Small diameter synthetic vascular grafts have yet to be clinically successful due to luminal narrowing from thrombosis and intimal hyperplasia. Current attempts to address this issue include the development of materials that support endothelialisation and protein modification to the material surfaces that reduce thrombosis. The extracellular matrix protein elastin has been found to be one of the least thrombogenic components of blood vessels, and its purified and recombinant forms have shown reduced thrombogenicity in both in vitro and in vivo models. Biomaterial coatings of elastin-like polypeptides (ELPs) recombinantly produced in the Woodhouse laboratory showed reduced fibrinogen adsorption, platelet adhesion, and platelet activity. However, the reason for their relative non-thrombogenicity is still not fully understood. In this work, the leukocyte response to ELP-coated materials was investigated. In particular, ELP1 and ELP4, which differ in molecular weight and sequence length, were physically adsorbed to a polyethylene terephthalate surface (MylarTM), yielding 0.22 ± 0.13 μg/cm2 and 0.37 ± 0.19 μg/cm2 surface coverage, respectively, as determined by the colorimetric assay, FastinTM Elastin. These surfaces were exposed to flowing citrated whole blood for surface and bulk evaluation of leukocyte activity using scanning electron microscopy and flow cytometry, respectively. Little leukocyte activation was observed on the surface of the controls, low-density polyethylene and uncoated MylarTM. In the bulk, tissue factor (TF) expression (monocytes: ELP1 = 38.6 ± 16.3 %, ELP4 = 33.9 ± 18.1 %) and platelet-leukocyte aggregates determined by CD61 (monocytes: ELP1 = 63.1 ± 17.1 %, ELP4 = 61.8 ± 16.8 %; granulocytes: ELP1 = 62.7 ± 17.0 %, ELP4 = 60.5 ± 20.1 %) were both decreased compared to uncoated MylarTM, while CD11b upregulation (monocytes: ELP1 = 18.7 ± 2.2 %, ELP4 = 19.7 ± 2.7 %; granulocytes: ELP = 21.4 ± 3.7 %, ELP4 = 22.0 ± 3.2 %) was increased. The statistical dependence of TF expression and platelet-monocyte aggregates was tested; however, no correlation was found. Overall, platelet-leukocyte aggregate formation was reduced and there were conflicting results with regards to the reduction of leukocyte activation for the ELP coatings on MylarTM. / Thesis (Master, Chemical Engineering) -- Queen's University, 2013-10-10 15:34:51.802

FibMat2.0 is a fibrinogen multilayer developed by AddBIO. Other proteins such as immunoglobulin G (IgG) and human serum albumin (HSA) can also be used to build multilayers with the same technique. The aim of this study of FibMat2.0 was to investigate if the manufacturing of the protein multilayer would induce an immune or haemostatic response in the body. The multilayers of IgG and HSA were also studied. Methods such as null ellipsometry, imaging of coagulation and the cone-and-plate setup were used to study immune reactions, activation of the coagulation cascade, and stability of the multilayers. Small amounts of plasma proteins were adsorbed to fibrinogen multilayers, but complement proteins adsorbed only to the IgG matrix and high molecular weight kininogen (HMWK) adsorbed only to the HSA monolayer. The imaging of coagulation method indicated that the titanium surface and the HSA monolayer activate surface induced coagulation rapidly, whereas fibrinogen and IgG multilayers demonstrated longer coagulation times. Platelets and a few white blood cells were bound to titanium surfaces and fibrinogen multilayers, but not to IgG multilayers or HSA monolayers. A conclusion in this study is that the surface of an implant can be coated with FibMat2.0 without any risks, but more studies are needed to better understand the interactions between the surfaces prepared in the present study and the immune and the haemostatic systems of the human body.

Aneurysms are a deadly asymptomatic cardiovascular disease that may occur especially where there are bends and bifurcations in the cerebral vasculature. A region where these features are especially prominent is the Circle of Willis (COW) in the brain, where aneurysms are known to occur. In the carotid artery, which feeds into the COW, the Reynolds number of blood flow is typically around 200-500. Even with such a low Reynolds number, turbulent-like flow, or tortuous flow, can occur due to bends, bifurcations and highly pulsatile flow which lower the effective Reynolds number where tortuous flow can occur. Highly pulsatile flow is unsteady flow that is high in magnitude and changes over time. Endothelial cells (ECs) line the inner wall of the blood vessel and experience the friction force of blood flow. This work is focused on designing a device that can expose ECs to forces they would undergo in an aneurysm-susceptible site. This is accomplished by exposing ECs to physiologically relevant Wall Shear Stress (WSS) and vibrations simultaneously. Vibrations in the body occur due to flow separation at the vessel wall, which leads to pressure changes. These pressure changes induce vibrations onto ECs. The fluid flow in the designed Parallel Plate Flow Chamber (PPFC) is laminar to induce a predictable WSS onto the cells, while the vibrations will induce a rapid cyclical force to simulate pressure fluctuations that may occur in vivo. The aneurysm-susceptible flow will simulate a more turbulent-like flow in the carotid artery; higher maximum WSS (around 2.2 Pa) with vibrations. The aneurysm-protective flow will have a lower WSS maximum (around 0.5 Pa). The PPFC, made of polycarbonate, is small and light enough to be conveniently vibrated using an electromagnetic vibration stage. The PPFC can be driven by a syringe or peristaltic pump, allowing for either steady or transient waveforms. The PPFC’s fluid domain will not change upon vibration, isolating the effect of vibration on the cells. Also, two side-by-side glass slide slots were included to allow for both protein and mRNA quantification from the same experiment, increasing experimental efficiency and flow-related consistency between the two cell areas. Simulations using ANSYS Fluent verified the flow field and WSS waveform on the cells for the designed geometry for 3D and 2D cases, as well as verified equal WSS values throughout all areas of ECs. Then, Particle Image Velocimetry (PIV) was done to verify the predicted flow rate in the machined PPFC given a steady flow rate driven by a syringe pump. Preliminary cell experiments were performed in an incubator under flow and vibration conditions to demonstrate cell survivability.

Entangled polymeric fluids subjected to shear display a stress plateau through a range of shear rates. The formation of this plateau is often attributed to an entanglement-disentanglement transition in scientific literature. However, to our best knowledge in situ studies recovering the intermediate scattering function of polymer dynamics under shear have until now never been performed. This thesis documents the successful development of a high viscosity shear device whose interaction with polarized neutrons is small enough to allow use for Neutron Spin Echo spectroscopy. Further, first measurements towards the direct observation of the variation of the degree of entanglement throughout increasing shear are documented, albeit yet for too short Fourier times to measure beyond Rouse dynamics.

Calcific aortic valve disease is highly prevalent, especially in the elderly. Currently, the exact mechanism of the calcification process is not completely understood, limiting our ability to prevent or cure the disease. Ex vivo investigations, however, have provided evidence that the aortic valve's biological response is sensitive to mechanical forces, including fluid shear stresses, leading to the hypothesis that adverse fluid shear stress environment play a role in leading to valve calcification. This thesis seeks to investigate this hypothesis. A method for performing experimental measurement of time-varying shear stress on aortic valve leaflets under physiologic flow conditions was first developed, based on the Laser Doppler Velocimetry technique, and was systematically validated. This method was then applied to both the aortic surface and the ventricular surface of a normal tricuspid the aortic valve, and then on a congenital bicuspid aortic valve, using suitable in vitro valve models and an in vitro pulsatile flow loop. It was found that in the tricuspid valve, the peak shear stress on the aortic surface under adult resting condition was approximately 15-19 dyn/cm². Aortic surface shear stresses were elevated during mid- to late-systole, with the development of the sinus vortex, and were low during all other instances. Aortic surface shear stresses were observed to increase with increasing stroke volume and with decreasing heart rate. On the ventricular surface, shear stresses had a systolic peak of approximately 64-71 dyn/cm² under adult resting conditions. During late systole, due to the Womersley effect, shear stresses were observed to reverse in direction to a substantial magnitude for a substantial period of time. Further, it was found that a moderately stenotic bicuspid aortic valve can experience excessive unsteadiness in shear stress experienced by its leaflets, most likely due to the turbulent forward flow resulting from the stenosis, and due to the skewed forward flow. To demonstrate that the measured shear stresses can have an effect on the aortic valve biology, ex vivo experiments were performed in specific to determine the effects of these various shear stress characteristics on the biological response of porcine aortic valve leaflets, using the cone and plate bioreactor. It was found that unsteady shear stress measured in the bicuspid valve resulted in increased calcium accumulation. Further, it was found that low shear stresses and high frequency shear stresses resulted in increased calcium accumulation. Thus, shear stress was found to affect aortic valve pathology, and low and unsteady fluid shear stresses can enhance pathology.