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Aerosol Delivery of Functionalized Gold Nanoparticles Target and Activate Dendritic Cells in a 3D Lung Cellular Model Kleanthis Fytianos, Savvina Chortarea, Laura Rodriguez-Lorenzo, Fabian Blank, Christophe von Garnier, Alke Petri-Fink, and Barbara Rothen-Rutishauser ACS Nano, Just Accepted Manuscript • DOI: 10.1021/acsnano.6b06061 • Publication Date (Web): 14 Dec 2016 Downloaded from http://pubs.acs.org on December 18, 2016
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Aerosol Delivery of Functionalized Gold Nanoparticles Target and Activate Dendritic Cells in a 3D Lung Cellular Model Author List Kleanthis Fytianos,1 Savvina Chortarea,1 Laura Rodriguez-Lorenzo,1 Fabian Blank,3 Christophe von Garnier,3 Alke Petri-Fink,1,2 Barbara Rothen-Rutishauser*1 1
Adolphe Merkle Institute, University of Fribourg, Fribourg, Switzerland Department of Chemistry, University of Fribourg, Fribourg, Switzerland 3 Respiratory Medicine, Inselspital, Universitiy of Bern, Bern, Switzerland 2
Corresponding author: Barbara Rothen-Rutishauser Adolphe Merkle Institute Chemin des Verdiers 4 CH-1700 Fribourg Switzerland Phone: +41 26 300 9502 Email:
[email protected] ABSTRACT Nanocarrier design combined with pulmonary drug delivery holds great promise for the treatment of respiratory tract disorders. In particular, targeting of dendritic cells that are key immune cells to enhance or suppress an immune response in the lung is a promising approach for the treatment of allergic diseases. Fluorescently-encoded polyvinyl-alcohol-coated gold nanoparticles, functionalized with either negative (COO–) or positive (-NH3+) surface charges, were functionalized with a DC-SIGN antibody on the particle surface, enabling binding to a dendritic cell surface receptor. A 3D co-culture model consisting of epithelial and immune cells (macrophages and dendritic cells) mimicking the human alveolar epithelial tissue barrier was then employed to assess the effects of aerosolized AuNPs. PVA-NH2 AuNPs showed higher uptake compared to their –COOH counterparts, with the highest uptake recorded in macrophages as shown by flow cytometry. None of the AuNPs induced cytotoxicity, necrosis or increased cytokine secretion, whereas only PVA-NH2 AuNPs induced higher apoptosis levels. DC-SIGN-AuNPs showed significantly increased uptake by MDDCs with subsequent activation compared to non-antibody-conjugated control AuNPs, independent of surface charge. Our results show that DC-SIGN conjugation to the AuNPs enhanced MDDC targeting and activation in a complex 3D lung cell model. These findings highlight the potential of immunoengineering approaches to the targeting and activation of immune cells in the lung by nanocarriers. Keywords: gold nanoparticles, dendritic cells, aerosol exposures, cellular uptake, immunomodulation
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The human body has a number of portals of entry available for drug administration, such as the lungs (via inhalation), the gastro-intestinal tract (via digestion), the blood vessels (via intravenous injection), and the skin (via transdermal application). The lung, with its huge internal surface of ca. 150 m2 (i.e. alveoli and airways),1 provides a promising, non-invasive portal of entry for aerosolized drugs. A number of inhaled drugs are currently applied to treat airway diseases, are in development, or have been considered for the treatment of various lung diseases. In 2004 there were about 25 drugs licensed for use in treating pulmonary diseases via inhalation, such as asthma, emphysema and chronic inflammation, chronic obstructive pulmonary disease (COPD), and lung cancer.2 In addition to the large lung surface, area a dense network of antigen-presenting cell populations, such as dendritic cells (DCs) and macrophages, are present within the lung parenchyma.3 DCs are among the most important immune cells in the lung, orchestrating innate and adaptive immune responses.4 Specific targeting of DCs via receptor–mediated mechanisms has previously been described as an innovative approach to drug delivery.5,6 Dendritic cell – specific intracellular adhesion molecule 3 (ICAM-3) – grabbing non-integrin (DC-SIGN) is a well-studied receptor that is found in abundance on the surface of mature DCs. It has a high affinity for mannose and is involved in the enhancement of antigen uptake, processing and presentation to Tcells in the draining of lymph nodes. Targeting DC-SIGN can lead to the development of immunomodulatory applications such as a change in pro-inflammatory cytokine secretion.6 The development of nanomaterials (i.e. a material with one, two, or three external dimensions in the nanoscale (
