scholarly journals Cytotoxicity analysis of biomass combustion particles in human pulmonary alveolar epithelial cells on an air–liquid interface/dynamic culture platform

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Shaorui Ke ◽  
Qi Liu ◽  
Xinlian Zhang ◽  
Yuhan Yao ◽  
Xudong Yang ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Indoor Air ◽  
Air Pollutants ◽  
Submerged Culture ◽  
Alveolar Epithelial Cells ◽  
Biomass Combustion ◽  
Lung Cells ◽  
Alveolar Epithelial ◽  
Air Liquid Interface ◽  
Scanning Electron

Abstract Background Exposure to indoor air pollution from solid fuel combustion is associated with lung diseases and cancer. This study investigated the cytotoxicity and molecular mechanisms of biomass combustion-derived particles in human pulmonary alveolar epithelial cells (HPAEpiC) using a platform that combines air–liquid interface (ALI) and dynamic culture (DC) systems. Methods HPAEpiC were cultured on the surface of polycarbonate (PC) membranes on the ALI–DC platform. The cells were sprayed with an aerosolized solution of biomass combustion soluble constituents (BCSCs) and simultaneously nourished with culture medium flowing beneath the permeable PC membranes. The ALI–DC method was compared with the traditional submerged culture approach. BCSC particle morphology and dosages deposited on the chip were determined for particle characterization. Flow cytometry, scanning electron microscopy, and transmission electron microscopy were used to investigate the apoptosis rate of HPAEpiC and changes in the cell ultrastructure induced by BCSCs. Additionally, the underlying apoptotic pathway was examined by determining the protein expression levels by western blotting. Results Scanning electron microscope images demonstrated that the sample processing and delivering approach of the ALI–DC platform were suitable for pollutant exposure. Compared with the submerged culture method, a significant decline in cell viability and increase in apoptosis rate was observed after BCSC exposure on the ALI–DC platform, indicating that the ALI–DC platform is a more sensitive system for investigating cytotoxicity of indoor air pollutants in lung cells. The morphology and ultrastructure of the cells were damaged after exposure to BCSCs, and the p53 pathway was activated. The Bcl-2/Bax ratio was reduced, upregulating caspase-9 and caspase-3 expression and subsequently inducing apoptosis of HPAEpiC. The addition of N-acetyl cysteine antioxidant significantly alleviated the cytotoxicity induced by BCSCs. Conclusion A novel ALI–DC platform was developed to study the cytotoxicity of air pollutants on lung cells. Using the platform, we demonstrated that BCSCs could damage the mitochondria, produce reactive oxygen species, and activate p53 in HPAEpiC, ultimately inducing apoptosis.

scholarly journals Cytotoxicity Analysis of Biomass Combustion Particles in Human Pulmonary Alveolar Epithelial Cells on an Air-Liquid Interface/Dynamic Culture Platform

2021 ◽  
Author(s):  
Shaorui Ke ◽  
Qi Liu ◽  
Xinlian Zhang ◽  
Yuhan Yao ◽  
Xudong Yang ◽  
...  
Keyword(s):  
Air Pollution ◽  
Electron Microscope ◽  
Epithelial Cells ◽  
Indoor Air ◽  
Liquid Interface ◽  
Alveolar Epithelial Cells ◽  
Biomass Combustion ◽  
Lung Cells ◽  
Alveolar Epithelial ◽  
Air Liquid Interface

Abstract BackgroundExposure to indoor air pollution from solid fuels combustion is associated with lung diseases and cancer. This study aimed to explore the cytotoxicity and molecular mechanisms of biomass combustion-derived particles in human pulmonary alveolar epithelial cells (HPAEpiC) using a platform that combines air-liquid-interface (ALI) and dynamic culture (DC) systems.MethodsHPAEpiC were cultured on the surface of polycarbonate (PC) membranes in ALI-DC platform. The top surface of cells was sprayed with a solution of biomass combustion soluble constituents (BCSCs) and simultaneously nourished with medium flowing from below PC membranes. BCSC particles' morphology and dosages deposited on the chip was determined for particle characterization. A flow cytometer, laser scanning confocal microscopy (LSCM), and transmission electron microscope (TEM) were used to investigate the apoptosis of HPAEpiC and the changes in the ultrastructure of HPAEpiC induced by BCSCs. Additionally, the underlying apoptotic pathway was studied through the determination of the protein expression levels by western blotting.ResultsScanning electron microscope (SEM) results demonstrated that the sample processing and delivering approach were very suitable for pollutant exposure of the platform. The decline in cell viability and increase in apoptosis rate after exposure to similar doses of BCSCs were more under ALI-DC conditions than under submerged conditions. This indicated that the ALI-DC platform is a superior system for investigating cytotoxicity of indoor air pollutants in lung cells. The morphology and ultrastructure of the cells were damaged after exposure to BCSCs. Our results demonstrated that the p53 pathway was activated after exposure of cells cultured using this platform to BCSCs. This further decreased the Bcl-2/Bax ratio thereby activating the expression of caspase-9 and caspase-3, which in turn induced apoptosis in HPAEpiC. Additionally, antioxidants (N-acetyl-cysteine; NAC) could significantly alleviate the cytotoxicity induced by BCSCs.ConclusionsA novel ALI-DC platform was successfully presented to study cytotoxicity of air pollution on lung cells. Based on the platform, the BCSCs was demonstrated that it could damage mitochondria, produce ROS, and activate p53 in HPAEpiC. Furthermore, BCSCs decreased the Bcl-2/Bax ratio and efficiently upregulated the expression of cleaved caspases-9 and − 3, ultimately inducing apoptosis.

scholarly journals TRIM72 is required for effective repair of alveolar epithelial cell wounding

2014 ◽  
Vol 307 (6) ◽  
pp. L449-L459 ◽  
Author(s):  
Seong Chul Kim ◽  
Thomas Kellett ◽  
Shaohua Wang ◽  
Miyuki Nishi ◽  
Nagaraja Nagre ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Molecular Mechanisms ◽  
Striated Muscle ◽  
Alveolar Epithelial Cell ◽  
Alveolar Epithelial Cells ◽  
Lung Cells ◽  
Alveolar Epithelial ◽  
High Tidal Volume Ventilation

The molecular mechanisms for lung cell repair are largely unknown. Previous studies identified tripartite motif protein 72 (TRIM72) from striated muscle and linked its function to tissue repair. In this study, we characterized TRIM72 expression in lung tissues and investigated the role of TRIM72 in repair of alveolar epithelial cells. In vivo injury of lung cells was introduced by high tidal volume ventilation, and repair-defective cells were labeled with postinjury administration of propidium iodide. Primary alveolar epithelial cells were isolated and membrane wounding and repair were labeled separately. Our results show that absence of TRIM72 increases susceptibility to deformation-induced lung injury whereas TRIM72 overexpression is protective. In vitro cell wounding assay revealed that TRIM72 protects alveolar epithelial cells through promoting repair rather than increasing resistance to injury. The repair function of TRIM72 in lung cells is further linked to caveolin 1. These data suggest an essential role for TRIM72 in repair of alveolar epithelial cells under plasma membrane stress failure.

scholarly journals An In Vitro Lung System to Assess the Proinflammatory Hazard of Carbon Nanotube Aerosols

2020 ◽  
Vol 21 (15) ◽  
pp. 5335
Author(s):  
Hana Barosova ◽  
Bedia Begum Karakocak ◽  
Dedy Septiadi ◽  
Alke Petri-Fink ◽  
Vicki Stone ◽  
...  
Keyword(s):  
Prolonged Exposure ◽  
Three Dimensional ◽  
Adverse Outcomes ◽  
Alveolar Epithelial Cells ◽  
Proinflammatory Response ◽  
Multiwalled Carbon ◽  
Alveolar Epithelial ◽  
Culture Model ◽  
Air Liquid Interface

In vitro three-dimensional (3D) lung cell models have been thoroughly investigated in recent years and provide a reliable tool to assess the hazard associated with nanomaterials (NMs) released into the air. In this study, a 3D lung co-culture model was optimized to assess the hazard potential of multiwalled carbon nanotubes (MWCNTs), which is known to provoke inflammation and fibrosis, critical adverse outcomes linked to acute and prolonged NM exposure. The lung co-cultures were exposed to MWCNTs at the air-liquid interface (ALI) using the VITROCELL® Cloud system while considering realistic occupational exposure doses. The co-culture model was composed of three human cell lines: alveolar epithelial cells (A549), fibroblasts (MRC-5), and macrophages (differentiated THP-1). The model was exposed to two types of MWCNTs (Mitsui-7 and Nanocyl) at different concentrations (2–10 μg/cm2) to assess the proinflammatory as well as the profibrotic responses after acute (24 h, one exposure) and prolonged (96 h, repeated exposures) exposure cycles. The results showed that acute or prolonged exposure to different concentrations of the tested MWCNTs did not induce cytotoxicity or apparent profibrotic response; however, suggested the onset of proinflammatory response.

Dépérissement et état hydrique des aiguilles chez le Picea abies

1988 ◽  
Vol 66 (9) ◽  
pp. 1693-1701
Author(s):  
P. M. Badot ◽  
J. P. Garrec ◽  
B. Millet ◽  
M. J. Badot ◽  
J. Mercier
Keyword(s):  
Electron Microscopy ◽  
Air Pollutants ◽  
Direct Action ◽  
Water Status ◽  
Major Elements ◽  
French Jura ◽  
Dieback Disease ◽  
Calcium And Magnesium ◽  
Spruce Needles ◽  
Scanning Electron

A study of the water status of spruce needles was conducted to characterize the physiology of the dieback disease of spruce in forests of the French Jura. Alterations of needles in the affected trees were observed with scanning electron microscopy. The water content of those trees was lower than that of healthy ones. Analyses of potassium, calcium, and magnesium levels in the needles have not revealed any deficiencies in major elements, but changes in mineral composition of the needles were observed in diseased trees. The direct action of air pollutants on foliage could explain the results. However, climatic, edaphic, and other factors could also be involved in the present dieback disease of forests. [Journal translation]

Mechanical force-induced signal transduction in lung cells

1999 ◽  
Vol 277 (4) ◽  
pp. L667-L683 ◽  
Author(s):  
Mingyao Liu ◽  
A. Keith Tanswell ◽  
Martin Post
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Fetal Lung ◽  
Muscle Cells ◽  
Research Field ◽  
Alveolar Epithelial Cells ◽  
Lung Cells ◽  
Airway Epithelial ◽  
Alveolar Epithelial ◽  
Physical Forces

The lung is a unique organ in that it is exposed to physical forces derived from breathing, blood flow, and surface tension throughout life. Over the past decade, significant progress has been made at the cellular and molecular levels regarding the mechanisms by which physical forces affect lung morphogenesis, function, and metabolism. With the use of newly developed devices, mechanical forces have been applied to a variety of lung cells including fetal lung cells, adult alveolar epithelial cells, fibroblasts, airway epithelial and smooth muscle cells, pulmonary endothelial and smooth muscle cells, and mesothelial cells. These studies have led to new insights into how cells sense mechanical stimulation, transmit signals intra- and intercellularly, and regulate gene expression at the transcriptional and posttranscriptional levels. These advances have significantly increased our understanding of the process of mechanotransduction in lung cells. Further investigation in this exciting research field will facilitate our understanding of pulmonary physiology and pathophysiology at the cellular and molecular levels.

Inner surface ultrastructure of the lungs of frog and lizard

1978 ◽  
Vol 36 (2) ◽  
pp. 504-505
Author(s):  
R. F. Bils ◽  
G. M. Hughes
Keyword(s):  
Epithelial Cells ◽  
Random Network ◽  
Lung Surfactant ◽  
Cell Types ◽  
Alveolar Epithelial Cells ◽  
Lung Cells ◽  
Alveolar Epithelial ◽  
Inner Surface ◽  
Mammalian Lung ◽  
Lung Capillaries

Mammalian lung cells and tissues have been studied extensively, resulting in a voluminous literature. In comparison, the ultrastructure study of the respiratory systems of the lower vertebrates is incomplete. Lung surfactant and the related osmiophilic lamellar inclusions were observed in bird lung by Lambson and Cohn and in cultured frog lung cells by Soloff and Burns. This report compares the inner surface structure, cell types and vascularization in the lungs of frog and lizard.Mature Rana pipiens and Lacerta viviparus lungs were fixed at normal volumes (approximately 0. 3 ml and 0. 05 ml, respectively) with cacodylate-buffered glutaraldehyde, post-fixed with OsO4 and processed for both TEM and SEM.Frog LungLooking outward from the central duct, the septa divide the respiratory regions into smaller and smaller compartments, ending in closed sacs or alveoli, ranging from 150 to 300 mμ in diameter (Fig. 1). The largest or primary septa are covered by ciliated epithelium, whereas a squamous microvillous epithelium covers the minor septa and alveolar walls (Fig. 2). In section, the microvillous epithelial cells appear as a combination of type 1 and type 2 alveolar epithelial cells of the mammalian lung. Capillaries appeared as a random network through the septa and alveolar walls.

Air Pollution-Induced Pulmonary Lesions using Sequential Examination by Sem, Lm, and Tem

1976 ◽  
Vol 34 ◽  
pp. 276-277
Author(s):  
D. Hyde ◽  
W. Tyler ◽  
D. Dungworth ◽  
J. Orthoefer
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Air Pollutants ◽  
Exposure Period ◽  
Beagle Dogs ◽  
Pulmonary Lesions ◽  
Critical Point Drying ◽  
Ambient Concentrations ◽  
Ambient Levels ◽  
Scanning Electron

Previous exposures of animals to air pollutants have mostly been for short terms and at levels considerably greater than ambient concentrations (1). This report, however, relates to a study in which beagle dogs were exposed for 68 months to a variety of air pollutants at high ambient levels. After a 33-36 month post-exposure period, the lungs from 71 beagle dogs were fixed via the airways with cacodylate-buffered glutaraldehyde-formaldehyde. Lungs of 6 dogs were examined from all but one of 8 experimental groups. Various levels of bronchi and bronchioles were identified under a dissecting microscope and bisected with razor blades. One-half of each airway was prepared for scanning electron microscopy (SEM) after dehydration by critical point drying with CO2 and coating with gold-palladium.

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