Interaction and Localization of Synthetic Nanoparticles in Healthy and Cystic Fibrosis Airway Epithelial Cells: Effect of Ozone Exposure

Ahmad, Shama; Raemy, David; Loader, Joan E.; Kailey, Jenai M.; Neeves, Keith B.; White, Carl W.; Ahmad, Aftab; Gehr, Peter; Rothen‐Rutishauser, Barbara

doi:10.1089/jamp.2011.0889

Cited by 14 publications

(9 citation statements)

References 46 publications

(57 reference statements)

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“…Thirty minutes before imaging, FITC-labeled fibrinogen was added (22). Confocal imaging was performed as described previously (8,23). A twophoton LSM 510 confocal microscope (Zeiss, Thornwood, NY) was used to obtain images and Z-stacks.…”

Section: Confocal Microscopymentioning

confidence: 99%

Tissue Factor Signals Airway Epithelial Basal Cell Survival via Coagulation and Protease-Activated Receptor Isoforms 1 and 2

Ahmad

Ahmad²,

Rancourt³

et al. 2013

Am J Respir Cell Mol Biol

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Tissue factor (TF) initiates the extrinsic coagulation cascade and is a high-affinity receptor for coagulation factor VII. TF also participates in protease-activated receptor (PAR)1 and PAR2 activation. Human epithelial basal cells were previously purified on the basis of TF expression. The purpose of this study was to determine if tracheobronchial epithelial basal cell-associated TF drives coagulation and/ or activates PARs to promote basal cell functions. We used human tracheobronchial tissues to isolate human airway epithelial cells using specific cell surface markers by flow cytometry and studied TF expression by immunostaining. TF-dependent fibrin network formation was observed by confocal and scanning electron microscopy. TF knockdown was done using short hairpin RNA, and TF mRNA was measured using quantitative RT-PCR. We found that 97 6 5% of firstpassage human tracheobronchial epithelial cells were basal cells, and 100% of these basal cells expressed TF. Basal cell-associated TF was active, but TF activity was dependent on added extrinsic coagulation cascade factors. TF inhibition caused basal cell apoptosis and necrosis. This was due to two parallel but interdependent TFregulated processes: failure to generate a basal cell-associated fibrin network and suboptimal PAR1 and PAR2 activity. The data indicate that membrane surface TF mediates airway epithelial basal cell attachment, which maintains cell survival and mitotic potential. The implications of these findings are discussed in the context of basal cell-associated TF activity in normal and injured tissues and of the potential for repair of airway epithelium in lung disease.Keywords: tissue factor; epithelium; clotting; fibrin; tracheobronchial Tissue factor (TF) is a transmembrane cell receptor and cofactor for factor VII (FVII) and serves biological functions beyond its established role in blood coagulation (1). TF may act as a transmembrane signaling molecule by activating and delivering coagulation factors for signaling, and the TF-coagulation FVIIa complex activates G protein-coupled receptors, including the protease-activated receptors (PARs) (2). Under pathologic conditions, TF can be expressed by circulating blood elements (monocytes, neutrophils, and platelets) and by an associated elevated number of cell-derived circulating microparticles. The role of TF in epithelial cell function is much less well understood than its role in the circulation and in hemostasis.In acute lung injury, increased coagulation and decreased fibrinolysis result in diffuse alveolar fibrin deposition, potentially increasing lung inflammation (3). Recent evidence indicates that the damaged alveolar epithelium expresses TF (4) and that FVII-activating protein expression is increased (5). Alveolar procoagulant microparticles are also elevated (6), and intrinsic elevation of tissue factor pathway inhibitor (TFPI) expression is insufficient to inhibit this procoagulant effect (7). Coagulation pathologies also occur in conducting airways. For example, the sulfur mustard ana...

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Section: Confocal Microscopymentioning

confidence: 99%

Tissue Factor Signals Airway Epithelial Basal Cell Survival via Coagulation and Protease-Activated Receptor Isoforms 1 and 2

Ahmad

Ahmad²,

Rancourt³

et al. 2013

Am J Respir Cell Mol Biol

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“…In recent years, the use of engineered nanoparticles (NPs) (defined as <100 nm in three dimensions according to ISO TS 27687:2008) has witnessed a strong rise in biomedical and pharmaceutical applications, specifically for targeted drug delivery [ 1 – 6 ], biosensing [ 7 ] and bio-medical imaging [ 8 ]. In order to develop optimal NPs for biomedical use, much attention is given to the understanding of the basic mechanism of NP interactions with cellular systems at the single cellular level [ 9 – 11 ].…”

Section: Introductionmentioning

confidence: 99%

Different endocytotic uptake mechanisms for nanoparticles in epithelial cells and macrophages

Kühn¹,

Vanhecke²,

Michen³

et al. 2014

Beilstein J. Nanotechnol.

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418

276

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SummaryPrecise knowledge regarding cellular uptake of nanoparticles is of great importance for future biomedical applications. Four different endocytotic uptake mechanisms, that is, phagocytosis, macropinocytosis, clathrin- and caveolin-mediated endocytosis, were investigated using a mouse macrophage (J774A.1) and a human alveolar epithelial type II cell line (A549). In order to deduce the involved pathway in nanoparticle uptake, selected inhibitors specific for one of the endocytotic pathways were optimized regarding concentration and incubation time in combination with fluorescently tagged marker proteins. Qualitative immunolocalization showed that J774A.1 cells highly expressed the lipid raft-related protein flotillin-1 and clathrin heavy chain, however, no caveolin-1. A549 cells expressed clathrin heavy chain and caveolin-1, but no flotillin-1 uptake-related proteins. Our data revealed an impeded uptake of 40 nm polystyrene nanoparticles by J774A.1 macrophages when actin polymerization and clathrin-coated pit formation was blocked. From this result, it is suggested that macropinocytosis and phagocytosis, as well as clathrin-mediated endocytosis, play a crucial role. The uptake of 40 nm nanoparticles in alveolar epithelial A549 cells was inhibited after depletion of cholesterol in the plasma membrane (preventing caveolin-mediated endocytosis) and inhibition of clathrin-coated vesicles (preventing clathrin-mediated endocytosis). Our data showed that a combination of several distinguishable endocytotic uptake mechanisms are involved in the uptake of 40 nm polystyrene nanoparticles in both the macrophage and epithelial cell line.

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“…A loss of transepithelial resistance was showed in CF cells exposed to O 3 suggesting the disruption of tight junctions ( Ahmad et al, 2011 ). However, these NP did not interfere with epithelial integrity as intact apical junctional complexes were reported after such exposure ( Ahmad et al, 2012 ; Jeannet et al, 2016 ). The observed modification of tight junction properties after O 3 but not NP exposure indicates a more pronounced susceptibility of CF cells to O 3 than to NP.…”

Section: Effects Of Air Pollution Exposure In Experimental Models Of ...mentioning

confidence: 94%

The Impact of Air Pollution on the Course of Cystic Fibrosis: A Review

et al. 2022

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Cystic fibrosis (CF) is a lethal and widespread autosomal recessive disorder affecting over 80,000 people worldwide. It is caused by mutations of the CFTR gene, which encodes an epithelial anion channel. CF is characterized by a great phenotypic variability which is currently not fully understood. Although CF is genetically determined, the course of the disease might also depend on multiple other factors. Air pollution, whose effects on health and contribution to respiratory diseases are well established, is one environmental factor suspected to modulate the disease severity and influence the lung phenotype of CF patients. This is of particular interest as pulmonary failure is the primary cause of death in CF. The present review discusses current knowledge on the impact of air pollution on CF pathogenesis and aims to explore the underlying cellular and biological mechanisms involved in these effects.

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Interaction and Localization of Synthetic Nanoparticles in Healthy and Cystic Fibrosis Airway Epithelial Cells: Effect of Ozone Exposure

Cited by 14 publications

References 46 publications

Tissue Factor Signals Airway Epithelial Basal Cell Survival via Coagulation and Protease-Activated Receptor Isoforms 1 and 2

Tissue Factor Signals Airway Epithelial Basal Cell Survival via Coagulation and Protease-Activated Receptor Isoforms 1 and 2

Different endocytotic uptake mechanisms for nanoparticles in epithelial cells and macrophages

The Impact of Air Pollution on the Course of Cystic Fibrosis: A Review

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