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Saarbrücken, Germany

Daum N.,Helmholtz Center for Infection Research | Kuehn A.,Helmholtz Center for Infection Research | Hein S.,Biopharmaceutics and Pharmaceutical Technology | Schaefer U.F.,Biopharmaceutics and Pharmaceutical Technology | And 2 more authors.
Methods in Molecular Biology

The blood-air barrier formed by the alveolar epithelium of the peripheral lung is crucial for the pulmonary delivery of drugs. Most existing in vitro models mimicking the blood-air barrier are represented by tumor cells or immortalized cells and lack biological relevance due to their genetic alterations and underexpressed essential physiological functions. However, the increasing interest of aerosol administration of medicines to the respiratory system requires the development and use of representative in vitro models. Thereby, human alveolar epithelial cells (hAEpC) are a suitable test system allowing standardized toxicity and transport studies for newly developed compounds and delivery systems. The isolation, purification, and cultivation of hAEpC are described as well as their possible application in the so-called Pharmaceutical Aerosol Deposition Device On Cell Cultures (PADDOCC) mimicking the complete inhalation process of a powder aerosol in vitro. © 2012 Springer Science+Business Media, LLC. Source

Herd H.,University of Utah | Herd H.,Helmholtz Center for Infection Research | Daum N.,Helmholtz Center for Infection Research | Jones A.T.,University of Cardiff | And 4 more authors.
ACS Nano

In order to engineer safer nanomaterials, there is a need to understand, systematically evaluate, and develop constructs with appropriate cellular uptake and intracellular fates. The overall goal of this project is to determine the uptake patterns of silica nanoparticle geometries in model cells, in order to aid in the identification of the role of geometry on cellular uptake and transport. In our experiments we observed a significant difference in the viability of two phenotypes of primary macrophages; immortalized macrophages exhibited similar patterns. However, both primary and immortalized epithelial cells did not exhibit toxicity profiles. Interestingly uptake of these geometries in all cell lines exhibited very different time-dependent patterns. A screening of a series of chemical inhibitors of endocytosis was performed to isolate the uptake mechanisms of the different particles. The results show that all geometries exhibit very different uptake profiles and that this may be due to the orientation of the nanoparticles when they interact with the cell surface. Additionally, evidence suggests that these uptake patterns initialize different downstream cellular pathways, dependent on cell type and phenotype. © 2013 American Chemical Society. Source

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