PBDEs Found in House Dust Impact Human Lung Epithelial Cell Homeostasis
<p>The general structure of polybrominated diphenyl ethers and the structure of the eight environmentally relevant congeners tested in this study.</p> "> Figure 2
<p>Viability of A549 cells after prolonged (24, 48 and 72 h) exposure to PBDEs at concentrations from 1.5 to 48 μg mL<sup>−1</sup> determined with the MTS assay; 0 μg mL<sup>−1</sup> represents untreated cells. The results are expressed as percentage of corresponding control, untreated cells and given as means ± SD (<span class="html-italic">n</span> = 6). The dashed frame highlights a time-dependent effect. & <span class="html-italic">p</span> < 0.05; # <span class="html-italic">p</span> < 0.01; * <span class="html-italic">p</span> < 0.0001 vs. untreated control.</p> "> Figure 3
<p>Cytotoxicity curves of BDE-99 and -209 at concentrations from 1.5 to 48 μg mL<sup>−1</sup> in A549 cells over 24, 48 and 72 h. The results of the MTS assay are given as means ± SD (<span class="html-italic">n</span> = 6).</p> "> Figure 4
<p>Prolonged exposure of A549 cells to a house dust sample extract. A549 cells were exposed to ∑PBDE in house dust at concentrations from 0.375 to 24 μg mL<sup>−1</sup> for 24, 48 and 72 h, and cell viability was determined with the MTS assay. The results are given as means ± SD (<span class="html-italic">n</span> = 6). The dashed frame highlights the concentration found in the house dust sample. & <span class="html-italic">p</span> < 0.05; # <span class="html-italic">p</span> < 0.01; vs. untreated control.</p> "> Figure 5
<p>LOAEL cytotoxicity of BDE-99 (7.5 μg mL<sup>−1</sup>) and BDE-209 (15 μg mL<sup>−1</sup>), alone and combined, in A549 cells treated for 24, 48 and 72 h. The results of the MTS assay are given as means ± SE (<span class="html-italic">n</span> = 6). # <span class="html-italic">p</span> < 0.01; <span>$</span> <span class="html-italic">p</span> < 0.001 vs. untreated control.</p> "> Figure 6
<p>Levels of LDH release after 4 h exposure of A549 cells to PBDE congeners and house dust sample extract. Triton (0.08%) was used as the positive control. The results are presented as percentage of LDH release and given as means ± SD (<span class="html-italic">n</span> = 3). # <span class="html-italic">p</span> < 0.01; <span>$</span> <span class="html-italic">p</span> < 0.001; * <span class="html-italic">p</span> < 0.0001 vs. untreated control.</p> "> Figure 7
<p>ROS levels after the 4 h exposure of A549 cells to PBDE congeners, house dust sample extract and H<sub>2</sub>O<sub>2</sub> (100 µM) as positive control. The results of DCF-fluorescence signal measurements are presented as relative fluorescence units (RFU) and given as means ± SD (<span class="html-italic">n</span> = 6). & <span class="html-italic">p</span> < 0.05; # <span class="html-italic">p</span> < 0.01; <span>$</span> <span class="html-italic">p</span> < 0.001; * <span class="html-italic">p</span> < 0.0001 vs. untreated control.</p> "> Figure 8
<p>GSH levels after the 4 h exposure of A549 cells to single PBDEs and house dust sample extract at 3, 6 and 12 µg mL<sup>−1</sup> and to tBOOH (100 µM) as positive control. The results of MCB-fluorescence measurements are presented as relative fluorescence units (RFU) and given as means ± SD (<span class="html-italic">n</span> = 6). & <span class="html-italic">p</span> < 0.05; # <span class="html-italic">p</span> < 0.01; <span>$</span> <span class="html-italic">p</span> < 0.001; * <span class="html-italic">p</span> < 0.0001 vs. untreated control.</p> "> Figure 9
<p>Mitochondrial membrane potential after 4 h exposure of A549 cells to single BDE congeners and house dust sample extract at 3, 6 and 12 µg mL<sup>−1</sup> and CCCP (50 µM) as positive control. The results are presented as relative fluorescence units (RFU) and given as means ± SD (<span class="html-italic">n</span> = 6). & <span class="html-italic">p</span> < 0.5, # <span class="html-italic">p</span> < 0.01, * <span class="html-italic">p</span> < 0.001 vs. untreated control.</p> "> Figure 10
<p>Percentage of different stages of apoptosis of A549 cells after 4 h exposure to single PBDE congener and house dust sample extract at 12 µg mL<sup>−1</sup> and PFA (0.08%, <span class="html-italic">v</span>/<span class="html-italic">v</span>) as a positive control. The results are presented as percentage of total apoptotic cells compared to vehicle control (2.4% methanol) and given as means ± SD (<span class="html-italic">n</span> = 6). * <span class="html-italic">p</span> < 0.001 vs. control.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Cell Culture
2.3. Measurement of Cell Viability (MTS)
2.4. Cell Membrane Integrity
2.5. Determination of Reactive Oxygen Species
2.6. Determination of Glutathione
2.7. Determination of Mitochondrial Membrane Potential
2.8. Apoptosis
2.9. In Silico Predictions of CYP Inhibition
2.10. Statistics
3. Results
3.1. Cell Viability
Cell Viability after Exposure to BDE-99 and -209 Mixture
3.2. Cell Status in the Presence of PBDEs
3.2.1. BDE-99, -153, -154 and -209 Affect Cell Membrane Integrity
3.2.2. PBDEs Induce ROS
3.2.3. PBDEs Decrease GSH Levels
3.2.4. BDE-100, -154 and -209 Reduce Mitochondrial Membrane Potential
3.2.5. BDE-28 Activates Apoptosis
3.3. Predicted CYP Enzyme Inhibition by PBDEs
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Time (h) | IC50 ± SD (µg mL−1) BDE-99 | IC50 ± SD (µg mL−1) BDE-209 |
---|---|---|
24 h | 9.8 ± 1.1 | 37.2 ± 2.9 |
48 h | 6.6 ± 1.1 | 13.8 ± 1.1 |
72 h | 5.3 ± 1.1 | 11.0 ± 1.1 |
CYP | BDE-28 | BDE-47 | BDE-99 | BDE-100 | BDE-153 | BDE-154 | BDE-183 | BDE-209 |
---|---|---|---|---|---|---|---|---|
1A2 | + | - | - | - | - | - | - | - |
2C19 | + | - | - | - | - | - | - | - |
2C9 | + | + | + | + | + | + | + | + |
2D6 | - | - | - | - | - | - | - | - |
3A4 | - | - | - | - | - | - | - | - |
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Zandona, A.; Jagić, K.; Dvoršćak, M.; Madunić, J.; Klinčić, D.; Katalinić, M. PBDEs Found in House Dust Impact Human Lung Epithelial Cell Homeostasis. Toxics 2022, 10, 97. https://doi.org/10.3390/toxics10020097
Zandona A, Jagić K, Dvoršćak M, Madunić J, Klinčić D, Katalinić M. PBDEs Found in House Dust Impact Human Lung Epithelial Cell Homeostasis. Toxics. 2022; 10(2):97. https://doi.org/10.3390/toxics10020097
Chicago/Turabian StyleZandona, Antonio, Karla Jagić, Marija Dvoršćak, Josip Madunić, Darija Klinčić, and Maja Katalinić. 2022. "PBDEs Found in House Dust Impact Human Lung Epithelial Cell Homeostasis" Toxics 10, no. 2: 97. https://doi.org/10.3390/toxics10020097