Hydrogels have attracted interest as biomaterials due to their similarity to native tissue and extracellular matrix as well as their versatility and tunability. Each of these characteristics allows hydrogels to be used in a wide variety of biomedical applications including drug delivery, tissue engineering, and regenerative medicine. Poly(oligoethylene glycol methacrylate) (POEGMA) has been shown to possess attractive biological and thermoresponsive properties, serving as an alternative to both poly(ethylene glycol) (PEG) and poly(N-isopropylacrylamide) (PNIPAM) depending on the number of ethylene oxide repeat units in the POEGMA side chain. Our group has shown the versatility of POEGMA and has successfully developed hydrazide- and aldehyde-functionalized polymer precursors that form an injectable in situ gelling hydrogel. By engineering the precursor polymer structure and crosslinking density (i.e. number of reactive functional groups in the precursor polymers), the properties of these hydrogels can be tuned. Herein, a hyperbranched structure was incorporated into POEGMA precursors to control the physical and biological properties of hydrogels independent of the chemistry while maintaining gel injectability. By varying the degree of branching (DoB) in these precursors, it was possible to tune the hydrogel properties based on reacting combinations of hyperbranched-linear and hyperbranched-hyperbranched precursor polymers. While it was feasible to tune the mechanical properties of the hyperbranched hydrogels based on the DoB, the hyperbranched-hyperbranched system showed diminished mechanical strength when compared to the hyperbranched-linear system. Overall, the mechanical properties of the whole hydrogel series were comparable to previously reported linear POEGMA hydrogels. In terms of swelling and degradation kinetics, the swelling and degradation rate in both acid-catalyzed conditions and in phosphate-buffered saline (PBS) at physiological temperature (37°C) correlated with DoB and polymer size. The precursor polymers showed minimal cytotoxicity in the presence of 3T3 mouse fibroblasts. Lastly, each of the hyperbranched hydrogels adsorbed higher quantities of protein compared to PEG-based hydrogels, but still relatively low amounts compared to other polymeric biomaterials. We have shown that it is possible to significantly tune the physicochemical properties by slightly changing the polymer precursor chemistry, namely by varying the amount of crosslinker and, thus, the degree of branching in the polymer network. Therefore, hyperbranched POEGMA offers a versatile platform to create tunable hydrogels based on polymer precursor structure for biomedical applications. / Thesis / Master of Applied Science (MASc)

Water-soluble fullerene containing polymers comprising of poly(2-(dimethylamino) ethyl methacrylate)-fullerene (PDMAEMA-C60) with targeting moieties, poly(oligo(ethylene glycol) methyl ether methacrylate)-C60 (POEGMA-C60), nanocrystalline cellulose-fullerene (NCC-C60) and NCC-C60-POEGMA were synthesized and their solution properties were investigated. PDMAEMA-C60 with galactose targeting moiety was prepared by atom transfer radical polymerization (ATRP) and atom transfer radical addition (ATRA) processes. The self-assembly of galactose functionalized PDMAEMA-C60 structure in aqueous solutions was investigated using dynamic light scattering (DLS) at different pHs. A smaller hydrodynamic radius (Rh) was observed at pH 10 than at pH 3 due to electrostatic repulsion at low pH values. In addition, free PDMAEMA chains induced the demicellization of self-assembled nanostructures caused by the formation of charge transfer complex between PDMAEMA and C60. A well-defined poly(di(ethylene glycol) methyl ether methacrylate–stat-oligo(ethylene glycol) methyl ether methacrylate)-block-poly(di(ethylene glycol) methyl ether methacrylate ((PMEO2MA-stat-POEGMA300)-b-PMEO2) was successfully synthesized at room temperature via a two-step ATRP process. The block copolymer exhibited two thermal transitions at ~ 30 and 45 oC, which was believed to be associated with the formation of micelles and larger aggregates. The Rh of the aggregates increased from 47 to 90 nm, the aggregation number increased from 76 to ~9800 and Rg/Rh increased from 0.75 to 1.2 within the temperature range of 34 to 45oC. Well-defined statistical (PMEO2MA-stat-POEGMA300)-C60 was synthesized via ATRP and ATRA. The lower critical solution temperature (LCST) of (PMEO2MA-stat-POEGMA300)-C60 increased with methanol content in water, exhibiting lower LCSTs than PMEO2MA-stat-POEGMA300 for all methanol/water compositions. Higher critical micelle concentration (CMC) and larger spherical micelles were observed for (PMEO2MA-stat-POEGMA300)-C60 with increasing methanol content. The Rh of the micelles remained constant at temperature below the LCST and increased dramatically at temperature greater than the LCST, and (Rg/Rh) increased from ~ 0.75 to ~ 1.0. Nanocrystalline cellulose (NCC) was modified with water-soluble C60-(β-cyclodextrin) and (PMEO2MA-stat-POEGMA300)-C60) through a radical coupling reaction. NCC-C60-(PMEO2MA-stat-POEGMA300) possessed thermal responsive behavior in water and ~3.5 oC hysteresis associated with the heating/cooling cycles. No observable damage to NCC occurred during the radical coupling reaction as determined by TEM. NCC-C60-(β-cyclodextrin) possessed a similar thermal degradation behavior as NCC except it possessed a broader temperature range. Both NCC-fullerene systems demonstrated a radical scavenging activity when screened with the 2,2-diphenyl-1-picrylhydrazyl (DPPH). In addition, the drug loading and delivery using PDMAEMA-C60 with targeting moieties was explored. Two model drugs, namely fluorescein and pyrene were employed to evaluate the location of drug in the self-assembled structure of PDMAEMA-C60. It was found that the hydrophobic drugs were partitioned between the PDMAEMA shells and the hydrophobic fullerene cores. The drug delivery profiles indicated that PDMAEMA-C60 is an efficient drug carrier, however, it was cytotoxic to cells. The gene transfection efficacy of PDMAEMA-C60 to different cell lines was investigated and the results demonstrated that PDMAEMA-C60 exhibited good gene transfection performance. However, the targeting selectivity to liver cells cannot be determined in both cases. This study demonstrates that nanostructures of stimuli-responsive fullerene polymers can be controlled and manipulated by changing the external environments. Several potential applications, such as in drug and gene delivery, and free radical scavenging can be further explored.

Water-soluble fullerene containing polymers comprising of poly(2-(dimethylamino) ethyl methacrylate)-fullerene (PDMAEMA-C60) with targeting moieties, poly(oligo(ethylene glycol) methyl ether methacrylate)-C60 (POEGMA-C60), nanocrystalline cellulose-fullerene (NCC-C60) and NCC-C60-POEGMA were synthesized and their solution properties were investigated. PDMAEMA-C60 with galactose targeting moiety was prepared by atom transfer radical polymerization (ATRP) and atom transfer radical addition (ATRA) processes. The self-assembly of galactose functionalized PDMAEMA-C60 structure in aqueous solutions was investigated using dynamic light scattering (DLS) at different pHs. A smaller hydrodynamic radius (Rh) was observed at pH 10 than at pH 3 due to electrostatic repulsion at low pH values. In addition, free PDMAEMA chains induced the demicellization of self-assembled nanostructures caused by the formation of charge transfer complex between PDMAEMA and C60. A well-defined poly(di(ethylene glycol) methyl ether methacrylate–stat-oligo(ethylene glycol) methyl ether methacrylate)-block-poly(di(ethylene glycol) methyl ether methacrylate ((PMEO2MA-stat-POEGMA300)-b-PMEO2) was successfully synthesized at room temperature via a two-step ATRP process. The block copolymer exhibited two thermal transitions at ~ 30 and 45 oC, which was believed to be associated with the formation of micelles and larger aggregates. The Rh of the aggregates increased from 47 to 90 nm, the aggregation number increased from 76 to ~9800 and Rg/Rh increased from 0.75 to 1.2 within the temperature range of 34 to 45oC. Well-defined statistical (PMEO2MA-stat-POEGMA300)-C60 was synthesized via ATRP and ATRA. The lower critical solution temperature (LCST) of (PMEO2MA-stat-POEGMA300)-C60 increased with methanol content in water, exhibiting lower LCSTs than PMEO2MA-stat-POEGMA300 for all methanol/water compositions. Higher critical micelle concentration (CMC) and larger spherical micelles were observed for (PMEO2MA-stat-POEGMA300)-C60 with increasing methanol content. The Rh of the micelles remained constant at temperature below the LCST and increased dramatically at temperature greater than the LCST, and (Rg/Rh) increased from ~ 0.75 to ~ 1.0. Nanocrystalline cellulose (NCC) was modified with water-soluble C60-(β-cyclodextrin) and (PMEO2MA-stat-POEGMA300)-C60) through a radical coupling reaction. NCC-C60-(PMEO2MA-stat-POEGMA300) possessed thermal responsive behavior in water and ~3.5 oC hysteresis associated with the heating/cooling cycles. No observable damage to NCC occurred during the radical coupling reaction as determined by TEM. NCC-C60-(β-cyclodextrin) possessed a similar thermal degradation behavior as NCC except it possessed a broader temperature range. Both NCC-fullerene systems demonstrated a radical scavenging activity when screened with the 2,2-diphenyl-1-picrylhydrazyl (DPPH). In addition, the drug loading and delivery using PDMAEMA-C60 with targeting moieties was explored. Two model drugs, namely fluorescein and pyrene were employed to evaluate the location of drug in the self-assembled structure of PDMAEMA-C60. It was found that the hydrophobic drugs were partitioned between the PDMAEMA shells and the hydrophobic fullerene cores. The drug delivery profiles indicated that PDMAEMA-C60 is an efficient drug carrier, however, it was cytotoxic to cells. The gene transfection efficacy of PDMAEMA-C60 to different cell lines was investigated and the results demonstrated that PDMAEMA-C60 exhibited good gene transfection performance. However, the targeting selectivity to liver cells cannot be determined in both cases. This study demonstrates that nanostructures of stimuli-responsive fullerene polymers can be controlled and manipulated by changing the external environments. Several potential applications, such as in drug and gene delivery, and free radical scavenging can be further explored.

Traditional methods for dermal wound closure such as sutures and staples are invasive and can result in soft tissue trauma, increasing the likelihood of localized inflammation and infections. Alternately, while tissue adhesive alternatives can effectively seal and adhere to the wounds, they can also present safety concerns relating to immunogenic responses and tissue toxicity. Herein, we fabricate injectable, adhesive, and cytocompatible poly(oligo(ethylene glycol) methyl ether methacrylate) (POEGMA)-dopamine (DA) hydrogels co-crosslinked via hydrazone and self-polymerized dopamine crosslinks that exhibit high water retention, improved tissue adhesiveness, and effective tissue regeneration properties. POEGMA-DA hydrogels exhibit independently tunable gelation properties based on their dual crosslinking mechanism, allowing for gelation as fast as 24 s (allowing for injection and rapid filling of irregularly-shaped wounds) while achieving relevant compressive moduli of up to 37 kPa and in vitro skin adhesion strengths of up to 1.2 kPa. The POEGMA-DA hydrogels induced no significant inflammation while demonstrating high interfacial adhesiveness in a stented skin excisional mouse model, enabling efficient dermal tissue regeneration by supporting collagen remodelling and enabling the regeneration of hair follicles, sebaceous glands, and blood vessels at the excision site over the 14-day study timeline. As such, injectable POEGMA-DA hydrogels represent a relevant non-toxic and adhesive alternative wound closure system for treating deep dermal wounds. / Thesis / Master of Applied Science (MASc) / Effective wound healing and subsequent tissue regeneration after a physical injury requires a moist sterile environment, the presence of oxygen, nutrients and enzymes, an efficient blood supply to the wound site, and a controlled inflammatory response to initiate the healing process. External methods of closing the wound to prevent infection aid in faster healing like sutures, staples, and liquid sealants which can result in infections and/or the stimulation of an inflammatory response that can hinder tissue restoration. Hydrogels, water-swellable polymer networks, represent an alternative solution that can both suppress infection while simultaneously promoting wound healing. Hydrogels have a similar structure to soft tissues like skin and can thus provide a supportive environment for cells to promote tissue regeneration and restore tissue structure and function. The swelling of hydrogels in water is highly beneficial for providing moisture at the wound site; however, this high degree of water retention also means they have a hard time sticking to tissues. To address this challenge, hydrogels can be modified with a component naturally derived from marine mussels that allows them to stick to their wet habitats, helping hydrogels to stick to the wound site while healing. In this thesis, mussel-inspired hydrogels are designed and can spontaneously gel and stick to a wound site to accelerate the restoration of the structure and function of skin. These biodegradable and injectable hydrogels are effective in accelerating wound closure with minimal evidence of scarring while suppressing negative inflammatory reactions and restoring the structure of skin by promoting the regeneration of hair follicles, sebaceous glands and blood vessels.

<p> Over the past decade, the development of surface-initiated living polymerization methods has brought a breakthrough to surface modification owing to their control ability. Surface-initiated atom transfer radical polymerization (si-ATRP), as the most popular one, has been widely employed to give novel polymer structures and functionalities to various surfaces for the purposes of tailoring surface properties, introducing new functions, or preparing so-called "smart surfaces", which can respond to external stimuli such as solvent type, pH, temperature, electric and magnetic fields etc. In this thesis, the mechanistic study of the si-ATRP was first carried out through modeling to gain good understanding of si-ATRP. Si-ATRP was then employed to prepare different types of polymer brushes to produce "smart surfaces". </p> <p> The kinetic model was developed using the method of moment. Combined with experimental data, a quantitative analysis was carried out for the si-ATRP mechanism. All information of grafted polymer chains, including active chain concentration, radical concentration, chain length, polydispersity, was illustrated. A new radical termination mechanism, termed as migration-termination, was proposed for si-ATRP. </p> <p> Si-ATRP was then employed to graft poly(oligo(ethylene glycol) methacrylate) (POEGMA) block poly(methyl methacrylate) (PMMA) brushes on silicon wafer surfaces. Simple solvent treatment gave nanoscale patterns via the phase segregation of POEGMA and PMMA segments. Various patterns including spherical aggregates, wormlike aggregates, stripe patterns, perforated layers and complete overlayers, were obtained by adjusting the upper block layer thickness. Furthermore, these nanopatterns had a unique stimuli-responsive property, i.e., switching between different morphologies reversibly after being treated with selective solvents. </p> <p> POEGMA-block-poly(2-(methacryloyloxy)ethyl trimethylammonium chloride) (PMETAC) brushes, having two hydrophilic segments, were synthesized by si-ATRP method. A variety of nanopatterns and their stimuli-responsive ability were observed. The adsorption behaviors of fibrinogen on these patterns were thoroughly studied by ellipsometry, water contact angel measurement, AFM and radio labelling method. </p> <p> A novel thermo-responsive copolymer, poly(2-(2-methoxyethoxy)ethyl methacrylate -co-oligo(ethylene glycol) methacrylate) (P(ME02MA-co-OEGMA)), was also grafted onto silicon wafers. Its thermo-responsive behavior and chain conformation in aqueous solution were studied by neutron reflectometry (NR). Both extended and collapsed brushes exhibited good protein adsorption resistance. </p> / Thesis / Doctor of Philosophy (PhD)