Vocal Fold Ion Transport and Mucin Expression Following Acrolein Exposure

Levendoski, Elizabeth Erickson; Sivasankar, M. Preeti

doi:10.1007/s00232-014-9651-2

Cited by 13 publications

(24 citation statements)

References 44 publications

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“…An untreated, sham challenge (PBS alone) was included as a control. Similar challenges have been utilized previously to investigate the adverse effects of clinically relevant insults on the vocal fold epithelial barrier and were specifically chosen for the purpose of the current investigation because these were hypothesized to reduce epithelial viability. Although the concentration of acrolein and pH of HCl is representative of what occurs in vivo, a lower concentration of H 2 O 2 has been used in previous studies in the vocal folds to mimic physiologic conditions .…”

Section: Methodsmentioning

confidence: 99%

“…Consequently, we tested whether three clinically relevant challenges would reduce viability as measured by epithelial metabolic activity and cell membrane integrity as compared to an untreated, sham challenge. Challenges included: acrolein, an environmentally pervasive pollutant; hydrochloric acid, the acidic component of laryngopharyngeal reflux; and hydrogen peroxide, a common reactive oxygen species . We hypothesized that these assays would detect reductions in epithelial viability following challenges as compared to sham.…”

Section: Introductionmentioning

confidence: 99%

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Utility of cell viability assays for use with ex vivo vocal fold epithelial tissue

2014

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Objective Ex vivo models are routinely used to investigate the barrier function of the vocal fold epithelium. However, there are limited reports on assays that can be used to investigate the effect of clinically-relevant challenges on vocal fold epithelial tissue viability. Our objective was to determine the utility of two assays routinely used in cell culture – a cellular metabolic activity assay and a cell membrane integrity assay - to investigate the viability of ex vivo porcine vocal fold epithelium. Study Design Prospective, ex vivo animal study Methods Porcine vocal folds were exposed to acrolein, hydrochloric acid, or hydrogen peroxide challenge. An untreated, sham challenge was included as a control. Assays including metabolic activity, cell membrane integrity, and histology were used to determine whether challenges reduced epithelial viability as compared to sham. Results Cell membrane integrity and metabolic activity assays detected reductions in viability following hydrochloric acid and hydrogen peroxide challenges but not acrolein challenge as compared to sham. No challenge produced significant changes in epithelial appearance as evidenced by light microscopy. Conclusion Metabolic activity and cell membrane integrity assays are valuable tools that can be used to evaluate the viability of ex vivo vocal fold epithelial tissue following clinically-relevant challenges. As viability is reduced, the ability of epithelial tissue to maintain its barrier function is compromised. Accurate assessment of viability may provide us clues into understanding mechanisms underlying vocal fold epithelial injury and disease.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Utility of cell viability assays for use with ex vivo vocal fold epithelial tissue

2014

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“…There is some research on the effects of acrolein exposure to the larynx. In excised porcine vocal folds, acute acrolein exposure reduced epithelial sodium ion transport, induced lipid peroxidation, and impaired epithelial integrity and barrier function . However, in vivo studies are needed to fully understand the mechanisms underlying pathological changes from specific pollutant exposures such as acrolein.…”

Section: Introductionmentioning

confidence: 99%

“…In excised porcine vocal folds, acute acrolein exposure reduced epithelial sodium ion transport, induced lipid peroxidation, and impaired epithelial integrity and barrier function. 14,15 However, in vivo studies are needed to fully understand the mechanisms underlying pathological changes from specific pollutant exposures such as acrolein. The underlying mechanisms for acrolein-induced inflammation in the distal airway have been investigated.…”

Section: Introductionmentioning

confidence: 99%

Subacute acrolein exposure to rat larynx in vivo

et al. 2018

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Objectives/Hypothesis Inhaled pollutants can contact vocal fold tissue and induce detrimental voice changes. Acrolein is a pollutant in cigarette smoke and can also be inhaled during the combustion of fossil fuels, animal fats, and plastics in the environment. However, the vocal fold pathological changes induced by acrolein and the underlying inflammatory pathways are not well understood. These biologic data are needed to understand why voice problems may result from pollutant exposure. Study Design In vivo prospective design with experimental and control groups. Methods Sprague‐Dawley male rats (N = 36) were exposed to acrolein (3 ppm) or filtered air (control) through a whole‐body exposure system for 5 hours/day, for 5 days/week, over 4 weeks. Histopathological changes, presence of edema, expression of proinflammatory cytokines and markers, and the phosphorylation of nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB) were investigated. Results Histological evaluation and quantification demonstrated that subacute acrolein exposure induced significant vocal fold edema. Acrolein exposure also induced epithelial sloughing and cell death. Quantitative polymerase chain reaction showed a significant upregulation of genes encoding interferon regulatory factor and chitinase‐3‐like protein 3. Western blot revealed a 76.8% increase in phosphorylation of NF‐κB P65 after subacute acrolein exposure. Conclusions These findings suggest that 4‐week exposures to 3 ppm acrolein induce vocal fold inflammation manifested as edema, related to the activation of NF‐κB signaling. The edema may underlie the voice changes reported in speakers exposed to pollutants. Level of Evidence NA Laryngoscope, 129:E313–E317, 2019

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“…This epithelial barrier can be adversely affected by pollutants found in ciagarettes. 9–12 Though no study has investigated the effect of CNTs on vocal fold epithelium, CNTs are reported to reduce epithelial resistance in a tracheobronchial epithelial cell line. 13 Fibroblasts are the most common cell type in the vocal folds and are important for synthesizing extracellular matrix (ECM) components such as collagen and elastin.…”

Section: Introductionmentioning

confidence: 99%

Acute Nanoparticle Exposure to Vocal Folds: A Laboratory Study

et al. 2017

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Objective Airway exposure to nanoparticles is common in occupational settings. Inhaled nanoparticles have toxic effects on respiratory tissue. Vocal folds are also at direct risk from inhaled nanoparticles. This study investigated the effects of single-walled carbon nanotubes (SWCNT), a type of nanoparticle, on vocal fold epithelium and fibroblasts. These cell types were selected for study since the epithelium is the outer layer of the vocal folds and fibroblasts are the most common cell type in connective tissue underlying the epithelium. Methods Native porcine vocal fold epithelium and cultured human vocal fold fibroblasts were exposed to SWCNTs (100 ng/mL) and control (no SWCNT), in vitro. Epithelial and fibroblast viability were measured using MTT assay. Epithelial barrier integrity was assessed with transepithelial resistance and sodium fluorescein permeability. Epithelial tight junctional protein occludin expression was measured with the Western blot. Gene expressions of the fibroblast specific protein 1 (FSP-1), α-smooth muscle actin (α-SMA), and collagen III (Col-III) were assessed using quantitative polymerase chain reaction. Results Transcriptional expression of genes encoding FSP-1 and Col-III were increased significantly following SWCNT exposure. There were no significant differences between control and SWCNT groups on any of the other measures. Conclusions SWCNT exposure induces vocal fold fibroblasts to a fibrotic phenotype. These data help us understand vocal fold defense mechanisms and lay the groundwork for studying the physiological effects of nanoparticle exposure in vivo.

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Vocal Fold Ion Transport and Mucin Expression Following Acrolein Exposure

Cited by 13 publications

References 44 publications

Utility of cell viability assays for use with ex vivo vocal fold epithelial tissue

Utility of cell viability assays for use with ex vivo vocal fold epithelial tissue

Subacute acrolein exposure to rat larynx in vivo

Acute Nanoparticle Exposure to Vocal Folds: A Laboratory Study

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