A method was developed to produce and incorporate a network of discrete, genipin-crosslinked gelatin microfibers around a pancreatic islet within a barium alginate microcapsule. This technique allows for the encapsulation of a porous fibrous matrix without the geometrical restrictions required for cellular aggregate seeding. Microfibers were produced from a novel vortex-drawn extrusion system with an alginate support matrix. Optimization culminated in a hydrated fiber diameter of 22.3 ± 0.4 μm, a 98% reduction in cross sectional area, while making the process more reliable and less labour intensive. The optimized microfibers were encapsulated at 40 vol% within 294 ± 4 μm 1.6% barium alginate microparticles by an electrostatic-mediated dropwise extrusion system. Pancreatic islets extracted from Sprague Dawley rats were encapsulated within the microparticles, and analyzed over a 21-day preliminary in vitro study. Acridine orange and propidium iodide fluorescent viability staining and light microscopy indicated a significant increase in viability for the fiber-laden particles relative to fiber-free control particles at days 7, 14, and 21. The fiber-laden system also reduced the incidence of disrupted islet cohesion from 31% to 8% at day 21, and showed evidence of islet-fiber adhesion. Preliminary investigations into insulin secretion and metabolic activity showed no significant difference between test and control groups. Further investigation into benefits of islet encapsulation within an extracellular matrix fiber network will be the subject of future studies with this body of work serving as a foundation. The system developed in this investigation could be developed into a modular scaffold system for tissue engineering beyond the field of islet research. / Thesis (Master, Chemical Engineering) -- Queen's University, 2011-08-18 15:05:50.917

In this thesis, the effectiveness of microparticle retention aid systems comprising of different cationic starches (tapioca and waxy maize), cationic flocculants (flocculant 1, linear with low charge density and flocculant 2, branched with medium charge density polyacrylamides) and anionic colloidal silica in improving retention and drainage of thermomechanical pulp (TMP) suspensions loaded with precipitated calcium carbonate (PCC) is studied. While starch is primarily added as dry strength agent in PCC-filled TMP suspensions, it also has a significant role in improving retention and drainage. Tapioca starch, which has both amylose and amylopectin, is found to be a better retention and drainage aid than waxy maize starch, which only contains amylopectin. In the absence of starch, both flocculants are ineffective in improving retention and drainage. With starch, both flocculant and silica are significant in enhancing retention and drainage further. Increasing the dosage of either flocculant or silica generally increases retention as well as drainage. Flocculant 2 is found to give slightly worse total and filler retention but better drainage compared to flocculant 1. The dosage of flocculant 2 is only half that of flocculant 1 though – for this reason overall flocculant 2 is deemed more effective in improving retention and drainage than flocculant 1. Split starch addition, where a portion of the starch is premixed with PCC and the rest added to the pulp, causes a slight decrease in both retention and drainage. Therefore, should the papermaker decide to use this approach, the starch should be split in a 25:75 ratio between PCC and the pulp to minimize the negative effects. Due to the cost benefits of using increased amounts of PCC, it is desirable to increase PCC content beyond the standard currently used (250 kg/t OD pulp for communications-grade paper). However, at very high dosages of PCC (500 kg/t OD pulp), the best combination identified in this work (tapioca starch, flocculant 2 and silica) is unable to maintain good retention and drainage. Increasing retention aid dosages may help retain more PCC, however this tactic can rapidly become uneconomical. A new approach is thus needed to achieve such highly-filled papers.

In this thesis, the effectiveness of microparticle retention aid systems comprising of different cationic starches (tapioca and waxy maize), cationic flocculants (flocculant 1, linear with low charge density and flocculant 2, branched with medium charge density polyacrylamides) and anionic colloidal silica in improving retention and drainage of thermomechanical pulp (TMP) suspensions loaded with precipitated calcium carbonate (PCC) is studied. While starch is primarily added as dry strength agent in PCC-filled TMP suspensions, it also has a significant role in improving retention and drainage. Tapioca starch, which has both amylose and amylopectin, is found to be a better retention and drainage aid than waxy maize starch, which only contains amylopectin. In the absence of starch, both flocculants are ineffective in improving retention and drainage. With starch, both flocculant and silica are significant in enhancing retention and drainage further. Increasing the dosage of either flocculant or silica generally increases retention as well as drainage. Flocculant 2 is found to give slightly worse total and filler retention but better drainage compared to flocculant 1. The dosage of flocculant 2 is only half that of flocculant 1 though – for this reason overall flocculant 2 is deemed more effective in improving retention and drainage than flocculant 1. Split starch addition, where a portion of the starch is premixed with PCC and the rest added to the pulp, causes a slight decrease in both retention and drainage. Therefore, should the papermaker decide to use this approach, the starch should be split in a 25:75 ratio between PCC and the pulp to minimize the negative effects. Due to the cost benefits of using increased amounts of PCC, it is desirable to increase PCC content beyond the standard currently used (250 kg/t OD pulp for communications-grade paper). However, at very high dosages of PCC (500 kg/t OD pulp), the best combination identified in this work (tapioca starch, flocculant 2 and silica) is unable to maintain good retention and drainage. Increasing retention aid dosages may help retain more PCC, however this tactic can rapidly become uneconomical. A new approach is thus needed to achieve such highly-filled papers. / Applied Science, Faculty of / Chemical and Biological Engineering, Department of / Graduate

In the papermaking industries spots in the finished product is a recurring problem. Billerud Korsnäs have in earlier studies identified poorly optimized retention systems as one of the reasons for these spots. Poorly optimized retention systems will allow for detrimental substances to flow freely in the system, which may cause agglomeration into larger particles that could end up as darker spots in the finished product. The aim of this thesis is to investigate a number of retention systems, consisting of a retention polymer and retention microparticles, and characterize the polymers.The retention aid systems task is to flocculate fibres, fines and fillers along with the colloidal material to improve process parameters. To investigate the retention systems three different retention polymers with different charge densities have been investigated alongside one microparticle. The parameters investigated in this thesis were the retention systems effect on drainage time, turbidity, charge demand and zeta potential. Pulp and white water from PM4 was used to imitate mill conditions.The results showed that the drainage time and turbidity was most effected by the retention aid systems. The polymer with the highest charge yielded the best results. Only minor effects could be detected on charge demand and Zeta potential.

Epithelial ovarian cancer is the fifth leading cause of cancer-related deaths in the United States. However, the mortality rate is relatively high, due to in part to the cancer being in an advance stage at diagnosis, since it is often asymptomatic at the early-stage with a ~94% of five-year survival rate if it is diagnosed at a localized stage (stage 1). Early detection of cancer would likely improve the survival rate. Scientists are searching for novel promising methods to detect ovarian cancer at an asymptomatic early stage; also, the method is cheap and user-friendly despite there are various techniques for ovarian cancer detection. Cancer antigen 125 (CA125), a type of serum biomarker that elevates ~50% of women with early-stage and ~80% of women with advanced-stage, is used mostly for screening epithelial ovarian cancer. However, the lack of sensitivity and specificity are known to be the main drawback of CA125. Finding new potential biomarkers that diagnose cancer at a localized stage will significantly reduce the mortality rate. Human epididymis protein 4 (HE4) is such a biomarker that has a higher sensitivity and specificity compared to all other known biomarkers, and recently it has been approved by food and drug administration (FDA) for clinical applications.In this project, we developed sandwich-type micro particles immunoassay for sensitive detection of HE4 biomarker in plasma. Here, we cross-link elemental particles to a specific functional group of the targeted biomolecules based on a covalent and non-covalent linking chemistry to improve the sensitivity and the selectivity of biomarker detection in which Fe3O4 and SiO2 microparticles were used to conjugate and purify the antibody-antigen in a media. The purified assay with the microparticles was analyzed with laser-induced breakdown spectroscopy (LIBS) for detection and quantization analysis of the HE4 biomarker. Furthermore, along with LIBS, Raman, Fourier transform infrared (FT-IR), and UV- Vis spectroscopic techniques were utilized to understand the conjugation dynamic and confirm the conjugation process.

Introduction: Cancer associated thrombosis is a well-recognized phenomenon that results in considerable patient morbidity and mortality. Malignancy conveys an increased risk for thrombosis and chemotherapy further elevates this risk. The pathophysiological mechanisms underlying this process remain poorly defined. Materials and Methods: A human acute monocytic leukemia cell line (THP-1) was treated with commonly used anthracycline chemotherapeutics at concentrations similar to those found in the plasma of cancer patients. Cells were analyzed for tissue factor (TF) mRNA, protein, and activity. Microparticle (MP) TF activity was also measured. Phosphatidylserine (PS) exposure on cells and MPs was analyzed by flow cytometry. PS levels on MPs was also evaluated in an annexin V capture assay. Results: Anthracycline treatment of THP-1 cells resulted in a concentration-dependent increase in cellular TF activity without a change in TF protein, which was associated with increased PS exposure on the cell surface and apoptosis. The increase in TF activity was abolished by annexin V or lactadherin indicating that PS exposure was required. Anthracycline treatment of THP-1 cells also increased the number of TF-positive MPs. Conclusion: Treatment of THP-1 cells with anthracyclines induces apoptosis and increases cellular TF activity. The increased activity required an increase in exposure of PS. Additionally, anthracyclines increase the release of TF-positive MPs from THP-1 cells. We propose that the increase in cellular TF activity in circulating leukemic cells, combined with increased numbers of TF-positive MPs, may contribute to thrombosis in cancer patients receiving chemotherapy.

Much is known about the atomic/molecular theories that govern adhesion as well as the macroscopic aspects and properties of adhesion. However, adhesion in the microparticle regime is poorly characterized. We report on experiments that use centrifugal force to remove polystyrene (PS) particles from the surface of a high-speed titanium rotor operated in vacuum. This unique rotor can apply forces far greater than other centrifugal force methods or related techniques using atomic force spectroscopy. The mode of attachment, whether particles were located on the windward side versus leeward side of the spinning rotor, time spent in vacuum prior to experiments, and surface imperfections all showed an effect on adhesion. Our observations show initial agreement with published results from atomic force spectroscopy experiments. We conclude that the liquid used to help suspend and apply the PS particles greatly influences the total adhesive forces present in the system. This in turn provides valuable clues as to the nature of the adhesive interaction.

No description available.

It is estimated that there are 5x1030 microorganisms on Earth and that approximately 50% live in unconsolidated sediment on the terrestrial subsurface. Subsurface disturbances caused by the constant search for natural resources and our dependence on groundwater make the abundance and diversity of these organisms a global concern. It is vital to many environmental fields, including bioremediation, water purification, and contaminant transport, that we understand how microorganisms and other colloidal particles attach to and detach from natural sediments and ultimately how they travel through porous media. Sticking efficiency (alpha) is a major component of most particle transport theories. It is defined as the ratio of particles that adhere to a collector surface compared to the total number of particles that collide with that surface. In this study, the Interaction Force Boundary Layer (IFBL) model was used to determine the sticking efficiencies of inorganic colloidal particles and Enterococcus faecalis cells against a silica glass collector surface. Sticking efficiencies were derived from intersurface potential energies that were determined from integrated force-distance data measured by Atomic Force Microscopy (AFM). Force data were measured in buffered aqueous solutions of varying pH and ionic strength to determine the influence of solution chemistry on particle removal from solution. Zeta-potentials were measured to determine the impact of particle and collector surface charge on force measurements. The results of this study indicate that alpha is strongly influenced by solution chemistry. The response of alpha to small changes in solution pH and ionic strength may be several orders of magnitude. Zeta-potential measurements imply that sticking efficiencies are strongly influenced by the electrical charges on both the particle and collector surfaces. Zeta-potentials of bacteria did not vary significantly with changing solution pH, but did respond to changing solution ionic strength. Historically, alpha has been very difficult to predict. This study is the first to report sticking efficiencies measured using AFM and the first to successfully apply the IFBL model to colloidal particles. Æ nThe incorporation of empirical nanoscale interactions into the measurement of alpha promises to more successfully describe particle adhesion and, thus, particle transport. / Ph. D.

Keratins are a family of proteins found within human hair, skin and nails, as well as a broad variety of animal tissue. Prior research suggests hydrogel constructs of keratin and keratin derivatives exhibit several mechanical and biological properties that support their use for tissue engineering and regenerative medicine applications. Microparticle formulations of these hydrogels are an intriguing delivery vehicle for drugs and cellular payloads for tissue engineering purposes due to the ability to exploit size, surface area, loading potential and importantly, non-invasive delivery (i.e. injection) of cells and biologics. Here we examine the water-in-oil emulsion synthesis procedure to produce keratin microparticles using an oxidized keratin derivative, keratose (KOS). Analyses of particle size, microstructure, and other characterization techniques were performed. Drug loading characteristics, release kinetics, and feasibility of use in two different microparticles was subsequently investigated, first using a model-drug and later testing an antibiotic payload on bacterial cultures to validate antibacterial applications. A suspension culture technique was developed to load bone marrow-derived mesenchymyal stromal cells (BM-MSCs), testing the capacity to maintain viability and express key protein-based factors in cell growth and development. Finally, we tested the in vitro effects of cell-loaded microparticles on the L6 skeletal muscle cell line to determine potentially beneficial outcomes for skeletal muscle tissue regeneration. Largely spherical particles with a porous internal structure were obtained, displaying hydrogel properties and forming viscoelastic gels with small differences between synthesis components (solvents, crosslinkers), generating tailorable properties. The uniquely fibrous microstructure of KOS particles may lend them to applications in rapid drug release or other payload delivery wherein a high level of biocompatibility is desired. Data showed an ability to inhibit bacterial growth in the emulsion-generated system, and thereby demonstrated the potential for a keratin-based microparticle construct to be used in wound healing applications. Dense cell populations were loaded onto particles. Particles maintained cell viability, even after freeze-thaw cycling, and provided a material substrate that supported cell attachment through the formation of focal adhesions. Finally, in vitro studies show that both KOS and BM-MSCs support varying aspects of skeletal muscle development, with combinatorial treatments of cell-loaded particles conferring the greatest growth responses. / Doctor of Philosophy / Keratins and keratin hydrogels may exhibit several properties that support their use for tissue engineering and regenerative medicine applications. Microparticle formulations of these hydrogels are an intriguing delivery vehicle for payloads for tissue engineering purposes. Here we examine the water-in-oil emulsion synthesis procedure to produce keratin microparticles that were analyzed based on drug loading characteristics. A suspension culture technique was developed to load bone marrow-derived mesenchymyal stromal cells (BM-MSCs). Finally, we tested these products to determine potentially beneficial outcomes for skeletal muscle tissue regeneration. Particles with a porous structure were obtained. The microstructure of these particles may lend them to applications in drug release or other payload delivery. Data showed an ability to load and unload specific drug payloads. Dense cell populations were loaded onto particles. Finally, studies show that both keratin and BM-MSCs support skeletal muscle development, with combinatorial treatments of cell-loaded particles conferring the greatest growth responses.