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Studies on the activities of serine proteases from Ficus carica / Ficus carica由来セリンプロテアーゼの活性に関する研究Nishimura, Kosaku 26 July 2021 (has links)
京都大学 / 新制・課程博士 / 博士(農学) / 甲第23433号 / 農博第2464号 / 新制||農||1086(附属図書館) / 学位論文||R3||N5348(農学部図書室) / 京都大学大学院農学研究科食品生物科学専攻 / (主査)教授 保川 清, 教授 谷 史人, 教授 橋本 渉 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM
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Alteration of cartilage-surface collagen fibers differs locally after immobilization of knee joints in rats / ラット膝関節不動後の軟骨表面のコラーゲン線維変化は領域により異なるNagai, Momoko 25 May 2015 (has links)
京都大学 / 0048 / 新制・課程博士 / 博士(人間健康科学) / 甲第19180号 / 人健博第28号 / 新制||人健||3(附属図書館) / 32172 / 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 高桑 徹也, 教授 市橋 則明, 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
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Regeneration of elastic fibers by three-dimensional culture on a collagen scaffold and the addition of latent TGF-β binding protein 4 to improve elastic matrix deposition / コラーゲン基材を用いた3次元培養系において、latent TGF-β binding protein 4は弾性線維再生を促進するAya, Rino 23 March 2016 (has links)
京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19570号 / 医博第4077号 / 新制||医||1013(附属図書館) / 32606 / 京都大学大学院医学研究科医学専攻 / (主査)教授 松田 秀一, 教授 開 祐司, 教授 妻木 範行 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM
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OPTIMIZED BIODEGRADABLE FIBRIN HYDROGELS AS IN VITRO MODELS OF WOUND HEALINGPatel, Hardika, 0000-0002-5048-0925 January 2022 (has links)
Skin is the largest organ of the body. Its integrity plays a crucial role in maintaining physiological homeostasis, protects against mechanical forces and infections, fluid imbalance, and thermal dysregulation. Numerous pathological states, such as diabetes mellitus, peripheral vascular disease, thermal injuries, or degloving lead to inadequate wound healing, necessitating medical intervention. Established wound healing techniques such as autologous and allogeneic skin grafts are inefficient due to the limited availability of donor tissues or probable immunogenic reactions. Current research in the field of tissue engineering aims to facilitate wound healing and restore skin functionality, focusing on key aspects of wound healing, such as extracellular matrix (ECM) reorganization, cell growth, and collagen synthesis/deposition. The research aims at developing and characterizing an in-vitro fibrin gel culture model system that stimulates the process of wound healing. The specific goal of this research is to investigate how the varied chemical composition of fibrin hydrogels can enhance fibroblast proliferation and promote accelerated collagen matrix formation, which is a significant step in tissue repair and regeneration.The fibrin gels are optimized by modulating the primary gel constituents (i.e. the concentrations of fibrin and thrombin). The ensuing hydrogels are characterized using Scanning Electron Microscope and compression testing to test for fiber size, porosity, elasticity, and mechanical properties. Cultured fibroblasts are used to investigate the effects of varying fibrin concentrations on cell-biomaterial
interactions, including cell proliferation, cellular infiltration, and network formation. Furthermore, matrix formation and maturation as a function of fibrinogen concentration as defined by collagen matrix deposition, are also studied.
Increasing the fibrinogen concentration, lead to an increase in elasticity and Young’s modulus, while a decrease in thrombin concentration generated a stronger fiber structure. Additionally, a decrease in fibrinogen concentration resulted in an increased proliferation rate of fibroblast cells, suggesting better cell adhesion and network formation within the gel substrate. These results were consistent and confirmed by quantifying a mature collagen matrix deposited by fibroblasts when subjected to ascorbic acid.
In summary, this research investigates how the varied chemical composition of fibrin hydrogels can enhance fibroblast proliferation and promote accelerated collagen matrix formation, which is a significant step in tissue repair and regeneration. / Bioengineering
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Characterization of Fat-Storing Cell Lines Derived From Normal and CCl<sub>4</sub>-Cirrhotic Livers. Differences in the Production of Interleukin-6Greenwel, P., Schwartz, M., Rosas, M., Peyrol, S., Grimaud, J. A., Rojkind, M. 01 December 1991 (has links)
Liver fat-storing cells (FSC) play an important role in collagen deposition. During the induction of liver cirrhosis, FSC lose their fat droplets, acquire an actin-rich cytoskeleton and transform into myofibroblasts. Myofibroblasts have been associated with increased collagen production in cirrhotic livers. Cultured FSC resemble myofibroblasts. However, it is not known whether regulation of collagen gene expression is similar in FSC obtained from normal or cirrhotic livers. In this communication, we describe the characterization of two fat-storing cell lines, one from normal (NFSC) and one from CCl4-cirrhotic liver (CFSC), obtained after spontaneous immortalization in culture. We studied the effect of serum and various growth factors on cell proliferation. We determined the production of collagen and fibronectin and we analyzed the presence of mRNA transcripts of collagens type I, III, and IV, fibronectin laminin, transforming growth factor-β and interleukin-6. We found that CFSC have a greater serum-dependency than NFSC. NFSC grow with a mixture of insulin and epidermal growth factor, whereas CFSC proliferate only with platelet-derived growth factor. Although we did not find significant differences in the expression of mRNAs for collagen type I, fibronectin and transforming growth factor-β, collagen and fibronectin synthesis was increased 2- and 1.5-fold respectively. NFSC contained 1.6- and 2.0-fold more type III collagen and laminin mRNAs, respectively, than CFSC. Neither cell line expressed type IV collagen mRNA. NFSC but not CFSC produced interleukin-6. These results suggest that, except for the lack of transcripts of collagen type IV, both cell lines resemble primary cultures of FSC. However, significant differences in cell proliferation and interleukin-6 production between the two cell lines were found. We suggest that these cell lines could be useful tools to study possible differences in regulation of matrix production by FSC.
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Stabilizing a FRET DNA Origami Sensor to Measure the Mechanical Properties of the Tumor Extracellular MatrixKolotka, Kelly L. January 2019 (has links)
No description available.
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Computational Bone Mechanics Modeling with Frequency Dependent Rheological Properties and CrosslinkingMoreno, Timothy G 01 March 2021 (has links) (PDF)
Bone is a largely bipartite viscoelastic composite. Its mechanical behavior is determined by strain rate and the relative proportions of its principal constituent elements, hydroxyapatite and collagen, but is also largely dictated by their geometry and topology. Collagen fibrils include many segments of tropocollagen in staggered, parallel sequences. The physical staggering of this tropocollagen allows for gaps known as hole-zones, which serve as nucleation points for apatite mineral. The distance between adjacent repeat units of tropocollagen is known as D-Spacing and can be measured by Atomic Force Microscopy (AFM). This D-Spacing can vary in length slightly within a bundle, but by an additional order of magnitude within the same specimen, and can significantly alter the proportion of hydroxyapatite. Previous researchers have built and refined a Finite Element Analysis “Complex Model” to capture the consequences of adjusting D-Spacing and the viscoelastic parameters. This will ultimately serve to elucidate and perhaps predict the mechanical consequences of biological events that alter these parameters. This study aims to further refine the model’s precision by accounting for crosslinking between fibrils, the presence of which serves to add mechanical strength. This study also looks to refine the currently used rheological models by way of frequency dependent parameters in the hopes of improving model accuracy over a wider frequency range.
Hormonal factors such as estrogen can significantly determine the composition of bone. Menopause marks a significant reduction in circulating estrogen and has been shown to factor heavily in the development of conditions like osteoporosis. Because sheep feature a hormonal cycle and skeletal structure similar to humans, three of six mature Columbia-Rambouillet ewes were randomly selected to undergo an ovariectomy, the remainder serving as sham-operated controls. Twelve months later twenty-five beam samples were harvested from their radius bones for mechanical analysis and other testing, including atomic force microscopy (AFM) and dynamic mechanical analysis (DMA). The data gleaned from these tests provide an experimental basis of comparison with The Complex Model.
A 2-D Finite Element Analysis model in Abaqus was first created by Miguel Mendoza, which enforced viscoelasticity and a realistic proportion and placement of hydroxyapatite and collagen. The viscoelasticity was modeled using a Standard Linear Solid involving springs and a dashpot element. Crosslinks of varying number and location were arranged within the former model configuration as node to surface tie-constraints to explore the treatment of the FEA Model as a more realistic assembly of parts. Frequencies utilized for this model included 1, 3, 9 and 12 Hz. This approach is referred to in this research as the Intermolecular Forces (IMF) Scheme.
The model was subsequently refined by Christopher Ha and Austin Cummings. The model was characterized by 2x100 unit half-cells, the lengths of which were randomly generated by a Python script. This script ingested the mean and standard deviation D-Spacing length to generate a model geometrically similar to a real specimen bearing those dimensions. A frequency dependent value for the dashpot element in the rheological model used for tropocollagen was developed using this latter FEA model, named the Complex Model. Dashpot values explored for this variable dashpot included 0.0125, 0.125, 0.3125, 0.45, 0.5875, 0.725, 0.8625 and 1.25 GPa-s, some values chosen for their high performance in past studies and others to further narrow the search for the best performing dashpot. All dashpot values were investigated over the previously stated frequencies in addition to 2, 5, 7 and 12 Hz. The best fit dashpot values were plotted against the frequencies in which they best performed and a polynomial trend line was fitted to establish an equation, and that equation was used to modify an existing user material subroutine for tropocollagen to provide an automatic frequency dependent dashpot value to Abaqus. This approach is referred to in this research as the Variable Dashpot (VD) Scheme.
Results for the IMF scheme generally performed poorly, with the fully tie-constrained model performing best with 0.77 and 0.024 for R2 and RMSE respectively. Of the randomized crosslink models, that with the lowest number (N=20) of randomly placed non-enzymatic crosslinks performed best with 0.81 and 0.051 for R2 and RMSE respectively. Increasing the number of randomized crosslinks reduced model fit, and the remaining three variants exhibited mean R2 and RMSE values of 0.66-0.67 and 0.052 respectively. For the VD scheme, models running custom modified variable dashpot UMATs yielded R2 and RMSE values of 0.87 and 0.012 for C2207, and 0.89 and 0.008 for C1809. This is a notable fit considering all other material property parameters are held constant throughout each frequency. In the rheological model, this research also found a striking difference between the frequency dependent viscous element values that made each model perform best. This indicates that differences in D-Spacing standard deviations between OVX and control may be associated with distinct strain-rate dependent mechanical responses.
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Biophysical study of the extracellular matrix for vascular physiology and cancer biology applicationsCortes Medina, Marcos G. January 2022 (has links)
No description available.
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Analysis of bendable osteochondral allograft treatment and investigations of articular cartilage wear mechanicsPetersen, Courtney A. January 2023 (has links)
Osteoarthritis is a highly prevalent, debilitating disease characterized by the wear and degradation of articular cartilage. While many surgical interventions exist, few are consistently effective and those that are effective are not necessarily suitable for all patients. The objective of this dissertation is to improve patient care through the development of a new surgical technique and through basic science studies which seek to better understand articular cartilage wear initiation. Four studies, which address this objective are summarized below.
Osteochondral allograft transplantation provides a safe and effective treatment option for large cartilage defects, but its use is limited partly due to the difficulty of matching articular surface curvature between donor and recipient. We hypothesize that bendable osteochondral allografts may provide better curvature matching for patella transplants in the patellofemoral joint. The finite element study presented in Chapter 2 investigates patellofemoral joint congruence for unbent and bendable osteochondral allografts, at various flexion angles. Finite element models were created for 12 femur-patella osteochondral allograft pairings. Two grooves were cut into the bony substrate of each allograft, allowing the articular layer to bend. Patellofemoral joints with either unbent (OCA) or permanently bent (BOCA) allografts were articulated from 40 to 70 degrees flexion and contact area was calculated. OCAs and BOCAs were then shifted 6 mm distally toward the tibia (S-OCA, S-BOCA) to investigate the influence of proximal-distal alignment on congruence. On average, no significant difference in contact area was found between native patellofemoral joints and either OCAs or BOCAs (p > 0.25), indicating that both types of allografts restored native congruence. This result provides biomechanical support in favor of an emerging surgical procedure. S-BOCAs resulted in a significant increase in contact area relative to the remaining groups (p < 0.02). The fact that bendable osteochondral allografts produced equally good results implies that these bendable allografts may prove useful in future surgical procedures, with the possibility of transplanting them with a small distal shift. Surgeons who are reluctant to use osteochondral allografts for resurfacing patellae based on curvature matching capabilities may be more amenable to adopting bendable osteochondral allografts.
The recent development of bendable osteochondral allografts provides the potential for improved osteoarthritis treatment for joints whose current treatment is unsatisfactory. One such joint is the carpometacarpal joint in the thumb. While the current standard of care for carpometacarpal osteoarthritis, ligament reconstruction and tendon interposition, can reduce pain in the joint, it does not restore full joint function and mobility. A proposed alternative includes using an osteochondral allograft harvested from the femoral trochlea in a donor knee, machining grooves in the bone to allow the allograft to bend, and replacing the trapezium with this bent osteochondral allograft [1,2]. Chapter 3 of this dissertation discusses adjustments to the original design of the bendable allograft and the design of a custom surgical tool to perform the proposed surgery. Specification changes of the allograft included an overall size reduction in order to better fit within the carpometacarpal joint, minimum bone thickness requirements to avoid bone cracking during the surgical procedure, and a reduction from three grooves to two grooves, which provided sufficient bending yet avoided fracture of the allograft. The surgical tool was designed to be a custom forceps device, whose primary features included (1) jaws with an angled face to match the angle of allograft bending and (2) insertion holes for the Kirschner wire and compression screws used to anchor the allograft in the bent position. These customizations allow the tool to be used to bend the allograft, fix it in the bent configuration, and place the allograft in its proper position in the hand during anchoring of the bent allograft to the native trapezium.
The final two studies presented in this dissertation focus on furthering our current understanding of wear and structure-function relationships of articular cartilage. We hypothesize that cartilage wears due to fatigue failure in reciprocating compression instead of reciprocating friction. Chapter 4 compares reciprocating sliding of immature bovine articular cartilage against glass in two testing configurations: (1) a stationary contact area configuration (SCA), which results in static compression, interstitial fluid depressurization and increasing friction coefficient during reciprocating sliding, and (2) a migrating contact area configuration (MCA), which maintains fluid pressurization and low friction while producing reciprocating compressive loading during reciprocating sliding. Contact stress, sliding duration, and sliding distance were controlled to be similar between test groups. SCA tests exhibited an average friction coefficient of μ=0.084±0.032, while MCA tests exhibited a lower average friction coefficient of μ=0.020±0.008 (p<10^(-4)).
Despite the lower friction, MCA cartilage samples exhibited clear surface damage with a significantly greater average surface deviation from a fitted plane after wear testing (R_q=0.125±0.095 mm) than cartilage samples slid in a SCA configuration (R_q=0.044±0.017 mm, p=0.002), which showed minimal signs of wear. Polarized light microscopy confirmed that delamination damage occurred between the superficial and middle zones of the articular cartilage in MCA samples. The greatest wear was observed in the group with lowest friction coefficient, subjected to cyclical instead of static compression, implying that friction is not the primary driver of cartilage wear. Delamination between superficial and middle zones imply the main mode of wear is fatigue failure under cyclical compression, not fatigue or abrasion due to reciprocating frictional sliding.
The final study of this dissertation, presented in Chapter 5, investigates the importance of collagen fibril distribution in articular cartilage computational models. Finite element models were created to approximate a bovine humeral head and replicate previous experimental loading conditions [3]. Five different finite element analyses were run, each using a different fibril distribution model. Three of the models used two, four, or eight discrete fibril bundles, while two models used continuous fibril distributions with either isotropic or depth-dependent ellipsoidal distributions.
Two primary findings arose from this investigation. The first was the discovery that as the fibril distribution became more isotropic, the strain throughout the tissue decreased, even though the contact area between the articular surface and rigid platen remained relatively equal across distribution models. This suggests that computational models which approximate the collagen fibrils with an isotropic distribution may be underestimating the strain through the depth of the tissue. The second primary finding was that in the discrete distribution model with two fibril bundles, which followed the classically described Benninghoff structure [4], the greatest magnitude of shear strain during compressive loading was observed in the middle zone. However, the highest magnitude of shear strain observed in the isotropic fibril distribution model occurred in the deep zone near the subchondral surface. The observed results suggest that the type of fibril distribution used to model collagen in articular cartilage plays a role in depth-dependent strain magnitude and strain distribution.
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Evaluation of Chitosan as a Cell Scaffolding Material for Cartilage Tissue EngineeringNettles, Dana Lynn 14 December 2001 (has links)
Current articular cartilage tissue engineering endeavors, using synthetic polymers as scaffolds, have been somewhat successful. However, the use of these materials has not yielded a satisfactory, functional replacement for articular cartilage. Therefore, this project focuses on an alternative to these materials, chitosan, which is a naturally occurring biopolymer. The first project objective was to fabricate and analyze bulk, porous chitosan scaffolds, based on total porosity, average pore diameter, mechanical integrity, and degradation susceptibility. Secondly, scaffolds were evaluated in terms of their ability to support neochondrogenesis, including assessments of cell attachment and viability, cell morphology, and the biosynthesis of proteoglycan and type-II collagen-rich extracellular matrix. Results indicated that chitosan scaffolds possessing an interconnecting, porous structure could be easily created through a simple freezing and lyophilization process, and these scaffolds did support neochondrogenesis. Results suggest chitosan may be a useful alternative to synthetic polymers for use in cartilage tissue engineering applications.
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