The field of nanomedicine has witnessed an incredible expansion, from a total market value in 2003 of $500 million expected to rise to $160 billion by 2015 (Global Industry Analysts, Inc.). The nanomedicine industry is forecasted to grow and have a significant impact on the economy, with sectors such as biomaterials, diagnostics and drug delivery expected to play a major role. This thesis gives a detailed account of the synthesis and characterisation of molecularly imprinted nanogels for drug delivery. Their toxicity and potential use as a targeted carrier to cancerous cells is evaluated. Initially an overview of nanomaterials and their uses in many areas such as agriculture, energy storage and technology are discussed. The impact of nanomaterials on the life sciences is examined; in particular their application in drug delivery is focussed upon. Chapters 2, 3 and 4 make up the results and discussion of this work. Chapter 2 focuses on developing the synthesis of the acrylamide based nanogels and, vitally, incorporating a suitable fluorescent tag in order to track the nanogels in vitro and in vivo. Fundamentally toxicity studies carried out on the nanogels, both in vitro and in vivo in Danio rerio (zebrafish) are reported in Chapter 3 to ensure the nanogels are biocompatible. Chapter 4 introduces an innovative approach, molecular imprinting, to incorporating a drug into the nanogels. The upload and release of Tamoxifen (a drug used to treat breast cancer) at reduced pH, was also analysed. Finally future development of the carrier is discussed and key issues that need to be addressed.

Chemical methods are generally used for the synthesis of active nanoparticles (metals, semi-metals, metal oxides, and etc) supported on high surface area materials. Chemical methods involve using strong solvents, harmful gases (H2 & CO), and high temperature techniques such as high boiling solvents, calcination and pyrolysis. The main drawbacks of using this approach, is the prevalence of chemical agents on nanomaterials which tends to negate its applications. Alternatively, photochemical and photothermal methods are widely being considered for the synthesis and design of nanomaterials. For these studies, the active nanomaterials incorporated within high surface area materials were prepared by the laser vaporization-controlled condensation (LVCC) technique or by the laser irradiation in solution (LIS) technique. The LVCC technique involves the irradiation of a solid target at the focal point of a laser beam (532 nm, 30 Hz) by the Nd: YAG laser inside a chamber that is sandwiched between two steel plates in the presence of high purity He. Whereas, the LIS technique involves the laser irradiation of chemical precursors in aqueous solvents using an unfocused beam. The LVCC technique was used for the preparation of carbonaceous and N-doped carbonaceous TiO2 support materials from MIL-125(Ti) and NH2-MIL-125(Ti) metal organic frameworks, Ge and GeO2 nanostructures, GeOx/PRGO nanocomposite, and the Fe3O4/PRGO nanocomposite. On the other hand, Pd supported on MIL-125(Ti) and NH2-MIL-125(Ti) nanocatalysts, GeO2/RGO, and the poly(ethylene glycol methacrylate-co-bisacrylamide) hydrogels were all prepared by the LIS technique.

The search for cellulosic based products as a viable alternative for petroleum-based products was the impetus for covalently crosslinking lignocellulosic fibers and nanocellulose whiskers with poly(methyl vinyl ether) co maleic acid (PMVEMA) - polyethylene glycol (PEG). The lignocellulosics used were ECF bleached softwood (pine) and ECF bleached birch kraft pulp. This thesis also tests the hypothesis that water absorption and retention can be improved by grafting PMVEMA-PEG to the surface of ECF bleached kraft pulp hardwood and softwood fibers via microwave initiated crosslinking. The crosslinking of the PMVEMA to hardwood and softwood kraft ECF bleached pulp fibers resulted in enhanced water absorbing pulp fibers where the PMVEMA is grafted onto the surface of the fibers. The crosslinking was initiated both thermally and via microwave irradiation and the water absorption and water retention was measured as the percent of grafted PMVEMA. This was the first application of microwave crosslinking of pulp fibers with the goal of creating water absorbing pulp fibers. Ultimately, the water absorption values ranged from 28.70 g water per g dry crosslinked pulp fiber (g/g) to 230.10 g/g and the water retention values ranged from 26% to 71% of the water retained that was absorbed by the crosslinked pulp fibers. The microwave initiated crosslinked fibers had comparable results to the thermally crosslinked fibers with a decreased reaction time, from 6.50 min (thermal) to 1 min 45 sec (microwave). Cellulose nanowhiskers, crystalline rods of cellulose, have been investigated due to their unique properties, such as nanoscale dimensions, low density, high surface area, mechanical strength, and surface morphology and available surface chemistry. Prior to this study, the crosslinking of cellulose whiskers with the matrix via solution casting of liquid suspensions of whiskers and matrix had not been explored. The hypothesis to be investigated was that incorporating cellulosic whiskers with the PMVEMA-PEG matrix and crosslinking the whiskers with the matrix would yield films that demonstrate unique properties when compared to prior work of crosslinking of PMVEMA-PEG to macroscopic ECF bleached kraft pulp fibers. Solution cast composites of cellulose nanowhiskers-PMVEMA-PEG were crosslinked at 135 °C for 6.5 min and analyzed for crosslinking, thermal stability, strength and mechanical properties, whisker dispersion, and water absorption and uptake rates. The whisker-composites demonstrated unique properties upon crosslinking the whiskers with PMVEMA-PEG, especially the elongation at break and tensile strength upon conditioning of the final materials at various relative humidities. In addition, the whiskers improved the thermal stability of the PMVEMA-PEG matrix. This is significant as methods of improving processing thermal stability are key to developing new materials that utilize cellulose whiskers, PMVEMA, and PEG. This thesis addresses the hypothesis that cellulose nanowhiskers that are crosslinked with a matrix can create new whisker-matrix composites that behave differently after crosslinking.

Nanoemulsions with extremely small droplet sizes (<100 nm) have shown several advantages over conventional emulsions. However, almost all nanoemulsions in usage are liquids that restrict their use in many soft materials. The aim of this thesis is to understand the formation and long-term stability of viscoelastic nanogels developed from liquid nanoemulsions. At first, gelation in 40 wt% canola oil-in-water nanoemulsions were investigated as a function of emulsifier type (anionic sodium dodecyl sulfate (SDS) or nonionic Tween 20) and concentration. Three different regimes of colloidal interactions were observed as a function of SDS concentration. 1) At low SDS concentration (0.5 – 2 times CMC) the counterion shell layer increased the effective volume fraction of the dispersed phase (eff) close to the random jamming, resulting in repulsive gelation. 2) At SDS concentration between 5 – 15 times CMC, micelle induced depletion attractions led to extensive droplet aggregation and gelation. 3) At very high SDS concentration, however, oscillatory structural forces (OSF) due to layered-structuring of excess micelles in the interdroplet regions led to loss of gelation. In repulsive gelation, reduction in droplet size coupled with the electrical double layer resulted in a linear increase of Gʹ. On the contrary, attractive nanoemulsions showed rapid increase in gel strength below a critical droplet radius, and was explained by transformation of OSF into depletion attraction. No gelation was seen in Tween 20 nanoemulsions, due to lack of repulsive interactions and weak depletion attraction. Next the influence of the dispersed phase volume fraction () on repulsive nanoemulsion gelation was investigated and the Gʹ values were modeled using empirical scaling law developed by Mason et al. (1995). It was found that an initial liquid regime transformed into glassy phase at a eff = g ~ 0.58, where droplets are entrapped in a cage of neighbouring droplets due to crowding. It was followed by jamming transition at a critical volume fraction (j), where droplet deformation led to large increase in elasticity. The model predicted j = 0.7, which is close to the predictions for repulsive polydispersed emulsions found in the literature. In the final phase long-term stability of the nanogels was evaluated until 90 days, during which the nanogels remained stable to creaming and coalescence. However, repulsive nanogels showed a significant decrease in Gʹ and the gels converted into flowable liquids over time. For attractive nanogels decrease in Gʹ was much less, although given enough time they would also transformed into weak gels. It was hypothesized that surface active compounds generated due to lipid oxidation altered interfacial charge cloud leading to loss of gel strength for repulsive nanogels. For attractive nanogels slippery bonds in the aggregates permitted rotational and translational diffusion of nanodroplets on the surface of each other leading to network compactness and a decrease in gel strength with time. Overall, it was concluded that it is possible to form nanogels from canola oil nanoemulsions using ionic emulsifiers. The gel strength and stability of the nanogels depends on emulsifier concentration, droplet size,  and the chemical stability of the oil used. More investigation is needed in order to improve the long-term stability of the nanogels. The nanogels possess high potential for use in low-fat foods, pharmaceuticals, and cosmetic products.

A very interesting class of nanoparticles extensively used for bio-applications is that of hydrogel particles, also called nanogels. There is an increasing interest in the design of hydrogel nanoparticles that have biofunctionality for applications in cell targeting, drug delivery, and biomedicine. The dissertation focuses on developing synthetic strategies for making diverse hydrogel nanoparticles customized to have desirable properties for various bio-applications. We have also investigated the potential of such nanoparticles as coatings for biomedical implants. Chapter 1 gives a brief introduction to hydrogel nanoparticles and the properties that make them attractive for various applications. The details of the syntheses of well defined, stable nanoparticles, commonly used in literature, are described in Chapter 2. Chapter 3 describes our synthesis of hollow sub-50 nm nanogels, which are otherwise difficult to synthesize based on the strategy discussed in Chapter 2. Chapter 4 also demonstrates how simple strategies borrowed from organic chemistry help in producing nanogels with multiple functionalities that are otherwise difficult to obtain, which also is an important advance over the synthetic methods discussed in Chapter 2. Chapter 5 describes how a general strategy based on photoaffinity labeling can yield materials with many applications ranging from optical materials, drug delivery, to biosensing. The latter part of the dissertation describes applications of various nanogels in biology especially as coatings that can control inflammation caused by biomaterials. Chapter 6 describes a method to functionalize flexible biomaterials with the nanogels, thus enabling in vivo investigations of the nanogels as potential coatings for controlling inflammation. Chapter 7 describes the biological studies performed (in collaboration with Garcia Group in the School of Mechanical Engineering at Georgia Tech) on various nanogels, aimed towards obtaining the most functional and efficient materials for implant applications. Chapter 8 describes application of hollow nanogels for covalently immobilizing biomolecules. This chapter also demonstrates how simple non-functional materials can be made unique and functional by means of traditional organic reactions. Finally, in order to broaden the applications of nanogel based materials.

Les nanogels hybrides constitués de polymères thermosensibles et de nanoparticules inorganiques stimulables telles que des nanoparticules magnétiques (NPMs) ou des nanobatônnets d’or (AuNRs) sont extrêmement intéressants pour des applications biomédicales. Leur matrice en polymère permet d’encapsuler et de libérer de grandes quantités de molécules actives, alors que les nanoparticules peuvent générer de la chaleur lorsqu’elles sont exposées à un champ magnétique alternatif (AMF) pour les NPMs, et à une irradiation proche infrarouge (NIR-L) pour les AuNRs. Ce manuscrit de thèse porte sur la synthèse et la caractérisation de nanogels biocompatibles, pH- et thermosensibles, à base de monomères en oligo (éthylène glycol) méthyl éther méthacrylate (OEGMAs), d’acide méthacrylique (MAA) et encapsulant des NPMs et/ou des AuNRs pour déclencher de manière contrôlée, par hyperthermie magnétique ou par photothermie, la libération d’une molécule anticancéreuse, la doxorubicine (DOX). Des nanogels hybrides magnétiques, plasmoniques et magnéto-plasmoniques ont été synthétisés. Ces nanogels ont un diamètre hydrodynamique entre 200 et 500 nm et une température de transition de phase volumique comprise entre 30 et 54 °C. Le comportement de gonflement-dégonflement des nanogels peut être induit par plusieurs stimuli (température, pH, AMF, NIR-L). Ces résultats démontrent que les MagNanoGels sont d’excellents nanovecteurs pour accroître l’internalisation cellulaire en augmentant la cytotoxicité de la DOX et qu’il est possible de déclencher à distance la libération intracellulaire de DOX sous AMF dans des conditions athermiques. Par ailleurs, les PlasMagNanoGels peuvent générer efficacement de la chaleur par photothermie pour une thermothérapie. En outre, les propriétés intrinsèques des NPMs, pour le ciblage magnétique et en tant qu’agents de contraste pour l’imagerie par résonance magnétique (MRI), font de ces nanogels des candidats idéaux pour une nouvelle approche thérapeutique (diagnostique et traitement) contre le cancer. / Hybrid nanogels, composed of thermoresponsive polymers and inorganic responsive nanoparticles, such as magnetic nanoparticles (NPMs) and gold nanorods (AuNRs) are highly interesting for biomedical applications. Their polymeric matrix makes them able to uptake and release high quantities of drugs, whereas nanoparticles can generate heat when exposed to an alternating magnetic field (AMF) for NPMs, and to a near-infrared light for AuNRs. This thesis manuscript focuses on the synthesis and the characterization of biocompatible, pH- and thermoresponsive nanogels, based on oligo(ethylene glycol) monomers (OEGMAs), methacrylic acid (MAA) and encapsulating NPMs and/or AuNR for remotely triggered doxorubicin (DOX, anticancer drug) release, by magnetic hyperthermia or phothothermia. Hybrid magnetic, plasmonic and magneto-plasmonic nanogels were synthesized. Theses nanogels have a hydrodynamic diameter between 200 and 500 nm and a volume phase transition temperature (VPTT) from 30 to 54°C. The nanogels’ swelling-deswelling behavior can be induced by several stimuli (temperature, pH, AMF, NIR-L). These results demonstrate that MagNanoGels are excellent nanocarriers for enhancing cellular internalization enhancing DOX cytotoxicity and that DOX release was significantly enhanced upon exposure to AMF in athermic conditions. In addition, PlasMagNanoGels can efficiently generate heat by photothermy for thermotherapy. Therefore, the intrinsic properties of NPMs for magnetic targeting and as contrast agents for Magnetic Resonance Imaging (MRI), make these nanogels ideal candidates for a new therapeutic approach (diagnosis and treatment) against cancer.

Le fascinant processus par lequel les signaux mécaniques sont transformés en réactions biochimiques dans la nature est appelé mécano-transduction. Le but de ma thèse a été de mimer la Nature en élaborant de nouveaux systèmes enzymatiques mécano-transductifs, i.e des matériaux capables de moduler une catalyse enzymatique lorsqu’ils sont sollicités mécaniquement. Nous avons d’abord étudié l’effet de l’étirement sur les chaînes constitutives de films multicouches de polyélectrolytes, matrices souvent utilisées pour le développement de biomatériaux intelligents. Dans le cadre d’une nouvelle stratégie axée sur la modulation mécanique de la conformation, nous avons ensuite élaboré des matrices étirables à base de poly(éthylène glycol)s. Nous avons en particulier développé de tout nouveaux revêtements covalents appelés nanogels qui se sont avérés être déposables sur le silicone étirable et fonctionnalisables avec différentes biomacromolécules, ouvrant ainsi de nouvelles routes biomimétiques. / The fascinating process by which mechanical signals are transformed into biochemical reactions in Nature is called mechanotransduction. The goal of my PhD was to mimic Nature by elaborating new types of mechanocatalytic materials, i.e materials able to modulate a catalytic activity when mechanically stimulated. We first aimed at understanding the impact of stretching on the structural properties of polyelectrolyte multilayers films, polymeric matrices often used for the design of smart biomaterials. Within the framework of a new strategy essentially relying on mechanically induced conformational changes, we then developed stretchable polymeric matrices based on poly(ethylene glycol)s. We more particularly designed new types of covalent coatings, called nanogels. We showed that these architectures were buildable on stretchable silicone and that they could be functionalized with different types of biomacromolecules; thus opening new biomimetic routes.

Oxidative stress is a pathophysiological condition defined by an increased production of reactive oxygen species (ROS), which can result in the growth arrest of cells followed by cell disintegration or necrosis. A number of small molecule antioxidants (e.g. curcumin, quercetin and resveratrol) are capable of directly scavenging ROS, thereby short-circuiting the self-propagating oxidative stress state. However, poor solubility and rapid 1st pass metabolism results in overall low bioavailability and acts as a barrier for its use as a drug to suppress oxidative stress efficiently. To overcome this limitation, these small molecule antioxidants were covalently conjugated into poly(β-amino ester) (PβAE) cross-linked networks to formulate prodrug gel microparticles and nanoparticles (nanogels). Being hydrolytically degradable in nature, these PβAE crosslinked systems released antioxidants in their original structural form in a sustained controlled fashion. Both quercetin and curcumin-PβAE nanogels showed prolonged suppression of cellular oxidative stress induced by H2O2. Curcumin PβAE nanogels also demonstrated protection against mitochondrial oxidative stress induced by H2O2 and polychlorinated biphenyls. Curcumin-PβAE gel microparticles were also developed as a platform to treat oral mucositis through a local antioxidant delivery route. The same synthesis chemistry was transferred to formulate resveratrol PβAE gel microparticles for topical applications, to treat UV radiation induced oxidative stress. Both formulations showed suppression of induced oxidative stress. An in vivo trial with curcumin-PβAE microparticles further showed relatively reduced the severity of induced oral mucositis (OM) in hamster check pouch as compared to placebo.

We have used a molecular modeling of both random and blocky sequence hydrogel networks of poly(N-vinyl-2-pyrrolidone-co-2-hydroxyethyl methacrylate) (P(VP-co-HEMA)) with a composition of VP:HEMA = 37:13 to investigate the effect of the monomeric sequence and the water content on the equilibrium structures and the mechanical and transport properties by full-atomistic molecular dynamics (MD) simulations. The degree of randomness of the monomer sequence for the random and the blocky copolymers, were 1.170 and 0.104, respectively, and the degree of polymerization was fixed at 50. The equilibrated density of the hydrogel was found to be larger for the random sequence than for the blocky sequence at low water contents (< 40 wt %), but this density difference decreased with increasing water content. The pair correlation function analysis shows that VP is more hydrophilic than HEMA and that the random sequence hydrogel is solvated more than the blocky sequence hydrogel at low water content, which disappears with increasing water content. Correspondingly, the water structure is more disrupted by the random sequence hydrogel at low water content but eventually develops the expected bulk-water-like structure with increasing water content. From mechanical deformation simulations, the stress-strain analysis showed that the VP is found to relax more efficiently, especially in the blocky sequence, so that the blocky sequence hydrogel shows less stress levels compared to the random sequence hydrogel. As the water content increases, the stress level becomes identical for both sequences. The elastic moduli of the hydrogels calculated from the constant strain energy minimization show the same trend with the stress-strain analysis. Ascorbic acid and D-glucose were used to study the effect of the monomeric sequence on the diffusion of small guest molecules within the hydrogels. By analyzing the pair correlation functions, it was found that the guest molecule has greater accessibility to the VP units than to the HEMA units with both monomeric sequences due to its higher hydrophilicity compared to the HEMA units. The monomeric sequence effect on the P(VP-co-HEMA) hydrogel is clearly observed with 20 wt % water content, but the monomeric sequence effect is significantly reduced with 40 wt % water content and disappears with 80 wt % water content. This is because the hydrophilic guest molecules are more likely to be associated with water molecules than with the polymer network at the high water content. By analyzing the mean square displacement, the displacement of the guest molecules and the inner surface area, it is also found that the guest molecule is confined in the system at 20 wt % water content, resulting in highly anomalous subdiffusion. Therefore, the diffusion of the guest molecules is directly affected by their interaction with the monomer units, the monomeric sequence and the geometrical confinement in the hydrogel at a low water content, but the monomeric sequence effect and the restriction on the diffusion of the guest molecule are significantly decreased with increasing the water content. We also investigated the de-swelling mechanisms of the surface-grafted poly(N-isopropylacrylamide) (P(NIPAAm)) brushes containing 1300 water molecules at 275 K, 290 K, 320 K, 345 K, and 370 K. We clearly observed the de-swelling of the water molecules for P(NIPAAm) above the lower critical solution temperature (LCST) (~305 K). Below the LCST, we did not observe the de-swelling of water molecules. Using the upper critical solution temperature (UCST) systems (poly(acrylamide) brushes) for comparison purposes, we did not observe the de-swelling of water molecules at a given range of temperatures. By analyzing the pair correlation functions and the coordination numbers, the de-swelling of the water molecules occurred distinctly around the isopropyl group of the P(NIPAAm) brush above the LCST because C(NIPAAm) does not offer sufficient interaction with the water molecules via the hydrogen bonding type of secondary interaction. We also found that the contribution of the N(NIPAAm)-O(water) pair is quite small because of the steric hindrance of the isopropyl group. By analyzing the change in the hydrogen bonds, the hydrogen bonds between polar groups and water molecules in the P(NIPAAm) brushes weaken with increasing temperature, which leads to the de-swelling of the water molecules out of the brushes above the LCST. Below the LCST, the change in the hydrogen bonds is not significant. Again, the contribution of the NH(NIPAAm)-water pairs is insignificant; the total number of hydrogen bonds is ~20, indicating that the interaction between the NH group and the water molecules is not significant due to steric hindrances. Lastly, we observed that the total surface area of the P(NIPAAm) brushes that is accessible to water molecules is decreased by collapsing the brushes followed by the de-swelling of water molecules above the LCST.

The use of hydrogel nanoparticles, or nanogels, as targeted delivery vehicles to cancer cells was described. The nanogels were synthesized by free radical precipitation polymerization, with poly(N-isopropylmethacrylamide) as the main monomer, and have a core/shell architecture. The nanogels were near 50 nm in radius, contained fluorescein for visualization, and had an amine-containing shell for bioconjugation, making these particles ideal for delivery studies. The nanogels were conjugated with the YSA (YSAYPDSVPMMSC) peptide, which is an ephrin mimic, allowing for uptake by the EphA2 (erythropoietin-producing hepatocellular) receptor. We have delivered YSA-conjugated nanogels to Hey cells and BG-1 cells, as evidenced by fluorescence microscopy. We have shown that the nanogels can encapsulate siGLO Red Transfection Indicator (siGLO) and deliver the siGLO to Hey cells in vitro. After successful delivery of the non-targeting siGLO, we delivered siRNA for knockdown of epidermal growth factor receptor (EGFR). We have shown protein knockdown from 24-120 h after nanogel delivery, as well as knockdown with different siRNA concentrations delivered to the cells. Furthermore, addition of taxol following EGFR knockdown suggests that the chemosensitivity of the Hey cells is increased. Successful in vitro delivery of the nanogels prompted in vivo studies with the nanogels. The nanogels were used to encapsulate silver nanoclusters for potential bioimaging applications. Targeting of the nanogels to MatrigelTM plugs in mice suggest that the particles hold promise as in vivo delivery agents.