An excised animal larynx model has been used in many studies to better understand the physiological and anatomical properties of the human larynx. One difference between an ex vivo model and an in vivo model is that ion loss occurs postmortem. To compensate for this in the excised model, researchers most commonly use a preservation method that includes completely submerging the specimen in isotonic saline (0.9% Na+Cl-) and then flash freezing it in liquid nitrogen. The flash freezing method allows researchers to maintain the integrity of the structures while also being able to gather specimens as they become available. Not enough research has been done to understand the effects of a preservation method on the outcomes of the study. Additionally, no common method has been established for preservation across studies to ensure that results are not being influenced by this variable. This prospective, mixed experimental design study includes three groups, a control group and two experimental groups. The control group consisted of 10 bench-mounted porcine larynges that were soaked in isotonic saline and flash frozen with liquid nitrogen. Prior to the experiment, the frozen larynges were thawed overnight before trials. The other two groups consisted of 10 bench-mounted porcine larynges each; these larynges were soaked in either isotonic saline or Ringer’s solution, a balanced fluid used in vivo to counteract dehydration. Larynges from these two groups were kept fresh and stored in a refrigerator overnight before trials. On the day of experimentation, each larynx was mounted on a bench top setup including three micropositioners to stabilize, adduct, and elongate the vocal folds. All the larynges were connected to a pseudolung via the trachea and humidified air was passed through to the vocal folds until phonation was achieved. Phonatory trials consisted of brief phonation followed by 5-minute desiccation intervals until phonation was no longer achieved. Phonation threshold flow (PTF), defined as the flow observed at the onset of phonation, was observed during each phonation trial; and flow values were compared within and between groups. Statistically significant differences were found between the Ringer’s group and the fresh saline group as well as between the Ringer’s group and the frozen saline group, indicating that PTF is influenced by the larynx preservation method.

Several studies involving excised animal larynges have been performed to simulate the structural and physiological properties of the human larynx. The most common way to preserve the laryngeal tissue being studied is by immersing it in a 0.9% isotonic saline solution and then flash freezing it. Isotonic saline is used empirically to replenish the potential ion loss that occurs postmortem. Each larynx is flash frozen so it can be used at a more convenient time while still maintaining the integrity of the tissue. However, the preservation methods found in previous studies tend to vary and no consensus had been reached about which method of preservation is ideal. This study sought to investigate the effects of solution and storage on phonation threshold pressure (PTP). Phonation threshold pressure is commonly used to investigate mucosal wave of the vocal folds, prephonatory glottal width, and vocal fold cover. This study involved a prospective, mixed experimental design with three groups, including a control group and two experimental groups. Each group consisted of 10 bench-mounted porcine larynges. The control group was immersed in 0.9% isotonic saline, flash frozen with liquid nitrogen within 24 hours postmortem, and thawed overnight before the experiment. The second group was immersed in 0.9% isotonic saline and the third group was submersed in Ringer's solution. Each of these groups was kept in their solution in a refrigerator for approximately 15 hours and was used for the experiment within 24 hours postmortem. Each larynx was mounted on a bench on a tabletop with three micropositioners to adduct and elongate the vocal folds. A pseudolung connected to the trachea directed humidified air to the vocal folds subglottally until phonation was achieved. The larynges in all three groups underwent these phonatory trials with 5-minute desiccation trials between each until phonation could no longer be achieved. Phonation threshold pressure was then observed and compared within groups and between groups. The signals were obtained using MATLAB. The results indicated that PTP was lowest for the frozen versus fresh groups. PTP values increased slightly for the frozen group, but the frozen group demonstrated less variability across specimens as compared to the fresh groups. Collectively, these results indicate that there are substantial differences between fresh and frozen specimens. These differences should be considered when designing tissue studies for purposes of generalization to human phonation.

Hydration of the vocal folds is important for the production of normal voice. Dehydration makes voice production more difficult and increases vocal effort. Laryngeal desiccation has been shown to increase phonation threshold pressure (PTP) and self-perceived phonatory effort (PPE) in females. Nebulized saline may reverse or offset this effect. However, few data exist regarding the effects of laryngeal desiccation and nebulized treatments in males. Further, the dose-response relationship between laryngeal desiccation and nebulized hydration treatments is unknown. This study examined the effects of two doses of nebulized isotonic saline following a laryngeal desiccation challenge in healthy male speakers. In a double-blinded, within-subjects design, 10 male college students (age range 18-26 years) attended two data collection sessions involving a 30-minute desiccation challenge followed by 3 mL or 9 mL of nebulized isotonic saline. PTP for the 10th and 80th fundamental frequency (F0) percentiles and PPE were collected before and after the desiccation challenge and at 5, 35, and 65 minutes after the nebulized treatment. PPE increased significantly following the laryngeal desiccation challenge (p < .01). Following nebulization, PPE decreased toward baseline for both doses of isotonic saline (p < .01), but failed to reverse the desiccation effect completely. No statistically significant changes in PTP occurred following the laryngeal desiccation challenge or subsequent treatments. Compared with previous research involving females, these results suggest males may respond differently to laryngeal desiccation and nebulized hydration treatments.

Vocal fold hydration is important for healthy function of the vocal mechanism. Vocal fold surface fluid protects the mucosa and facilitates efficient vocal fold oscillation. Dry air exposure, mouth breathing, insufficient intake of liquids, and behavioral factors may contribute to laryngeal dehydration. Singers are believed to be particularly at risk for voice problems related to dehydration due to environmental and voice use factors. Laryngeal desiccation and nebulized hydration treatments have been shown to influence phonation threshold pressure (PTP) and self-perceived phonatory effort (PPE) in females. However, little research exists exploring the effects of hydration in males. Additionally, few studies have examined the dose-response relationship of hydration treatments. This investigation examined the effects of a laryngeal desiccation challenge and two different doses of nebulized isotonic saline on voice production in trained male singers. In a double-blind, within-subjects repeated measures crossover investigation, 10 male singers (ages 18 to 24) received a 30 minute laryngeal desiccation challenge followed by either 3 mL or 9 mL of nebulized isotonic saline on two consecutive weeks. PTP, PPE, and self-perceived mouth and throat dryness were sampled during the following observations: predesiccation, post-desiccation, and at 5, 35, and 65 minutes post-nebulization. No differences in PTP were observed after desiccation or nebulized treatment. PPE, however, rose significantly after desiccation and returned near baseline after treatment. No significant differences between dosages were observed.

An excised animal larynx model has been used in many studies to better understand the physiological and anatomical properties of the human larynx. One difference between an ex vivo model and an in vivo model is that ion loss occurs postmortem. To compensate for this in the excised model, researchers most commonly use a preservation method that includes completely submerging the specimen in isotonic saline (0.9% Na+Cl-) and then flash freezing it in liquid nitrogen. The flash freezing method allows researchers to maintain the integrity of the structures while also being able to gather specimens as they become available. Not enough research has been done to understand the effects of a preservation method on the outcomes of the study. Additionally, no common method has been established for preservation across studies to ensure that results are not being influenced by this variable. This prospective, mixed experimental design study includes three groups, a control group and two experimental groups. The control group consisted of 10 bench-mounted porcine larynges that were soaked in isotonic saline and flash frozen with liquid nitrogen. Prior to the experiment, the frozen larynges were thawed overnight before trials. The other two groups consisted of 10 bench-mounted porcine larynges each; these larynges were soaked in either isotonic saline or Ringers solution, a balanced fluid used in vivo to counteract dehydration. Larynges from these two groups were kept fresh and stored in a refrigerator overnight before trials. On the day of experimentation, each larynx was mounted on a bench top setup including three micropositioners to stabilize, adduct, and elongate the vocal folds. All the larynges were connected to a pseudolung via the trachea and humidified air was passed through to the vocal folds until phonation was achieved. Phonatory trials consisted of brief phonation followed by 5-minute desiccation intervals until phonation was no longer achieved. Phonation threshold flow (PTF), defined as the flow observed at the onset of phonation, was observed during each phonation trial; and flow values were compared within and between groups. Statistically significant differences were found between the Ringers group and the fresh saline group as well as between the Ringers group and the frozen saline group, indicating that PTF is influenced by the larynx preservation method.