scholarly journals In Vitro Cytotoxicity Evaluation of the Magnéli Phase Titanium Suboxides (TixO2x−1) on A549 Human Lung Cells

2019 ◽  
Vol 20 (1) ◽  
pp. 196 ◽  
Author(s):  
Veno Kononenko ◽  
Damjana Drobne
Keyword(s):  
Photocatalytic Activity ◽  
Tio2 Nanoparticles ◽  
Human Lung ◽  
A549 Cells ◽  
Cellular Interactions ◽  
Biological Interactions ◽  
Lysosomal Membranes ◽  
Human Lung Cells ◽  
Magnéli Phase ◽  
Magneli Phase

The use of titanium suboxides, known as Magnéli phase TiOx, is expected to increase in the near future due to their desirable properties. In order to use Magnéli phase TiOx nanoparticles safely, it is necessary to know how nanoparticles interact with biological systems. In this study, the cytotoxicity of three different Magnéli TiOx nanoparticles was evaluated using human lung A549 cells and the results were compared with hazard data on two different TiO2 nanoparticles whose biological interactions have already been extensively studied. After A549 cells were exposed to nanoparticles, the metabolic activity was measured by the Resazurin assay, the amount of cellular proteins was measured by the Coomassie Blue assay, and lysosomal integrity was measured by the Neutral Red Uptake assay. In order to investigate possible modes of particle actions, intracellular Ca2+ level, reactive oxygen species (ROS) production, and photo-oxidative disruptions of lysosomal membranes were assessed. All experiments were performed in serum-containing and in serum-deprived cell culture mediums. In addition, the photocatalytic activity of Magnéli TiOx and TiO2 nanoparticles was measured. The results show that Magnéli TiOx nanoparticles increase intracellular Ca2+ but not ROS levels. In contrast, TiO2 nanoparticles increase ROS levels, resulting in a higher cytotoxicity. Although Magnéli TiOx nanoparticles showed a lower UV-A photocatalytic activity, the photo-stability of the lysosomal membranes was decreased by a greater extent, possibly due to particle accumulation inside lysosomes. We provide evidence that Magnéli TiOx nanoparticles have lower overall biological activity when compared with the two TiO2 formulations. However, some unique cellular interactions were detected and should be further studied in line with possible Magnéli TiOx application. We conclude that Magnéli phase nanoparticles could be considered as low toxic material same as other forms of titanium oxide particles.

scholarly journals TiO2 nanoparticles generate superoxide and alter gene expression in human lung cells

RSC Advances ◽  
2019 ◽  
Vol 9 (43) ◽  
pp. 25039-25047 ◽  
Author(s):  
Dhanya T. Jayaram ◽  
Ashwath Kumar ◽  
Linda E. Kippner ◽  
Po-Yi Ho ◽  
Melissa L. Kemp ◽  
...  
Keyword(s):  
Gene Expression ◽  
Fluorescence Microscopy ◽  
Tio2 Nanoparticles ◽  
Human Lung ◽  
Lung Cells ◽  
Rna Seq ◽  
Nanoparticle Exposure ◽  
Human Lung Cells

Human lung cells have a multi-generational response to TiO2 nanoparticle exposure determined by RNA-Seq and fluorescence microscopy.

scholarly journals The Hag Protein of Moraxella catarrhalis Strain O35E Is Associated with Adherence to Human Lung and Middle Ear Cells

2003 ◽  
Vol 71 (9) ◽  
pp. 4977-4984 ◽  
Author(s):  
Melissa M. Holm ◽  
Serena L. Vanlerberg ◽  
Darren D. Sledjeski ◽  
Eric R. Lafontaine
Keyword(s):  
Cell Lines ◽  
Middle Ear ◽  
Moraxella Catarrhalis ◽  
Human Lung ◽  
Primary Cultures ◽  
A549 Cells ◽  
Pcr Analysis ◽  
Nucleotide Sequence Data ◽  
Human Lung Cells ◽  
Conjunctival Cells

ABSTRACT Previous studies have demonstrated that the Moraxella catarrhalis surface antigen UspA1 is an adhesin for Chang human conjunctival cells. The present report demonstrates that lack of UspA1 expression does not affect the adherence of strain O35E to A549 human lung cells or primary cultures of human middle ear epithelial (HMEE) cells. These results imply that another molecule mediates the adherence of M. catarrhalis to these two cell lines. To identify this adhesin, strain O35E was mutagenized with a transposon and 1,000 mutants were screened in a microcolony formation assay using A549 cells. Nine independent isolates exhibited an 8- to 19-fold reduction in adherence and contained a transposon in the same locus. Nucleotide sequence data and PCR analysis indicated that the transposons were inserted in different locations in the gene encoding the surface protein Hag. Quantitative assays using one representative transposon mutant, O35E.TN2, showed considerably decreased binding to A549 as well as HMEE cells. However, this mutant adhered at wild-type levels to Chang conjunctival cells. These findings suggest that the M. catarrhalis Hag protein is an adhesin for cell lines derived from human lung and middle ear tissues.

scholarly journals Aptamer Profiling of A549 Cells Infected with Low-Pathogenicity and High-Pathogenicity Influenza Viruses

Viruses ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1028 ◽  
Author(s):  
Kevin M. Coombs ◽  
Philippe F. Simon ◽  
Nigel J. McLeish ◽  
Ali Zahedi-Amiri ◽  
Darwyn Kobasa
Keyword(s):  
Influenza A ◽  
Human Lung ◽  
A549 Cells ◽  
Influenza Viruses ◽  
Post Translational Modification ◽  
Emerging Pathogens ◽  
Influenza A Viruses ◽  
Human Lung Cells ◽  
Wide Range ◽  
High Pathogenicity

Influenza A viruses (IAVs) are important animal and human emerging and re-emerging pathogens that are responsible for yearly seasonal epidemics and sporadic pandemics. IAVs cause a wide range of clinical illnesses, from relatively mild infections by seasonal strains, to acute respiratory distress during infections with highly pathogenic avian IAVs (HPAI). For this study, we infected A549 human lung cells with lab prototype A/PR/8/34 (H1N1) (PR8), a seasonal H1N1 (RV733), the 2009 pandemic H1N1 (pdm09), or with two avian strains, an H5N1 HPAI strain or an H7N9 strain that has low pathogenicity in birds but high pathogenicity in humans. We used a newly-developed aptamer-based multiplexed technique (SOMAscan®) to examine >1300 human lung cell proteins affected by the different IAV strains, and identified more than 500 significantly dysregulated cellular proteins. Our analyses indicated that the avian strains induced more profound changes in the A549 global proteome compared to all tested low-pathogenicity H1N1 strains. The PR8 strain induced a general activation, primarily by upregulating many immune molecules, the seasonal RV733 and pdm09 strains had minimal effect upon assayed molecules, and the avian strains induced significant downregulation, primarily in antimicrobial response, cardiovascular and post-translational modification systems.

scholarly journals Influence of the physicochemical features of TiO2 nanoparticles on the formation of a protein corona and impact on cytotoxicity

RSC Advances ◽  
2020 ◽  
Vol 10 (72) ◽  
pp. 43950-43959
Author(s):  
Ozge Kose ◽  
Marion Stalet ◽  
Lara Leclerc ◽  
Valérie Forest
Keyword(s):  
Tio2 Nanoparticles ◽  
Human Lung ◽  
Protein Corona ◽  
Lung Cells ◽  
Human Lung Cells ◽  
Potential Relationship ◽  
Physicochemical Features

Characterization of the formation of the protein corona of TiO2 nanoparticles as a function of the main nanoparticle properties and investigation of potential relationship with the cytotoxicity nanoparticles induce in vitro in human lung cells.

scholarly journals Quantitative Proteomic Analyses of Influenza Virus-Infected Cultured Human Lung Cells

2010 ◽  
Vol 84 (20) ◽  
pp. 10888-10906 ◽  
Author(s):  
Kevin M. Coombs ◽  
Alicia Berard ◽  
Wanhong Xu ◽  
Oleg Krokhin ◽  
Xiaobo Meng ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Protein Function ◽  
Human Lung ◽  
Isotope Labeling ◽  
Morphological Changes ◽  
A549 Cells ◽  
Host Cells ◽  
Human Influenza Virus ◽  
Human Lung Cells ◽  
Cell Immunity

ABSTRACT Because they are obligate intracellular parasites, all viruses are exclusively and intimately dependent upon host cells for replication. Viruses, in turn, induce profound changes within cells, including apoptosis, morphological changes, and activation of signaling pathways. Many of these alterations have been analyzed by gene arrays, which measure the cellular “transcriptome.” Until recently, it has not been possible to extend comparable types of studies to globally examine all the host cellular proteins, which are the actual effector molecules. We have used stable isotope labeling by amino acids in cell culture (SILAC), combined with high-throughput two-dimensional (2-D) high-performance liquid chromatography (HPLC)/mass spectrometry, to determine quantitative differences in host proteins after infection of human lung A549 cells with human influenza virus A/PR/8/34 (H1N1) for 24 h. Of the 4,689 identified and measured cytosolic protein pairs, 127 were significantly upregulated at >95% confidence, 153 were significantly downregulated at >95% confidence, and a total of 87 proteins were upregulated or downregulated more than 5-fold at >99% confidence. Gene ontology and pathway analyses indicated differentially regulated proteins and included those involved in host cell immunity and antigen presentation, cell adhesion, metabolism, protein function, signal transduction, and transcription pathways.

Antagonistic effect of co-exposure to short-multiwalled carbon nanotubes and benzo[a]pyrene in human lung cells (A549)

2019 ◽  
Vol 35 (6) ◽  
pp. 445-456
Author(s):  
Mansour Rezazadeh Azari ◽  
Yousef Mohammadian ◽  
Habibollah Peirovi ◽  
Meisam Omidi ◽  
Fariba Khodagholi ◽  
...  
Keyword(s):  
Carbon Nanotubes ◽  
Multiwalled Carbon Nanotubes ◽  
Human Lung ◽  
A549 Cells ◽  
In Vitro Toxicity ◽  
Ros Generation ◽  
Exposure Condition ◽  
Multiwalled Carbon ◽  
Human Lung Cells ◽  
Starting Point

In theenvironment, co-exposure to short-multiwalled carbon nanotubes (S-MWCNTs) and polycyclic aromatic compounds (PAHs) has been reported. In the co-exposure condition, the adsorption of PAHs onto MWCNTs may reduce PAHs toxic effect. The objective of this study was to investigate the cytotoxicity of S-MWCNTs and benzo[a]pyrene (B[a]P) individually, and in combination in human lung cell lines (A549). The adsorption of B[a]P onto MWCNTs was measured spectrometrically. In vitro toxicity was assessed through cell viability, reactive oxygen species (ROS) generation, apoptosis, and 8-hydroxy-2′-deoxyguanosine (8-OHdG) generation experiments. The S-MWCNTs demonstrated cytotoxicity through the generation of ROS, apoptosis, and 8-OHdG in A549 cells. Co-exposure to S-MWCNTs and B[a]P demonstrated a significant reduction in ROS generation and apoptosis compared with the sum of their separate toxic effects at the same concentrations. Decreasing the bioavailability of B[a]P by MWCNT interaction is the probable reason for the antagonistic effects of the co-exposure condition. The findings of this study will contribute to a better understanding of the health effects of co-exposures to air pollutants and could be a starting point for modifying future health risk assessments.

scholarly journals Rapid and Efficient Testing of The Toxicity of Graphene-Related Materials in Primary Human Lung Cells

2021 ◽  
Author(s):  
Javier Frontiñán-Rubio ◽  
Viviana Jehová-González ◽  
Ester Vázquez ◽  
Mario Durán-Prado
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Human Lung ◽  
Lung Tumor ◽  
A549 Cells ◽  
Tumor Cell Line ◽  
Screening Methods ◽  
Suitable Model ◽  
Human Lung Cells ◽  
Graphene Derivatives

Abstract Background Graphene and its derivative materials are manufactured by numerous companies and research laboratories, during which processes they can come into contact with their handlers’ physiological barriers—for instance, their respiratory system. Despite their potential toxicity, these materials have even been used in face masks to prevent COVID-19 transmission. The increasingly widespread use of these materials requires the design and implementation of appropriate, versatile, and accurate toxicological screening methods to guarantee their safety. Murine models are adequate, though limited when exploring different doses and lengths of exposure—as this increases the number of animals required, contrary to the Three R’s principle in animal experimentation. This article proposes an in vitro model using primary, non-transformed normal human bronchial epithelial (NHBE) cells as an alternative to the most widely used model to date, the human lung tumor cell line A549. The model has been tested with three graphene derivatives—graphene oxide (GO), few-layer graphene (FLG), and small FLG (sFLG). Results We observed a cytotoxic effect (necrosis and apoptosis) at early (6- and 24-hour) exposures, which intensified after seven days of contact between cells and the graphene-related materials (GRMs)—with cell death reaching 90% after a 5 µg/mL dose. A549 cells are more resistant to necrosis and apoptosis, yielding values less than half those of NHBE cells at low concentrations of GRMs (between 0.05 and 5 µg/mL). Indeed, GRM-induced cell death in NHBE cells is comparable to that induced by toxic compounds such as diesel exhaust particles on the same cell line. Conclusions We propose NHBE as a suitable model to test GRM-induced toxicity, allowing refinement of the dose concentrations and exposure timings for better-designed in vivo mouse assays.

scholarly journals Grouping of Poorly Soluble Low (Cyto)Toxic Particles: Example with 15 Selected Nanoparticles and A549 Human Lung Cells

Nanomaterials ◽  
2019 ◽  
Vol 9 (5) ◽  
pp. 704
Author(s):  
Veno Kononenko ◽  
David B. Warheit ◽  
Damjana Drobne
Keyword(s):  
Human Lung ◽  
A549 Cells ◽  
Test System ◽  
Alveolar Type ◽  
In Vitro Test ◽  
Human Lung Cells ◽  
Poorly Soluble ◽  
Lysosomal System ◽  
Biological Endpoint

Poorly soluble, low (cyto)toxic particles (PSLTs) are often regarded as one group, but it is important that these particles can be further differentiated based on their bioactivity. Currently, there are no biological endpoint based groupings for inhaled nanoparticles (NPs) that would allow us to subgroup PSLTs based on their mode of action. The aim of this study was to group NPs based on their cytotoxicity and by using the in vitro response of the endo-lysosomal system as a biological endpoint. The endo-lysosomal system is a main cellular loading site for NPs. An impaired endo-lysosomal system in alveolar type II cells may have serious adverse effects on the maintenance of pulmonary surfactant homeostasis. The 15 different NPs were tested with human lung adenocarcinoma (A549) cells. The highly soluble NPs were most cytotoxic. With respect to PSLTs, only three NPs increased the cellular load of acid and phospholipid rich organelles indicating particle biopersistence. All the rest PSLTs could be regarded as low hazardous. The presented in vitro test system could serve as a fast screening tool to group particles according to their ability to interfere with lung surfactant metabolism. We discuss the applicability of the suggested test system for bringing together substances with similar modes-of-action on lung epithelium. In addition, we discuss this approach as a benchmark test for the comparative assessment of biopersistence of PSLTs.

scholarly journals Mammalian Adaptive Mutations of the PA Protein of Highly Pathogenic Avian H5N1 Influenza Virus

2015 ◽  
Vol 89 (8) ◽  
pp. 4117-4125 ◽  
Author(s):  
Reina Yamaji ◽  
Shinya Yamada ◽  
Mai Q. Le ◽  
Mutsumi Ito ◽  
Yuko Sakai-Tagawa ◽  
...  
Keyword(s):  
Amino Acid ◽  
Mammalian Cells ◽  
H5n1 Virus ◽  
Human Lung ◽  
A549 Cells ◽  
Amino Acid Substitutions ◽  
Highly Pathogenic ◽  
Adaptive Mutations ◽  
Human Lung Cells ◽  
H5n1 Influenza

ABSTRACTHighly pathogenic H5N1 influenza A viruses continue to circulate among avian species and cause sporadic cases of human infection. Therefore, the threat of a pandemic persists. However, the human cases of H5N1 infection have been limited mainly to individuals in close contact with infected poultry. These findings suggest that the H5N1 viruses need to acquire adaptive mutations to gain a replicative advantage in mammalian cells to break through the species barrier. Many amino acid mutations of the polymerase complex have been reported to enhance H5N1 virus growth in mammalian cells; however, the mechanism for H5N1 virus of adaptation to humans remains unclear. Here, we propose that the PA of an H5N1 influenza virus isolated from a human in Vietnam (A/Vietnam/UT36285/2010 [36285]) increased the ability of an avian H5N1 virus (A/chicken/Vietnam/TY31/2005 [Ck/TY31]) to grow in human lung epithelial A549 cells. The five PA amino acid substitutions V44I, V127A, C241Y, A343T, and I573V, which are rare in H5N1 viruses from human and avian sources, enhanced the growth capability of this virus in A549 cells. Moreover, these mutations increased the pathogenicity of the virus in mice, suggesting that they contribute to adaptation to mammalian hosts. Intriguingly, PA-241Y, which 36285 encodes, is conserved in more than 90% of human seasonal H1N1 viruses, suggesting that PA-241Y contributes to virus adaptation to human lung cells and mammalian hosts.IMPORTANCEMany amino acid substitutions in highly pathogenic H5N1 avian influenza viruses have been shown to contribute to adaptation to mammalian hosts. However, no naturally isolated H5N1 virus has caused extensive human-to-human transmission, suggesting that additional, as-yet unidentified amino acid mutations are needed for adaptation to humans. Here, we report that five amino acid substitutions in PA (V44I, V127A, C241Y, A343T, and I573V) contribute to the replicative efficiency of H5N1 viruses in human lung cells and to high virulence in mice. These results are helpful for assessing the pandemic risk of isolates and further our understanding of the mechanism of H5N1 virus adaptation to mammalian hosts.

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