Browsing by Subject "Microparticles"
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Item Development and evaluation of enzymatically-degradable hydrogel microparticles for pulmonary delivery of nanoparticles and biologics(2012-12) Wanakule, Prinda 1985-; Roy, KrishnenduThe emerging class of biologic drugs, including proteins, peptides, and gene therapies, are widely administered by injection, despite potential systemic side effects. Rational design of targeted carriers that can be delivered non-invasively, with reduced side effects, is essential for the success of these therapies, as well as for the improvement of patient compliance and quality of life. One potential approach is to take advantage of specific physiological cues, such as enzymes, which would trigger drug release from a drug carrier. Enzymatic cleavage is highly specific and could be tailored for certain diseased tissues where specific enzymes are up regulated. Enzymatically-degradable hydrogels, which incorporate an enzyme- cleavable peptide into the network structure, have been extensively reported for releasing drugs for tissue engineering applications. These studies showed that a rapid response and corresponding drug release occurs upon enzyme exposure, whereas minimal degradation occurs without enzyme. Recently, Michael addition reactions have been developed for the synthesis of such enzymatically-degradable hydrogels. Michael addition reactions occur under mild physiological conditions, making them ideally suited for polymerizing hydrogels with encapsulated biologic drugs without affecting its bioactivity, as in traditional polymerization and particle synthesis. The focus of my research was to create enzymatically-degradable hydrogel microparticles, using Michael addition chemistry, to evaluate for use as an inhalable, disease-responsive delivery system for biologic drugs and nanoparticles. In this dissertation, I utilize bioconjugation and Michael addition chemistries in the design and development of enzymatically-degradable hydrogels, which may be tailored to a multitude of disease applications. I then introduce a new method of hydrogel microparticle, or microgel, synthesis known as the Michael Addition During Emulsion (MADE) method. These microgel carriers were evaluated in vitro, and found to exhibit triggered release of encapsulated biologic drugs in response to enzyme, no significant cytotoxic effects, and the ability the avoid rapid clearance by macrophages. Lastly, in vivo studies in mice were conducted, and microgels were found to exhibit successful delivery to the deep lung, as well as prolonged pulmonary retention after intratracheal aerosol delivery. In conclusion, a new class of enzymatically-degradable microgels were successfully developed and characterized as a versatile and promising new system for pulmonary, disease-responsive delivery of biologic drugs.Item Glycomic insights into microvesicle biogenesis(2011-08) Batista, Bianca Stella; Stein, David S.; Mahal, Lara K.; Iyer, Vishwanath R.; Sullivan, Christopher S.; Liu, Hung-WenCells can mediate intercellular communication by the secretion and uptake of microvesicles, nano-sized membranous particles that carry signaling molecules, antigens, lipids, mRNA and miRNA between cells. The biological function of these vesicles is dependent upon their composition and cellular origin which is regulated by mechanisms that are not well understood. Based on their molecular content, microvesicles may play a role in immune regulation, cancer progression, the spread of infectious agents and numerous other important normal and pathogenic processes. The proteomic content of microvesicles from diverse sources has been intensely studied. In contrast, little is known about their glycomic content. The glycosylation pattern of a protein or lipid plays a key role in determining its functional properties in several ways. Glycans can determine the trafficking of a protein to particular regions of the cell as well as the protein’s half life. In addition, the glycan-dervied oligomerization of glycolipids and glycoproteins is a known mechanism for the activation of receptors and recognition of ligands on the surface of the cell. Glycomic analysis may thus provide valuable insights into microvesicle function. I utilized lectin microarray technology to compare the glycosylation patterns of microvesicles derived from a variety of biological sources. When compared to cellular membranes, microvesicles were enriched in high mannose, polylactosamine, α2-6 sialic acid, and complex N-linked glycans but exclude terminal blood group A and B antigens. The polylactosamine signature in microvesicles from different cell lines derives from distinct glycoprotein cohorts. After treatment of Sk-Mel-5 cells with lactose to inhibit lectin-glycan interactions, secretion of microvesicle resident proteins was severely reduced. Taken together, this work provides evidence for a role of glycosylation in microvesicle-directed protein sorting.Item High Resolution Study of Micro-Meter Particle Detachment and Resuspension on Different Surfaces(2012-08-16) Kassab, Asmaa 1983-In an effort to understand the resuspension phenomena, interactions of spherical micro-meter particles (glass beads (GB) and Stainless steel (SS)) were investigated experimentally on different surfaces (glass, ceramic, hardwood, metal and chemical agent resistant coated metal (CARC)). Particles were deposited on the lower surface of a 10 cm square wind tunnel by gravitational settling. Air flows were imposed from an open entrance at average velocities up to 16 m/s. Individual particle trajectories obtained by high-speed imaging reveal three different types of motion: rolling/bouncing, immediate liftoff and complex motion. Surface roughness significantly affects the particle initial motion prior to liftoff. The majority of particle trajectories from the glass substrate were parallel to the surface with complex motion, covering 25% of the total distance traveled in rolling/bouncing motion before liftoff. Hardwood substrates took the longest time for initial particle movement (t >1 s) causing a more rapid liftoff. The ceramic substrate showed the most rolling/bouncing motion, for 80% of the particles. Additionally, single layer detachment showed that the detachment percentage initially follow an exponentially increasing trend for a period of ~ 1 s, followed by a plateau phase for a period of 5 s. Changing velocity, substrate and particle size significantly affects GB particle detachment. Furthermore, detachment from the metal substrate was consistently higher than the CARC substrates. However, particle density is not a significant difference in the bigger particle size studied. Initial 3-D particle tracking showed that particles seem to travel in a constant angle to the left rather than going straight in the flow direction. A detachment mode model showed that the detachment by direct liftoff required a much higher speed than rolling motion with a minimum of 14 m/s for both GB70 and SS70 on glass and metal surface, and the velocity increased to 21 m/s for the smaller particle. Incorporating the different types of particle motion prior to liftoff into resuspension models, and how their relative contributions change with different particle and substrate materials, can potentially yield improved predictive capabilities.Item Intelligent delivery via enzyme active hydrogels(2009-12) Marek, Stephen Richard; Peppas, Nicholas A., 1948-Advances in medical treatment are leading away from generalized care towards intelligent systems or devices which can sense and respond to their environment. With these devices, the burden of monitoring and dosing for treatment can be removed from the doctor (or the patient) and be placed on the device itself. Implicit closed-loop control systems will allow the device to respond to its environment and release therapeutic agent in response to a specific stimulus. Environmentally responsive hydrogels show great promise in being incorporated in such an intelligent device, such as pH-responsive hydrogels which can swell and deswell in response to changes in the pH of the media. Thus, pH changes can be exploited for controlled and intelligent drug delivery when used in combination with these pH-responsive hydrogels. In this work, heterogeneous, thermal-redox initiated free-radical polymerizations were developed to synthesize novel pH-responsive hydrogels, microparticles, and nanogels. The specific disease of interest was type I diabetes, which requires daily doses of insulin both at a basal amount and either a postprandial or preprandial bolus in order to maintain blood glucose levels within safe limits. To allow pH-responsive hydrogels to be sensitive to glucose, glucose oxidase was incorporated which oxidizes glucose to gluconic acid. A novel inverse-emulsion polymerization method was developed for the synthesis of poly[2-(diethylaminoethyl methacrylate)-grafted-polyethylene glycol monoethyl ether monomethacrylate] (P(DEAEM-g-PEGMMA)) nanogels (100-400 nm) for intelligent insulin delivery. The new polymerization method allowed the incorporation of hydrophilic components, such as glucose oxidase and catalase, as well as PEG surface tethers of lengths 400 Da up to 2000 Da. Surface tethers successfully decreased the surface charge of the nanogels. Insulin loading and release was determined for microparticles which were able to imbibe substantial amounts of insulin from solution when swollen, entrap the insulin when collapsed, and then release the insulin in response to either a pH or glucose stimulus.