The objective of this research was to study the catalytic graphitization of cellulose fibers coated with iron oxide nanoparticles. Bleached cellulose fibers and iron oxide nanoparticles coated cellulose fibers were pyrolyzed at five elevated temperatures. The crystallographic structures of carbon-encapsulated iron oxide nanoparticles were then investigated by the following techniques: Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD), Raman Spectroscopy, Transmission Electron Microscopy (TEM), and Selected-Area Electron Diffraction (SAED). The graphitization of cellulosic fibers was enhanced by the presence of iron oxide nanoparticles. Moreover, iron oxide nanoparticles deposited on cellulosic fiber samples pyrolyzed above 800°C produced graphitic structures. TEM and XRD were performed to identify and characterize the phase transitions of carbon-encapsulated iron oxide nanoparticles after pyrolysis treatment at four temperatures: 500°C, 800°C, 1000°C, and 1600°C. TEM of samples pyrolyzed at or above 800°C showed resulting units were core-shell structures consisting of dark grains and a light matrix with graphitic structure.

This study examines the use of unmodified magnetite nanoparticles (Fe3O4 NPs) for selective extraction and enrichment of the catecholamines dopamine (DA), noradrenaline (NE), and adrenaline (E), prior to analysis using capillary electrophoresis with UV detection. Coordination between Fe3+ on-the-surface Fe3O4 NPs and the catechol moiety of catecholamines enables Fe3O4 NPs to capture catecholamines from an aqueous solution. We obtained maximum loading of catecholamines on the NP surface by adjusting the pH of the solution to 7.0. In addition, catecholamine loading on the Fe3O4 NPs increased in conjunction with NP concentrations. Ligand exchange found H3PO4 to be efficient in the removal of adsorbed catecholamines on the NP surface. Adding 1.2% poly(diallyldimethylammonium chloride) to the background electrolyte caused efficient separation of the liberated catecholamines with baseline resolution within 20 min. Under optimal extraction and separation conditions, the limit of detections at a signal-to-noise ratio of 3 for E, NE, and DA were 9 nM, 8 nM, and 10 nM, respectively. Significantly, we successfully used the combination of a phenylboronate-containing spin column and the proposed method to determine the concentrations of NE and DA in urine and the content of NE in Portulaca oleracea L. leaves.

An essential component of developing successful neural stem cell (NSC)-based therapies involves the establishment of methodologies to noninvasively monitor grafted NSCs within brain tissues in real time. In this context, ex vivo labeling with ultrasmall superparamagnetic iron oxide (USPIO) particles has been shown to enable efficient tracking of transplanted NSCs via magnetic resonance imaging (MRI). However, whether and how USPIO labeling affects the intrinsic biology of NSCs is not thoroughly understood, and remains an active area of investigation. Here, we perform a comprehensive examination of rat NSC survival and regenerative function upon labeling with the USPIO, Molday ION Rhodamine B (MIRB), which allows for dual magnetic resonance and optical imaging. After optimization of labeling efficiency, two specific doses of MIRB (20 and 50 mu g/mL) were chosen and were followed for the rest of the study. We observed that both MIRB doses supported the robust detection of NSCs, over an extended period of time in vitro and in vivo after transplantation into the striata of host rats, using MRI and post hoc fluorescence imaging. Both in culture and after neural transplantation, the higher 50 mu g/mL MIRB dose significantly reduced the survival, proliferation, and differentiation rate of the NSCs. Interestingly, although the lower 20 mu g/mL MIRB labeling did not produce overtly negative effects, it increased the proliferation and glial differentiation of the NSCs. Additionally, application of this dose also changed the morphological characteristics of neurons and glia produced after NSC differentiation. Importantly, the transplantation of NSCs labeled with either of the two MIRB doses upregulated the immune response in recipient animals. In particular, in animals receiving the 50 mu g/mL MIRB-labeled NSCs, this immune response consisted of an increased number of CD68(+)-activated microglia, which appeared to have phagocytosed MIRB particles and cells contributing to an exaggerated MRI signal dropout in the animals. Overall, these results indicate that although USPIO particles, such as MIRB, may have advantageous labeling and magnetic resonance-sensitive features for NSC tracking, a further examination of their effects might be necessary before they can be used in clinical scenarios of cell-based transplantation.

The main challenge in disease treatment is no more the discovery of new therapeutic drugs, but to provide targeted delivery of therapeutic drugs to specific sites without incurring systemic toxicity effects. An efficient way of reducing the toxicity is by encapsulating the drug with a biodegradable matrix that can provide controlled release of the drug along with local heating of the drug. Local heating can be obtained by incorporating magnetic iron oxide particles that heat upon exposure to AC electromagnetic fields. The magnetic iron oxide nanoparticles are also gaining much attention as MRI contrast agents. Thus it would be of potential benefit if a drug delivery system is designed to encapsulate the drug as well as the magnetic iron oxide nanoparticles within a biodegradable matrix, thereby providing a dual modal imaging and therapeutic delivery system. The key step in the design of a dual modal drug delivery system is the encapsulation of the magnetic iron oxide nanoparticles with polymer of choice. The magnetic iron oxide nanoparticles were encapsulated into a robust poly (styrene-co-vinylbenzylchloride-co-divinylbenzene) (PSVBDVB) to study these synthetic variations upon encapsulation with a polymer. The next step to the design of drug delivery system was to replace the PSVBDVB polymer by a biocompatible and biodegradable polymer- Poly (lactide-co-glycolide) (PLGA). The PLGA composites containing the Fe@FeOx core shell nanoparticles and the drug analog [Ru(bpy) dye] was prepared by oil-in water emulsion solvent evaporation technique. The local heating of the PLGA composites was also achieved by irradiating the Fe@FeOx nanoparticles with 2.45 GHz microwave radiations. Higher Ru(bpy) dye release from the composites by locally heating the sample with 2.45 GHz microwave pulse compared to externally heating the composite sample was achieved. The final step was the design of controlled release drug delivery system with dual modal imaging and therapeutic capabilities. To obtain narrow sized PLGA composites the Fe@FeOx nanoparticles were replaced by chloroform based ferrofluid. The ferrofluid was synthesized by novel thermolysis technique. The release of the dye from the PLGA composites when placed in the Rf induction coil was determined by fluorescence spectroscopy and a linear increase in the fluorescent intensity was observed with time. Also, the controlled release of the dye from the composites was achieved by a pulsed Rf treatment. Magnetic resonance imaging was also performed using the PLGA composites which showed enhancement in the T2-weighted image contrast and thus negligible reduction in the contrast capabilities of the iron oxide particles (R2 = 58.7 s-1mM-1). The PLGA composites containing the drug analog and the iron oxide nanoparticles thus constitute a controlled release drug delivery system with dual modal imaging and therapeutic capabilities.

In this dissertation, research efforts mainly focused on exploring the applications of superparamagnetic iron oxide nanoparticles (SPIONs) in MR imaging and nanocatalysis via surface functionalization. A dopamine-based surface-functionalization strategy was established. The Simanek dendrons (G1 to G3), oligonucleotides and amino acids were loaded onto SPION surfaces via this approach to develop pH-sensitive MRI contrast agents, specific-DNA MR probes and a biomimetic hydrolysis catalyst. Dendron-SPION conjugates (G1 to G3) have good aqueous solubilities and high transverse relaxivities (>300 s-1*mM-1). They also showed interesting strong pH-sensitive R2 and R2* relaxivities, which were governed by the clustering states of dendron-SPIONs in different pH environments. Values of R2m and R2* m/R2m varied by over an order of magnitude around pH 5. The efficient cell-uptake (~3 million/cell) and low cytotoxicity of G1 to G3-SPIONs were demonstrated on HeLa cell cultures. The strong R2* effects were observed indicating the SPION clustering in HeLa cells. Two SPION-oligonuleotide conjugates were synthesized by coupling two half-match oligonucleotides onto domapine-capped SPIONs via SPDP linkers. They served as MR probes to detect a single-strand DNA with the same sequence to miRNA-21 based on the change of R2 values due to the DNA-bridged SPION clustering. The detection limit of the DNA could reach to 16.5 nM. A biomimetic hydrolysis nanocatalyst (i.e., Fe2O3-Asp-His complex) was developed by loading Asp and His-dopamine derivatives onto SPIONs. Paraoxon and nitrophenyl acetate were hydrolyzed under a mild condition (neutral pH, 37 °C) catalyzed by the Fe2O3-Asp-His complex. The two amino acids Asp and His cooperated with each other on the SPION surfaces to catalyze hydrolysis reactions. This catalyst could be recycled by a magnet and reused for four times without a significant loss of catalytic activity.

Title: Nanoparticles based on 3d metal oxides - correlation of structure and magnetism Author: RNDr. Simona Kubíčková Department: Institute of Physics CAS, v.v.i. Supervisor: doc. RNDr. Jana Kalbáčová Vejpravová, Ph.D., Institute of Physics CAS, v.v.i. Abstract: The thesis is focused on the correlation of the magnetic response of iron oxide nanoparticles (NPs) with their internal structure. Several complementary methods were used and compared that bring insight into the relative crystallinity of the investigated NPs. The main goal was devoted to the elucidation of the origin of the so-called spin canting angle determined by In-field Mössbauer Spectroscopy (IFMS) by examination of samples with different internal structure. It has been observed that the IFMS is not an unambiguous method to study the surface effects in the NPs as the IFMS is sensitive only to the average value of all spins and does not distinguish between the surface and core effects. Moreover, the IFMS was performed on the epsilon phase of the iron(III) oxide NPs in order to inspect the peculiar behavior of this phase in an external magnetic field. Keywords: iron oxide nanoparticles, magnetism, In-field Mössbauer Spectroscopy, spin canting

Cell surface proteins play an important role in understanding cell-cell communication, cell signaling pathways, cell division and molecular pathogenesis in various diseases. Commonly used biotinylation regents for cell surface mapping have shown some potential drawbacks such as crossing the cell membrane, difficult recovery of biotinylated proteins from streptavidin/avidin beads, interference from endogenous biotin and nonspecific nature of streptavidin. With aim to solve these problems, we introduced sulfo-N-hydroxysuccinimidyl (NHS) ester functionalized magnetic nanoparticles containing cleavable groups to label solvent exposed primary amine groups of proteins. Silica coated iron oxide magnetic nanoparticles (Fe3O4@SiO2 MNPs) were linked to NHS ester groups via a cleavable disulfide bond. Additionally, the superparamagnetic properties of Fe3O4@SiO2 MNPs facilitate efficient separation of the labeled peptides and removal of the detergent without any extra step of purification. In the last step, the disulfide bond between the labeled peptides and MNPs was cleaved to release the labeled peptides. The disulfide linked NHS ester modified Fe3O4@SiO2 MNPs were tested using a small peptide, and a model protein (bovine serum albumin) followed by liquid chromatography-tandem mass spectrometry analysis (LC-MS/MS) of labeled peptides. In the next step, disulfide linked, NHS ester modified Fe3O4@SiO2 MNPs (150 nm) successfully labeled the solvent exposed cell surface peptides of Saccharomyces cerevisae. Electron microscopic analysis confirmed the cell surface binding of NHS ester modified Fe3O4@SiO2 MNPs. Mass spectrometric analysis revealed the presence of 30 unique proteins containing 56 peptides. Another MNPs based labeling reagent was developed to target solvent exposed carboxyl acid residues of peptides and proteins. The surface of Fe3O4@SiO2 MNPs was modified with free amine groups via a disulfide bond. Solvent exposed carboxyl groups of ACTH 4-11 and BSA were labeled by using1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) chemistry. Upon cleaving the disulfide bond, labeled peptides were analyzed by LC-MS/MS. The MNPs containing labeling reagents offers specific labeling under physiological conditions and rapid magnetic separation of labeled peptides prior to mass spectrometric analysis. The ability of large Fe3O4@SiO2 MNPs to specifically attach to cell surface makes them a potential candidate to study the surface of variety of different cell types and complex proteins surrounded by lipid bilayer.

Health-care associated infections (HAIs) on medical implant surfaces present a unique challenge to physicians due to their existence in the biofilm phenotype which defends the pathogen from antibiotics and the host’s own immune system. A 2004 study in the U.S. showed that 2 to 4% of implanted devices become infected and must be treated via surgical explantation—a process that is both expensive and dangerous for the patient. A potential, alternative strategy to antibiotics and surgery is to use heat delivered wirelessly by a magnetic coating. This thermal treatment strategy has the potential to kill these HAIs directly on the implanted surface and without the patient requiring surgery. This thesis introduces an iron oxide nanoparticle composite coating that is wirelessly heated using energy converted from an alternating magnetic field. Iron oxide nanoparticle composites are demonstrated to be remotely heated in both hydrophilic and hydrophobic polymer composites. In designing the composite coating, multiple parameters were investigated for how they impact the normalized heating rate of the material. Specifically, the amount of iron in the coating, the coating thickness, the polymer type, and the orientation of the coating relative to the applied magnetic field were investigated. Power output was shown to increase proportionally with iron loading whereas nearly two times the amount of power output was observed for the same coatings positioned parallel to magnetic field lines versus those positioned perpendicular—a result believed to be due to magnetic shielding from neighboring particles. Microscope slides coated with 226 µm of composite delivered up to 10.9 W cm⁻² of power when loaded with 30.0% Fe and positioned parallel in a 2.3 kA m⁻¹AMF. Pseudomonas aeruginosa biofilms were grown directly on these coatings and heated for times ranging from 1 to 30 min and temperatures from 50 to 80 °C. Less than one order of magnitude of cell death was observed for temperatures less than 60 °C and heat shock times less than 5 min. Up to six orders of magnitude reduction in viable bacteria were observed for the most extreme heat shock (80 °C for 30 min). Introducing this wirelessly heated composite into the body has the potential to kill harmful bacteria but at the risk of thermally damaging the surrounding tissue and organs if the treatment is not designed and predicted intelligently. Thermal energy will propagate differently depending on the surrounding heat sink, with convective heat sinks (i.e. those due to blood flow) requiring much more power to reach the same surface temperature than a conduction-only heat sink. To study how heat is transferred in biological tissues, a robust, poly(vinyl alcohol) tissue phantom was developed that can be poured to accommodate any geometry, is volume stable in water and under thermal stress, and can be modified with inert particle fillers to adjust its thermal conductivity from 0.475 to 0.795 W m⁻¹°C⁻¹. In vitro heat transfer was measured through this hydrogel tissue phantom with at least 10 °C of temperature rise, penetrating 5 mm of tissue in less than 120 sec for an 80 °C boundary condition. A computational model was used to solve three-dimensional energy transfer through a combined fluid mimic/tissue mimic heat sink spanning the same surface boundary condition. The model was validated with experimental models using a custom designed heat transfer station. This scenario is applicable in the instance where the same coating is subject to starkly different heat sinks: half subject to convective heat loss, half to conductive heat loss. Based on these conditions, a magnetic coating would need to be designed that has a power gradient up to 15 times larger on the fluid half versus the other.

Aquesta tesi engloba la síntesi i la caracterització estructural i magnètica de dos tipus de nanopartícules polymagnètiques: estructures nucli-escorça (core|shell, CS) i tipus ceba (onion-like). El primer sistema està format per un nucli-MnO|escorça-Mn 3O4 ( -Mn2O3) amb doble inversió, on el nucli i l'escorça mostren un comportament AFM i FiM, respectivament . Es de neix com estructuralment inversa ja que l'AFM es localitza al nucli i el FiM a l'escorça; a més, també es troba magnèticament invertida, és a dir, la temperatura de Néel de l'AFM presenta valors més elevats que la temperatura de Curie del FiM. Les nanopartícules nucli-escorça de MnO|Mn3O4 ( -Mn2O3) s'han obtingut a través de la passivació controlada de l'escorça de nanopartícules de MnO prèviament sintetitzades. Aquest procés permet controlar tant la grandària del nucli com el gruix de l'escorça. Es va con rmar que la composició de l'escorça un cop passivada depèn de la grandària inicial de les nanopartícules; conseqüentment, les nanopartícules més grans estan formades principalment per Mn3O4. No obstant, a mesura que es disminueix la grandària, la densitat de defectes augmenta obtenint, d'aquesta manera, una escorça més estable formada per la fase -Mn2O3. D'altra banda, nuclis AFM de MnO relativament petits poden induir un efecte magnètic de proximitat (magnetic proximity e ects) a l'escorça de FiM -Mn2O3 tot mantenint el seu ordre magnètic molt per sobre de la seva temperatura de Curie, TC; a més, aquest sistema presenta un augment de la temperatura de Néel de l'AFM. El segon sistema es basa en la síntesi de nanopartícules d'òxid de manganès i ferro del tipus nucli-escorça i ceba. Nanopartícules de dos òxids de ferro diferents (FeO|Fe3O4 CS i Fe3O4 monofàsica) s'han utilitzat com a llavors per a la posterior deposició d'òxid de manganès. A partir de les llavors nucli-escorça d'òxid de ferro s'han sintetitzat dos tipus de nanopartícules ceba (tres-components FeO|Fe 3O4|Mn3O4 i quatre-components FeO|Fe3O4|MnO|Mn3O4). D'altra banda, nanopartícules d'òxid de ferro monofàsiques han estat utilitzades com a llavors per a dipositar una capa na de manganès al seu voltant amb l'objectiu d'incentivar l'interdifusió del manganès-ferro i formar nanopartícules nucli-escorça de MnxFe3 ��xO4|FexMn3 ��xO4 amb una interfase graduada. S'ha observat que l'òxid de manganès creix epitaxialment en els plans (111) sobre les cares truncades del llavors cúbiques d'oxid de ferro. Finalment, nanopartícules nucli-escorça de MnxFe3 ��xO4|FexMn3 ��xO4 formades per una estructura tou-FiM/dur- FiM amb una composició gradual a l'interfase demostren un bon l'acoblament magnètic entre ambdues, tova i dura, fases FiM. / This thesis deals with the synthesis and magnetic and structural characterization of two di erent systems based in polymagnetic nanoparticles with core|shell (CS) and onion-like architectures. The rst system is formed by a double inverted core-MnO|shell- Mn3O4 ( -Mn2O3) where core and shell display an antiferromagnetic (AFM) and ferrimagnetic (FiM) behavior, respectively. It is de ned as structurally inverted since the AFM is placed in the core and the FiM in the shell (in contrast to conventional ferromagnetic( FM)/AFM CS systems); in addition, it is, also, magnetically inverted because the Néel temperature of the AFM is larger than the Curie temperature of the FiM (contrarily to standard exchange bias systems). MnO|Mn3O4 ( -Mn2O3) CS nanoparticles have been synthesized through the controlled shell passivation of pre-made MnO nanoparticles. This procedure allows reaching a good control over the nal core size and shell thickness. It was con rmed that the passivated shell composition depends on the nanoparticle size, where the larger nanoparticles presenting mainly Mn3O4. However, when the size diminishes, the density of defects in the MnO core increases and consequently -Mn2O3 is the more stable shell phase. Besides, small AFM MnO cores can induce a magnetic proximity e ect to the FiM -Mn2O3 shell, maintaining its magnetic order well above its Curie temperature, TC. Moreover, surface e ects in the MnO core can also lead an increase of the Néel temperature of the AFM. Further, given the AFM/FiM exchange coupling the system exhibits large coercivities and loop shifts along the eld axis, i.e., exchange bias. The second type of system comprises the synthesis of CS and onion-like nanoparticles based in manganese and iron oxides. Two di erent iron oxide nanoparticles (FeO|Fe3O4, AFM|FiM, CS and single phase FiM Fe3O4) have been used as seeds for the posterior manganese oxide deposition. From iron oxide CS seeds two di erent onion-like nanoparticles (three-components FeO|Fe3O4|Mn3O4 and four-components FeO|Fe3O4|MnO|Mn3O4) have been synthesized. The temperature dependence of the magnetization of these onion nanoparticles exhibits several magnetic transitions, in concordance with the presence of diverse magnetic phases. In addition, single phase iron oxide seeds were employed to deposit a manganese thin shells at high temperatures forcing a manganese-iron interdi usion to form the nal MnxFe3 �xO4|FexMn3 �xO4 CS nanoparticles with a graded interphase. The structural results show that the (111) planes of the manganese oxide grow epitaxially onto the (111) planes of the truncated faces of the initial cubic iron oxide seeds. Finally, the CS MnxFe3 �xO4|FexMn3 �xO4 nanoparticle, formed by soft-FiM|hard-FiM structure with a graded interphase composition, shows a strong exchange coupling between the hard and soft FiM phases.

The first study reports the development of a reusable, single-step system for the detection of specific substrates using oxidase-functionalized Fe3O4 nanoparticles (NPs) as a bienzyme system and using amplex ultrared (AU) as a fluorogenic substrate. In the presence of H2O2, the reaction pH between Fe3O4 NPs and AU was similar to the reaction of oxidase and the substrate. The catalytic activity of Fe3O4 NPs with AU was nearly unchanged following modification with poly(diallyldimethylammonium chloride) (PDDA). Based on these features, we prepared a composite of PDDA-modified Fe3O4 NPs and oxidase for the quantification of specific substrates through the H2O2-mediated oxidation of AU. By monitoring fluorescence intensity at 587 nm of oxidized AU, the minimum detectable concentrations of glucose, galactose, and choline were found to be 3, 2, and 20 £gM using glucose oxidase-Fe3O4, galactose oxidase-Fe3O4, and choline oxidase-Fe3O4 composites, respectively. The identification of glucose in blood was selected as the model to validate the applicability of this proposed method. The second study follows the first one. Using the catalytic activity of Fe3O4 NPs with AU to detect four kinds of neurotransmitter, such as dopamine, L-DOPA, adrenaline (epinephrine) and noradrenaline (norepinephrine). Because of there is specific interaction between Fe3O4 NPs and catecholamines (CAs), the Fe3O4 NPs will form CAs-Fe3O4 NPs composites in presence of CAs. The CAs on the Fe3O4 NPs surface must shelter the reaction between AU and H2O2, cause the fluorescence to be turned-off. The CAs just like a inhibitor, to inhibit the catalytic activity of Fe3O4 NPs. Therefore, we could use this inhibited system to detect the CAs compound concentration in the real sample.