Proteomic Analysis of Vocal Fold Fibroblasts Exposed to Cigarette Smoke Extract: Exploring the Pathophysiology of Reinke's Edema*

Gugatschka, Markus; Darnhofer, Barbara; Grossmann, Tanja; Schittmayer, Matthias; Hortobagyi, David; Kirsch, Andrijana; Karpf, E; Brčić, Luka; Birner‐Gruenberger, Ruth; Karbiener, Michael

doi:10.1074/mcp.ra119.001272

Cited by 23 publications

(25 citation statements)

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“…Although vibration had no effect, we found a significant upregulation of HA concentrations after CSE exposure. This might be a protective reaction of hVFF and corroborates previous data from our group 13 . In a clinical study, Woo described that the application of HYAL into RE diminished VF mass, highlighting the role of HA in RE 19 .…”

Section: Discussionsupporting

confidence: 90%

“…This might be a protective reaction of hVFF and corroborates previous data from our group. 13 In a clinical study, Woo described that the application of HYAL into RE diminished VF mass, highlighting the role of HA in RE. 19 COL1A1 and FN1 are important extracellular matrix components and provide structural strength to tissue.…”

Section: Discussionmentioning

confidence: 99%

“…Generation of CSE medium was performed as described previously. 13 Briefly, a 100 mL Erlenmeyer flask with a 5 mm polypropylene tube was guided through the flasks' rubber cap and connected to a 200 μm borosilicate glass frit at its lower end. The lateral opening of the Erlenmeyer flask was connected to a 60 mL syringe via a silicone tube.…”

Section: Cell Culturementioning

confidence: 99%

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Exploring the Pathophysiology of Reinke's Edema: The Cellular Impact of Cigarette Smoke and Vibration

et al. 2020

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Objectives To explore the isolated or combined effects of cigarette smoke extract (CSE) and vibration on human vocal fold fibroblasts (hVFF) in an in vitro setting in order to elucidate their influence in the pathophysiology of Reinke's edema (RE). Study design Immortalized hVFF were exposed to CSE or control medium under static or vibrational conditions. A phonomimetic bioreactor was used to deliver vibrational patterns to hVFF over a period of 5 days. Methods Cytotoxicity was quantified using a lactate dehydrogenase assay. We employed reverse transcription–quantitative polymerase chain reaction, enzyme‐linked immunosorbent assay, and Magnetic Luminex(R) assays (R&D Systems, Minneapolis, MN) to assess the influence on extracellular matrix production, fibrogenesis, inflammation, and angiogenesis. Results We observed significant changes induced by CSE alone (hyaluronic acid, matrix metalloproteinase 1, Interleukin‐8, cyclooxygenase [COX]1, COX2, vascular endothelial growth factor [VEGF]D), as well as settings in which only the combination of CSE and vibration led to significant changes (transforming growth factor beta 1, VEGFA, VEGFC). Also, CSE‐induced levels of COX2 were only significantly reduced when vibration was applied. Conclusion We were able to explore the cellular effects of CSE and vibration on hVFF by employing a phonomimetic bioreactor. Whereas cigarette smoke is generally accepted as a risk factor for RE, the role of vibration remained unclear as it is difficult to study in humans. Our data showed that some genes and proteins in the pathophysiological context of RE were only affected when CSE in combination with vibration was applied. Level of Evidence NA Laryngoscope, 131:E547–E554, 2021

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

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Exploring the Pathophysiology of Reinke's Edema: The Cellular Impact of Cigarette Smoke and Vibration

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“…Proangiogenic factors. A previous investigation of the effect of cigarette smoke extract on VFF by our group showed a significant upregulation of vascular endothelial growth factors (VEGF) A and C [42]. In order to evaluate potential changes on VEGF A and C following cytokine treatment and vibration, these two factors were included in the present study.…”

Section: Selection Of Genes and Moleculesmentioning

confidence: 99%

In vitro mechanical vibration down-regulates pro-inflammatory and pro-fibrotic signaling in human vocal fold fibroblasts

et al. 2020

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Introduction Voice rest following phonotrauma or phonosurgery has a considerable clinical impact, but clinical recommendations are inconsistent due to inconclusive data. As biopsies of the vocal folds (VF) for molecular biology studies in humans are unethical, we established a new in vitro model to explore the effects of vibration on human vocal fold fibroblasts (hVFF) in an inflammatory and normal state, which is based on previously published models. Methods By using a phonomimetic bioreactor we were able to apply predefined vibrational stress patterns on hVFF cultured under inflammatory or normal conditions. Inflammatory and pro-fibrotic stimuli were induced by interleukin (IL)1β and transforming growth factor (TGF)β1, respectively. Mechanical stimulation was applied four hours daily, over a period of 72 hours. Outcome measurements comprised assessment of extracellular matrix (ECM)-related components, angiogenic factors, and inflammatory and fibrogenic markers on gene expression and protein levels. Results Under inflammatory conditions, the inflammatory cytokine IL11, as well as the myofibroblast marker alpha smooth muscle actin (α-SMA) were significantly reduced when additional vibration was applied. The desirable anti-fibrotic ECM component hyaluronic acid was increased following cytokine treatment, but was not diminished following vibration. Conclusion Our experiments revealed the effect of vibrational stress on hVFF in an inflammatory state. Elevated levels of certain pro-inflammatory/pro-fibrotic factors could be mitigated by additional vibrational excitation in an in vitro setting. These findings corroborate clinical studies which recommend early voice activation following an acute event.

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“…Whole search criteria were used as follows: trypsin, max. Missed cleavage sites: 1; search mode: MS/MS ion search with decoy database search comprised; precursor mass tolerance +/− 10ppm; product mass tolerance +/− 0.8 Da; acceptance parameters for identification: 1 % target FDR ( 30 ).…”

Section: Methodsmentioning

confidence: 99%

Identification and Immunological Characterization of Somatic Proteins from Adults of Toxocara cati by Proteomics Technique

Soleymani¹,

Grunberger²,

Abnous³

et al. 2021

IJPA

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Background: Toxocara cati is considered as one of the main etiological agents of toxocariasis with global and regional importance. As there is no information on proteomics of T. cati, herein, we reported the results obtained by proteomic analysis of somatic proteins extract, using a mass spectrometry (LC–MS/MS) approach. Methods: Somatic extract fractions were separated by two-dimensional SDSPAGE and were electro blotted on to PVDF membranes for immunoblot analysis, then collected the immunogenic spots which response of antibodies of the paratenic hosts (mice) to the antigens ( Mashhad, 2017), and analyzed by LC–MS/MS. The LC-MS/MS data were analyzed by Mascot database, Taxonomy Toxocara, and common contaminants, in Omics Center, Biotechnology Medical University of Graz (Austria, 2018). Result: The protein spots were isolated between 15–140 kDa ranges using 3–10 non-linear IPG strips and Brilliant Blue Coomassie. Ten proteins were characterized as immunogenic proteins, seven of them were identified and three of them were unknown proteins. Conclusion: This study provided additional information about the somatic antigens of T. cati, which can lead to the development of new strategies for novel immuno-modulators, drug targets, subunit vaccines and immunodiagnostic kits for toxocariasis.

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Proteomic Analysis of Vocal Fold Fibroblasts Exposed to Cigarette Smoke Extract: Exploring the Pathophysiology of Reinke's Edema*

Cited by 23 publications

References 63 publications

Exploring the Pathophysiology of Reinke's Edema: The Cellular Impact of Cigarette Smoke and Vibration

Exploring the Pathophysiology of Reinke's Edema: The Cellular Impact of Cigarette Smoke and Vibration

In vitro mechanical vibration down-regulates pro-inflammatory and pro-fibrotic signaling in human vocal fold fibroblasts

Identification and Immunological Characterization of Somatic Proteins from Adults of Toxocara cati by Proteomics Technique

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