Philosophiae Doctor - PhD / The prevalence of obesity amongst South Africans is alarming, with more than 29% of men and 56% of women considered to be obese. Angiogenesis, a process for development of new blood vessels play a major role in growth and survival of the adipose tissues. Pharmacological inhibitors of angiogenesis are therefore a sensible strategy to reduce excess body weight. Current anti-obesity drugs have limitations because of their lack of selectivity and specificity, which lead to undesirable side effects and reduced drug efficacy. Future anti-obesity therapeutic strategies should be target-specific, with minimal toxicity towards healthy tissues will be more appropriate for obesity treatment. Targeted nano-therapeutic agents are currently being developed to overcome the drawbacks associated with conventional drug therapies. The nano-based delivery vehicles that specifically target diseased cells are appealing as they could reduce drug toxicity towards healthy tissues and be more effective at lower dosages. The main aim of this study was to develop a receptor-mediated nanotherapy that specifically targets the white adipose tissue vasculature and trigger the death of these cells through apoptosis. The 14 nm gold nanoparticles (AuNPs) were synthesized using theTurkevich method following reduction of gold aurate by sodium citrate salt. Different chemistries were used to functionalise the AuNPs for biological application by conjugating with either vascular targeting peptide or pro-apoptotic peptide on their surface or both. The nanomaterials were characterised by UV-Vis, Zeta potential and transmission electron microscopy (TEM). The sensitivity and specificity of various AuNP conjugates were tested in vitro on colon and breast cancer cell lines. A human (Caco-2) cell line that expresses the receptor for the adipose homing peptide was chosen as an in vitro model system. Cellular toxicity and uptake of the nanoparticles was evaluated using the WST-1 assay, Inductively Coupled Plasma-Optical Emission Spectra (ICP-OES) and TEM. The induction of apoptosis following exposure to the nanoparticles was examined by Western blot and flow cytometric analysis. The anti-proliferative activity of the targeted therapeutic nanoparticles on the cells was more pronounced on the cells expressing the receptor for the adipose homing peptide. The uptake of unfunctionalised AuNPs was higher compared to functionalised nanoparticles, but this did not impair cell viability. The activity of the therapeutic peptide was retained and enhanced following conjugation to AuNPs as shown by Western blot and flow cytometric analysis. The nanotherapy under study demonstrated receptor mediated targeting, and enhanced activity on the cells expressing the receptor. However, the therapeutic and efficacy of the targeted nanotherapy still need to be tested in animal models of obesity to confirm the treatment specificity.

The operating temperature of energy conversion and electronic devices affects their efficiency and efficacy. In many devices, however, the reference values of the thermal properties of the materials used are no longer applicable due to processing techniques performed. This leads to challenges in thermal management and thermal engineering that demand accurate predictive tools and high fidelity measurements. The thermal conductivity of strained, nanostructured, and ultra-thin dielectrics are predicted computationally using solutions to the Boltzmann transport equation. Experimental measurements of thermal diffusivity are performed using transient grating spectroscopy. The thermal conductivities of argon, modeled using the Lennard-Jones potential, and silicon, modeled using density functional theory, are predicted under compressive and tensile strain from lattice dynamics calculations. The thermal conductivity of silicon is found to be invariant with compression, a result that is in disagreement with previous computational efforts. This difference is attributed to the more accurate force constants calculated from density functional theory. The invariance is found to be a result of competing effects of increased phonon group velocities and decreased phonon lifetimes, demonstrating how the anharmonic contribution of the atomic potential can scale differently than the harmonic contribution. Using three Monte Carlo techniques, the phonon-boundary scattering and the subsequent thermal conductivity reduction are predicted for nanoporous silicon thin films. The Monte Carlo techniques used are free path sampling, isotropic ray-tracing, and a new technique, modal ray-tracing. The thermal conductivity predictions from all three techniques are observed to be comparable to previous experimental measurements on nanoporous silicon films. The phonon mean free paths predicted from isotropic ray-tracing, however, are unphysical as compared to those predicted by free path sampling. Removing the isotropic assumption, leading to the formulation of modal ray-tracing, corrects the mean free path distribution. The effect of phonon line-of-sight is investigated in nanoporous silicon films using free path sampling. When the line-of-sight is cut off there is a distinct change in thermal conductivity versus porosity. By analyzing the free paths of an obstructed phonon mode, it is concluded that the trend change is due to a hard upper limit on the free paths that can exist due to the nanopore geometry in the material. The transient grating technique is an optical contact-less laser based experiment for measuring the in-plane thermal diffusivity of thin films and membranes. The theory of operation and physical setup of a transient grating experiment is detailed. The procedure for extracting the thermal diffusivity from the raw experimental signal is improved upon by removing arbitrary user choice in the fitting parameters used and constructing a parameterless error minimizing procedure. The thermal conductivity of ultra-thin argon films modeled with the Lennard-Jones potential is calculated from both the Monte Carlo free path sampling technique and from explicit reduced dimensionality lattice dynamics calculations. In these ultra-thin films, the phonon properties are altered in more than a perturbative manner, referred to as the confinement regime. The free path sampling technique, which is a perturbative method, is compared to a reduced dimensionality lattice dynamics calculation where the entire film thickness is taken as the unit cell. Divergence in thermal conductivity magnitude and trend is found at few unit cell thick argon films. Although the phonon group velocities and lifetimes are affected, it is found that alterations to the phonon density of states are the primary cause of the deviation in thermal conductivity in the confinement regime.

>Magister Scientiae - MSc / South Africa is one of two countries responsible for the production of approximately 80% of the world’s Zr. The increase in mining activity has detrimental effects on the environment, especially crop plants, as more pollutants are leached into the soil. Consequently, it is necessary to understand how plants respond to this form of abiotic stress. Therefore, this study focused on determining the physiological and biochemical responses of two genotypes of Brassica napus L (Agamax and Garnet) in response to Zr stress. The levels of cell death, lipid peroxidation and ROS were higher in Garnet, whereas the chlorophyll content was higher in Agamax. Furthermore, native PAGE analysis detected seven SOD isoforms and seven APX isoforms in Agamax, compared to 6 SOD isoforms and 7 APX isoforms in Garnet. The results thus indicate that Agamax is tolerant to Zr-induced stress, whereas Garnet is sensitive. An assay for the rapid quantification of Zr within plant samples was subsequently developed, which revealed that Agamax retained the bulk of the Zr within its roots, whereas Garnet translocated most of the Zr to its leaves. The ability of Agamax to sequester Zr in its roots comes forth as one of the mechanisms which confers greater tolerance to Zr-induced stress. As a consequence, our study sought to use the optical, physical and chemical properties of quantum dots to image the uptake and translocation of Zr in B. napus genotypes. ICPOES was also performed to quantify Zr levels in various plant organs. Data from the ICPOES revealed varying patterns of uptake and translocations between Garnet and Agamax. These patterns were similarly shown in IVIS Lumina images, tracing the transport of QD/Zr conjugates. This method ultimately proved to be successful in tracing the uptake of Zr, and could essentially be a useful tool for targeting and imaging a number of other molecules.

The most common treatment used for cancer is chemotherapy. Chemotherapeutic agents have a greater affinity for rapidly dividing cells which is a characteristic of tumour cells. Although anti-cancer agents have their advantages in providing anti-cancer effects, they can be seen as highly toxic molecules posing a threat to normal healthy tissue within the human body. However, these toxic therapies need to be delivered to tumour sites without damaging healthy tissue. Liposomes can serve as a delivery system for these toxic molecules and be delivered to the tumour site via the EPR effect. Hence, liposomes that fuse with tumour cell line membranes are advantageous in delivering payloads of drugs directly into the tumour cell without damaging normal healthy tissue. The aim of the study was to formulate an optimised liposome preparation in order to enhance cellular uptake by MCF-7, Caco-2 and C3A cancer cell lines via membrane fusion. The optimal liposome formulation was aimed to be prepared utilising a statistical design approach in order to determine the ranges of the parameters that were furthermost optimal in formulating an ideal liposome preparation. The primary screening design was conducted using a 24-1 fractional factorial design that took into account the four parameters that were used to determine the optimisation of the liposomal preparation. The four variables used in the liposome preparation were the phospholipid type (PS or DOPE), the concentration of cholesteryl hemisuccinate (CHEMS) (10 – 40 %), the concentration of PEG2000-PE (0.5 – 4 %) and liposome size (100 or 200 nm). Liposomes were prepared using thin film hydration method and characterisation for size and zeta potential was carried out using photon correlation spectroscopy (PCS). Visual characterisation of liposome size was carried out using atomic force microscopy (AFM). Liposomes were exposed the cancer cell lines with visualisation and uptake being measured using fluorescent microscopy and flow cytometry, respectively. An optimal liposome preparation was prepared following the statistical design method. The optimal liposome preparation consisted of phospholipid type PS, 22.91 % of CHEMS, 4 % of PEG2000-PE and a liposome size of 200 nm. AFM analysis has shown that optimal liposome sizes ranged between 130 and 170 nm. Flow cytometry analysis indicated high level of liposome uptake with actual values falling below the predicted values set out by the statistical design. Fluorescence microscopy captured images of the fluorescent liposomes concentrated on the membrane of cells. The objective of the study was to determine from literature which variables would be desirable in preparing an optimal non-targeted liposome preparation. This was achieved by identifying four such variables and utilising them in a statistical design approach which was screened in order to determine the ideal parameters in preparing the optimised liposome batch. Therefore, from the results obtained it can be concluded that the aim of the study were met by preparing an optimal liposome preparation that has the ability to fuse with the tumour cell line membranes.

Nanoindentation has brought in many features of research over the past decade. This novel technique is capable of producing insights into the small ranges of deformation. This special point has brought a lot of focus in understanding the deformation behavior under the indenter. Nickel, iron, tungsten and copper-niobium alloy system were considered for a surface deformation study. All the samples exhibited a spectrum of residual deformation. The change in behavior with indentation and the materials responses to deformation at low and high loads is addressed in this study. A study on indenter geometry, which has a huge influence on the contact area and subsequently the hardness and modulus value, has been attempted. Deformation mechanisms that govern the plastic flow in materials at low loads of indentation and their sensitivity to the rate of strain imparted has been studied. A transition to elastic, plastic kind of a tendency to an elasto-plastic tendency was seen with an increase in the strain rate. All samples exhibited the same kind of behavior and a special focus is drawn in comparing the FCC nickel with BCC tungsten and iron where the persistence of the elastic, plastic response was addressed. However there is no absolute reason for the inconsistencies in the mechanical properties observed in preliminary testing, more insights can be provided with advanced microscopy techniques where the study can be focused more to understand the deformation behavior under the indenter. These experiments demonstrate that there is a wealth of information in the initial stages of indentation and has led to much more insights into the incipient stages of plasticity.

Toxicity of nano-scaled Al2O3, SiO2, TiO2 and ZnO to bacteria (Bacillus subtilis, Escherichia coli and Pseudomonas fluorescens) was examined and compared to that of their respective bulk (micro-scaled) counterparts. All nanoparticles (NPs) but TiO2 showed higher toxicity than their bulk counterparts. Toxicity of released metal ions was differentiated from that of the oxide particles. ZnO was the most toxic among the three NPs, causing 100% mortality to the three tested bacteria. TEM images showed attachment of NPs to the bacteria, suggesting that the toxicity was affected by bacterial attachment. The effects of oxide NPs on bacteria cells and bacterial surface biomolecules were studied by FTIR spectroscopy to provide a better understanding of their cytotoxicity. Lipopolysaccharide (LPS) and lipoteichoic acid could bind to oxide NPs through hydrogen bonding and ligand exchange, but the cytotoxicity of NPs seemed largely related to the function-involved or structural changes to proteins and phospholipids. The three NPs decreased the intensity ratio of β-sheets/α-helices, indicating protein structure change, which may affect cell physiological activities. The phosphodiester bond of L-α-Phosphatidyl-ethanolamine (PE) was broken by ZnO NPs, forming phosphate monoesters and resulting in the highly disordered alkyl chain. Such damage to phospholipid molecular structure may lead to membrane rupture and cell leaking, which is consistent with the fact that ZnO is the most toxic of the three NPs. LPS and PE are amphiphilic biomolecules that are major constituents of the outer membrane of Gram-negative bacteria. Their micelles and vesicles were studied as model cell membranes to evaluate NP effects on membrane construction. The adsorption of polysaccharides on Al2O3 and TiO 2 NPs dispersed LPS vesicles and micelles. LPS coated Al2O 3 NPs, while it caused the aggregation of TiO2 NPs according to atom force microscopy images. Desorption from the two NPs was slow due to the firm hydrogen bonding. For PE, Al2O3 NPs induced large multilamillar vesicles, while ZnO NP converted vesicles to tiny aggregates due to molecular structure breakup. PE stability in solution was disturbed by adding NPs, but its stability was enhanced by increasing pH. The electrostatic force was the determining factor for the vesicle stability.

Colloidal Microcapsules (MCs), i.e. capsules stabilized by nano-/microparticle shells are highly modular inherently multi-scale constructs with applications in many areas of material and biological sciences e.g. drug delivery, encapsulation and microreactors. These MCs are fabricated by stabilizing emulsions via self-assembly of colloidal micro/nanoparticles at liquid-liquid interface. In these systems, colloidal particles serve as modular building blocks, allowing incorporation of the particle properties into the functional capabilities of the MCs. As an example, nanoparticles (NPs) can serve as appropriate antennae to induce response by external triggers (e.g. magnetic fields or laser) for controlled release of encapsulated materials. Additionally, the dynamic nature of the colloidal assembly at liquid-liquid interfaces result defects free organized nanostructures with unique electronic, magnetic and optical properties which can be tuned by their dimension and cooperative interactions. The physical properties of colloidal microcapsules such as permeability, mechanical strength, and biocompatibility can be precisely controlled through the proper choice of colloids and preparation conditions for their. This thesis illustrates the fabrication of stable and robust MCs through via chemical crosslinking of the surface engineered NPs at oil-water interface. The chemical crosslinking assists NPs to form a stable 2-D network structure at the emulsion interface, imparting robustness to the emulsions. In brief, we developed the strategies for altering the nature of chemical interaction between NPs at the emulsion interface and investigated their role during the self-assembly process. Recently, we have fabricated stable colloidal microcapsule (MCs) using covalent, dative as well as non-covalent interactions and demonstrated their potential applications including encapsulation, size selective release, functional devices and biocatalysts.

Mesoporous materials are being widely used in the chemical industry in various environmentally friendly separation processes and as catalysts. Our research can be broadly described as an effort to understand the behavior of fluids confined in such materials. More specifically we try to understand the influence of state variables like temperature and pore variables like size, shape, connectivity and structural heterogeneity on both the dynamic and equilibrium behavior of confined fluids. The dynamic processes associated with the approach to equilibrium are largely unexplored. It is important to look into the dynamic behavior for two reasons. First, confined fluids experience enhanced metastabilities and large equilibration times in certain classes of mesoporous materials, and the approach to the metastable/stable equilibrium is of tremendous interest. Secondly, understanding the transport resistances in a microscopic scale will help better engineer heterogeneous catalysts and separation processes. Here we present some of our preliminary studies on dynamics of fluids in ideal pore geometries. The tool that we have used extensively to investigate the relaxation dynamics of fluids in pores is the dynamic mean field theory (DMFT) as developed by Monson [P. A. Monson, J. Chem. Phys., 128, 084701 (2008)]. The theory is based on a lattice gas model of the system and can be viewed as a highly computationally efficient approximation to the dynamics averaged over an ensemble of Kawasaki dynamics Monte Carlo trajectories of the system. It provides a theory of the dynamics of the system consistent with the thermodynamics in mean field theory. The nucleation mechanisms associated with confined fluid phase transitions are emergent features in the calculations. We begin by describing the details of the theory and then present several applications of DMFT. First we present applications to three model pore networks (a) a network of slit pores with a single pore width; (b) a network of slit pores with two pore widths arranged in intersecting channels with a single pore width in each channel; (c) a network of slit pores with two pore widths forming an array of ink-bottles. The results illustrate the effects of pore connectivity upon the dynamics of vapor liquid phase transformations as well as on the mass transfer resistances to equilibration. We then present an application to a case where the solid-fluid interactions lead to partial wetting on a planar surface. The pore filling process in such systems features an asymmetric density distribution where a liquid droplet appears on one of the walls. We also present studies on systems where there is partial drying or drying associated with weakly attractive or repulsive interactions between the fluid and the pore walls. We describe the symmetries exhibited by the lattice model between pore filling for wetting states and pore emptying for drying states, for both the thermodynamics and dynamics. We then present an extension of DMFT to mixtures and present some examples that illustrate the utility of the approach. Finally we present an assessment the accuracy of the DMFT through comparisons with a higher order approximation based on the path probability method as well as Kawasaki dynamics.

Increasing motivation for the development of nanotechnology with applications in sensing, nanotheranostics, combinatorial therapy and drug/protein delivery have brought a broad spectrum of multifunctional polymeric materials. With interest in obtaining more efficient methods for fast and accurate diagnostics, nanostructures able to rapidly obtain large sets of data for analyte sensing are necessary. Likewise, interests in, not only obtaining accurate diagnostics, but also being able to concurrently and accurately provide therapy has inspired us to develop such technologies. We particularly focus on understanding the assembly and disassembly processes of polymeric nanostructures in response to biologically relevant stimuli. We are interested in mimicking natural sensing events and take advantage of the differential binding affinity of a set of receptors to generate analyte-specific patterns to use as sensors. Receptors developed in our laboratories have demonstrated impressive recognition capabilities for proteins. When a set of polyelectrolytes and surfactants with different hydrophobic and electronic properties, as well as the guest molecule (transducer) with different characteristics, are combined, fluorescence patterns for proteins can be generated. Creating patterns using protein-induced disassembly not only provides the opportunity to have a new method for sensing analytes that are not electronically complementary to the fluorescent transducers, but also reduces the synthetic complexity even further, since these are assembled from its components noncovalently. A versatile and efficient delivery vehicle should have a tunable particle size, provide protection and stability to the cargo to prevent premature release before approaching the target site and should provide ease of cargo incorporation strategies. This ease of incorporation becomes more challenging when we are talking about incorporating molecules with different characteristics. Here we demostrate the versatility of self-crosslinked polymeric nanogels on the incorporation of lipophilic small molecules on its interior and hydrophilic proteins at the surface. Also, different approaches of protein incorporation can be obtained using these polymeric nanogels, which provide differences in protein release and cell internalization. These technologies were found to be potential candidates for applications such as nanotheranostics, combinatorial therapy and protein delivery.

The self–assembly of block copolymers (BCPs) into well–ordered nanoscopic arrays holds promise for new technological breakthroughs as templates and scaffolds for the fabrication of nanostructured materials. It is essential to establish convenient approaches to control the morphology of BCPs, since some applications require addressability, the BCP microdomains must be perfectly aligned and oriented. The theme of this thesis is the use of external forces, specifically thermal and solvent annealing, to guide the self–assembly of BCPs to obtain microphase separated morphologies for different applications. Three representative BCP systems, having lamellar, cylindrical and spherical microdomains are discussed. First, the self–assembly of lamella–forming brush block copolymers (BrBCPs) having polylactide (PLA) and polystyrene (PS) side chains were studied in the bulk and in thin films. The domain size increased approximately linearly with the molecular weight of the backbone, which indicated that the backbone was in an extended conformation that was confirmed theoretically. In situ small angle x–ray scattering (SAXS) measurements indicated that the self–assembly of the BrBCPs was quite rapid, due to the rigid nature of the backbone chain, Second, the directed self–assembly of cylinder–forming polystyrene–block–poly(ethylene oxide)s (PS–b–PEOs) in thin films were investigated. The polymer–surface interactions were tuned by hydroxyl end–functionalized polymers, so that the orientation of the PS–b–PEO microdomains was controlled during thermal annealing. Cylindrical PEO microdomains embedded in a PS matrix orientated normal to the silicon substrates were observed over a wide processing window when the substrates were modified by PS– b–PEO BCPs. PS–b–PEOs with an o–nitrobenzyl ester junction point (PS–hν –PEOs) were used to fabricate nanoscopic dot and line patterns having long–range lateral order. The cylindrical BCP microdomains were oriented perpendicular or parallel to the silicon substrates by varying the solvent annealing conditions. The third BCP system examined in this study was a sphere–forming polystyrene–block–polydimethylsiloxane (PS–b–PDMS). Solvent annealing in N–methyl–2–pyrrolidone was used to direct the self–assembly of the spherical microdomains into high areal density arrays on flat Si substrates, PS modified substrates and lithographically patterned substrates, respectively.