Effects of Larynx Preservation Method on Phonation Threshold Flow in an Excised Porcine Benchtop Model
Abstract
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.