Journal Article

Combining analytical techniques to assess the translocation of diesel particles across an alveolar tissue barrier in vitro

Petri-Fink, Alke, He, Ruiwen, Taladriz-Blanco, Patricia, Balog, Sandor, Gunasingam, Gowsinth, Rothen-Rutishauser, Barbara

2024

Overview

Background During inhalation, airborne particles such as particulate matter ≤ 2.5 μm (PM 2.5 ), can deposit and accumulate on the alveolar epithelial tissue. In vivo studies have shown that fractions of PM 2.5 can cross the alveolar epithelium to blood circulation, reaching secondary organs beyond the lungs. However, approaches to quantify the translocation of particles across the alveolar epithelium in vivo and in vitro are still not well established. In this study, methods to assess the translocation of standard diesel exhaust particles (DEPs) across permeable polyethylene terephthalate (PET) inserts at 0.4, 1, and 3 μm pore sizes were first optimized with transmission electron microscopy (TEM), ultraviolet-visible spectroscopy (UV-VIS), and lock-in thermography (LIT), which were then applied to study the translocation of DEPs across human alveolar epithelial type II (A549) cells. A549 cells that grew on the membrane (pore size: 3 μm) in inserts were exposed to DEPs at different concentrations from 0 to 80 µg.mL − 1 ( 0 to 44 µg.cm − 2 ) for 24 h. After exposure, the basal fraction was collected and then analyzed by combining qualitative (TEM) and quantitative (UV-VIS and LIT) techniques to assess the translocated fraction of the DEPs across the alveolar epithelium in vitro. Results We could detect the translocated fraction of DEPs across the PET membranes with 3 μm pore sizes and without cells by TEM analysis, and determine the percentage of translocation at approximatively 37% by UV-VIS (LOD: 1.92 µg.mL − 1 ) and 75% by LIT (LOD: 0.20 µg.cm − 2 ). In the presence of cells, the percentage of DEPs translocation across the alveolar tissue was determined around 1% at 20 and 40 µg.mL − 1 (11 and 22 µg.cm − 2 ), and no particles were detected at higher and lower concentrations. Interestingly, simultaneous exposure of A549 cells to DEPs and EDTA can increase the translocation of DEPs in the basal fraction. Conclusion We propose a combination of analytical techniques to assess the translocation of DEPs across lung tissues. Our results reveal a low percentage of translocation of DEPs across alveolar epithelial tissue in vitro and they correspond to in vivo findings. The combination approach can be applied to any traffic-generated particles, thus enabling us to understand their involvement in public health.

• Publisher: BioMed Central,BioMed Central Ltd,BMC
• Subject: A549 Cells / Air Pollutants - analysis / Air Pollutants - toxicity / Air pollution / Alveolar Epithelial Cells - drug effects / Alveolar Epithelial Cells - metabolism / Biomedical and Life Sciences / Biomedicine / Chemical properties / Diesel motor exhaust gas / Diesel particles (DEPs) / Environmental aspects / Environmental Health / Ethylenediaminetetraacetic acid / Health aspects / Humans / Lock-in thermography (LIT) / Methods / Microscopy, Electron, Transmission / Nanotechnology / Particle Size / Particles / Particulate Matter - analysis / Particulate Matter - toxicity / Pharmacology/Toxicology / Physiological aspects / Pneumology/Respiratory System / Polyethylene terephthalate / Polyethylene Terephthalates - chemistry / Polyethylene Terephthalates - toxicity / Public Health / Pulmonary alveoli / Pulmonary Alveoli - drug effects / Pulmonary Alveoli - metabolism / Spectrum analysis / Translocation / Transmission electron microscopy (TEM) / Ultraviolet – visible (UV-VIS) / Vehicle Emissions - analysis / Vehicle Emissions - toxicity