The Acute Toxicity of Mineral Fibres: A Systematic In Vitro Study Using Different THP-1 Macrophage Phenotypes
Abstract
:1. Introduction
2. Results and Discussion
2.1. Cell–Fibre Interaction Imaging and Fibre Surface Characterisation
Mineral Fibre | Impurities | Fibre Length (µm) | Fibre Width (µm) |
---|---|---|---|
Chrysotile (Balangero, Turin, Italy) | Antigorite, balangeroite, calcite, clinochlore, diopside, dolomite, magnetite, microcline, plagioclase, talc, mackinawite, hematite, ilmenite, lepidocrocite, Fe-Ni sulphide, Fe-Mg carbonate | * Min: 4.02 Mean: 34.7 ** Max: 188 | Min: 0.18 Mean: 0.59 Max: 1.17 |
Crocidolite (UICC) | Hematite, magnetite, quartz, talc, lizardite, calcite, siderite, minnesotaite | Min: 2.52 Mean: 16.1 Max: 131 | Min: 0.23 Mean: 0.64 Max: 1.98 |
Erionite (Jersey, Nevada, USA) | Clinoptilolite, iron-rich nanoparticles, iron oxides/hydroxides, nontronite | Min: 3.23 Mean: 9.39 Max: 55.0 | Min: 0.25 Mean: 0.55 Max: 6.70 |
2.2. Acute Toxicity of Mineral Fibres
2.3. ROS Production and DNA Damage
2.4. Intracellular Dissolution of Fibres and Metal Release
2.5. Inflammatory Response
3. Materials and Methods
3.1. Chemicals
3.2. Mineral Fibres
3.3. Cell Culture
3.4. Cell–Fibre Interaction Imaging and Fibre Surface Characterisation
3.5. Cell Viability with Calcein Green-AM
3.6. Cytotoxicity Assessment by MTT and LDH
3.7. Apoptosis Detection by Confocal Microscopy
3.8. ROS Intracellular Detection
3.9. Evaluation of DNA Damage
3.10. Intracellular Silicate Quantification
3.11. Intracellular Trace Metal Quantification
3.12. Extracellular Release of Inflammatory Cytokines
3.13. RNA Extraction, cDNA Synthesis and qPCR Analyses
3.14. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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GENE | GenBank (a.n.) | Forward | Reverse | Size (bp) |
---|---|---|---|---|
IL-1β | NM_000576.3 | TGATGGCTTATTACAGTGGCAATG | GTAGTGGTGGTCGGAGATTCG | 140 |
IL-6 | NM_001318095.2 | CAGATTTGAGAGTAGTGAGGAAC | CGCAGAATGAGATGAGTTGTC | 194 |
TNF-α | NM_000594.4 | GTGAGGAGGACGAACATC | GAGCCAGAAGAGGTTGAG | 113 |
IL-8 | NM_000584.4 | AATTCATTCTCTGTGGTATC | CCAGGAATCTTGTATTGC | 127 |
MCP-1 | NM_002982 | CTTCTGTGCCTGCTGCTC | CTTGCTGCTGGTGATTCTTC | 156 |
HPRT-1 | NM_000194.3 | GGTCAGGCAGTATAATCCAAAG | TTCATTATAGTCAAGGGCATATCC | 144 |
Crocidolite | Chrysotile | Erionite | Ref. | |
---|---|---|---|---|
Biodurability | *** (66 yrs) | * (0.3 yrs) | *** (181 yrs) | [44] |
Cell death | Mainly apoptosis | Apoptosis and cell lysis | Apoptosis and cell lysis | |
ROS production | *** | ** | ** | |
Redox-active metal release | Fe | Mg, Fe, Ni, Cr, Co | Al | |
DNA damage | *** | *** | *** | |
Cytokine release | *** | *** | *** | |
Inflammatory gene upregulation | *** | ** | ||
Cation-exchange capacity (CEC) | *** | [9] |
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Mirata, S.; Almonti, V.; Di Giuseppe, D.; Fornasini, L.; Raneri, S.; Vernazza, S.; Bersani, D.; Gualtieri, A.F.; Bassi, A.M.; Scarfì, S. The Acute Toxicity of Mineral Fibres: A Systematic In Vitro Study Using Different THP-1 Macrophage Phenotypes. Int. J. Mol. Sci. 2022, 23, 2840. https://doi.org/10.3390/ijms23052840
Mirata S, Almonti V, Di Giuseppe D, Fornasini L, Raneri S, Vernazza S, Bersani D, Gualtieri AF, Bassi AM, Scarfì S. The Acute Toxicity of Mineral Fibres: A Systematic In Vitro Study Using Different THP-1 Macrophage Phenotypes. International Journal of Molecular Sciences. 2022; 23(5):2840. https://doi.org/10.3390/ijms23052840
Chicago/Turabian StyleMirata, Serena, Vanessa Almonti, Dario Di Giuseppe, Laura Fornasini, Simona Raneri, Stefania Vernazza, Danilo Bersani, Alessandro F. Gualtieri, Anna Maria Bassi, and Sonia Scarfì. 2022. "The Acute Toxicity of Mineral Fibres: A Systematic In Vitro Study Using Different THP-1 Macrophage Phenotypes" International Journal of Molecular Sciences 23, no. 5: 2840. https://doi.org/10.3390/ijms23052840