNS, directly contacting the basal lamina in vascular sites that are devoid of astrocyte endfeet. I demonstrate a high percentage of microglia are associated with the vasculature during the first week of postnatal development, which is concomitant with a peak in microglial colonization of the cortex and recruitment to synapses. I find that as microglia colonize the cortex, juxtavascular microglia are highly motile along vessels and become largely stationary as the brain matures. 2-photon live imaging in adult mice reveals that these vascular-associated microglia in the mature brain are stable and stationary for several weeks. Further, a decrease in microglia motility along the vasculature is tightly correlated with the expansion of astrocyte endfeet along the vasculature. Finally, I provide evidence that the timing of these microglia-vascular interactions during development is regulated by the microglial fractalkine receptor (CX3CR1). Together, these data support a model by which microglia use the vasculature as a scaffold to migrate and colonize the developing brain and the timing of these associations is modulated by CX3CR1. This migration along the vasculature becomes restricted as astrocyte vascular endfoot territory expands and, upon maturation, vascular-associated microglia become largely stationary.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21684号 / 医博第4490号 / 新制||医||1036(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 松田 道行, 教授 松田 秀一, 教授 椛島 健治 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Blunt carotid artery injury (BCAI), resulting primarily from automobile accidents, is a major contributor to the high mortality and morbidity rates associated with carotid artery dissection. More work is needed to characterize carotid artery injury mechanisms, quantify stages of damage, and elucidate failure modalities as a result of this type of injury. The present study examines the structure and mechanics of the carotid artery in the circumferential and axial directions by employing uniaxial tensile testing, high speed videography, interruption testing, scanning electron microscopy (SEM), histological analysis, real-time environmental SEM assessment, and atomic force microscopy (AFM). Results are as follows: (i) the carotid artery exhibits anisotropic, viscoelastic behavior; (ii) intimal failure precedes ultimate tissue failure, and the layers in order of increasing strength are intima, adventitia, and media; (iii) tissue damage accumulates as strain level increases, and failure occurs as a result of void nucleation, void growth, and void coalescence.

ABSTRACT Implementation of Physiologic Pressure Conditions in a Blood Vessel Mimic Bioreactor System Kevin Mark Okarski Tissue engineering has traditionally been pursued as a therapeutic science intended for restoring or replacing diseased or damaged biologic tissues or organs. Cal Poly’s Blood Vessel Mimic Laboratory is developing a novel application of tissue engineering as a tool for the preclinical evaluation of intravascular devices. The blood vessel mimic (BVM) system has been previously used to assess the tissue response to deployed stents, but under non-physiologic conditions. Since then, efforts have been made to improve the vessel and bioreactor’s ability to emulate in vivo conditions. The ability to tissue engineer constructs similar to their native tissue counterparts is heavily reliant upon controlling the environment and mechanical stimuli the construct is exposed to. Mimicking physiologic conditions influences cellular growth, proliferation, and differentiation. Two important mechanical stimuli are cyclic strain and wall shear stress. Previous work sought to improve these factors within the BVM bioreactor and resulted in the implementation of pulsatile perfusion and increased fluid viscosity. These previous bioreactor design modifications generated pulsatile pressures of approximately 80 mmHg and a wall shear stress of 6.4 dynes/cm2. However, physiologic pressure waveforms were not achieved. Studies in this thesis were carried out to implement an effective means of establishing a more physiologic pressure wave within the bioreactor that is accurate, consistent, and easily adjustable. As a result of conducting the present studies, modifications to the bioreactor system were made that uphold the overall goals of efficacy and efficiency. The desired pressure wave was created by setting the degree of pump tubing occlusion on the 3-roller peristaltic pump head and using a water column to backpressure the bioreactor chamber. Maintaining a desired backpressure within the system necessitated the development of a new bioreactor chamber with increased extraluminal leak pressure resistance. The opportunity was also used to further improve upon the bioreactor chamber design to allow for 360° rotation to reduce cell sedimentation. Modifications to the bioreactor system required quantitative evaluation to assess their impact upon local flow dynamics to the tissue construct. A system model was created and evaluated using computational modeling. Through the work performed in this thesis, pulsatile pressure waves of approximately 120/80 mmHg were successfully established within the bioreactor. The ability to accurately model physiologic pressures will ultimately help yield tissue constructs more similar to native tissues – both healthy and pathological. The newly designed bioreactor chamber and computational model for the system will be helpful tools for implementing or evaluating future bioreactor developments or improvements. While the main objective of the thesis has been completed by creating a system capable of emulating physiologic pressure fluctuations, there still remains room for further improvements in back-pressuring and scaling the system, refining the rotational bioreactor chamber design, and building upon the complexity and accuracy of the computational model.

“Blood vessel mimics” (BVMs) are tissue-engineered constructs that serve as in vitro preclinical testing models for intravascular devices. The Cal Poly Tissue Engineering lab specifically uses BVMs to test the cellular response to stent implantation. PLGA scaffolds are electrospun in-house using the current “Standard Protocol” and used as the framework for these constructs. The performance of BVMs greatly depends on material and mechanical properties of the scaffolds. It is desirable to create BVMs with reproducible properties so that they can be consistent models that ultimately generate more reliable results for intravascular device testing. Reproducibility stems from the consistency of the scaffolds. Thus, scaffolds with consistent material and mechanical properties are necessary for creating reproducible BVMs. The aim of this thesis was to characterize the reproducibility of the electrospun PLGA scaffolds using fiber diameter measurements and compliance testing. Initial work in this investigation involved designing and testing several experimental electrospinning protocols to obtain smaller fiber diameters, which have been shown to elicit more ideal cellular responses. The most successful protocol in that regard was then analyzed for the reproducibility of fiber diameters and compared to the reproducibility of the Standard Protocol. After determining that the Standard Protocol produced scaffolds with more consistent fibers, a large-scale reproducibility study was performed using this protocol. In this expanded study, both fiber diameter and compliance were analyzed and used to characterize the scaffolds. It was established that the scaffolds demonstrated inconsistent mean fiber diameter and mean compliance. The current standard electrospinning protocol therefore does not create PLGA scaffolds with statistically reproducible properties. Future modifications should be made to the electrospinning parameters in order to reduce variability between the scaffolds and future studies should be performed to determine the acceptable range of properties.

Drosophila melanogaster has a tubular heart called the dorsal vessel, which is composed of contractile cardiomyocytes and hemolymph filtering pericardial cells. During larval development the dorsal vessel (heart) grows in size, and the luminal space inside the heart expands, however it has not been clear which cells are responsible for laying the extracellular matrix (ECM) during this expansion. Hemocytes (white blood cells), pericardial cells and cells of the fat body are candidate cell types that may secrete ECM for assembly during the growth of the heart lumen. With gene knock-down techniques we are exploring whether hemocytes participate in assembly of the heart ECM at this location. Additionally, studies of fluorescently tagged hemocytes in intact larvae reveal that hemocytes aggregate around pericardial cells of the dorsal vessel in 3rd instars. Confocal studies of dissected larval hearts indicate that hemocytes aggregate within infoldings of basement membrane associated with pericardial cells. Hemocyte-pericardial cell association could indicate that hemocytes take up proteins that are produced by pericardial cells and deliver them to other locations or that there might be a previously unidentified hematopoietic site at the Drosophila larval heart. / Thesis / Master of Science (MSc)

Over the past three decades, the field of tissue engineering has witnessed significant advancements. However, a persistent challenge is the development of an approach to generate rapidly perfused vascular networks at scale to support engineered tissues of appreciable size and able to adapt to changing needs. Current techniques able to create perfusable channels such as 3D printing are resource intensive and have not overcome the inherent tradeoff between vessel resolution and assembly time, limiting their utility and scalability for engineering tissues. Here we present two sacrificial self-assembly techniques that collectively develop microvascular networks and can anastomose to a variety of engineered forms. The first is vasculogenic cellular self-assembly, which leverages the innate ability of endothelial and sacrificial support cells to spontaneously form a capillary network, which we term CAMEO, or Controlled Apoptosis in Multicellular Tissues for Engineered Organogenesis. By varying the removal timing of the support cells, we determine fibroblasts are necessary for the initial vascular morphogenesis in our engineered system, and that this initial support period is sufficient for the endothelial cells to form a perfusable vasculogenic network and enhance the function of primary hepatocyte aggregates. The second is a flexible and scalable technique we term SPAN – Sacrificial Percolation of Anisotropic Networks. It uses microvascular-scale sacrificial fibers that make contacts to span a volume above a percolation density threshold and are then degraded. The resulting interconnected anisotropic voids form a perfusable fluidic network within minutes. We show that SPAN relieves hypoxia compared to bulk gels only, and the resulting voids created by SPAN can be endothelialized in a scalable way. These simple platforms can generate conduits with length scales spanning arterioles to capillaries within constructs. We show that both techniques can be used in combination with common tissue engineering processes, paving the way for rapid assembly of adaptable and perfusable tissues. / 2026-01-17T00:00:00Z

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