IN VITRO EVALUATION OF CYTOTOXICITY, OXIDATIVE STRESS, DNA DAMAGE AND INFLAMMATION INDUCED BY DIESEL EXHAUST PARTICLES IN HUMAN A549 LUNG CELLS AND MURINE RAW 264.7 MACROPHAGES
Objective: The aim of the present study was to examine the in vitro oxidative stress, cytotoxicity, inflammation and DNA damage induced by Diesel Exhaust Particles (DEPs).
Methods: Alveolar macrophages (murine RAW 264.7 cells) and cultured type II epithelium cells (human A549 lung cells) were exposed to various concentrations of Diesel Exhaust Particles for 24 h. The experiment was evaluated for cell viability, oxidative stress, cytotoxicity, inflammation and DNA damage parameters.
Results: The results showed that overall both cell lines had similar patterns in response to the oxidative stress, cytotoxicity, inflammation and DNA damage parameters induced by DEPs. Vehicle control showed no changes compared to the control. Both cells showed significant changes at the dose of 20 Î¼g/mL and significant changes were found in cytotoxicity, oxidative stress, DNA damage and inflammation indexes.
Conclusion: Hence, exposure to DEPs resulted in doseâ€dependent toxicity in cultured A549 cells and RAW264.7cells and was closely correlated to increased inflammation and oxidative stress.
2. Bayram H, Devalia JL, Sapsford RJ, Ohtoshi T, Miyabara Y, Sagai M, et al. The effect of diesel exhaust particles on cell function and release of inflammatory mediators from human bronchial epithelial cells in vitro. Am J Respir Cell Mol Biol 1998;18:441-8.
3. Donaldson K, Tran L, Jimenez LA, Duffin R, Newby DE, Mills N, et al. Combustion-derived nanoparticles: a review of their toxicology following inhalation exposure. Part Fibre Toxicol 2005;2:1-14.
4. Westerholm RM, Jocob A, Li H. Chemical and biological characterization of particulate, semivolantile and gaz-phase associated compounds in diluted heavy-duty diesel exhaust: a comparison of three different semivolatile-phase samplers. Environ Sci Technol 1991;25:332-8.
5. Brooks A, Wolf R, Royer R, Clark C, Sanchez A, Mcclellan RO. Deposition and biological availability of diesel particles and their associated mutagenic chemicals. Environ Int 1987;5:263-7.
6. Durga M, Devasena T, Murthy PB. Toxicity of exhaust nanoparticles. Afr J Pharm Pharmaco 2013;7:318-31.
7. Krivoshto IN, Richards JR, Albertson TE, Derlet RW. The toxicity of diesel exhaust: implications for primary care. JABFM 2008;21(1):55-62.
8. Boland S, Baeza-Squiban A, Fournier T, Houcine O, Gendron MC, Chevrier M, et al. Diesel exhaust particles are taken up by human airway epithelial cells in vitro and alter cytokine production. Am J Physiol 1999;276:604-13.
9. Boland S, Baeza-Squiban A, Bonvallot V, Houcine O, Pain C, Meyer M, et al. Similar cellular effects induced by diesel exhaust particles from a representative diesel vehicle recovered from filters and Standard Reference Material 1650. Toxicol In vitro 2001;15:379-85.
10. Lehmann AD, Blank F, Baum O, Gehr P, Rothen-Rutishauser BM. Diesel exhaust particles modulate the tight junction proteinoccludin in lung cells in vitro. Part Fibre Toxicol 2009;6(26):1-14.
11. Ohtsuka K, Sato K, Watanabe H, Kimura M, Asakura H, Abo T. Unique order of the lymphocyte subset induction in the liver and intestine of mice during Listeria monocytogenes infection. Cell Immunol 1995;161:112â€24.
12. Marie C, Remy P, Casanova S, Dhawan A, Lazzaroni A, Guillard JC, et al. Impact of titanium dioxide nanoparticle dispersion state and dispersion method on their toxicity towards a549 lung cells and escherichia coli bacteria. J Translational Toxicology 2014;1:10-20.
13. Sagai M, Saito H, Ichinose T, Kodama M, Mori Y. Biological effects of diesel exhaust particles. I. In vitro production of superoxide and in vivo toxicity in mouse. Free Radic Biol Med 1993;14:37â€“47.
14. Nemmar A, Zia S, Subramaniyan D, Al-Amri I, Al Kindi MA, Ali BH. Interaction of diesel exhaust particles with human, rat and mouse erythrocytes in vitro. Cell Physiol Biochem 2012;29:163-70.
15. Carmichael J, William G, De Graff, Adi F, Gazdar, Minna JD, Mitchell JB. Evaluation of a Tetrazolium-based Semiautomated Colorimetric Assay: assessment of chemosensitivity testing. Cancer Res 1987;47:936-42.
16. Oberdorster G, Yu CP. Lung dosimetryâ€“considerations for non inhalation studies. Exp Lung Res 1999;25:1â€“6.
17. Nel A, Xia T, Madler L, Li N. Toxic potential of materials at the nanolevel. Sci 2006;311:622â€7.
18. Oberdorster G, Oberdorster E, Oberdorster J. Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect 2005;113:823â€39.
19. Totlandsdal AI, Cassee FR, Schwarze P, Refsnes M, Lag M. Diesel exhaust particles induce CYP1A1 and pro-inflammatory responses via differential pathways in human bronchial epithelial cells. Part Fibre Toxicol 2010;7:41.
20. Redington AE. Modulation of nitric oxide pathways: therapeutic potential in asthma and chronic obstructive pulmonary disease. Eur J Pharmacol 2006;533:263-76.
21. Shvedovaa A, Kisin E, Murray AR, Johnson VJ, Gorelik O, Arepalli S, et al. Inhalation vs. aspiration of single-walled carbon nanotubes in C57BL/6 mice: inflammation, fibrosis, oxidative stress, and mutagenesis. Am J Physiol Lung Cell Mol Physiol 2008;295:552â€“65.
22. Rhaman I, Mulier B, Gilmour PS, Watchorn T, Donaldson K, Jeffery PK, et al. Oxidant-mediated lung epithelial cell tolerance: the role of intracellular glutathione and nuclear factorkappa B. Biochem Pharmacol 2001;62:787-94.
23. Moller P, Jacobsen NR, Folkmann JK, Danielsen PH, Mikkelsen L, Hemmingsen JG, Vesterdal LK, Forchhammer L, Wallin H, Loft S. Role of oxidative damage in toxicity of particulates. Free Radic Res 2010;44:1-46.
24. Cachon BF, FirminB, Verdin A, Ayi-fanou L, Billet S, Cazier F, et al. Proinflammatory effects and oxidative stress within human bronchial epithelial cells exposed to atmospheric particulate matter (PM2.5 and PM>2.5) collected from Cotonou, Benin. Environmental Pollution 2014;185:340â€“51.
25. Bruce A, Alexander J, Julian L, Martin R, Keith R, Peter W. Apoptosis: programmed cell death eliminates unwanted cells. Molecular biology of the cell. 5th ed. Garland Science; 2008. p. 11-15.
26. Bourgeois B, Owens. The influence of Hurricanes Katrina and Rita on the inflammatory cytokine response and protein expression in A549 cells exposed to PM2.5 collected in the Baton Rougeâ€“Port Allen industrial corridor of Southeastern Louisiana in 2005. Toxicol Mech Methods 2014;24:1â€“23.