M. Preeti Sivasankar

Quantification of acute vocal fold epithelial surface damage with increasing time and magnitude doses of vibration exposure.

Because the vocal folds undergo repeated trauma during continuous cycles of vibration, the epithelium is routinely susceptible to damage during phonation. Excessive and prolonged vibration exposure is considered a significant predisposing factor in the development of vocal fold pathology. The purpose of the present study was to quantify the extent of epithelial surface damage following increased time and magnitude doses of vibration exposure using an in vivo rabbit phonation model. Forty-five New Zealand white breeder rabbits were randomized to nine groups and received varying phonation time-doses (30, 60, or 120 minutes) and magnitude-doses (control, modal intensity phonation, or raised intensity phonation) of vibration exposure. Scanning electron microscopy and transmission electron microscopy was used to quantify the degree of epithelial surface damage. Results revealed a significant reduction in microprojection density, microprojection height, and depth of the epithelial surface with increasing time and phonation magnitudes doses, signifying increased epithelial surface damage risk with excessive and prolonged vibration exposure. Destruction to the epithelial cell surface may provide significant insight into the disruption of cell function following prolonged vibration exposure. One important goal achieved in the present study was the quantification of epithelial surface damage using objective imaging criteria. These data provide an important foundation for future studies of long-term tissue recovery from excessive and prolonged vibration exposure.

Influence of Obligatory Mouth Breathing, During Realistic Activities, on Voice Measures

OBJECTIVE Low humidity environments and mouth breathing may contribute to superficial vocal fold dehydration. The purpose of this study was to investigate the effects of obligatory mouth breathing, during daily activities in low- and high-humidity environments, on voice measures. The activities included 15 minutes of obligatory mouth breathing alone, during loud reading and during exercise. The effects of mouth breathing and humidity were compared in subjects who either reported or did not report vocal worsening after heavy voice use. STUDY DESIGN Prospective, between-group, repeated-measures design. METHODS Sixty-three healthy adults with normal respiratory function and perceptually normal voice participated in this study. Thirty-one subjects reported symptoms of voice worsening with heavy voice use. Thirty-two subjects who did not report these symptoms participated as controls. Phonation threshold pressure and perceived phonatory effort were measured at baseline and after each obligatory mouth breathing challenge. Ambient humidity was set to either low or high humidity. RESULTS Obligatory mouth breathing in loud reading and exercise significantly increased phonation threshold pressure when compared with mouth breathing alone. This increase in phonation threshold pressure was observed at low and high humidity, in both subject groups. There were no significant effects for perceived phonatory effort. CONCLUSIONS Obligatory mouth breathing during loud reading and exercise negatively impact phonation threshold pressure. Future investigations that include longer challenge durations, and subjects with voice disorders, are needed to elucidate the underlying mechanisms for increases in phonation threshold pressure.

Reducing the Negative Vocal Effects of Superficial Laryngeal Dehydration With Humidification

Objectives: Environmental humidification is a simple, cost-effective method believed to reduce superficial laryngeal drying. This study sought to validate this belief by investigating whether humidification treatment would reduce the negative effects of superficial laryngeal dehydration on phonation threshold pressure (PTP). Phonation threshold pressure data analysis may be vulnerable to bias because of lack of investigator blinding. Consequently, this study investigated the extent of PTP analysis reliability between unblinded and blinded investigators. Methods: Healthy male and female adults were assigned to a vocal fatigue (n = 20) or control group (n = 20) based on their responses to a questionnaire. PTP was assessed after 2 hours of mouth breathing in low humidity (dehydration challenge), following a 5-minute break in ambient humidity, and after 2 hours of mouth breathing in high humidity (humidification). Results: PTP significantly increased following the laryngeal dehydration challenge. After humidification, PTP returned toward baseline. These effects were observed in both subject groups. PTP measurements were highly correlated between the unblinded and blinded investigator. Conclusions: Humidification may be an effective approach to decrease the detrimental voice effects of superficial laryngeal dehydration. These data lay the foundation for future investigations aimed at preventing and treating the negative voice changes associated with chronic, surface laryngeal drying.

Effects of phonation time and magnitude dose on vocal fold epithelial genes, barrier integrity, and function.

To investigate the effects of increasing time and magnitude doses of vibration exposure on transcription of the vocal folds junctional proteins, structural alterations, and functional tissue outcomes.

Role for Ion Transport in Porcine Vocal Fold Epithelial Defense to Acid Challenge

Objective. The vocal fold epithelium is routinely exposed to gastric contents, including acid and pepsin, during laryngopharyngeal reflux events. The epithelium may possess intrinsic defenses to reflux. The first objective of the current study was to examine whether vocal fold epithelial ion transport is one potential mechanism of defense to gastric contents. The second objective was to determine whether ion transport in response to gastric contents is associated with the secretion of bicarbonate. Study Design. Prospective design in excised porcine larynges. Setting. Laboratory. Subjects and Methods. Porcine vocal folds (N = 56) were exposed on the luminal surface to acid, pepsin, or sham challenges. Ion transport at baseline and following challenge exposure was measured using electrophysiological techniques. To examine specific ion transport mechanisms, vocal folds were pretreated with either a sodium channel blocker or bicarbonate channel blocker. Results. Within 60 seconds of acid but not pepsin exposure, there was a significant increase in ion transport. This rapid increase in ion transport was transient and related to bicarbonate secretion. Conclusion. The current data suggest that porcine vocal folds immediately increase bicarbonate secretion following exposure to acid. Bicarbonate secretion may act to neutralize acid. These findings contribute to the identification of the mechanisms underlying vocal fold defense to reflux and offer implications for the development of treatments for reflux-induced vocal fold injury.

The Interaction of Surface Hydration and Vocal Loading on Voice Measures.

OBJECTIVES Vocal loading tasks provide insight regarding the mechanisms underlying healthy laryngeal function. Determining the manner in which the larynx can most efficiently be loaded is a complex task. The goal of this study was to determine if vocal loading could be achieved in 30 minutes by altering phonatory mode. Owing to the fact that surface hydration facilitates efficient vocal fold oscillation, the effects of environmental humidity on vocal loading were also examined. This study also investigated whether the detrimental effects of vocal loading could be attenuated by increasing environmental humidity. METHODS Sixteen vocally healthy adults (8 men, 8 women) completed a 30-minute vocal loading task in low and moderate humidity. The order of humidities was counterbalanced across subjects. The vocal loading task consisted of reading with elevated pitch and pressed vocal quality and low pitch and pressed and/or raspy vocal quality in the presence of 65 dB ambient, multi-talker babble noise. RESULTS Significant effects were observed for (1) cepstral peak prominence on soft sustained phonation at 10th and 80th pitches, (2) perceived phonatory effort, and (3) perceived tiredness ratings. No loading effects were observed for cepstral peak prominence on the rainbow passage, although fundamental frequency on the rainbow passage increased post loading. No main effect was observed for humidity. CONCLUSIONS Following a 30-minute vocal loading task involving altering laryngeal vibratory mode in combination with increased volume. Also, moderate environmental humidity did not significantly attenuate the negative effects of loading.

A Review of Vocal Loading Tasks in the Voice Literature

Vocal loading tasks (VLTs) are used to assess range of function and vulnerability of the laryngeal system. The manner in which VLTs compromise the laryngeal mechanism is multifaceted and only partially understood. This paper presents a scoping review of the literature on VLTs and their effects on the healthy voice. This review paper details the varied nature of VLT duration, task type, and additional factors including hydration and ambient noise that have been used to compromise the larynx. In addition, the voice measures used to assess the effects of VLTs on the healthy larynx are discussed. This study details the current state of the literature, draws conclusions from our current knowledge, and provides directions for further research.

Quantifying the effects of altering ambient humidity on ionic composition of vocal fold surface fluid.

Vocal fold surface fluid (VFSF) is important in hydration and defense of underlying epithelial cells. The objective of this study was to quantify changes in the ionic composition of VFSF after altering the humidity of inhaled air. We tested the hypothesis that low humidity exposure would increase the concentration of VFSF sodium (Na+) and chloride (Cl−) ions but that high humidity exposure would decrease the concentration of VFSF Na+ and Cl− ions as compared to the low humidity challenge.

Vocal Loading and Environmental Humidity Effects in Older Adults

The goal of this study was to investigate the effects of a vocal loading challenge that combined prolonged speaking, in child-directed voice within a noisy environment, in older adults. An additional goal was to determine whether increased environmental humidity would attenuate the negative effects of this vocal loading challenge. METHOD Thirteen healthy subjects (five males and eight females; >65 years) completed a vocal loading challenge. The challenge involved 45 minutes of child-directed speech in the presence of 65 dB multitalker babble background noise. Subjects completed this challenge in both low humidity and moderate humidity in counterbalanced order. Vocal function was assessed before and after the challenge using phonation threshold pressure (PTP), cepstral peak prominence (CPP), low/high spectral ratio (LHR), perceived phonatory effort (PPE), and perceived vocal tiredness. RESULTS Negative changes were observed in the aging larynx following the 45-minute vocal loading challenge. Measures of PTP, PPE, and perceived vocal tiredness demonstrated statistically significant loading effects. Increased ambient humidity significantly alleviated the negative changes observed in PPE and perceived vocal tiredness. Increased humidity significantly improved CPP measures both before and after the vocal loading challenge. CONCLUSIONS This study furthers our understanding of how older adults respond to a vocal loading challenge of prolonged nonhabitual speech in a noisy environment. Our data suggest that the aging voice is negatively affected by prolonged loud speaking and that humidification may be beneficial in reducing some of these negative effects.

Acute Acrolein Exposure Induces Impairment of Vocal Fold Epithelial Barrier Function.

Acrolein is a ubiquitous pollutant abundant in cigarette smoke, mobile exhaust, and industrial waste. There is limited literature on the effects of acrolein on vocal fold tissue, although there are clinical reports of voice changes after pollutant exposures. Vocal folds are responsible for voice production. The overall objective of this study was to investigate the effects of acrolein exposure on viable, excised vocal fold epithelial tissue and to characterize the mechanism underlying acrolein toxicity. Vocal fold epithelia were studied because they form the outermost layer of the vocal folds and are a primary recipient of inhaled pollutants. Porcine vocal fold epithelia were exposed to 0, 50, 100, 500, 900 or 1300 μM of acrolein for 3 hours; the metabolic activity, epithelial resistance, epithelial permeability, tight junction protein (occludin and claudin 3) expression, cell membrane integrity and lipid peroxidation were investigated. The data demonstrated that acrolein exposure at 500 μM significantly reduced vocal fold epithelial metabolic activity by 27.2% (p≤0.001). Incubation with 100 μM acrolein caused a marked increase in epithelial permeability by 130.5% (p<0.05) and a reduction in transepithelial electrical resistance (TEER) by 180.0% (p<0.001). While the expression of tight junctional protein did not change in acrolein-treated samples, the cell membrane integrity was significantly damaged with a 45.6% increase of lipid peroxidation as compared to controls (p<0.05). Taken together, these data provide evidence that acute acrolein exposure impairs vocal fold epithelial barrier integrity. Lipid peroxidation-induced cell membrane damage may play an important role in reducing the barrier function of the epithelium.