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Investigating the Effects of an MMP-inhibitory Biomaterial on the Host Inflammatory Response using an Air Pouch Mouse ModelPatel, Ritesh 13 January 2011 (has links)
An earlier approach to restore homeostatic levels of ECM degrading matrix metalloproteases (MMPs) by the Sefton Lab utilized hydroxamate-based MMP inhibitory (MI) beads. While the MI beads delayed ECM degradation in the context of skin wound healing, they caused elevated cell infiltration in a subcutaneous implant model. The primary goal of this project was to further investigate this finding using an air pouch implant model in mice and a different control group – methacrylic acid-based (MAA) beads. Exudate analysis indicated that the MI beads, implanted subcutaneously with gelatin discs, elicited a similar biological response as the MAA beads. Exudates corresponding to both biomaterials had similar cell counts and chemokine levels, which were greater than those corresponding to the control used earlier, poly-methyl methacrylate-based (PMMA) beads. Further, both MI and MAA beads activated infiltrating macrophages in the classical manner, and influenced the activity of an MMP8 catalytic domain in a similar manner.
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"Role of SRY-related HMG box (SOX)-7 in Skeletal Muscle Development" and "Effect of an extracellular matrix on skeletal and cardiac muscle development"Ebadi, Diba 01 November 2011 (has links)
A complex network of transcription factors, which are regulated by signalling molecules, is responsible in coordinating the formation of differentiated skeletal and cardiac myocytes from undifferentiated stem cells. The present study aims to understand and compare the transcriptional regulation of skeletal and/or cardiac muscle development in the absence of Sox7 or in the presence of a collagen-based matrix in P19 embyonal carcinoma (EC) and mouse embryonic stem (ES) cells.
First, knock-down of Sox7 , by shRNA, in muscle inducing conditions (+DMSO) and in the absence of RA (-RA), decreased muscle progenitor transcription factor and myogenic regulatory factor (MRF) levels, suggesting that Sox7 is necessary for myogenesis. However, knock-down of Sox7 in the presence of RA (+RA) and DMSO increased expression of muscle progenitor markers and MRFs, suggesting that Sox7 is inhibitory for myogenesis +RA. Furthermore, Sox7 overexpression enhanced myogenesis -RA, but inhibited myogenesis and enhanced neurogenesis +RA. These results suggest an important interplay between RA signalling and Sox7 function during P19 differentiation.
Second, Q-PCR analysis showed that compared to the mouse ES cells differentiated on the regular TC plates, differentiation on the collagen matrices had a higher expression of skeletal and cardiac precursors, MRFs and terminal differentiation markers. Collagen alone enhanced myotube formation. The enhanced collagen matrix, containing the oligosaccharide sialyl LewisX (sLeX), specifically enhanced cardiomyogenesis.
These studies have added to our understanding of the transcriptional regulation of premyogenic mesoderm factors and the role of Sox7 in this process. In addition these studies provide a vision for possible use of biomaterials in directed differentiation of stem cells for the purpose of cell therapy.
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Estudio de la interacción de bacterias implicadas en la formulación de placa dentro-bacteriana con superficies de titanio comercialmente puro in vitro y su asociación con la peri-implantitis.Rodríguez Hernández, Ana Guadalupe 22 December 2009 (has links)
La peri-implantitis puede poner en riesgo la integridad funcional de un implante dental, ya que dependiendo de la severidad del daño, éste puede ocasionar la pérdida irremediable del implante. Dicho padecimiento está asociado directamente con la formación de la placa dento-bacteriana sobre la superficie del implante.En este trabajo de tesis se estudiarón, por un lado las propiedades de superficie de 6 tipos diferentes de tratamientos utilizados en implantes dentales (tales como: la rugosidad, la mojabilidad y la energía superficial (Capítulo 2) y su influencia en el comportamiento de adhesión de dos cepas bacterianas Streptococcus sanguinis y Lactobacillus salivarius. La primera de ellas muy importante en la formación de la placa dento-bacteriana por ser una de las bacterias que colonizan la superficie del implante inmediatamente después de ser colocado, facilitando la incorporación de otras bacterias periodontopatogénicas, la segunda, tiene la función de mantener el equilibrio de la placa dento-bacteriana. Y por otro lado, la repercusión que tiene la adhesión y colonización de estas bacterias sobre las superficies de titanio.Para esto, se realizaron dos tipos de ensayos de manera general, los primeros en sus medios rutinarios de cultivo para cada cepa bacteriana (Capítulo 3) y los segundos en una saliva artificial modificada (Capítulo 4). Los resultados observados en ambos casos evidenciaron, el daño ocasionado por la presencia de bacterias en contacto directo con las superficies de titanio, los cuales fueron evaluados detalladamente en el capítulo 5. / Peri-implantitis is a soft tissue dental implant inflammation, which may increase implant failure risk, depending direct on severity illness. This device affection is correlated with implant surface bacterial plaque formation. In this thesis work we studied in one hand, titanium surface properties of 6 different kind of treatments commonly used on dental dental implants (such as: roughness, wettability and surface energy) and its adhesion influence on 2 bacteria strains Streptococcus sanguinis and Lactobacillus salivarius (the first one is implied on dental plaque formation, because these bacteria colonize implant surface immediately after implantation, leading and facilitating the periodontopatogenic bacteria coaggregation, whereas the second strain, has to maintain dental plaque equilibrium). And in the other hand, the last part of this work was focused on the bacterial adhesion effect on titanium surfaces. In general we made two sort of experiments: in rutinary culture media (specific for each strain) and in modified artificial saliva (Chapters 3 and 4 respectively). Our results showed a titanium surface damage because of bacterial attachement on that, which were detailly studied in chapter 5
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Creation of an optimized acellular scaffold for improved vascular engineeringNagao, Ryan Joseph 14 July 2014 (has links)
Engineering a complex tissue that exceeds 100-200 [mu]m requires a vascular connection. Methods to enhance vascularization include the delivery of angiogenic factors, and the use of scaffolds that encourage vascular ingrowth. However, these techniques rely on the host to vascularize the construct upon implantation, which is often too slow to provide nutrients to the entire construct. Hence, recent research has focused on creating de novo vascular networks prior to implantation. Such technologies would enable faster anastomosis with the host vascular system, as well as fully perfused constructs that can increase cell viability. Many techniques have been investigated to create de novo vascular networks with varying levels of success. Our approach was to utilize native vascular extracellular matrix (ECM) obtained from decellularizing highly vascularized tissue as a substrate for re-endothelialization and thus to create a three-dimensional vascular bed for ultimate use with various implant and tissue engineering applications. We have demonstrated a method of chemical decellularization that effectively removes cellular material while leaving behind an organized patent vascular network down to the capillary scale. Standard histological methods, DNA quantification, as well as vascular corrosion casting demonstrated this efficacy. Subsequent subcutaneous implantation then explantation of the scaffold at 7 and 28 days was used to assess the immunogenicity of the graft by analyzing the presence of immune cells. This scaffold was then re-endothelialized with human dermal microvascular endothelial cells (HDMECs) and conditioned with peristaltic flow for 60 hours to help improve vascular patency. Cellular distribution was determined qualitatively by first incubating the HDMECs with gold nanotracers, then imaging their presence upon implantation through ultrasound-guided photoacoustic (US/PA) imaging. Following the culture process, the scaffolds were analyzed for vascular patency through vascular corrosion casting, and cellular phenotype through histological methods---demonstrating a decrease in vascular damage. The re-endothelialized scaffolds were then assessed for functional vascular performance by perfusing whole blood through them. Results demonstrated better blood clearance in re-endothelialized scaffolds compared to scaffolds without cells. These results point to the ability of the optimized acellular (OA) scaffold to be used in future experiments focused on vascular and tissue engineering. / text
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Cellular Response to Surface Wettability Gradient on Microtextured SurfacesPlaisance, Marc Charles 18 August 2015 (has links)
Objective: Topography, chemistry, and energy of titanium (Ti) implants alter cell response through variations in protein adsorption, integrin expression, and downstream cell signaling. However, the contribution of surface energy on cell response is difficult to isolate because altered hydrophilicity can result from changes in surface chemistry or microstructure. Our aim was to examine a unique system of wettability gradients created on microstructured Ti on osteoblast maturation and phenotype.
Method: A surface energy gradient was created on sand-blasted/acid-etched (SLA) Ti surfaces. Surfaces were treated with oxygen plasma for 2 minutes, and then allowed to age for 1, 12, 80, or 116 hours to generate a wettability gradient. Surfaces were characterized by contact angle and SEM. MG63 cells were cultured on SLA or experimental SLA surfaces to confluence on TCPS. Osteoblast differentiation (IBSP, RUNX2, ALP, OCN, OPG) and integrin subunits (ITG2, ITGA5, ITGAV, ITGB1) measured by real-time PCR (n=6 surfaces per variable analyzed by ANOVA/Bonferroni’s modified
Student’s t-test).
Result: After plasma treatment, SLA surface topography was retained. A gradient of wettability was obtained, with contact angles of 32.0° (SLA116), 23.3° (SLA80), 12.5° (SLA12), 7.9° (SLA1). All surfaces were significantly more hydrophilic than the original SLA surface (126.8°). Integrin expression was affected by wettability. ITGA2 was higher on wettable surfaces than on SLA, but was highest on SLA1. ITGAV and ITGB1 were decreased on hydrophilic surfaces, but ITGA5 was not affected. IBSP, RUNX2, and ALP increased and OPG decreased with increasing wettability. OCN decreased with increasing wettability, but levels on the most wettable surface were similar to SLA.
Conclusion: Here we elucidated the role of surface energy on cell response using surfaces with the same topography and chemistry. The results show that osteoblastic maturation was regulated in a wettability-dependent manner and suggest that the effects are mediated by integrins.
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Non-metallic biomaterials for bone substitutes and resorbable biomaterials on orthopaedics / Μη μεταλλικά βιο-υλικά για τα υποκατάστατα οστών και βιο-απορροφημένα βιο-υλικά στην ορθοπεδικήBaciu, Diana-Elena 29 June 2007 (has links)
Thanks to recent advances in science and engineering, the field of biomaterials stands poised to increase the effectiveness and longevity of established devices as well as to provide new options to biomedical engineers who work at designing future products.
From its beginning, the field of bioengineering has focused on providing the best artificial devices - hearing aids, artificial limbs and other prostheses - to replace body parts that are missing, broken, or dysfunctional.
Regeneration of body parts requires a biomaterial with a structure, components and chemical signals that allow the body ’s tissue cells to recognize, respond to, and remodel the material without rejecting it as foreign.
Bone, cartilage and the major load bearing joints of the body all suffer degenerative changes with age and trauma. This area of research focus will seek solutions to the problems of osteoporosis, the fixation of implants in bone and the replacement of damaged bone and cartilage. This will be achieved through the development of non-metallic biomaterials and resorbable biomaterials that provide appropriate load bearing characteristics and the potential to interact suitably with the biology.
The non-metallic materials for bone substitutes serve as scaffolds and may have modified surfaces to encourage natural tissue growth or the ability to be seeded with the hosts own cells before implantation. This will have applications for both bone and cartilage substitute materials. These bone substitutes biomaterials are the second-most implanted of all materials.
Resorbable biomaterials, on the other hand, gradually disappear from the body as a result of hydrolysis, because are made from molecules similar to those in the human body, which resorb while the tissue is healing. This eliminates the need for a second surgery.
The goal of this thesis is to explain the usefulness of these biomaterials in medical applications and especially in orthopaedics, focusing on the latest acquisitions.
The first chapter makes an introduction in biomaterials, with emphasis on orthopaedic biomaterials.
The second chapter contains information about: (1) the bone characteristics (anatomy and mechanics), in order to understand the basis for tissue engineered therapies and how damaged bones heal, (2) the non-metallic biomaterials (polymers, biodegradable polymers, ceramics and composites) for bone substitutes, giving examples of modern biomaterials used today and (3) the principles involved in the Modern Cementing Technique.
The third chapter is a review of the chemistry of the polymers used in bioresorbable biomaterials, including synthesis and degradation, describe how properties can be controlled by proper synthetic controls such as copolymer composition, highlight special requirements for processing and handling, and presents in detail some of the commercial resorbable biomaterials. / Αυτή η διατριβή εξηγεί τη χρησιμότητα των μη μεταλλικών βιο-υλικών για τα υποκατάστατα οστών και των βιο-απορροφημένων βιο-υλικών στις ιατρικές εφαρμογές και ειδικά στις ορθοπεδικές, που εστιάζουν στις πιό πρόσφατες αποκτήσεις.
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DEPOSITION AND CHARACTERIZATION OF MESOPOROUS SILICA COATINGS ON MAGNESIUM ALLOYSAl Hegy, Afrah 17 March 2014 (has links)
In recent years, magnesium and magnesium alloys have received much attention as a new biomaterial in orthopaedic applications due to their biodegradability, biocompatibility, and their mechanical properties that are similar to natural bone tissue. The most common problem associated with magnesium as a biomaterial is low corrosion resistance in physiological solutions. This decreases the mechanical integrity of the implants in the early stages of healing and has a negative impact on the overall biocompatibility. The main goal of this study was to create a multi-layered coating consisting of a silica sol-gel under-layer to protect the substrate from corrosion in body fluids and a mesoporous silica top-layer to enhance the bioactivity of the coated implant material.
The results indicate that the deposited multi-layered coating enhances both the bioactivity and the corrosion resistance of the material.
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Silicon complexes in silicon doped calcium phosphate biomaterialsGillespie, Paul Andrew 03 January 2008 (has links)
The silicon complexes in silicon doped calcium phosphate bioceramics have been studied using $^{29}$Si magic angle spinning nuclear magnetic resonance spectroscopy. The replacement of phosphorus by silicon in these materials requires a charge compensation mechanism which is difficult to study by many experimental techniques due to the small amount of silicon added. Producing these materials using an isotopically enriched source of silicon made the use of NMR spectroscopy feasible. Three different materials have been studied: a multiphase material commercially available under the trade name Skelite$^{\rm TM}$ composed of predominantly a silicon stabilized $\alpha$-tricalcium phosphate ($\alpha$-TCP) phase as well as a silicon doped hydroxyapatite (HA) phase, a single phase Si-HA material and a single phase silicon stabilized $\alpha$-TCP material. Slight changes to the material production method were first introduced to accommodate the switch to an isotopically enriched silicon source. Characterization of the enriched materials was carried out using Rietveld refinement of X-ray powder diffraction spectra and X-ray fluorescence spectroscopy to confirm that these materials were similar to the previously studied, non-enriched, materials in terms of the silicon contents, Ca/(P+Si) molar ratios and lattice parameters. NMR Spectroscopy showed that in all three materials, the silicon formed Q$^1$ structures in which two silicate tetrahedra joined together by sharing an oxygen, creating an oxygen vacancy which compensated the substitution of two silicon for phosphorus. This is the first observation of this charge compensation mechanism in Si-HA and may explain the interesting phase evolution previously found in the system studied in this work in which the Si-HA transforms to silicon stabilized $\alpha$-TCP upon sintering. / Thesis (Master, Physics, Engineering Physics and Astronomy) -- Queen's University, 2007-12-18 14:45:38.721
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Novel radiation sensors based on bio/nanomaterialsAhmadi, Morteza January 2013 (has links)
X-ray sensors are essential to many applications which are not limited to diagnostics and imaging technologies. Such sensors are extensively used in industry, medicine, research and space technology for applications such as safety, security, quality control, imaging and treatment. Depending on the effect of the radiation on the matter employed in the sensor, different types of X-ray sensors are fabricated. However, available techniques of X-ray detection have been under development due to specific shortcomings such as finite life time, low sensitivity, and post-processing requirements. This thesis is focused on design, fabrication and characterization of novel radiation sensors based on bio/nanomaterials.
Bacteriorhodopsin (BR), a proton pump protein in the cell membrane of Halobacterium Salinarum, has been used to fabricate a sensor to measure dose and dose rate of X-ray beam in the kilovoltage and megavoltage energy range. The mass attenuation coefficients, effective atomic numbers and electron densities of BR and its comprising amino acids have been calculated for 1 keV-100 GeV photons to better understand the interaction of BR with X-ray photons.
A theoretical formulation for calculating the change in the conductivity of nanoparticles under radiation is also provided. In particular, the dependence of radiation induced conductivity to irradiated particle size is given. In addition to that, an X-ray sensor based on thin film of bismuth sulfide has been fabricated using laser micromachining and chemical deposition techniques. This sensor has been characterized under a diagnostic X-ray machine with kilovoltage energy beam.
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Angiogenesis in Patches and Injectable Biomaterials for Cardiac RepairChiu, Loraine 11 December 2012 (has links)
Treatment of cardiac diseases involves transplantation of donor hearts, since the damaged heart has limited self-regeneration potential. An alternative treatment option has emerged as engineered cardiac tissues, grown in vitro by cultivation of cardiac cells on biomaterials, have comparable properties to native myocardium and can be implanted for cardiac repair. Major current limitations are a viable cell source and adequate vascularization to support cell survival. In this thesis, two proangiogenic biomaterials, a scaffold and a hydrogel, were developed to achieve vascularization in vitro and in vivo for cardiac repair. Scaffold patches are suitable for repairing congestive heart failure or congenital malformations, while injectable biomaterials allow minimally-invasive treatment post-myocardial infarction (MI). In the first aim, a collagen scaffold with covalently immobilized vascular endothelial growth factor (VEGF) was developed, and improved cell mobilization, survival and proliferation when used for free wall repair in adult rats. This increased angiogenesis, which aided in retaining the biomaterial size to allow tissue growth. In the second aim, a collagen-chitosan hydrogel with encapsulated thymosin β4 (Tβ4) was developed to 1) recruit cells from the heart epicardium for repair post-MI in vivo, and 2) guide capillary outgrowths from arteries and veins to form oriented capillary structure for in vitro cardiac tissue engineering. Results showed that the encapsulation of Tβ4 into collagen-chitosan hydrogels led to cell outgrowths from rat or mouse cardiac explants in vitro. A portion of the recruited cells were CD31-positive endothelial cells (ECs) that formed tubes. The hydrogel was injected in vivo to increase vascularization and number of cardiomyocytes within the infarct area post-MI, which improved left ventricular wall thickness. Tβ4-hydrogel also promoted the outgrowth of capillaries from vascular explants that followed the direction of the hydrogel-coated grooves of a micropatterned polydimethylsiloxane (PDMS) substrate. These capillary outgrowths eventually formed a vascular bed for engineering vascularized cardiac tissues. This thesis presents two bioinstructive biomaterials with sustained and localized delivery of angiogenic molecules to be used for in situ cardiac repair based on improved vascularization. The use of cell-free bioactive materials overcomes limitations of cell isolation and expansion as required for cell therapies or implantation of engineered tissues.
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