H2O2 Concentration in Exhaled Breath Condensate Increases After Phonotrauma: A Promise of Noninvasive Monitoring?



      The present study was designed to observe the concentration of hydrogen peroxide (H2O2) in exhaled breath condensate (EBC) after induced phonotrauma.


      Thirty-five participants were randomly assigned to one of two conditions (1) Vocal demand and (2) Control. Participants in the experimental group (vocal demand) were asked to read aloud some texts during 1 hour, at 85-90 dB. Inflammation (H2O2 from exhaled breath condensate), acoustic, aerodynamic, and subjective measures were obtained at four time points: before vocal demand (baseline), immediately after baseline, 4-hour after baseline, and 24 hours after baseline. The same acquisition process was implemented for subjects in control group, except that they were not asked to engage in any vocal demand tasks at all.


      As for biological samples, a significant effect for group was observed. Higher values were found for participants in experimental condition. Significant differences were observed for within contrasts in the experimental group, namely 4 hours against baseline, 4 hours against immediately post, and 24 hours against 4 hours. Instrumental outcomes did not show significant differences across the different conditions at any time points. Self-reported measures (vocal fatigue and sensation of muscle tension) showed a significant main effect for group and main effect for condition.


      Intense vocal demand causes an increase in the concentration of H2O2 obtained from EBC at four hours after baseline, which is compatible with the generation of an inflammatory process in the vocal folds (phonotrauma). Moreover, the increase in the sensation of vocal fatigue and muscle tension after demand tasks seems to be an immediate reaction that did not match in time with the increment of H2O2 concentration.

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        • Gray S.
        • Titze I
        Histologic investigation of hyperphonated canine vocal cords.
        Annal Otol Rhinol Laryngol. 1988; 97: 381-388
        • Verdolini K
        • Rosen CA
        • Branski RC
        • et al.
        Shifts in biochemical markers associated with wound healing in laryngeal secretions following phonotrauma: a preliminary study.
        Annal Otol Rhinol Laryngol. 2003; 112: 1021-1025
        • Rousseau B
        • Suehiro A
        • Echemendia N
        • et al.
        Raised intensity phonation compromises vocal fold epithelial barrier integrity.
        Laryngoscope. 2011; 121-2: 346-351
        • Kojima T
        • Van Deusen M
        • Jerome WG
        • et al.
        Quantification of acute vocal fold epithelial surface damage with increasing time and magnitude doses of vibration exposure.
        PlOS One. 2014; 9-3: e91615
        • Li NY
        • Heris HK
        • Mongeau L
        Current understanding and future directions for vocal fold mechanobiology.
        J Cytol Molecular Biol. 2013; 1: 001
        • Li NY
        • Vodovotz Y
        • Kim KH
        • et al.
        Biosimulation of acute phonotrauma: an extended model.
        Laryngoscope. 2011; 121-11: 2418-2428
        • YK Li
        Three-dimensional arrayed amino aerogel biochips for molecular recognition of antigens.
        Biomaterials. 2011; 32-30: 7347-7354
        • Branski RC
        • Verdolini K
        • Sandulache V
        • et al.
        Vocal fold wound healing: a review for clinicians.
        J Voice. 2006; 20-3: 432-442
        • Mittal M
        • Siddiqui MR
        • Tran K
        • et al.
        Reactive oxygen species in inflammation and tissue injury.
        Antioxidants Redox Signal. 2014; 20-7: 1126-1167
        • Branski RC
        • Rosen CA
        • Verdolini K
        • et al.
        Markers of wound healing in vocal fold secretions from patients with laryngeal pathology.
        Annal Otol Rhinol Laryngol. 2004; 113-1: 23-29
        • Rousseau B
        • Kojima T
        • Novaleski CK
        • et al.
        Recovery of vocal fold epithelium after acute phonotrauma.
        The Laryngoscope. 2017; 204 - 2: 93-104
        • Verdolini K
        • Rosen CA
        • Branski RC
        Classification Manual for Voice Disorders-I.
        1st ed. Psychology Press, 2005
        • Verdolini Abbott K
        • Li NY
        • Branski RC
        • et al.
        Vocal exercise may attenuate acute vocal fold inflammation.
        J Voice. 2012; 26 (e1-13): 814
        • Davis MD
        • Montpetit AJ
        Exhaled breath condensate: an update.
        Immunol Allergy Clin North Am. 2018; 38-4: 667-678
        • Hunt J
        Exhaled breath condensate: an overview.
        Immunol Allergy Clin North Am. 2007; 27-4: 587-596
        • Rahimpour E
        • Khoubnasabjafari M
        • Jouyban-Gharamaleki V
        • et al.
        Non-volatile compounds in exhaled breath condensate: review of methodological aspects.
        Anal Bioanal Chem. 2018; 410-25: 6411-6440
        • Araneda OF
        • García C
        • Lagos N
        • et al.
        Lung oxidative stress as related to exercise and altitude. Lipid peroxidation evidence in exhaled breath condensate: a possible predictor of acute mountain sickness.
        Eur J Appl Physiol. 2005; 95: 383-390
        • Valenzuela OF
        • Encina MP
        Design and evaluation of a device for collecting exhaled breath condensate.
        J Bras Pneumol. 2009; 35-1: 69-72
        • Araneda OF
        • Contreras-Briceño F
        • Cavada G
        • et al.
        Swimming versus running: effects on exhaled breath condensate pro-oxidants and pH.
        Eur J Appl Physiol. 2018; 118-11: 2319-2329
        • Contreras-Briceño F
        • Espinosa-Ramirez M
        • Viscor G
        • et al.
        Humidity prevents the exercise-induced formation of hydrogen peroxide and nitrite in exhaled breath condensate in recreational cyclists.
        Eur J Appl Physiol. 2020; 120-10: 2339-2348
        • Araneda OF
        • et al.
        Exhaled breath condensate analysis after long distance races.
        Int J Sports Med. 2012; 33-12: 955-961
        • Nourooz-Zadeh J
        • Tajaddini-Sarmasi J
        • Wolff SP
        Measurement of plasma hydroperoxide concentrations by the ferrous oxidation-xylenol orange assay in conjunction with triphenylphosphine.
        Anal Biochem. 1994; 1: 403-409
        • Araneda OF
        • Urbina-Stagno R
        • Tuesta M
        • et al.
        Increase of pro-oxidants with no evidence of lipid peroxidation in exhaled breath condensate after a 10-km race in non-athletes.
        Physiol Biochem. 2014; 70-1: 107-115
        • Połomska J
        • Bar K
        • Sozańska B
        Exhaled breath condensate-a non-invasive approach for diagnostic methods in asthma.
        J Clin Med. 2021; 10-12: 2697
        • Duman B
        • Borekci S
        • Akdeniz N
        • et al.
        Inhaled corticosteroids' effects on biomarkers in exhaled breath condensate and blood in patients newly diagnosed with asthma who smoke.
        J Asthma. 2021; 14: 1-8
        • Jin Z
        • Zhang W
        • Zhu M
        • et al.
        Assessment of ventilator-associated pneumonia by combining 8-isoprostane and nitric oxide levels in exhaled breath condensate with the clinical pulmonary infection score.
        J Int Med Res. 2020; 48-5300060520
        • Guatura SB
        • Martinez JA
        • Santos Bueno PC
        • Santos ML
        Increased exhalation of hydrogen peroxide in healthy subjects following cigarette consumption.
        Sao Paulo Med J. 2000; 6: 93-98
        • Tuesta M
        • Alvear M
        • Carbonell T
        • et al.
        Effect of exercise duration on pro-oxidants and pH in exhaled breath condensate in humans.
        J Physiol Biochem. 2016; 72-2: 353-360
        • Contreras-Briceño F
        • Espinosa-Ramirez M
        • Hevia G
        • et al.
        Reliability of NIRS portable device for measuring intercostal muscles oxygenation during exercise.
        J Sports Sci. 2019; 37-23: 2653-2659
        • Remacle A
        • Morsomme D
        • Berrué E
        • et al.
        Vocal impact of a prolonged reading task in dysphonic versus normophonic female teachers.
        J Voice. 2012; 26-6 (e1-13): 820
        • Laukkanen AM
        • I
        • Lepp.nen K
        • et al.
        Acoustic measures and self-reports of vocal fatigue by female teachers.
        J Voice. 2008; 22: 283-289
        • Järvinen K
        • Laukkanen A-M
        Vocal loading in speaking a foreign language.
        Folia Phoniatr Logop. 2015; 67-1: 1-7
        • Herndon N
        • Sundarrajan A
        • Sivasankar M
        • et al.
        Respiratory and laryngeal function in teachers: pre- and postvocal loading challenge.
        J Voice. 2019; 33-3: 302-309
        • Heman-Ackah YD
        • Heuer RJ
        • Michael DD
        • et al.
        Cepstral peak prominence: a more reliable measure of dysphonia.
        Ann Otol Rhinol Laryngol. 2003; 112: 324-333
        • Heman-Ackah YD
        • Michael DD
        • Goding GS
        The relationship between cepstral peak prominence and selected parameters of dysphonia.
        J Voice. 2002; 16: 20-27
        • Awan SN
        • Roy N
        • Dromey C.
        Estimating dysphonia severity in continuous speech: application of a multi-parameter spectral/cepstral model.
        Clin Linguist Phon. 2009; 23: 825-841
        • Awan SN
        • Roy N
        • Jetté ME
        • et al.
        Quantifying dysphonia severity using a spectral/cepstral-based acoustic index: comparisons with auditory-perceptual judgements from the CAPE-V.
        Clin Linguist Phon. 2010; 24: 742-758