>Magister Scientiae - MSc / Cancer is one of the leading causes of death, worldwide. Mortality tolls are estimated to reach approximately 13.1 million in 2030. These statistics suggest that current therapeutic strategies are not effective. This is partly due to the fact that the drugs used in the treatment of cancer lack selectivity and specificity, which lead to undesirable side effects and reduced drug efficacy. There is therefore a need for alternative therapeutic approaches. In view of this, the therapeutic goal of chemotherapy has shifted towards targeted drug delivery systems, which have been successfully demonstrated using nanotechnology. The nano-based drug 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 gold nanorods (AuNRs) capable of inducing cell death in cancer cells specifically. Selectivity of the AuNRs (denoted as AGK) for cancer cells was achieved by conjugating the AuNRs to a peptide (Adipose Homing Peptide or AHP) that has high affinity and specificity for a cell surface receptor (prohibitin or PHB) that is expressed on some cancer cells. Cell death was achieved through conjugating the AuNRs to a pro-apoptotic peptide, D(KLAKLAK)2. Spherical AuNPs (AuNSs) conjugated with AHP and D(KLAKLAK)2, capable of selectively inducing apoptosis in cancer cells that express PHB, was previously reported. However, in this study the AuNSs were replaced with AuNRs. AuNRs has the ability to absorb light in the near infrared (NIR) light spectrum and converts this light energy into heat. This property of AuNRs has been used in several studies to demonstrate the application of AuNRs for the treatment of cancer using photothermal therapy (PTT). Consequently, the AuNRs described in this study can also be used for PTT. These AuNRs can induce cell death through the target specific delivery of the pro-apoptotic peptide D(KLAKLAK)2 as well as through PTT. The study showed that three human cancer cell lines (PC-3, Caco-2 and U-87) express PHB. The cytotoxicity testing of AGK AuNPs on PC-3 cells showed that these AuNRs could induce apoptosis in these cells without exposure to a NIR light source. The study also shows that AuNRs conjugated with the targeting peptide only (denoted as AG) can induce cell death in Caco-2 through PTT. This study demonstrates the potential of the AuNRs described in this study for application in the targeted elimination of cancer cells through the selective induction of PTT and apoptosis.

The objectives of this thesis were to optimize the synthesis and surface coating of metallic and semiconductor nanoparticles, to understand how these materials interact with cells and physiological systems and to investigate how they can be used to deliver thermal therapy for medical applications. Reproducible high-yield synthesis of gold nanorods and surface coating with a variety of polymers and silica was optimized. Using gold nanorods as a model system, the relationship between particle surface chemistry, surface charge and cellular uptake was studied, as well as the toxicity of nanoparticles of different surface chemistry. Low toxicity in vitro was encouraging and was confirmed in vivo by intravenously injecting Sprague-Dawley rats with semiconductor quantum dots of various surface coatings. Low toxicity was found during biochemical, haematological and pathological assessment, and these results indicate that applications of nanoparticles should be further investigated. One such application is the use of near infrared absorbing gold nanorods in remotely activated hyperthermia. It was shown that gold nanorods act synergistically with the chemotherapeutic cisplatin to improve cytotoxicity, and reduce the required cytotoxic drug dose to 33% of the unheated amount. Due to the success of hyperthermia treatment in vitro, continuing and future work involves the use of gold nanorods ex vivo on excised human corneas in a novel application to weld corneal tissue for improved wound closure following cataract surgery.

This thesis describes the synthesis and assembly of metal and semiconductor nanoparticles (NPs). The two research topics include i) hetero-assembly of metal and semiconductor NPs, ii) effect of ionic strength on homo-assembly of gold nanorods (GNRs). First, we present hetero-assembly of GNRs and semiconductor quantum dots (QDs) in a chain using biotin-streptavidin interaction. We synthesized alloyed CdTeSe QDs and modified them with mercaptoundecanoic acid to render them water-soluble and to attach streptavidin. We synthesized GNRs by a seed-mediated method and selectively modified the ends with biotin. Hetero-assembly of QDs and GNRs depended on the size, ligands, and ratio of QDs and GNRs. Second, we controlled the rate of homo-assembly of GNRs by varying the ionic strength of the DMF/water solution. The solubility of polystyrene on the ends of GNRs depended on the ionic strength of the solution, which correlated with the rate of assembly of GNRs into chains.

This thesis describes the synthesis and assembly of metal and semiconductor nanoparticles (NPs). The two research topics include i) hetero-assembly of metal and semiconductor NPs, ii) effect of ionic strength on homo-assembly of gold nanorods (GNRs). First, we present hetero-assembly of GNRs and semiconductor quantum dots (QDs) in a chain using biotin-streptavidin interaction. We synthesized alloyed CdTeSe QDs and modified them with mercaptoundecanoic acid to render them water-soluble and to attach streptavidin. We synthesized GNRs by a seed-mediated method and selectively modified the ends with biotin. Hetero-assembly of QDs and GNRs depended on the size, ligands, and ratio of QDs and GNRs. Second, we controlled the rate of homo-assembly of GNRs by varying the ionic strength of the DMF/water solution. The solubility of polystyrene on the ends of GNRs depended on the ionic strength of the solution, which correlated with the rate of assembly of GNRs into chains.

The objectives of this thesis were to optimize the synthesis and surface coating of metallic and semiconductor nanoparticles, to understand how these materials interact with cells and physiological systems and to investigate how they can be used to deliver thermal therapy for medical applications. Reproducible high-yield synthesis of gold nanorods and surface coating with a variety of polymers and silica was optimized. Using gold nanorods as a model system, the relationship between particle surface chemistry, surface charge and cellular uptake was studied, as well as the toxicity of nanoparticles of different surface chemistry. Low toxicity in vitro was encouraging and was confirmed in vivo by intravenously injecting Sprague-Dawley rats with semiconductor quantum dots of various surface coatings. Low toxicity was found during biochemical, haematological and pathological assessment, and these results indicate that applications of nanoparticles should be further investigated. One such application is the use of near infrared absorbing gold nanorods in remotely activated hyperthermia. It was shown that gold nanorods act synergistically with the chemotherapeutic cisplatin to improve cytotoxicity, and reduce the required cytotoxic drug dose to 33% of the unheated amount. Due to the success of hyperthermia treatment in vitro, continuing and future work involves the use of gold nanorods ex vivo on excised human corneas in a novel application to weld corneal tissue for improved wound closure following cataract surgery.

Diabetes mellitus is a potentially lethal disease that affects 7.6 percent of American people. In the US, it is recognized as the 6th leading cause of death. Failure to control blood glucose levels (BGL) in patients with either type of diabetes can lead to other serious complications as well, such as loss of limb, blindness and other health problems. Controlling and monitoring the BGL in post-op and intensive care patients in the hospital is also vital to their health. Currently the most reliable method of monitoring BGL is through an invasive procedure which monitors the amount of glucose in blood directly. A non-invasive glucose sensor would drastically improve the treatment of sensitive patients, and serve to improve the quality of diabetic patients' lives. This glucose sensor is strongly based upon the mercury sensor developed by F.E. Hernandez and his colleagues. Glucose is used as a reducing agent to reduce mercury from Hg2+ to Hg0, which will form amalgams with the gold nanorods in solution. The change in aspect ratio of gold nanorods leads to a change in the UV-Visible spectrum of the solution. The blue shift seen was measured and correlated with the glucose concentration of the system. The system was then tested varying conditions such as pH, temperature, gold nanorod concentration, and mercury concentration. A preliminary study of the kinetics of the reaction was also done. The results showed a limit of detection of 1.58x10-13 and a linear dynamic range covering the concentrations of human tear glucose levels that are currently cited in the literature.

Nanomaterials such as carbon nanotubes, gold nanorods, magnetic nanocrystals, and binary nanocrystal superlattices have exciting potential applications. However, before these ideas can be applied, it is imperative to fully understand the materials synthesis. Multiwall carbon nanotubes were synthesized in supercritical toluene using cobaltocene, nickelocene, ferrocene, or metal nanocrystals as catalysts. Toluene served as both the solvent and carbon source for nanotube growth. The reaction was optimized by introducing supplemental carbon sources; either hexane or ethanol increased the yield relative to pure toluene and catalytic amounts of water minimized carbon filament and amorphous carbon formation. Gold nanorods were synthesized by the colloidal seed-mediated, surfactantassisted approach using cetyltrimethylammonium bromide (CTAB) obtained from ten different suppliers. The gold nanorod yield depended strongly on the CTAB used: with the same recipe, three of the CTABs produced only spherical particles, whereas the other CTABs produced nanorods with nearly 100% yield. Inductively coupled plasma mass spectrometry revealed a trace iodide impurity in the CTABs that did not yield nanorods. Further experiments introducing potassium iodide to the nanorod synthesis verified the detrimental effect of iodide on nanorod formation. Multifunctional colloidal core-shell nanoparticles of magnetic nanocrystals or gold nanorods coated with a fluorescent dye (Tris(2,2 -bipyridyl)dichlororuthenium(II) hexahydrate) doped silica shells were also synthesized. The as-prepared magnetic nanocrystals were initially hydrophobic and silica coated using a microemulsion approach, while the gold nanorods were hydrophilic and silica coated using a Stöber process. These colloidal heterostructures have the potential to be used as dual-purpose tags, exhibiting a fluorescent signal that could be combined with either dark-field optical contrast or enhanced contrast in magnetic resonance imaging. Binary superlattices (BSLs) of large iron oxide and small gold nanocrystals were assembled by slow evaporation of colloidal dispersions on tilted substrates. SEM and grazing incidence small angle X-ray scattering (GISAXS) confirmed the BSLs were simple hexagonal AB2 superlattices with long range order. GISAXS also revealed that the superlattice was slightly contracted perpendicular to the substrate as a result of solvent drying during the deposition process. Additionally, in some BSLs nearly periodic superlattice dislocations consisting of inserted half-planes of gold nanocrystals were observed. / text

Cette thèse comporte trois parties principales : la première concerne la synthèse et la fonctionnalisation des nanoparticules (NPs) d’or de forme sphérique et cylindrique. Les NPs d’or sont obtenues par réduction d’un sel métallique. En faisant varier certains paramètres de synthèse tels que la concentration en sels et/ou la concentration en réducteur, on peut facilement ajuster la taille de NPs sphériques. Bien qu’il existe diverses méthodes permettant de préparer des NPs cylindriques, il subsiste un problème de reproductibilité basée sur la pureté des produits de synthèse. Les différentes analyses de CTAB nous ont permis de proposer des solutions à ce problème. La fonctionnalisation de la surface des NPs d’or permet d’apporter à ces dernières de nouvelles propriétés tout en conservant leurs propriétés intrinsèques. Le but de cette fonctionnalisation est de modifier la surface des NPs en fonction des applications visées. Nous présentons ainsi différentes études de fonctionnalisation par des polyélectrolytes (PEI et PSS), des couches oxydes de SiO2, et de TiO2 et par le TDBC. La réponse optique des NPs d’or étant sensible à l’indice de réfraction du milieu environnant, il est possible de suivre et de quantifier cette fonctionnalisation par spectroscopie d’absorption. Dans une seconde partie, nous avons développé une voie de synthèse simple permettant de préparer le cœur coquille Au@TDBC sans ajout supplémentaire de sels ou de bases et à température ambiante. Etant donné qu’il est possible de moduler la taille des particules, l’optimisation du couplage fort entre les transitions électroniques du TDBC et les modes plasmon de résonances des Nps d’Au a été obtenu correspondant à une énergie de Rabi de 220 meV valeur qui n’a pas encore été obtenue avec un tel système. Dans une troisième partie, nous avons développé une nouvelle approche basée sur la méthode de Stöber pour fonctionnaliser les NPs avec des couches d’oxyde de SiO2 et de TiO2. L’utilisation des systèmes Au@TiO2 est une perspective intéressante en photocatalyse car le contact entre le métal et le semi-conducteur devrait entrainer une nette augmentation de l’efficacité photocatalytique. En effet, le métal agit comme un réservoir de photoélectrons améliorant le transfert de charges interfaciales tout en retardant la recombinaison des paires électrons-trous photo excités du semi-conducteur. / This thesis has three main parts: the first part relates to the synthesis and the functionalization of spherical gold nanoparticles (NPs) and nanorods. Gold NPs are obtained by chemical reduction of gold salt. By varying some synthesis parameters such as gold salt concentration and or reducing agent concentration, we can easily adjust the size of gold NPs. Different methods can be used to prepare gold nanorods, but there is a problem of reproducibility. This problem is based on the purity of products used to prepare gold nanorods. Different analysis of CTAB allowed us to propose solutions to this problem. Functionalization brings to gold NPs new properties. The aim of the functionalization is to modify the surface of gold NPs based on the intended applications. We present various functionalization of Au NPs by polyelectrolytes (PEI and PSS), oxides (SiO2 and TiO2) and TDBC. The optical response of Au NPs is sensitive to the refractive indexe of the surrounding medium, it is then possible to monitor and quantify this functionalization by absorption spectroscopy. In the second part of this work, we have developed a simple synthetic route for preparing Au@TDBC core shell without additional salts or bases at room temperature. Since it is possible to vary the particle size, optimization of the strong coupling between the electronic transitions of TDBC and the resonance plasmon of Au Nps obtained correspond to Rabi energy of 220 meV value which has not yet been achieved with such a system. In the third part, we have developed a new approach based on the Stöber method to functionalize the NPs with an oxide layer of SiO2 and TiO2. The use of Au@TiO2 core shell system is an interesting perspective in photocatalysis because the contact between metal and semiconductor should cause a marked increase in the photocatalytic efficiency. Indeed, the metal acts as a reservoir of photoelectrons improving the interfacial charge transfer while retarding the recombination of electron-hole pair of the semiconductor.

Embora exista considerável progresso na área de biossensores, ainda é primordial aprimorar muitos desses dispositivos. Este trabalho tem por objetivo contribuir para o contínuo crescimento dos biossensores a base de enzimas e de moléculas de DNA, sendo dois biossensores eletroquímicos descritos. O primeiro trata-se de um biossensor enzimático utilizando tirosinase (Tyr) imobilizada por intermédio de cistamina (CYS) e glutaraldeído (GA) para a detecção de compostos fenólicos. Eletrodos de carbono impressos (SPCE) foram modificados com nanobastões de ouro (AuNRs) em filme de poli(amido amina) (PAMAM) geração 4 para o favorecimento da transferência direta de elétrons (DET) entre o eletrodo e o sítio ativo da enzima. Para caracterizar os AuNRs e AuNRs-PAMAM, espectroscopia de absorção no UV-Visível, espalhamento de luz dinâmico (DLS) e microscopia eletrônica de varredura (SEM) foram empregadas. As etapas do biossensor foram estudadas por voltametria cíclica e linear, amperometria e microscopia de força atômica (AFM) na presença de dois analitos: catecol (CAT) e dopamina (DA). A faixa linear para CAT foi de 2,8 a 30,3 μmol L-1 com um limite de detecção (LD) de 1,0μmol L-1 enquanto para a DA, a faixa linear foi de 27,8 a 448,7 μmol L-1 e LD de 10,0 μmol L-1. Além de apresentar ótima resposta frente a possíveis interferentes, o sensor também mostrou excelente desempenho em amostras reais, que atrelados aos testes de repetibilidade e reprodutibilidade mostram a estabilidade e acurácia do biossensor. O segundo sensor, trata-se de um sensor de DNA impedimétrico em eletrodo de ouro para a detecção da mutação c.68_69del relacionada à predisposição ao câncer de mama. A imobilização da sequência de captura (SH-ssDNA) no eletrodo ocorreu pela ligação ouro-enxofre (Au-S) e o modelo de hibridização escolhido foi a hibridização direta. O genossensor distinguiu eficientemente entre as sequências alvo (tarDNA) e não-complementar (ncsDNA), apresentando faixa linear de 1,0 a 200,0 nmol L-1 e LD de 0,14 nmol L-1. Os resultados sugerem que ambos biossensores têm potencial e que as estratégias propostas são promissoras para o desenvolvimento de outros biossensores. / Despite a considerable progress in the area of biosensors, it is still crucial to improve most of these sensors. This study aims to contribute to the ongoing growth of enzyme- and DNA-based biosensors, being described two electrochemical biosensors. The first one is an enzyme-based biosensor with immobilized Tyrosinase (Tyr), through cystamine (CYS) and glutaraldehyde (GA), for detection of phenolic compounds. Screen-printed carbon electrodes (SPCE) were modified by gold nanorods (AuNRs) stabilized with poly(amide amine) PAMAM generation 4 to facilitate direct electron transfer (DET) between electrode and enzyme active site. AuNRs and AuNRs-PAMAM were characterized using UV-Visible absorption spectroscopy, dynamic light scattering (DLS) e scanning electron microscopy (SEM). Biosensor stages were studied by cyclic and linear voltametry, amperommetry and atomic force microscopy (AFM) and tested agains two analytes: catechol (CAT) and dopamine (DA). Detection limit (LD) for CAT is 1 μmol L-1 and linear range from 2.8 to 30.3 μmol L-1, for DA, LD is 10.0 μmol L-1 and linear range 27.8 to 448.7 μmol L-1. Besides, the biosensor shows great response in the presence of interferents, it also had an excellent performance in real samples that along with repeatability and reproducibility tests indicate stability and accuracy of the biosensor. The second sensor is an impedimetric DNA sensor prepared on gold electrode to detect c.68_69del mutation related to breast cancer predisposition. Capture sequence (HS-ssDNA) immobilization occurred due to gold-sulfur bond (Au-S) and direct hybridization was the chosen hybridization model. The genosensor was able to distinguishing between target sequence (tarDNA) and non-complementary sequence (ncsDNA) and linear range and LD were found to be 1.0 to 200.0 nmol L-1 and 0.14 nmol L-1, respectively. Results suggest both biosensors have potential and proposed strategies are promising for other biosensors development.

- An extensive summary in French is available in Appendix E on page 189 -