<div><br></div><div> Today, autoinjectors are developed for more viscous drug solutions, which require larger forces for actuating the syringe and impose larger stresses on the drug solution during the administration of autoinjectors. We developed experimentally validated high-fidelity simulations to investigate the liquid jet formation, liquid slosh and cavitation during the insertion process of an autoinjector. </div><div> </div><div> The jet formed due to an acceleration-deceleration motion of syringe is found to be governed by the interplay between inertial, viscous, surface tension and gravitational forces. A scaling for the jet velocity and a criterion for the jet breakup in a simplified geometry are proposed.</div><div> </div><div> When the syringe accelerates and decelerates during the insertion, liquid slosh occurs and there is a vehement motion of the air-liquid interface. Here, we quantified the area of air-liquid interface and hydrodynamic strain rate, which increase with the air gap size, syringe velocity, tilt angle and inner wall hydrophobicity, and decrease with the solution viscosity and hardly change with the liquid column height and surface tension. The strain rate is not sufficient to unfold the protein and the air-liquid interface is more likely to cause protein aggregation.</div><div> </div><div> In a spring-driven autoinjector, the plunger is actuated by the impact of a driving rod, which generates a strong pressure wave and can cause cavitation inception. The cavtiation bubbles can be impeded by the syringe walls and form a re-entrant jet shooting toward the syringe wall. During the process, the protein molecules are focused in the jet, pushed toward the syringe wall and spread across the wall, which can be the reason for the protein aggregation and adsorption on the syringe walls. The impedance effects of the wall decreases with the wall distance and increases with the maximum bubble size. The maximum bubble radius also increases with the liquid column size and nucleus size and decreases with the air gap pressure. Since inertia effects dominate in the cavitation process, the liquid viscosity and surface tension hardly changes the cavitation bubble dynamics. Small bubbles can also form in the bulk, which may generate aggregates in the bulk solution. Bubbles in the cavitation bubble cloud may coalesce with nearby bubbles and induce a higher pressure at the collapse (up to 1000 bar). This high pressure can potentially generate hydroxyl radicals that oxidize the protein molecules.</div><div> </div><div> The current study presents a detailed picture of fluid flows in autoinjectors and provide recommendations for mitigating the liquid slosh and cavitation generated in syringes. The results can be combined with future experiments to understand the implications of fluid flows on protein drugs and the performance of autoinjectors.</div>

Partitioning of poly(ethyleneglycol) (PEG) molecules in 2-D and 3-D systems is presented as a self-assembly approach for controlling non-specific adsorption of proteins at interfaces. Lateral restructuring of multi-component Langmuir monolayers to accommodate adsorbing proteins was investigated as a model 2-D system. Ferritin adsorption to monolayers containing cationic, nonionic, and PEG bearing phospholipids induced protein sized binding pockets surrounded by PEG rich regions. The number, size, and distribution of protein imprint sites were controlled by the molar ratios, miscibility, and lateral mobility of the lipids. The influence of PEG chain length on the ternary monolayer restructuring and protein distribution was also investigated using DSPE-PEGx (x= 7, 16, 22). Monolayer miscibility analysis demonstrated that longer PEG chains diminished the condensed phase formation for a fixed ratio of lipids. Thus, incorporation of longer PEG chains, intended to diminish protein adsorption outside of the imprint sites of cationic / non-ionic lipids, leads to dramatic changes in monolayer phase behavior and protein distribution in this 2-D system. The assembly of PEG-amphiphiles at elastomer surfaces and subsequent protein adsorption was investigated as a model 3-D system. Polydimethylsiloxane (PDMS) substrates were modified with block copolymers comprised of PEG and PDMS segments by two methods: (1) the block copolymer was mixed with PDMS during polymerization; (2) the block copolymer diffused into solvent swollen PDMS monoliths. Hydrophilic surfaces resulted for both approaches that, for 600 D block copolymer, exhibited up to 85% reduction in fibrinogen adsorption as compared to native PDMS. Higher MW block copolymers (up to 3000 D) resulted in less hydrophilic surfaces and greater protein adsorption, presumably due to diffusion limitations of copolymer in the PDMS monolith. All modified PDMS surfaces were dynamic and restructured when cycled between air and water. PDMS transparency also decreased with increase in block copolymer concentration for both methods, limiting this modification protocol for applications requiring high polymer transparency. The 2-D system presents a bottom-up approach, where adsorbing protein constructs the binding site, while the 3-D system presents a top down approach, where protein-binding elements may be introduced into the PEG-bearing polymer for fabrication of surfaces with controlled protein adsorption.

<p>  </p> <p>The potential of lignocellulose for producing fermentable sugars as feedstock to manufacture fuels, chemicals, and materials for decarbonization remains untapped due to costly logistics and conversion processes. Pelleting technology offers a solution by addressing logistical issues and impacting downstream conversion, though it comes with its own costs. An overview of recent advances in pelleting technologies and their impact on bioconversion highlights the importance of understanding variables and product attributes. The interplay between pelleting and pretreatment processes, considering various feedstocks, is crucial for future design. Practical considerations such as energy consumption, costs, and environmental impacts must not be overlooked, along with exploration of cutting-edge technologies and strategies in this field. This work further presents a comprehensive investigation into the recalcitrance of pelleted corn stover to enzymatic digestion prior to any pretreatment.</p> <p>The potential of lignocellulose for producing fermentable sugars as feedstock to manufacture fuels, chemicals, and materials for decarbonization remains untapped due to costly logistics and conversion processes. Pelleting technology offers a solution by addressing logistical issues and impacting downstream conversion, though it comes with its own costs. An overview of recent advances in pelleting technologies and their impact on bioconversion highlights the importance of understanding variables and product attributes. The interplay between pelleting and pretreatment processes, considering various feedstocks, is crucial for future design. Practical considerations such as energy consumption, costs, and environmental impacts must not be overlooked, along with exploration of cutting-edge technologies and strategies in this field. </p> <p>This dissertation further presents a comprehensive investigation into the recalcitrance of pelleted corn stover to enzymatic digestion prior to any pretreatment. One aspect focuses on the role of high moisture pelleting in enhancing the enzymatic digestibility of corn stover before pretreatment, along with the relevant substrate characteristics. The pelleting process increases the digestibility of unpretreated corn stover, resulting in a glucan conversion increase from 8.2% to 15.5% at a 5% solid loading using 1 FPU Cellic® CTec2 per gram of solids. Under the same enzymatic hydrolysis conditions, the conversion of glucan remains higher for pelleted corn stover compared to its non-pelleted counterpart, even though both samples underwent identical milling processes and passed through the same screen to minimize particle impact. Compositional analysis reveals that loose and pelleted corn stover have similar non-dissolvable compositions, albeit with differences in their extractives. Using microcrystalline cellulose (Avicel) as a substrate, the presence of corn stover extractives results in a lower sugar yield compared to using citrate buffer instead, particularly for extractives from pelleted corn stover. This indicates a more negative impact of pelleted corn stover extractives on the activity of employed enzyme, CTec2. However, pelleted corn stover still shows increased overall glucan conversion compared to loose corn stover, suggesting improved digestibility of non-dissolvable components after milling and washing. The improvement in digestibility of pelleted material can be attributed to factors such as reduced particle size, enhanced substrate accessibility, and hydrolysis of cross-linking structures induced by the pelleting process. These findings offer valuable insights for the development of processing strategies aimed at sustainable and efficient utilization of lignocellulose.</p> <p>Furthermore, this dissertation delves into the profound impact of extractives on enzymatic hydrolysis, prompting a thorough examination of the composition and characteristics of extractives derived from pelleted corn stover, as well as their effects on enzymatic conversion. In contrast to previous reports, it is discovered that water extractable materials actually enhance the enzymatic hydrolysis of extractive-free stover, while the enzyme activities diminish when using microcrystalline cellulose as a substrate. This divergent behavior of extractives is attributed to the presence of lignin, which may interact with inhibitory compounds like phenolics, thereby mitigating the detrimental effects of soluble inhibitors or insoluble lignin, or both. These findings significantly advance our fundamental understanding of the intrinsic behavior of extractives and contribute to the optimization of schemes for efficient and cost-competitive enzymatic conversion of lignocellulose. </p>

No description available.

The adsorption of cytochrome c onto mesoporous silica (MCM-41) was investigated in this study. MCM-41 was synthesized and characterized by different methods. The pore size of MCM-41 was calculated from each method and all were in agreement with each other. Result from SAXS method showed a well ordered 2D hexagonal structure of MCM- 41. To investigate the effect of pH on adsorption process, different buffers were used with various pH in the range from 3.8 to 10.7. It was observed that the maximum adsorption occurs at pH near the isoelectric point of cytochrome c. The surface charges of cytochrome c and MCM-41 play an essential role for the process of adsorption. Desorption of cytochrome c from MCM-41 was investigated as well. Pure water and buffers with pH 7.1 and 10.7 were used to study the desorption. The result shows that desorption of cytochrome c from MCM-41 takes place at a pH above its isoelectric point.

Integration of living cells with novel microdevices requires the development of innovative technologies for manipulating cells. Chemical surface patterning has been proven as an effective method to control the attachment and growth of diverse cell populations. Patterning polyelectrolyte multilayers through the combination of layer-by-layer self-assembly technique and photolithography offers a simple, versatile and silicon compatible approach that overcomes chemical surface patterning limitations, such as short-term stability and low protein adsorption resistance. In this study, direct photolithographic patterning of PAA/PAAm and PAA/PAH polyelectrolyte multilayers was developed to pattern mammalian neuronal, skeletal and cardiac muscle cells. For all studied cell types, PAA/PAAm multilayers behaved as a negative surface, completely preventing cell attachment. In contrast, PAA/PAH multilayers have shown a cell-selective behavior, promoting the attachment and growth of neuronal cells (embryonic rat hippocampal and NG108-15 cells) to a greater extent, while providing a little attachment for neonatal rat cardiac and skeletal muscle cells (C2C12 cell line). PAA/PAAm multilayer cellular patterns have also shown a remarkable protein adsorption resistance. Protein adsorption protocols commonly used for surface treatment in cell culture did not compromise the cell attachment inhibiting feature of the PAA/PAAm multilayer patterns. The combination of polyelectrolyte multilayer patterns with different adsorbed proteins could expand the applicability of this technology to cell types that require specific proteins either on the surface or in the medium for attachment or differentiation, and could not be patterned using the traditional methods.

No description available.

Mucous membranes are covered with mucus, a viscoelastic hydrogel that plays an essential role in their protection from shear and pathogens. The viscoelasticity of mucus is owing to mucins, a group of densely glycosylated proteins. Mucins can interact with a wide range of surfaces; thus, there is big interest in exploring and manipulating such interactions for biomedical applications. This thesis presents investigations of mucin interactions with hydrophobic surfaces in order to identify the key features of mucin lubricity, as well as describes the development of materials that are optimized to interact with mucins.   In Paper I we investigated the domains which make mucins outstanding boundary lubricants. The results showed that the hydrophobic terminal domains of mucins play a crucial role in the adsorption and lubrication on hydrophobic surfaces. Specifically, protease digestion of porcine gastric mucins and salivary mucins resulted in the cleavage of these domains and the loss of lubricity and surface adsorption. However, a “rescue” strategy was successfully carried out by grafting hydrophobic phenyl groups to the digested mucins and enhancing their lubricity. This strategy also enhanced the lubricity of polymers which are otherwise bad lubricants.   In Paper II we developed mucoadhesive materials based on genetically engineered partial spider silk proteins. The partial spider silk protein 4RepCT was successfully functionalized with six lysines (pLys-4RepCT), or the Human Galectin-3 Carbohydrate Recognition Domain (hGal3-4RepCT). These strategies were aiming to either non-specific electrostatic interactions between the positive lysines and the negative mucins, or specific binding between the hGal3 and the mucin glycans. Coatings, fibers, meshes and foams were prepared from the new silk proteins, and the adsorption of porcine gastric mucins and bovine submaxillary mucins was measured, demonstrating enhanced adsorption.   The work presented demonstrates how mucin-material interactions can provide us with valuable information for the development of new biomaterials. Specifically, mucin-based and mucin-inspired lubricants could provide desired lubrication to a wide range of surfaces, while our new silk based materials could be valuable tools for the development of mucosal dressings. / Slemhinnor täckts av slem, en viskoelastisk hydrogel som spelar en viktig roll för att skydda mot mekanisk nötning och patogener. Muciner, en grupp av tätt glykosylerade proteiner, spelar en viktig roll i viskoelasticiteten av slem. Eftersom muciner kan interagera med diverse ytor är det av stort intresse att utforska och manipulera sådana interaktioner för biomedicinska tillämpningar. Denna avhandling presenterar undersökningar av mucininteraktioner med hydrofoba ytor för att identifiera de viktigaste egenskaperna hos mucinsmörjning, samt beskriver utveckling av material som optimerades för att interagera med muciner.   I Artikel I undersökte vi de domäner som bidrar till  mucinernas enastående kapacitet som smörjmedel. Resultaten visade att mucinernas hydrofoba terminaldomäner spelar en avgörande roll vid adsorption och smörjning på hydrofoba ytor. Mer specifikt, proteasklyvning av svinmagemuciner och salivmuciner resulterade i klyvningen av dessa domäner och förlust av smörjning och ytadsorption. Genom att länka hydrofobiska fenylgrupper till de uppbrutna mucinerna, lyckades deras smörjningsegenskaper förbättras. Denna strategi förbättrade också smörjningsegenskaper hos andra polymerer som annars har  dåliga smörjningsegenskaper.   I Artikel II utvecklade vi mukoadhesiva material baserade på genetiskt modifierade partiella spindelsilkeproteiner. Spindelsilkeproteinet 4RepCT funktionaliserades framgångsrikt med tillsats av sex lysiner (pLys-4RepCT), eller den mänskliga Galectin-3 karbohydrat igenkänningsdomänen (hGal3-4RepCT). Syftet med dessa strategier var antingen att öka ospecifika elektrostatiska interaktioner mellan de positiva lysinerna och de negativa mucinerna, eller den specifika bindningen mellan hGal3 och mucin-glykanerna. Beläggningar, fibrer, nät och skum framställdes från de nya silkeproteinerna. Efter att adsorption av svinmagsmuciner och bovina submaxillära muciner uppmätts, visade de nya silkeproteinerna förbättrad mucin adsorption.   Detta arbete visar hur interaktioner mellan mucin-material kan bidra med värdefull information för utvecklingen av nya biomaterial. Mucinbaserade och mucininspirerade smörjmedel kan ge önskad smörjning till ett brett spektrum av ytor, medan vår nya silkesbaserad material kan vara ett värdefullt verktyg för utvecklingen av slemhinneförband. / <p>QC 20190412</p>

<p>I put department up there as Biomedical Engineering. The full title should be: School of Biomedical Engineering.</p> / <p>The work presented in this thesis was focused on the surface modification of biomaterials with combinations of polyethylene glycol (PEG) and bioactive molecules (protein anticoagulants) for improved blood compatibility. Since the fate of biomaterials in contact with blood depends significantly on plasma protein-surface interactions, the objective of this work was to reduce non-specific protein adsorption using PEG and to promote specific protein interactions that could inhibit clot formation using protein anticoagulants as modifiers.</p> <p>Two anticoagulant molecules were used in this work: hirudin, a specific inhibitor of thrombin and corn trypsin inhibitor (CTI), a specific inhibitor of clotting factor XIIa. Gold, used as a model substrate, was modified with PEG and anticoagulant molecules using two methods referred to as sequential and direct. In the sequential method PEG was first immobilized on the surface and then the bioactive molecule was attached (conjugated) to the PEG. In the direct method, a PEG-bioactive molecule conjugate was first formed and then immobilized on the surface. Surfaces were characterized by contact angle, ellipsometry and x-ray photoelectron spectroscopy (XPS). Uptake of the bioactive molecules was measured by radiolabeling. Biointeraction studies included plasma protein adsorption, bioactivity assays using chromogenic substrates and clotting time assays. For PEG-hirudin and PEG-CTI surfaces (both direct and sequential) the protein resistance was similar to that of the PEG-alone surfaces. Despite having a lower density of bioactive molecule (both hirudin and CTI), the sequential surfaces showed superior bioactivity compared to the direct ones.</p> <p>To determine the optimal ratio of free PEG and bioactive molecule-PEG conjugate on the surface (best combination of protein resistance and bioactivity), PEG-CTI was immobilized on gold substrate with varying ratio of conjugated to free PEG using both direct and sequential methods. As the ratio increased, protein resistance was maintained while specific interactions (bioactivity) increased. The optimal composition appeared to be where all PEG molecules are conjugated to a CTI molecule.</p> <p>In the final part of this project, PEG and CTI were immobilized on polyurethane as a material with applicability to medical device construction. A sequential method was developed for this substrate. Comparison of the PEG-CTI surface with PEG only or CTI only surfaces indicated that the combination of PEG-CTI was effective both in reducing non-specific protein adsorption and promoting the specific interactions of CTI with its target plasma protein, factor XIIa. In fact, the presence of PEG improved CTI interactions with FXIIa compared with CTI only surfaces. Thus, sequential attachment of PEG and CTI may be effective for modifying polyurethane surfaces used in blood-contacting medical devices.</p> / Doctor of Philosophy (PhD)

<p>Medical devices used for diagnosis and treatment often involve the exposure of the patient’s blood to biomaterials that are foreign to the body, and blood-material contact may trigger coagulation and lead to thrombotic complications. Therefore, the risk of thrombosis and the issue of blood compatibility are limitations in the development of biomaterials for blood-contacting applications. The objective of this research was to develop a dual strategy for surface modification of polydimethylsiloxane (PDMS) to prevent thrombosis by (1) grafting polyethylene glycol (PEG) to inhibit non-specific protein adsorption, and (2) covalently attaching an antithrombin-heparin (ATH) covalent complex to the distal end of the PEG chains to inhibit coagulation at the surface.</p> <p>Surface characterization via contact angle measurements confirmed reductions in hydrophobicity for the modified surfaces and x-ray photoelectron spectroscopy (XPS) indicated that heparin and ATH were present. The predisposition of PDMS to induce blood coagulation was investigated, and advantages of ATH over heparin in inhibiting coagulation on PDMS were demonstrated. Studies of protein interactions using radiolabelling and Western blotting demonstrated the ability of PEG-modified surfaces to resist non-specific protein adsorption, and the ability of ATH- and heparin-modified surfaces to specifically bind AT present in plasma, thereby providing anticoagulant activity. Through specific interactions with the pentasaccharide sequence on the heparin moiety, the ATH-modified surfaces bound AT more efficiently than the heparin-modified surfaces. Thromboelastography (TEG) was used to evaluate further the anticoagulant potential of the ATH-modified surfaces. It was found that coagulation occurred at a slower rate on the ATH-modified surfaces compared to unmodified PDMS, and the resulting clot was mechanically weaker. By creating a surface with bioinert and bioactive properties, non-specific protein adsorption was reduced and anticoagulation at the surface through specific protein binding was promoted. This dual PEO/ATH modification strategy may therefore offer an improved approach for the minimization of thrombosis on PDMS and biomaterial surfaces more generally.</p> / Master of Applied Science (MASc)