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  • Description - Work Package 3 - Uptake and Biodistribution of Nanoparticles in Cells and Tissues
    produced particles Nanoparticles shall be labeled by fluorescent markers on their surface It is intended to take advantage of the fact that biomolecules can firmly bind to particles corona formation Components of the lung surfactant or of the lung lining fluid will be tested as candidates for the development of such post production labels 2 Biokinetic behavior of nanoparticles and subcellular localization in vitro Uptake of nanoparticles will be investigated using well defined cell cultures and co cultures of up to three lung cell types to model basic cellular interactions in the lung Emphasis will be put on the dynamics of particle uptake as this is important to determine the intracellular particle dose at the Lowest Observed Adverse Effect Levels LOAEL The LOAEL is defined as particle dose causing first signs of damage e g due to oxidative effects 3 Biokinetic behavior during uptake of nanoparticles in vivo The behavior of nanoparticles in the lung shall be investigated shortly after application i e after minutes to hours These experiments shall elucidate the interaction of nanoparticles with surfactant components in vivo c f APQ In this context a possible agglomeration of particles the role of alveolar macrophages and or epithelial cells for particle uptake as well as the possible penetration of particles from the lung compartment into circulation will be investigated Studies will be conducted with normal rats In parallel the established animal model of asthmatic mice ovalbumin treated will be studied to investigate whether the asthmatic lung differs from the normal state with respect to nanoparticles biokinetic behavior or toxicity The disposition of surface modified particles applied to the lungs via inhalation or intratracheal instillation and also to the gastro intestinal tract will be investigated Tissue samples retrieved in the course of toxicological analyses conducted in WP 4 will be

    Original URL path: http://www.nanogem.de/cms/nanogem/front_content.php?idcat=146&lang=11 (2016-02-14)
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  • Description - Work Package Q - Changes of nanomaterials over their life cycle degree of agglomeration, formation of adducts and surface conditioning
    tests using cell culture models the dispersion will be done in cell culture medium with and without serum For in vivo studies dispersion will be done for instance in instillation medium and furthermore for comparing to in vivo inhalation we will disperse nanoparticles in broncho alveolar lavage fluid BALF For all these different dispersions we will develop adopt standard operating SOP s which also will be made publicly available via nanoGEM and DaNa webpages later in the project In addition we will use a set of complementary measurement methods to study the degree of agglomeration In particular we will use hydrodynamic methods based on fractionation analytical ultracentrifugation AUC BASF optical methods via particle tracking NTA IBE BIG optical methods based on light scattering DLS UdS BfR optical methods via laser diffraction BASF using electron microscopy via kryopreparation BTS BASF sedimentation analysis BIG Figure 2 Example for analyzing agglomeration nanosilver via NanoSight Tracking analysis Depicted on the left is a size distribution plot and on the right a picture from the movie showing the scattered light from nanoparticles Source IBE R D gGmbH BIG at FH Dortmund Figure 3 Following sedimentation over time under conditions used for cell culture to follow agglomeration showing measured and modelled data The insert depicts particles under phase contrast Source IBE R D gGmbH BIG at FH Dortmund 2 Changes on the surface of the nanoparticles via adsorption of biomolecules Our goal is a systematic analysis of the particles surface via different complementary methods to determine the binding of biomolecules lipids proteins As outlined above for objective 1 we will apply different dispersion media Furthermore for selected particles and scenarios we are interested to determine the binding strength and the temporal evolution of the corona In particular we will apply gel based analysis of protein corona

    Original URL path: http://www.nanogem.de/cms/nanogem/front_content.php?idcat=148&lang=11 (2016-02-14)
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