Synthesis and Surface Modification of Inorganic Nanoparticles for Application in Physics and Medicine

The core focus of this cumulative thesis is the synthesis, the characterization, and the polymer coating or the surface modification of different types of inorganic nanoparticles (NPs), e.g., semiconductor, magnetic, plasmonic, and titanium oxide NPs. These NPs are used in the field of physics, biotechnology, and in nanomedicine or life sciences for both diagnosis and therapy. The applications of these NPs depend on their unique properties, which are correlated to their size, shape, and the material composition. The colloidal stability of these nanocrystals or NPs in different media (e.g. organic, water, cell culture media) was achieved by means of capping agents or by wrapping suitable ligands or surfactants around the core of the NPs. The colloidal NPs that were synthesised during this research work were capped with hydrophobic ligands (e.g. oleic acid, oleylamine, etc.) to keep them stable in the organic media, e.g., toluene, chloroform, etc. The phase transfer from organic to aqueous is a mandatory step prior to their use in the few desired applications, especially when these NPs are exposed to aqueous medium or cell media. This is carried out by wrapping the NPs with an amphiphilic polymer, i.e., poly(isobutylene-alt-maleic anhydride) (Mw= 6000 Da) that is grafted with hydrophobic side chains of dodecylamine. The mentioned four types of produced NPs were: (i) Semiconductor NPs which include the hydrophobic cadmium sulfide (CdS) quantum dots (QDs) that are used: for organic scintillation neutrino detection experiments; for PPO (2, 5-diphenyloxazole) styrene based plastic scintillator detectors; for time resolved spectral measurement, and for fluorescence studies with different surface coatings; additionally, water soluble CdS, manganese doped CdS, and zinc sulphide (ZnS) with and without manganese doping were synthesized and engineered to run several experiments on nanomaterials’ (NMs) behavior in environmental media, e.g., river and lake water; (ii) magnetic NPs (MNPs) that include core only (iron oxide, e.g. magnetite) and core shell composite iron oxide magnetic NPs combined with cobalt and manganese ferrites; (iii) plasmonic NPs such as gold and silver NPs that were used in combination with iron-oxide NPs (4 nm each) for toxicity screening and dose determination assays, and (vi) titanium dioxide iv (TiO2) NPs with different sizes and shapes (i.e. cube, rods, plates, and bipyramids), which were used for in vivo experiments: To evaluate the bio-distribution, organ accumulation, biological barrier passage, and potential organ toxicity after a single intravenous administration of TiO2 NPs, and to assess the influence of the TiO2 NPs shape and geometry on the mentioned effects. Furthermore TiO2 NPs were also used to perform few more in vivo studies to investigate: (i) The effect of biological environment (e.g. lung lining liquid, saliva, gastric/intestinal fluids) on NPs’ behaviour and toxicity, using complex co-culture systems for the intestine and alveoli, (ii) the effect of NPs on the activation of the inflammasome, and (iii) the influence of NPs on the maturation and activation of dendritic cells. In addition to above mentioned experiments for synthesis and surface modification another study was carried out with the aim to transfer three different types of NPs (i.e. plasmonic, fluorescent and magnetic) in aqueous phase to be employed in hydrogels, aerogels, and heterogels applications. In this study bimetallic (gold-copper) plasmonic nanocubes, fluorescent (cadmium selenide/CdS) core shell nanorods and magnetic iron oxide (Fe3O4) nanospheres were successfully transferred to the aqueous phase irrespective of their different sizes ranging from 5-40 nm in at least one dimension. All water soluble NPs were cleaned by means of gel electrophoresis or by ultracentrifugation to get rid of micelles (empty polymer) followed by sterilization for all in vivo studies. The qualitative and quantitative analyses all of these NPs were performed by means of different characterization techniques, e.g., ultraviolet-visible spectroscopy, fluorescence spectroscopy, dynamic light scattering, zeta potential measurements gel electrophoresis, transmission electron microscopy, inductively coupled plasma mass spectrometry, and the X-ray diffraction analysis

The impact of species and cell type on the nanosafety profile of iron oxide nanoparticles in neural cells

Background: While nanotechnology is advancing rapidly, nanosafety tends to lag behind since general mechanistic insights into cell-nanoparticle (NP) interactions remain rare. To tackle this issue, standardization of nanosafety assessment is imperative. In this regard, we believe that the cell type selection should not be overlooked since the applicability of cell lines could be questioned given their altered phenotype. Hence, we evaluated the impact of the cell type on in vitro nanosafety evaluations in a human and murine neuroblastoma cell line, neural progenitor cell line and in neural stem cells. Acute toxicity was evaluated for gold, silver and iron oxide (IO) NPs, and the latter were additionally subjected to a multiparametric analysis to assess sublethal effects. Results: The stem cells and murine neuroblastoma cell line respectively showed most and least acute cytotoxicity. Using high content imaging, we observed cell type-and species-specific responses to the IONPs on the level of reactive oxygen species production, calcium homeostasis, mitochondrial integrity and cell morphology, indicating that cellular homeostasis is impaired in distinct ways. Conclusions: Our data reveal cell type-specific toxicity profiles and demonstrate that a single cell line or toxicity end point will not provide sufficient information on in vitro nanosafety. We propose to identify a set of standard cell lines for screening purposes and to select cell types for detailed nanosafety studies based on the intended application and/or expected exposure

Halbleiternanopartikel-modifizierte Elektrode zum Nachweis von Substraten von NADH-abhängigen Enzymreaktionen

Es wurde ein Elektrodensystem entwickelt, das aufbauend auf Halbleiternanopartikeln (so genannte Quantenpunkte) die sensitive Detektion des Enzymkofaktors NADH (nicotinamide adenine dinucleotide) erlaubt. Kolloidale halbleitende CdSe/ZnS-Nanokristalle sind durch ein Dithiol über Chemisorption an Gold gebunden. Das Stromsignal kann durch die Beleuchtung der Quantenpunkt modifizierten Oberfläche beeinflusst werden. Durch Photoanregung entstehen Elektron-Loch- Paare in den Nanopartikeln, die als anodischer oder kathodischer Photostrom detektiert werden können. Die Immobilisierung der Nanokristalle ist durch amperometrische Photostrom- und Quarzmikrowaage-Messungen (quartz crystal microbalance) verifiziert. Diese Studie zeigt, dass CdSe/ZnS-Quantenpunktmodifizierte Elektroden eine konzentrationsabhängige NADH-Detektion im Bereich von 20μM bis 2mM bei relativ niedrigem Potential (um 0V vs Ag/AgCl, 1 M KCl) ermöglichen. Somit können solche Elektroden in Kombination mit NADH-produzierenden Reaktionen für die lichtgesteuerte Analyse der entsprechenden Substrate des Biokatalysators genutzt werden. Es wird gezeigt, dass mit einem solchen Elektrodensystem und Photostrommessungen ein Glukosenachweis möglich ist.An electrode system based on semiconductive nanoparticles (so called quantum dots) was developed which allows the sensitive detection of the enzyme cofactor NADH (nicotinamide adenine dinucleotide). Colloidal semiconductive CdSe/ZnS nanocrystals are bound to gold via a dithiol compound by chemisorption. The current signal can be influenced by illumination of the quantum dot-modified electrode surface. Because of photoexcitation electron-holepairs are generated in the nanoparticles which can be detected as anodic or cathodic photocurrent. The immobilisation of the nanocrystals is verified by photocurrent and quarz crystal microbalance (QCM) measurements. This study shows that CdSe/ZnS-quatum dot-modified electrodes provide a concentration-dependent detection of NADH in the range of 20μM up to 2mM at relatively low overpotentials (around 0V vs Ag/ AgCl, 1 M KCl). Such electrodes can be used in combination with NADH-producing reactions for the lighttriggered analysis of the corresponding substrate of the biocatalyst. The detection of glucose with such an electrode system and photocurrent measurements is shown

Lichtgesteuerter bioelektrochemischer Sensor basierend auf CdSe/ZnS-Quantum Dots

Diese Studie beschäftigt sich mit der Untersuchung der Sauerstoffsensitivität von QD-Elektroden auf Basis von CdSe/ZnS-Nanopartikeln. Das Verhalten des sauerstoffabhängigen Photostroms wurde dabei in Abhängigkeit des pH-Wertes und des Potentials untersucht. Auf Grundlage dieser Sauerstoffabhängigkeit wurde die Enzymaktivität von GOD über Photostrommessungen evaluiert. Für die Konstruktion eines photobioelektrochemischen Sensors, der durch Beleuchtung der entsprechenden Elektrodenfläche ausgelesen werden kann, wurden Multischichten auf die CdSe/ZnS-modifizierten Elektroden aufgetragen. Die Layer-by-Layer Deposition von GOD mit Hilfe des Polyelektrolyten PAH zeigte, dass eine Sensorkonstruktion möglich ist. Die Sensoreigenschaften dieser Elektroden werden drastisch durch die Menge an immobilisiertem Enzym auf der Quantum Dot-Schicht beeinflusst. Durch die Präparation von vier Bilayern [GOD/PAH]4 an CdSe/ ZnS Elektroden kann ein schnell ansprechbarer Sensor für Konzentrationen zwischen 0.1 – 5 mM Glukose hergestellt werden. Dies eröffnet neue Möglichkeiten für die Multianalytdetektion mit nichtstrukturierten Sensorelektroden, lokalisierten Enzymen und räumlich aufgelöster Auslesung durch Licht.This study reports on the oxygen sensitivity of quantum dot electrodes modified with CdSe/ZnS nanocrystals. The photocurrent behaviour is analysed in dependence on pH and applied potential by potentiostatic and potentiodynamic measurements. On the basis of the influence of the oxygen content in solution on the photocurrent generation the enzymatic activity of GOD is evaluated in solution. In order to construct a photobioelectrochemical sensor which can be read out by illuminating the respective electrode area multilayers were build up on the CdSe/ZnS-modified electrodes. The layer-by-layer deposition of GOD by means of the polyelectrolyte PAH show that a sensor construction is possible. The sensing properties of such kind of electrodes are drastically influenced by the amount and density of the enzyme on top of the quantum dot layer. By depositing 4 bilayers [GOD/ PAH]4 on the CdSe/ZnS electrode a fast responding sensor for the concentration range 0.1mM-5mM glucose can be prepared. This opens the door to a multianalyte detection with a non-structured sensing electrode, localized enzymes and spatial read-out by light

Subcellular Carrier-Based Optical Ion-Selective Nanosensors

In this review, two carrier systems based on nanotechnology for real-time sensing of biologically relevant analytes (ions or other biological molecules) inside cells in a non-invasive way are discussed. One system is based on inorganic nanoparticles with an organic coating, whereas the second system is based on organic microcapsules. The sensor molecules presented within this work use an optical read-out. Due to the different physicochemical properties, both sensors show distinctive geometries that directly affect their internalization patterns. The nanoparticles carry the sensor molecule attached to their surfaces whereas the microcapsules encapsulate the sensor within their cavities. Their different size (nano and micro) enable each sensors to locate in different cellular regions. For example, the nanoparticles are mostly found in endolysosomal compartments but the microcapsules are rather found in phagolysosomal vesicles. Thus, allowing creating a tool of sensors that sense differently. Both sensor systems enable to measure ratiometrically however, only the microcapsules have the unique ability of multiplexing. At the end, an outlook on how more sophisticated sensors can be created by confining the nano-scaled sensors within the microcapsules will be given

Mechanistic insights and selected synthetic routes of atomically precise metal nanoclusters

Abstract During the last few decades, noble metal nanoclusters (NCs) have become an exciting building block in the field of nanoscience. With their ultrasmall size that ranges between 1 and 2 nm, NCs fill the gap between atoms and nanoparticles (NPs), and they show significantly different physicochemical properties compared to their bulk counterparts, such as molecule‐like HOMO‐LUMO discrete electronic transitions, photoluminescence, etc. These properties made NCs potential candidates in various applications, including catalysis, chemical/bioimaging, biomedicine, sensing, and energy conversion. Controlling the size of NPs, which usually exhibit a degree of polydispersity, has been a significant challenge for nano‐scientists. However, metal NCs with atomic precision pave the way to accurately fabricate NPs based on an atom‐by‐atom assembly. This Perspective is directed to the community of nano‐scientists interested in the field of NCs and summarizes the most commonly used synthetic routes of atomically precise metal NCs. Moreover, this Perspective provides an understanding of the different techniques used to control the size of metal NCs with insights on switching the surface ligands from phosphine to thiol. This Perspective also explains the role of physicochemical parameters in different synthetic routes such as high‐temperature route, CO‐directed route, solid‐state route, ligand‐exchange‐induced size/structure transformation (LEIST), etc. We finally give a brief outlook on future challenges of currently used synthetic routes with some suggestions to improve them

Synthesis and perspectives of complex crystalline nano‐structures

Research on inorganic colloidal nanocrystals has moved from the synthesis of simple structures, such as spherical nanoparticles, to more elaborate particles with shapes such as rods, stars, discs, and branched nanocrystals, and recently to nanoparticles that are composed out of sections of different materials. Nanocrystal heterostructures represent a convenient approach to the development of nanoscale building blocks, as they merge sections with different functionality in the same particle, without the need of inorganic cross-linkers. The present article gives an overview of synthesis strategies to complex nanocrystals and will highlight their structural properties, as well as discuss some envisaged applications

Synthesis and Functionalization of Monodisperse Near-ultraviolet and Visible Excitable Multifunctional Eu3+, Bi3+:REVO4 Nanophosphors for Bioimaging and Biosensing Applications

Near-ultraviolet and visible excitable Eu- and Bi-doped NPs based on rare earth vanadates (REVO4, RE = Y, Gd) have been synthesized by a facile route from appropriate RE precursors, europium and bismuth nitrate, and sodium orthovanadate, by homogeneous precipitation in an ethylene glycol/water mixture at 120 °C. The NPs can be functionalized either by a one-pot synthesis with polyacrylic acid (PAA) or by a Layer-by-Layer approach with poly(allylamine hydrochloride) (PAH) and PAA. In the first case, the particle size can also be tuned by adjusting the amount of PAA. The Eu- Bi-doped REVO4 based nanophosphors show the typical red luminescence of Eu(III), which can be excited through an energy transfer process from the vanadate anions, resulting in a much higher luminescence intensity in comparison to the direct excitation of the europium cations. The incorporation of Bi into the REVO4 structure shifts the original absorption band of the vanadate anions towards longer wavelengths, giving rise to nanophosphors with an excitation maximum at 342 nm, which can also be excited in the visible range. The suitability of such nanophosphors for bioimaging and biosensing applications, as well as their colloidal stability in different buffer media of biological interest, their cytotoxicity, their degradability at low pH, and their uptake by HeLa cells have been evaluated. Their suitability for bioimaging and biosensing applications is also demonstrated.European Union 267226Ministerio de Economía y Competitividad MAT2014-54852-

Magnetically triggered release of molecular cargo from iron oxide nanoparticle loaded microcapsules

Iron oxide nanocube-modified microcapsules as a platform for magnetically triggered molecular release