Health assessment and medical surveillance of workers exposed to combustion nanoparticles are challenging. The aim was to evaluate the feasibility of using exhaled breath condensate (EBC) from healthy volunteers for (1) assessing the lung deposited dose of combustion nanoparticles and (2) determining the resulting oxidative stress by measuring hydrogen peroxide (H2O2) and malondialdehyde (MDA).
Methods:
Fifteen healthy nonsmoker volunteers were exposed to three different levels of sidestream cigarette smoke under controlled conditions. EBC was repeatedly collected before, during, and 1 and 2 hr after exposure. Exposure variables were measured by direct reading instruments and by active sampling. The different EBC samples were analyzed for particle number concentration (light-scattering–based method) and for selected compounds considered oxidative stress markers.
Results:
Subjects were exposed to an average airborne concentration up to 4.3×105 particles/cm3 (average geometric size ∼60–80 nm). Up to 10×108 particles/mL could be measured in the collected EBC with a broad size distribution (50th percentile ∼160 nm), but these biological concentrations were not related to the exposure level of cigarette smoke particles. Although H2O2 and MDA concentrations in EBC increased during exposure, only H2O2 showed a transient normalization 1 hr after exposure and increased afterward. In contrast, MDA levels stayed elevated during the 2 hr post exposure.
Conclusions:
The use of diffusion light scattering for particle counting proved to be sufficiently sensitive to detect objects in EBC, but lacked the specificity for carbonaceous tobacco smoke particles. Our results suggest two phases of oxidation markers in EBC: first, the initial deposition of particles and gases in the lung lining liquid, and later the start of oxidative stress with associated cell membrane damage. Future studies should extend the follow-up time and should remove gases or particles from the air to allow differentiation between the different sources of H2O2 and MDA.
Get full access to this article
View all access options for this article.
References
1.
DelfinoRJ, SioutasC, and MalikS: Potential role of ultrafine particles in associations between airborne particle mass and cardiovascular health. Environ Health Perspect., 2005; 113:934–946.
2.
AillonKL, XieY, El-GendyN, BerklandCJ, and ForrestML: Effects of nanomaterial physicochemical properties on in vivo toxicity. Adv Drug Deliv Rev., 2009; 61:457–466.
3.
JonesCF, and GraingerDW: In vitro assessments of nanomaterial toxicity. Adv Drug Deliv Rev., 2009; 61:438–456.
RiedikerM, Schubauer-BeriganMK, BrouwerDH, NelissenI, KoppenG, FrijnsE, ClarkKA, HoeckJ, LiouS-H, HoSF, BergamaschiE, and GibsonR: A road map toward a globally harmonized approach for occupational health surveillance and epidemiology in nanomaterial workers. J Occup Environ Med., 2012; 54:1214–1223.
6.
NelA, XiaT, MadlerL, and LiN: Toxic potential of materials at the nanolevel. Science., 2006; 311:622–627.
7.
DonaldsonK, BeswickPH, and GilmourPS. Free radical activity associated with the surface of particles: a unifying factor in determining biological activity?. Toxicol Lett., 1996; 88:293–298.
8.
RiedikerM, CascioWE, GriggsTR, HerbstMC, BrombergPA, NeasL, WilliamsRW, and DevlinRB: Particulate matter exposure in cars is associated with cardiovascular effects in healthy young men. Am J Respir Crit Care Med., 2004; 169:934–940.
9.
LeeW, and ThomasPS: Oxidative stress in COPD and its measurement through exhaled breath condensate. Clin Transl Sci., 2009; 2:150–155.
10.
AyresJG, BormP, CasseeFR, CastranovaV, DonaldsonK, GhioA, HarrisonRM, HiderR, KellyF, KooterIM, MaranoF, MaynardRL, MudwayI, NelA, SioutasC, SmithS, Baeza-SquibanA, ChoA, DugganS, and FroinesJ: Evaluating the toxicity of airborne particulate matter and nanoparticles by measuring oxidative stress potential—a workshop report and consensus statement. Inhal Toxicol., 2008; 20:75–99.
11.
BrookRD, RajagopalanS, PopeCA3rd, BrookJR, BhatnagarA, Diez-RouxAV, HolguinF, HongY, LuepkerRV, MittlemanMA, PetersA, SiscovickD, SmithSCJr, WhitselL, and KaufmanJD: Particulate matter air pollution and cardiovascular disease: an update to the scientific statement from the American Heart Association. Circulation., 2010; 121:2331–2378.
12.
TsaiDH, AmyaiN, Marques-VidalP, WangJL, RiedikerM, MooserV, PaccaudF, WaeberG, VollenweiderP, and BochudM: Effects of particulate matter on inflammatory markers in the general adult population. Part Fibre Toxicol., 2012; 9:24.
13.
TsaiDH, RiedikerM, WuerznerG, MaillardM, Marques-VidalP, PaccaudF, VollenweiderP, BurnierM, and BochudM: Short-term increase in particulate matter blunts nocturnal blood pressure dipping and daytime urinary sodium excretion. Hypertension., 2012; 60:1061–1069.
14.
RiedikerM, CascioWE, GriggsTR, HerbstMC, BrombergPA, NeasL, WilliamsRW, and DevlinRB: Particulate matter exposure in cars is associated with cardiovascular effects in healthy, young men. Am J Respir Crit Care Med., 2004; 169:934–940.
15.
RiedikerM, DevlinRB, GriggsTR, HerbstMC, BrombergPA, WilliamsRW, and CascioWE: Cardiovascular effects in patrol officers are associated with fine particulate matter from brake wear and engine emissions. Part Fibre Toxicol., 2004; 1:2.
16.
ConwayJ: Lung imaging—two dimensional gamma scintigraphy, SPECT, CT and PET. Adv Drug Deliv Rev., 2012; 64:357–368.
17.
MutluGM, GareyKW, RobbinsRA, DanzigerLH, and RubinsteinI: Collection and analysis of exhaled breath condensate in humans. Am J Respir Crit Care Med., 2001; 164:731–737.
18.
MontuschiP, and BarnesPJ: Analysis of exhaled breath condensate for monitoring airway inflammation. Trends Pharmacol Sci., 2002; 23:232–237.
19.
GoldoniM, CaglieriA, De PalmaG, AcampaO, GergelovaP, CorradiM, ApostoliP, and MuttiA: Chromium in exhaled breath condensate (EBC), erythrocytes, plasma and urine in the biomonitoring of chrome-plating workers exposed to soluble Cr(VI). J Environ Monit., 2010; 12:442–447.
20.
MontuschiP: Analysis of exhaled breath condensate in respiratory medicine: methodological aspects and potential clinical applications. Ther Adv Respir Dis., 2007; 1:5–23.
21.
GoldoniM, CatalaniS, De PalmaG, ManiniP, AcampaO, CorradiM, BergonziR, ApostoliP, and MuttiA: Exhaled breath condensate as a suitable matrix to assess lung dose and effects in workers exposed to cobalt and tungsten. Environ Health Perspect., 2004; 112:1293–1298.
22.
LärstadM, LjungkvistG, OlinAC, and TorenK: Determination of malondialdehyde in breath condensate by high-performance liquid chromatography with fluorescence detection. J Chromatogr B Analyt Technol Biomed Life Sci., 2001; 766:107–114.
23.
BergamaschiE: Human biomonitoring of engineered nanoparticles: an appraisal of critical issues and potential biomarkers. J Nanomater., 2012;Article ID 564121.
24.
MuttiA, and CorradiM: Recent developments in human biomonitoring: non-invasive assessment of target tissue dose and effects of pneumotoxic metals. Med Lav., 2006; 97:199–206.
25.
CorradiM, GergelovaP, and MuttiA: Use of exhaled breath condensate to investigate occupational lung diseases. Curr Opin Allergy Clin Immunol., 2010; 10:93–98.
26.
CaglieriA, GoldoniM, AcampaO, AndreoliR, VettoriMV, CorradiM, ApostoliP, and MuttiA: The effect of inhaled chromium on different exhaled breath condensate biomarkers among chrome-plating workers. Environ Health Perspect., 2006; 114:542–546.
27.
BrodingHC, MichalkeB, GoenT, and DrexlerH: Comparison between exhaled breath condensate analysis as a marker for cobalt and tungsten exposure and biomonitoring in workers of a hard metal alloy processing plant. Int Arch Occup Environ Health., 2009; 82:565–573.
28.
FalgayracG, Cherot-KornobisN, de BrouckerV, HuloS, EdmeJ-L, SobaszekA, and PenelG: Noninvasive molecular identification of particulate matter in lungs by Raman microspectrometry. J Raman Spectrosc., 2011; 42:1484–1487.
29.
PinheiroT, BarreirosMA, AlvesLC, FelixPM, FrancoC, SousaJ, and AlmeidaSM: Particulate matter in exhaled breath condensate: a promising indicator of environmental conditions. Nucl Instrum Methods Phys Res B., 2011; 269:2404–2408.
30.
BarregardL, SaellstenG, AnderssonL, AlmstrandAC, GustafsonP, AnderssonM, and OlinAC: Experimental exposure to wood smoke: effects on airway inflammation and oxidative stress. Occup Environ Med., 2008; 65:319–324.
31.
LiuL, PoonR, ChenL, FrescuraA-M, MontuschiP, CiabattoniG, WheelerA, and DalesR: Acute effects of air pollution on pulmonary function, airway inflammation, and oxidative stress in asthmatic children. Environ Health Perspect., 2009; 117:668–674.
32.
RomieuI, Barraza-VillarrealA, Escamilla-NunezC, AlmstrandAC, Diaz-SanchezD, SlyPD, and OlinAC: Exhaled breath malondialdehyde as a marker of effect of exposure to air pollution in children with asthma. J Allergy Clin Immunol., 2008; 121:903–909.
33.
TengY, SunP, ZhangJ, YuR, BaiJ, YaoX, HuangM, AdcockIM, and BarnesPJ: Hydrogen peroxide in exhaled breath condensate in patients with asthma a promising biomarker?. Chest., 2011; 140:108–116.
34.
SyslovaK, KacerP, KuzmaM, NajmanovaV, FenclovaZ, VlckovaS, LebedovaJ, and PelclovaD: Rapid and easy method for monitoring oxidative stress markers in body fluids of patients with asbestos or silica-induced lung diseases. J Chromatogr B Analyt Technol Biomed Life Sci., 2009; 877:2477–2486.
35.
van der VaartH, PostmaDS, TimensW, and Ten HackenNHT: Acute effects of cigarette smoke on inflammation and oxidative stress: a review. Thorax., 2004; 59:713–721.
36.
GuilleminM: Setting up and use of an experimentation chamber. Arch Mal Prof., 1975; 36:421–428.
37.
NowakD, KaluckaS, BialasiewiczP, and KrolM: Exhalation of H2O2 and thiobarbituric acid reactive substances (TBARS) by healthy subjects. Free Radic Biol Med., 2001; 30:178–186.
38.
PerretV, HuynhCK, DrozPO, DucTV, and GuilleminM: Assessment of occupational exposure to diesel fumes—parameter optimization of the thermal coulometric measurement method for carbon. J Environ Monit., 1999; 1:367–372.
39.
MillerMR, HankinsonJ, BrusascoV, BurgosF, CasaburiR, CoatesA, CrapoR, EnrightP, van der GrintenCPM, GustafssonP, JensenR, JohnsonDC, MacIntyreN, McKayR, NavajasD, PedersenOF, PellegrinoR, ViegiG, and WangerJ: Standardisation of spirometry. Eur Respir J., 2005; 26:319–338.
40.
ZappacostaB, PersichilliS, MormileF, MinucciA, RussoA, GiardinaB, and De SoleP: A fast chemiluminescent method for H2O2 measurement in exhaled breath condensate. Clin Chim Acta., 2001; 310:187–191.
41.
Navas DíazA, García SanchezF, and González GarcíaJA: Hydrogen peroxide assay by using enhanced chemiluminescence of the luminol-H2O2-horseradish peroxidase system: comparative studies. Anal Chim Acta., 1996; 327:161–165.
42.
SauvainJJ, SetyanA, WildP, TacchiniP, LaggerG, StortiF, DeslarzesS, GuilleminM, RossiMJ, and RiedikerM: Biomarkers of oxidative stress and its association with the urinary reducing capacity in bus maintenance workers. J Occup Med Toxicol., 2011; 6:1–13.
43.
CrossCE, VandervlietA, O'NeillCA, LouieS, and HalliwellB: Oxidants, antioxidants, and respiratory-tract lining fluids. Environ Health Perspect., 1994; 102:185–191.
44.
PryorWA, StoneK, CrossCE, MachlinL, and PackerL: Oxidants in cigarette-smoke—radicals, hydrogen-peroxide, peroxynitrate, and peroxynitrite. Ann N Y Acad Sci., 1993; 686:12–28.
45.
DalyBJ, SchmidK, and RiedikerM: Contribution of fine particulate matter sources to indoor exposure in bars, restaurants, and cafes. Indoor Air., 2010; 20:204–212.
46.
EffrosRM, PetersonB, CasaburiR, SuJ, DunningM, TordayJ, BillerJ, and ShakerR: Epithelial lining fluid solute concentrations in chronic obstructive lung disease patients and normal subjects. J Appl Physiol., 2005; 99:1286–1292.
47.
Gallego-UrreaJA, TuoriniemiJ, PallanderT, and HassellovM: Measurements of nanoparticle number concentrations and size distributions in contrasting aquatic environments using nanoparticle tracking analysis. Environ Chem., 2009; 7:67–81.
48.
GschwindS, HagendorferH, FrickDA, and GuentherD: Mass quantification of nanoparticles by single droplet calibration using inductively coupled plasma mass spectrometry. Anal Chem., 2013; 85:5875–5883.
49.
MullerWJ, HessGD, and SchererPW: A model of cigarette-smoke particle deposition. Am Ind Hyg Assoc J., 1990; 51:245–256.
50.
AlmstrandAC, LjungstromE, LausmaaJ, BakeB, SjovallP, and OlinAC: Airway monitoring by collection and mass spectrometric analysis of exhaled particles. Anal Chem., 2009; 81:662–668.
51.
LeTT, SaveynP, HoaHD, and Van der MeerenP: Determination of heat-induced effects on the particle size distribution of casein micelles by dynamic light scattering and nanoparticle tracking analysis. Int Dairy J., 2008; 18:1090–1096.
52.
GuaturaSB, MartinezJAB, BuenoPCdS, and dos SantosML: Increased exhalation of hydrogen peroxide in healthy subjects following cigarette consumption. Sao Paulo Med J., 2000; 118:93–98.
53.
HuY, ZhangZ, and YangC: The determination of hydrogen peroxide generated from cigarette smoke with an ultrasensitive and highly selective chemiluminescence method. Anal Chim Acta., 2007; 601:95–100.
54.
KnoblochH, BecherG, DeckerM, and ReinholdP: Evaluation of H2O2 and pH in exhaled breath condensate samples: methodical and physiological aspects. Biomarkers., 2008; 13:319–341.
55.
WangY, ArellanesC, and PaulsonSE: Hydrogen peroxide associated with ambient fine-mode, diesel, and biodiesel aerosol particles in Southern California. Aerosol Sci Technol., 2012; 46:394–402.
56.
FortezaR, SalatheM, MiotF, and ConnerGE: Regulated hydrogen peroxide production by Duox in human airway epithelial cells. Am J Respir Cell Mol Biol., 2005; 32:462–469.
57.
DoniecZ, NowakD, TomalakW, PisiewiczK, and KurzawaR: Passive smoking does not increase hydrogen peroxide (H2O2) levels in exhaled breath condensate in 9-year-old healthy children. Pediatr Pulmonol., 2005; 39:41–45.
58.
EsterbauerH, SchaurRJ, and ZollnerH: Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic Biol Med., 1991; 11:81–128.
59.
Medina-NavarroR, Mercado-PichardoE, Hernandez-PerezO, and HicksJJ: Identification of acrolein from the ozone oxidation of unsaturated fatty acids. Hum Exp Toxicol., 1999; 18:677–682.
60.
RundellKW, SleeJB, CavistonR, and HollenbachAM: Decreased lung function after inhalation of ultrafine and fine particulate matter during exercise is related to decreased total nitrate in exhaled breath condensate. Inhal Toxicol., 2008; 20:1–9.
61.
SpiraA, BeaneJ, ShahV, LiuG, SchembriF, YangXM, PalmaJ, and BrodyJS: Effects of cigarette smoke on the human airway epithelial cell transcriptome. Proc Natl Acad Sci U S A., 2004; 101:10143–10148.
62.
WrightWR, ParzychK, CrawfordD, MeinC, MitchellJA, and Paul-ClarkMJ: Inflammatory transcriptome profiling of human monocytes exposed acutely to cigarette smoke. PLoS ONE., 2012; 7:e30120.
63.
BarretoM, VillaMP, OlitaC, MartellaS, CiabattoniG, and MontuschiP: 8-Isoprostane in exhaled breath condensate and exercise-induced bronchoconstriction in asthmatic children and adolescents. Chest., 2009; 135:66–73.
64.
LucidiV, CiabattoniG, BellaS, BarnesPJ, and MontuschiP: Exhaled 8-isoprostane and prostaglandin E2 in patients with stable and unstable cystic fibrosis. Free Radic Biol Med., 2008; 45:913–919.
65.
MontuschiP, ParisD, MelckD, LucidiV, CiabattoniG, RaiaV, CalabreseC, BushA, BarnesPJ, and MottaA: NMR spectroscopy metabolomic profiling of exhaled breath condensate in patients with stable and unstable cystic fibrosis. Thorax., 2012; 67:222–228.
66.
MottaA, ParisD, MelckD, de LaurentiisG, ManiscalcoM, SofiaM, and MontuschiP: Nuclear magnetic resonance-based metabolomics of exhaled breath condensate: methodological aspects. Eur Respir J., 2012; 39:498–500.
67.
MontuschiP, MoresN, TroveA, MondinoC, and BarnesPJ: The electronic nose in respiratory medicine. Respiration., 2013; 85:72–84.
68.
MalerbaM, and MontuschiP: Non-invasive biomarkers of lung inflammation in smoking subjects. Curr Med Chem., 2012; 19:187–196.
69.
BofanM, MoresN, BaronM, DabrowskaM, ValenteS, SchmidM, TroveA, ConfortoS, ZiniG, CattaniP, FusoL, MautoneA, MondinoC, PagliariG, D'AlessioT, and MontuschiP: Within-day and between-day repeatability of measurements with an electronic nose in patients with COPD. J Breath Res., 2013; 7:017103.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
