Determinants of biological activity of nanomaterials in innate immunity cells: the role of aspect ratio, environmental contaminants and protein corona

As defined by the European Union, “ ’Nanomaterial’ (NM) means a natural, incidental or manufactured material containing particles, in an unbound state or as an aggregate or agglomerate, where, for 50 % or more of the particles in the number size distribution, one or more external dimensions is in the size range 1 nm-100 nm ” (2011/696/UE). Given their peculiar physico-chemical features, nanostructured materials are largely used in many industrial fields (e.g. cosmetics, electronics, agriculture, biomedical) and their applications have astonishingly increased in the last fifteen years. Nanostructured materials are endowed with very large specific surface area that, besides making them very useful in many industrial processes, renders them very reactive towards the biological systems and, hence, potentially endowed with significant hazard for human health. For these reasons, in recent years, many studies have been focused on the identification of toxic properties of nanostructured materials, investigating, in particular, the mechanisms behind their toxic effects as well as their determinants of toxicity. This thesis investigates two types of nanostructured TiO2 materials, TiO2 nanoparticles (NP), which are yearly produced in tonnage quantities, and TiO2 nanofibres (NF), a relatively novel nanomaterial. Moreover, several preparations of MultiWalled Carbon Nanotubes (MWCNT), another nanomaterial widely present in many products, are also investigated.- Although many in vitro and in vivo studies have characterized the toxic properties of these materials, the identification of their determinants of toxicity is still incomplete. The aim of this thesis is to identify the structural determinants of toxicity, using several in vitro models. Specific fields of investigation have been a) the role of shape and the aspect ratio in the determination of biological effects of TiO2 nanofibres of different length; b) the synergistic effect of LPS and TiO2 NP on the expression of inflammatory markers and the role played therein by TLR-4; c) the role of functionalization and agglomeration in the biological effects of MWCNT. As far as biological effects elicited by TiO2 NF are concerned, the first part of the thesis demonstrates that long TiO2 nanofibres caused frustrated phagocytosis, cytotoxicity, hemolysis, oxidative stress and epithelial barrier perturbation. All these effects were mitigated by fibre shortening through ball-milling. However, short TiO2 NF exhibited enhanced ability to activate acute pro-inflammatory effects in macrophages, an effect dependent on phagocytosis. Therefore, aspect ratio reduction mitigated toxic effects, while enhanced macrophage activation, likely rendering the NF more prone to phagocytosis. These results suggest that, under in vivo conditions, short NF will be associated with acute inflammatory reaction, but will undergo a relatively rapid clearance, while long NF, although associated with a relatively smaller acute activation of innate immunity cells, are not expected to be removed efficiently and, therefore, may be associated to chronic inflammatory responses. As far as the relationship between the effects of TiO2 NP and LPS, investigated in the second part of the thesis, are concerned, TiO2 NP markedly enhanced macrophage activation by LPS through a TLR-4-dependent intracellular pathway. The adsorption of LPS onto the surface of TiO2 NP led to the formation of a specific bio-corona, suggesting that, when bound to TiO2 NP, LPS exerts a much more powerful pro-inflammatory effect. These data suggest that the inflammatory changes observed upon exposure to TiO2 NP may be due, at least in part, to their capability to bind LPS and, possibly, other TLR agonists, thus enhancing their biological activities. Finally, the last part of the thesis demonstrates that surface functionalization of MWCNT with amino or carboxylic groups mitigates the toxic effects of MWCNT in terms of macrophage activation and capability to perturb epithelial barriers. Interestingly, surface chemistry (in particular surface charge) influenced the protein adsorption onto the MWCNT surface, allowing to the formation of different protein coronae and the tendency to form agglomerates of different size. In particular functionalization a) changed the amount and the type of proteins adsorbed to MWCNT and b) enhanced the tendency of MWCNT to form large agglomerates. These data suggest that the different biological behavior of functionalized and pristine MWCNT may be due, at least in part, to the different tendency to form large agglomerates, which is significantly influenced by their different capability to interact with proteins contained in biological fluids. All together, these data demonstrate that the interaction between physico-chemical properties of nanostructured materials and the environment (cells + biological fluids) in which these materials are present is of pivotal importance for the understanding of the biological effects of NM. In particular, bio-persistence and the capability to elicit an effective inflammatory response are attributable to the interaction between NM and macrophages. However, the interaction NM-cells is heavily influenced by the formation at the nano-bio interface of specific bio-coronae that confer a novel biological identity to the nanostructured materials, setting the basis for their specific biological activities.

CFD Modellierung einer partikelbelasteten Kühlmittelströmung im Sumpf und in der Kondensationskammer

Der Bericht beschreibt die Arbeiten zur CFD-Modellentwicklung zur Beschreibung des Fasertransportes in einer Wasserströmung, die im Unterauftrag der Hochschule Zittau/Görlitz erfolgten. Während die experimentellen Arbeiten zu dieser Thematik in Zittau durchgeführt wurden, lag der Schwerpunkt der theoretischen Arbeiten in Rossendorf. Im Arbeitspunkt EZ 1 des Projektantrages ist die Erweiterung der Einzeleffektuntersuchungen vorgesehen. Die entsprechenden Modellansätze zum Partikeltransport sind im Kapitel 3.1. beschrieben. Die Modellanpassung und Validierung ist in 3.2 und 3.3 dargestellt. Der Fasertransport in einer Wasserströmung wird durch Jet-Phänomene bestimmt. Untersuchungen dazu sind im EZ3.1 des Projektantrages: „3D-Phänomene infolge Blasenmitriss“ vorgesehen und die Modellansätze und der Vergleich zu Experimenten in den Kapiteln 4.1 bis 4.3 dargestellt. Des Weiteren wird der Einfluss auf den Ausgleich der Temperatur für den Fall untersucht, dass der Jet kälter als die Wasservorlage im Tank ist. Dieser Abschnitt entspricht damit der EZ3.2 des Antrages: „3D-Phänomene infolge Temperaturdifferenzen. Im Kapitel 4.4 wird auf die Strömungsvorgänge in der Zittauer Strömungswanne eingegangen und damit der Punkt EZ4 des Antrages: Integraluntersuchungen bearbeitet. Kapitel 5 beschreibt die Entwicklung eines Sieb-Modells, das die Faser-Kompaktierung berücksichtigt und auf der Darcy-Gleichung basiert. Die Modellparameter werden an Experimenten in Zittau justiert. Diese Experimente wurden für verschiedene Materialien durchgeführt und mit deren Hilfe ein Koeffizientenkatalog erstellt. Das Modell wurde in den CFD-Code CFX implementiert und anhand einiger Anwendungsbeispiele demonstriert.

CFD-Analyses on the behaviour of mineral wool in the reactor sump

The investigation of insulation debris generation, transport and sedimentation becomes important with regard to reactor safety research for PWR and BWR, when considering the long-term behaviour of emergency core cooling systems during all types of loss of coolant accidents. A joint research project on such questions is being performed in cooperation between the University of Applied Sciences Zittau/Görlitz and the Forschungszentrum Dresden-Rossendorf. The project deals with the experimental investigation of particle transport phenomena in coolant flow and the development of CFD models for its description. While the experiments are performed at the University at Zittau/Görlitz, the theoretical modelling efforts are concentrated at Forschungszentrum Dresden-Rossendorf. In the current presentation the basic concepts for CFD modelling are described and feasibility studies are presented. On the example of a complex flow situation at plunging jet conditions the model capabilities are demonstrated.

CFD analyses of fibre transport and fibre deposition at plunging jet conditions

The investigation of insulation debris generation, transport and sedimentation becomes important with regard to reactor safety research for PWR and BWR, when considering the long-term behaviour of emergency core cooling systems during all types of loss of coolant accidents. A joint research project on such questions is being performed in cooperation between the University of Applied Sciences Zittau/Görlitz (HSZG) and the Helmholtz-Zentrum Dresden-Rossendorf (HZDR). The project deals with the experimental investigation of particle transport phenomena in coolant flow and the development of CFD models for its description (see [10-12]). While the experiments are performed at the University at Zittau/Görlitz, the theoretical modelling efforts are concentrated in Rossendorf. In the current paper, the basic concepts for CFD modelling are described and feasibility studies are presented. The model capabilities are demonstrated via complex flow situations, where a plunging jet agitates insulation debris. © Carl Hanser Verlag, München.

Experimental investigation and CFD simulation of insulation debris transport phenomena in water flow

The investigation of insulation debris generation, transport and sedimentation becomes important with regard to reactor safety research for PWR and BWR, when considering the long-term behavior of emergency core cooling systems during all types of loss of coolant accidents (LOCA). The insulation debris released near the break during a LOCA incident consists of a mixture of disparate particle population that varies with size, shape, consistency and other properties. Some fractions of the released insulation debris can be transported into the reactor sump, where it may perturb/impinge on the emergency core cooling systems. Open questions of generic interest are the sedimentation of the insulation debris in a water pool, its possible re-suspension and transport in the sump water flow and the particle load on strainers and corresponding pressure drop. A joint research project on such questions is being performed in cooperation between the University of Applied Sciences Zittau/Görlitz and the Forschungszentrum Dresden-Rossendorf. The project deals with the experimental investigation of particle transport phenomena in coolant flow and the development of CFD models for its description. While the experiments are performed at the University at Zittau/Görlitz, the theoretical modeling efforts are concentrated at Forschungszentrum Dresden-Rossendorf. In the presentation the basic concepts for CFD modeling are described and feasibility studies including the conceptual design of the experiments are presented.

CFD-modeling and experiments of insulation debris transport phenomena in water flow

The investigation of insulation debris generation, transport, and sedimentation becomes more important with regard to reactor safety research for pressurized water reactors and boiling water reactors when considering the long-term behavior of emergency core coolant systems during all types of loss-of-coolant accidents (LOCAs). The insulation debris released near the break during a LOCA incident consists of a mixture of disparate particle populations that varies with size, shape, consistency, and other properties. Some fractions of the released insulation debris can be transported into the reactor sump, where it may perturb/impinge on the emergency core cooling systems. Open questions of generic interest are, for example, the particle load on strainers and corresponding pressure drop, the sedimentation of the insulation debris in a water pool, and its possible resuspension and transport in the sump water flow. A joint research project on such questions is being performed in cooperation with the University of Applied Sciences Zittau/Görlitz. The project deals with the experimental investigation and the development of computational fluid dynamics (CFD) models for the description of particle transport phenomena in coolant flow. While the experiments are performed at the University of Applied Sciences Zittau/Görlitz, the theoretical work is concentrated at Forschungszentrum Dresden-Rossendorf. In the current paper the basic concepts for CFD modeling are described and feasibility studies including the conceptual design of the experiments are presented.

Experimental investigation and CFD simulation of the behaviour of mineral wool in the reactor sump

The investigation of insulation debris generation, transport and sedimentation becomes important with regard to reactor safety research for PWR and BWR, when considering the long-term behavior of emergency core cooling systems during all types of loss of coolant accidents (LOCA). The insulation debris released near the break during a LOCA incident consists of a mixture of disparate particle population that varies with size, shape, consistency and other properties. Some fractions of the released insulation debris can be transported into the reactor sump, where it may perturb/impinge on the emergency core cooling systems. Open questions of generic interest are the sedimentation of the insulation debris in a water pool, its possible re-suspension and transport in the sump water flow and the particle load on strainers and corresponding pressure drop. A joint research project on such questions is being performed in cooperation between the University of Applied Sciences Zittau/Gorlitz and the Forschungszentrum Dresden-Rossendorf. The project deals with the experimental investigation of particle transport phenomena in coolant flow and the development of CFD models for its description. While the experiments are performed at the University at Zittau/Gorlitz, the theoretical modeling efforts are concentrated at Forschungszentrum Dresden-Rossendorf. In the current paper the basic concepts for CFD modeling are described and feasibility studies including the conceptual design of the experiments are presented. Copyright © 2008 by ASME.

Application of CFD to model fast pyrolysis of biomass

The article deals with the CFD modelling of fast pyrolysis of biomass in an Entrained Flow Reactor (EFR). The Lagrangian approach is adopted for the particle tracking, while the flow of the inert gas is treated with the standard Eulerian method for gases. The model includes the thermal degradation of biomass to char with simultaneous evolution of gases and tars from a discrete biomass particle. The chemical reactions are represented using a two-stage, semi-global model. The radial distribution of the pyrolysis products is predicted as well as their effect on the particle properties. The convective heat transfer to the surface of the particle is computed using the Ranz-Marshall correlation.

CFD modelling of the fast pyrolysis of an in-flight cellulosic particle subjected to convective heat transfer

The pyrolysis of a freely moving cellulosic particle inside a 41.7mgs -1 source continuously fed fluid bed reactor subjected to convective heat transfer is modelled. The Lagrangian approach is adopted for the particle tracking inside the reactor, while the flow of the inert gas is treated with the standard Eulerian method for gases. The model incorporates the thermal degradation of cellulose to char with simultaneous evolution of gases and vapours from discrete cellulosic particles. The reaction kinetics is represented according to the Broido–Shafizadeh scheme. The convective heat transfer to the surface of the particle is solved by two means, namely the Ranz–Marshall correlation and the limit case of infinitely fast external heat transfer rates. The results from both approaches are compared and discussed. The effect of the different heat transfer rates on the discrete phase trajectory is also considered.

CFD modelling of insulation debris transport phenomena in water flow

The investigation of insulation debris generation, transport and sedimentation becomes important with regard to reactor safety research for PWR and BWR, when considering the long-term behaviour of emergency core cooling systems during all types of loss of coolant accidents. A joint research project on such questions is being performed in cooperation between the University of Applied Sciences Zittau/Görlitz and the Forschungszentrum Dresden-Rossendorf. The project deals with the experimental investigation of particle transport phenomena in coolant flow and the development of CFD models for its description. While the experiments are performed at the University at Zittau/Görlitz, the theoretical modelling efforts are concentrated at Forschungszentrum Dresden-Rossendorf. In the current paper the basic concepts for CFD modelling are described and feasibility studies are presented. © Carl Hanser Verlag.