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Kinetics of cell attachment and spreading on hard and soft substratesRedmann, Anna-Lena January 2019 (has links)
A very important aspect for the functioning of an organism is that cells adapt their behaviour to external stimuli. They continuously interact with their environment, and biochemical and physical cues can activate cellular signalling, which leads to changes in cell behaviour such as proliferation and shape. Understanding cells' interactions with their environment is also important for understanding diseases. For example mechanosensing, which is the sensing of the cell's mechanical environment, has been associated with cancer development. In order for a cell to be able to sense its mechanical environment, it needs to form attachments to the environment. In my thesis, I have worked on three different tasks: the development of a new measurement technique and the study of initial cell adhesion and of cell spreading. When a cell from suspension first comes into contact with a substrate, it forms initial attachment bonds with proteins on the substrate surface. These bonds are mediated through integrins, which are transmembrane heterodimers, binding to the cell's environment on one side and to the cell's cytoskeleton on the other side. I study this initial cell attachment by measuring the force needed to detach cells, called cell adhesion strength. For these experiments I built a detachment device, which allows the detachment of cells from a substrate by vibrating the substrate in liquid. The device combines cell incubation, detachment and imaging. I measured the dependence of initial integrin bond formation on external factors such as incubation temperature and substrate stiffness. Once initial integrin bonds are formed, many different proteins are recruited to the adhesion site in order to form stronger adhesions. Amongst these proteins are signalling proteins, which direct the behaviour of the cell as a whole. One of the first cellular reactions to a substrate after initial integrin binding is cell spreading. This can be seen by the cell changing its shape from spherical to dome-like on the substrate. Because cell spreading is a very early response of a cell to a substrate, the onset time of spreading can be used as a quantitative measure for the time it takes the cell to sense a substrate and signal shape change. In my work, I look at the distribution of the time of initial cell spreading in a population of cells. I measure this distribution under different growth conditions such as pH, change of incubation medium from DMEM to PBS, substrate stiffness and incubation temperature. In my detachment experiments, I observe that vibration accelerates cell spreading in those cells which remain on the substrate. This is a connection between the detachment experiments and the cell spreading experiments and it shows how cells react to external forces. By changing the medium temperature in the cell detachment and cell spreading experiments, I am able to analyse the kinetics of these two processes. I use a signalling network model to analyse the internal cellular signalling path that leads from a spherical to a spread cell.
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Wet adhesion properties of oilseed proteins stimulated by chemical and physical interactions and bondingLiu, Haijing January 1900 (has links)
Doctor of Philosophy / Department of Grain Science and Industry / X. Susan Sun / The ecological and public health liabilities related with consuming petroleum resources have inspired the development of sustainable and environmental friendly materials. Plant protein, as a byproduct of oil extraction, has been identified as an economical biomaterial source and has previously demonstrated excellent potential for commercial use. Due to the intrinsic structure, protein-based materials are vulnerable to water and present relatively low wet mechanical properties. The purpose of this study focuses on increasing protein surface hydrophobicity through chemical modifications in order to improve wet mechanical strength. However, most of the water sensitive groups (WSG), such as amine, carboxyl, and hydroxyl groups, are also attributed to adhesion. Therefore, the goal of this research is to reduce water sensitive groups to an optimum level that the modified soy protein presents good wet adhesion and wet mechanical strength.
In this research, we proposed two major approaches to reduce WSG: 1). By grafting hydrophobic chemicals onto the WSGs on protein surface; 2). By interacting hydrophobic chemicals with the WSGs. For grafting, undecylenic acid (UA), a castor oil derivative with 11-carbon chain with a carboxyl group at one end and naturally hydrophobic, was used. Carboxyl groups from UA reacted with amine groups from protein and converted amines into ester with hydrophobic chains grafting on protein surface. The successful grafting of UA onto soy protein isolate (SPI) was proved by both Infrared spectroscopy (IR) and ninhydrin test. Wood adhesive made from UA modified soy protein had reached the highest wet strength of 3.30 ± 0.24 MPa with fiber pulled out, which was 65% improvement than control soy protein. Grafting fatty acid chain was verified to improve soy protein water resistance.
For interaction approach, soy oil with three fatty acid chains was used to modify soy protein. Soy oil was first modified into waterborne polyurethanes (WPU) to improve its compatibility and reactivity with aqueous protein. The main forces between WPU and protein were hydrogen bonding, hydrophobic interactions, and physical entanglement. Our results showed that WPU not only increased protein surface hydrophobicity with its fatty acid chains but also enhanced the three-dimensional network structure in WPU-SPI adhesives. WPU modification had increased wet adhesion strength up to 3.81 ± 0.34 MPa with fiber pulled out compared with 2.01 ± 0.46 MPa of SPI. Based on IR and thermal behavior changes observed by DSC, it was inferred that a new crosslinking network formed between WPU and SPI.
To exam if the UA and WPU technologies developed using soy protein are suitable for other plant proteins, we selected camelina protein because camelina oil has superior functional properties for jet fuels and polymers. Like soy protein, camelina protein is also highly water sensitive. However, simply applied UA and WPU to camelina protein following the same methods used for soy proteins, we did not obtain the same good adhesion results compared to what we achieved with soy protein. After protein structure analysis, we realized that camelina protein is more compact in structure compared to soy protein that made it weak in both dry and wet adhesion strength. Therefore, for camelina protein, we unfolded its compact structure with Polymericamine epichlorohydrine (PAE) first to improve flexible chains with more adhesion groups for future reaction with UA or WPU. PAE with charged groups interacted camelina protein through electrostatic interaction and promoted protein unfolding to increase reactivity within protein subunits and between protein and wood cells. Therefore, the wet adhesion strength of camelina protein was improved from zero to 1.30 ± 0.23 MPa, which met the industrial standard for plywood adhesives in terms of adhesion strength. Then the wet adhesion strength of camelina protein was further improved after applying UA and WPU into the PAE modified camelina protein. In addition, we also found PAE unfolding significantly improved the dry adhesion strength of camelina protein from 2.39 ± 0.52 to 5.39 ± 0.50 MPa with 100% wood failure on two-layer wood test.
Camelina meal which is even more economical than camelina protein was studied as wood adhesive. Through a combination of PAE and laccase modification method, the wet adhesion strength of camelina meal was improved as high as 1.04 ± 0.19MPa, which also met industrial standards for plywood adhesives.
The results of this study had proven successful modification of oilseed protein to increase water resistance and wet mechanical strength. We have gained in-depth understanding of the relationship between protein structure and wet adhesion strength. The successful modification of plant proteins meeting the industrial needs for bio-adhesives will promote the development of eco-friendly and sustainable materials.
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A novel approach to measurement of the adhesion strength of a single cell on a substrateColbert, Marie-Josee January 2005 (has links)
No abstract provided / Thesis / Master of Science (MSc)
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A New Technique to Study Temperature Effects on Ice Adhesion Strength for Wind Turbine MaterialsGouni, Rajkiran January 2011 (has links)
No description available.
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Design of multifunctional materials with controlled wetting and adhesion propertiesChanda, Jagannath 29 March 2016 (has links) (PDF)
Ice accretion on various surfaces can cause destructive effect of our lives, from cars, aircrafts, to infrastructure, power line, cooling and transportation systems. There are plenty of methods to overcome the icing problems including electrical, thermal and mechanical process to remove already accumulated ice on the surfaces and to reduce the risk of further operation. But all these process required substantial amount of energy and high cost of operation. To save the global energy and to improvement the safety issue in many infrastructure and transportation systems we have to introduce some passive anti-icing coating known as ice-phobic coating to reduce the ice-formation and ice adhesion onto the surface. Ice-phobic coatings mostly devoted to utilizing lotus-leaf-inspired superhydrophobic coatings. These surfaces show promising behavior due to the low contact area between the impacting water droplets and the surface.
In this present study we investigate systematically the influence of chemical composition and functionality as well as structure of surfaces on wetting properties and later on icing behavior of surfaces. Robust anti-icing coating has been prepared by using modified silica particles as a particles film. Polymer brushes were synthesized on flat, particle surfaces by using Surface initiated ATRP. We have also investigated the effect of anti-icing behavior on the surfaces by varying surface chemistry and textures by using different sizes of particles. This approach is based on the reducing ice accumulation on the surfaces by reducing contact angle hysteresis. This is achieved by introducing nano to micro structured rough surfaces with varying surface chemistry on different substrates.
Freezing and melting dynamics of water has been investigated on different surfaces by water vapour condensation in a high humidity (80%) condition ranging from super hydrophilic to super hydrophobic surfaces below the freezing point of water. Kinetics of frost formation and ice adhesion strength measurements were also performed for all samples. All these experiments were carried out in a custom humidity and temperature controlled chamber. We prepared a superhydrophobic surface by using Poly dimethyl siloxane (PDMS) modified fumed silica which display very low ice-adhesion strength almost 10 times lower than the unmodified surface. Also it has self-cleaning behavior after melting of ice since whole ice layer was folded out from the surface to remove the ice during melting. Systematic investigation of the effect of three parameters as surface energy, surface textures (structure, geometry and roughness) and mechanical properties of polymers (soft and stiff) on icing behavior has also been reported.
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Adesão metal-polímero: dispositivos de medição e correlações físico-químicasGasparin, Alexandre Luis January 2011 (has links)
Uma maneira de quantificar a resistência da interface de materiais compósitos é medindo a tensão necessária para separar o filme do substrato. Esta tensão é frequentemente usada como um parâmetro de projeto para desenvolver estes materiais. Entretanto, muitos métodos de medição da adesão não são capazes de eliminar as falhas de coesão das falhas de adesão filme/substrato. Neste trabalho um novo método de medição de adesão é proposto com o objetivo de eliminar a interferência da falha de coesão por cisalhamento, inerente ao filme delaminado, da medida da tensão normal de adesão feita por método padrão. O novo método provou ser eficiente e seus resultados mostraram serem mais precisos que os do método normalizado. Foram testados os seguintes compostos obtidos por evaporação de cobre usando canhão de elétrons através do processo de deposição física de vapor (PVD), formando um filme metálico sobre quatro substratos poliméricos: polipropileno (PP), poliamida 6 (PA 6), poliestireno de alto impacto (HIPS) e poli(tereftalato) de etileno (PET). A análise de espectroscopia de retroespalhamento Rutherford foi utilizada para caracterizar os filmes e o ângulo de contato para caracterizar a interface. As superfícies dos polímeros foram modificadas através de flambagem e lixamento para validar os resultados do novo método. Finalmente os polímeros delaminados através dos métodos padrão e proposto foram observados por microscopia óptica e eletrônica de varredura (MEV), comprovando assim, que somente no método novo ocorre a separação da interface metal-polímero livre da influência da falha coesiva do filme de cobre. / One of the methods to quantify the interface strength of composites is to measure the tensile stress necessary to separate a film from the substrate surface. Such value is often used as a project parameter to develop the composite. However, most of methods cannot avoid the interference of the cohesion bulk failures from the film/substrate adhesion measures. In this work a new method is proposed in order to eliminate the influence of the cohesion shear failure inherent to the delaminated film of the normal adhesion stress measured by the standard method. The new method has proved to work and their results have become more accurate than the standard pull-off method. The experiment consisted in delaminate a copper film deposited by physical vapor evaporation (PVD) through electron gun on four polymers: polypropylene (PP), polyamide 6 (PA 6), high impact polystyrene (HIPS) and polyethylene terephthalate (PET). The Rutherford backscattering spectrometry was used to characterize the films and the contact angle analysis to characterize the interfaces. The polymer surfaces have also been modified to verify the adhesion strengths of the copper film through sanding and flaming processes to validate the new method. Finally the substrates delaminated were analyzed for both methods, standard and proposed, through optical and scanning electron microscopies, proving that only the new method is effective in pulling-off the metal/polymer interface without the cohesive failure influence of the copper film.
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Cell Adhesion and Migration on NDGA Cross-Linked Fibrillar Collagen Matrices for Tendon Tissue EngineeringRioja, Ana Ysabel 01 January 2012 (has links)
Tendons, essential tissues that connect muscles to bones, are susceptible to rupture/degeneration due to their continuous use for enabling movement. Often surgical intervention is required to repair the tendon; relieving the pain and fixing the limited mobility that occurs from the damage. Unfortunately, post-surgery immobilization techniques required to restore tendon properties frequently lead to scar formation and reduced tendon range of motion. Our ultimate goal is to create an optimal tendon prosthetic that can stabilize the damaged muscle-bone connection and then be remodeled by resident cells from the surrounding tissues over time to ensure long-term function. To achieve this, we must first understand how cells respond to and interact with candidate replacement materials.
The most abundant extracellular matrix (ECM) protein found in the body, collagen, is chosen as the replacement material because it makes up the majority of tendon dry mass and it can be remodeled by cell-based homeostatic processes. Previous studies found that Di-catechol nordihydroguaiaretic acid (NDGA) cross-linked fibers have greater mechanical strength than native tendons; and for this reason this biomaterial could be used for tendon replacement.
This work focuses on investigating the behavior of fibroblasts on NDGA cross-linked and uncross-linked collagen samples to determine if cross-linking disrupts the cell binding sites affecting cell spreading, attachment, and migration. The in-vitro platform was designed by plasma treating 25 mm diameter cover slips that were exposed to 3-aminopropyl-trimetoxysilane/toluene and glutaraldehyde/ethanol solutions. The collagen solution was then dispensed onto the glutaraldehyde-coated cover slip and incubated for fibrillar collagen matrix formation. The collagen matrices were submerged in NDGA cross-linking solution for 24 hours to ensure the surface was completely cross-linked. Collagen films were made by allowing the uncross-linked gels to dry overnight before and after NDGA treatment, resulting in a more compacted structure.
A spinning disk device was employed to quantify the ability of cells to remain attached to the collagen samples when exposed to hydrodynamic forces. To avoid any cell-cell interaction and focus on cell-surface interactions, 50-100 cells/mm2 were seeded carefully on each sample. Temporal studies demonstrated that cell adhesion strength and spreading area reached steady-state by 4 hr. Adhesion and spreading studies along with migration experiments demonstrated that NDGA treatment affects cellular behavior on films, partially reducing adhesion strength, migration, and spreading area. However, on the cross-linked gels which are less dense, the only change in cell behavior observed was in migration speed.
We hypothesize that these differences are due to the collapsing of the collagen films. This compaction suggests a less open organization and could be allowing the collagen fibers to form more inter-chain bonds as well as bonds with the small NDGA cross-linker; while NDGA treatment of the fully hydrated gels may rely more on NDGA polymerization to span the greater distance between collagen fibrils. From these results, we can determine that the chemical/physical masking of the adhesion sites by NDGA on collagen films affects cellular behavior more than the masking that occurs in the cross-linked gels. Although this study shows an effect in cell behavior on the cross-linked films, it also demonstrates that cells can adhere and migrate to this NDGA biomaterial supporting the idea that this biomaterial can be utilized for tendon replacement.
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Adesão metal-polímero: dispositivos de medição e correlações físico-químicasGasparin, Alexandre Luis January 2011 (has links)
Uma maneira de quantificar a resistência da interface de materiais compósitos é medindo a tensão necessária para separar o filme do substrato. Esta tensão é frequentemente usada como um parâmetro de projeto para desenvolver estes materiais. Entretanto, muitos métodos de medição da adesão não são capazes de eliminar as falhas de coesão das falhas de adesão filme/substrato. Neste trabalho um novo método de medição de adesão é proposto com o objetivo de eliminar a interferência da falha de coesão por cisalhamento, inerente ao filme delaminado, da medida da tensão normal de adesão feita por método padrão. O novo método provou ser eficiente e seus resultados mostraram serem mais precisos que os do método normalizado. Foram testados os seguintes compostos obtidos por evaporação de cobre usando canhão de elétrons através do processo de deposição física de vapor (PVD), formando um filme metálico sobre quatro substratos poliméricos: polipropileno (PP), poliamida 6 (PA 6), poliestireno de alto impacto (HIPS) e poli(tereftalato) de etileno (PET). A análise de espectroscopia de retroespalhamento Rutherford foi utilizada para caracterizar os filmes e o ângulo de contato para caracterizar a interface. As superfícies dos polímeros foram modificadas através de flambagem e lixamento para validar os resultados do novo método. Finalmente os polímeros delaminados através dos métodos padrão e proposto foram observados por microscopia óptica e eletrônica de varredura (MEV), comprovando assim, que somente no método novo ocorre a separação da interface metal-polímero livre da influência da falha coesiva do filme de cobre. / One of the methods to quantify the interface strength of composites is to measure the tensile stress necessary to separate a film from the substrate surface. Such value is often used as a project parameter to develop the composite. However, most of methods cannot avoid the interference of the cohesion bulk failures from the film/substrate adhesion measures. In this work a new method is proposed in order to eliminate the influence of the cohesion shear failure inherent to the delaminated film of the normal adhesion stress measured by the standard method. The new method has proved to work and their results have become more accurate than the standard pull-off method. The experiment consisted in delaminate a copper film deposited by physical vapor evaporation (PVD) through electron gun on four polymers: polypropylene (PP), polyamide 6 (PA 6), high impact polystyrene (HIPS) and polyethylene terephthalate (PET). The Rutherford backscattering spectrometry was used to characterize the films and the contact angle analysis to characterize the interfaces. The polymer surfaces have also been modified to verify the adhesion strengths of the copper film through sanding and flaming processes to validate the new method. Finally the substrates delaminated were analyzed for both methods, standard and proposed, through optical and scanning electron microscopies, proving that only the new method is effective in pulling-off the metal/polymer interface without the cohesive failure influence of the copper film.
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Adesão metal-polímero: dispositivos de medição e correlações físico-químicasGasparin, Alexandre Luis January 2011 (has links)
Uma maneira de quantificar a resistência da interface de materiais compósitos é medindo a tensão necessária para separar o filme do substrato. Esta tensão é frequentemente usada como um parâmetro de projeto para desenvolver estes materiais. Entretanto, muitos métodos de medição da adesão não são capazes de eliminar as falhas de coesão das falhas de adesão filme/substrato. Neste trabalho um novo método de medição de adesão é proposto com o objetivo de eliminar a interferência da falha de coesão por cisalhamento, inerente ao filme delaminado, da medida da tensão normal de adesão feita por método padrão. O novo método provou ser eficiente e seus resultados mostraram serem mais precisos que os do método normalizado. Foram testados os seguintes compostos obtidos por evaporação de cobre usando canhão de elétrons através do processo de deposição física de vapor (PVD), formando um filme metálico sobre quatro substratos poliméricos: polipropileno (PP), poliamida 6 (PA 6), poliestireno de alto impacto (HIPS) e poli(tereftalato) de etileno (PET). A análise de espectroscopia de retroespalhamento Rutherford foi utilizada para caracterizar os filmes e o ângulo de contato para caracterizar a interface. As superfícies dos polímeros foram modificadas através de flambagem e lixamento para validar os resultados do novo método. Finalmente os polímeros delaminados através dos métodos padrão e proposto foram observados por microscopia óptica e eletrônica de varredura (MEV), comprovando assim, que somente no método novo ocorre a separação da interface metal-polímero livre da influência da falha coesiva do filme de cobre. / One of the methods to quantify the interface strength of composites is to measure the tensile stress necessary to separate a film from the substrate surface. Such value is often used as a project parameter to develop the composite. However, most of methods cannot avoid the interference of the cohesion bulk failures from the film/substrate adhesion measures. In this work a new method is proposed in order to eliminate the influence of the cohesion shear failure inherent to the delaminated film of the normal adhesion stress measured by the standard method. The new method has proved to work and their results have become more accurate than the standard pull-off method. The experiment consisted in delaminate a copper film deposited by physical vapor evaporation (PVD) through electron gun on four polymers: polypropylene (PP), polyamide 6 (PA 6), high impact polystyrene (HIPS) and polyethylene terephthalate (PET). The Rutherford backscattering spectrometry was used to characterize the films and the contact angle analysis to characterize the interfaces. The polymer surfaces have also been modified to verify the adhesion strengths of the copper film through sanding and flaming processes to validate the new method. Finally the substrates delaminated were analyzed for both methods, standard and proposed, through optical and scanning electron microscopies, proving that only the new method is effective in pulling-off the metal/polymer interface without the cohesive failure influence of the copper film.
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Vliv povrchu na pevnost lepeného spoje / Effect of surface on bonded jointTrhoň, Vojtěch January 2011 (has links)
This master’s thesis is divided into two parts. In the first part are these topics: bonding theory, treatment of bonding surface, types of adhesives, adhesives in the transportation industry and construction and stress of bonded joints. In the second part of this thesis is experimental evaluation of the effect of surface of material on strength of bonded joint.
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