Interstitial protein alterations in rabbit vocal fold with scar

Fibrous and interstitial proteins compose the extracellular matrix of the vocal fold lamina propria and account for its biomechanic properties. Vocal fold scarring is characterized by altered biomechanical properties, which create dysphonia. Although alterations of the fibrous proteins have been confirmed in the rabbit vocal fold scar, interstitial proteins, which are known to be important in wound repair, have not been investigated to date. Using a rabbit model, interstitial proteins decorin, fibromodulin, and fibronectin were examined immunohistologically, two months postinduction of vocal fold scar by means of forcep biopsy. Significantly decreased decorin and fibromodulin with significantly increased fibronectin characterized scarred vocal fold tissue. The implications of altered interstitial proteins levels and their affect on the fibrous proteins will be discussed in relation to increased vocal fold stiffness and viscosity, which characterizes vocal fold scar.     

Hyaluronic Acid: Its Role in Voice

The extracellular matrix (ECM), once regarded simply as a structural scaffold, is now recognized as an important modulator of cellular behavior and function. One component that plays a prominent role in this process is hyaluronic acid (HA)--a molecule found in many different tissues. Research into the roles of HA indicates that it plays a key role in tissue viscosity, shock absorption, and space filling. Specifically, research into the role of HA in laryngology indicates that it has profound effects on the structure and viscosity of vocal folds. This article provides an introduction to the structure and biological functions of HA and its importance in voice. In addition, an overview of the pharmaceutical applications of HA is discussed.     

Voice source characteristics in Mongolian "throat singing" studied with high-speed imaging technique, acoustic spectra, and inverse filtering.

Mongolian "throat singing" can be performed in different modes. In Mongolia, the bass-type is called Kargyraa. The voice source in bass-type throat singing was studied in one male singer. The subject alternated between modal voice and the throat singing mode. Vocal fold vibrations were observed with high-speed photography, using a computerized recording system. The spectral characteristics of the sound signal were analyzed. Kymographic image data were compared to the sound signal and flow inverse filtering data from the same singer were obtained on a separate occasion. It was found that the vocal folds vibrated at the same frequency throughout both modes of singing. During throat singing the ventricular folds vibrated with complete but short closures at half the frequency of the true vocal folds, covering every second vocal fold closure. Kymographic data confirmed the findings. The spectrum contained added subharmonics compared to modal voice. In the inverse filtered signal the amplitude of every second airflow pulse was considerably lowered. The ventricular folds appeared to modulate the sound by reducing the glottal flow of every other vocal fold vibratory cycle.     

Vestibular vocal fold behavior during phonation in unilateral vocal fold paralysis

Voice quality in patients with vocal fold paralysis can be affected by several factors, such as the position of the paralyzed vocal fold, its degree of atrophy, the configuration of its free edge, and the level differences between both vocal folds. Depending on the related vocal deficiency the patient will attempt to compensate using different maneuvers, such as increment of vocal tract and neck muscle contraction to improve glottal closure. This is probably one of the reasons why ventricular folds are frequently requested. The objective of this study is to analyze the behavior of the homolateral and contralateral vestibular folds to delineate patterns of vestibular motion during sustained phonation, in cases of unilateral vocal fold paralysis.     

Voice Source Characteristics in Mongolian “Throat Singing” Studied with High-Speed Imaging Technique, Acoustic Spectra, and Inverse Filtering

Mongolian “throat singing” can be performed in different modes. In Mongolia, the bass-type is called Kargyraa. The voice source in bass-type throat singing was studied in one male singer. The subject alternated between modal voice and the throat singing mode. Vocal fold vibrations were observed with high-speed photography, using a computerized recording system. The spectral characteristics of the sound signal were analyzed. Kymographic image data were compared to the sound signal and flow inverse filtering data from the same singer were obtained on a separate occasion. It was found that the vocal folds vibrated at the same frequency throughout both modes of singing. During throat singing the ventricular folds vibrated with complete but short closures at half the frequency of the true vocal folds, covering every second vocal fold closure. Kymographic data confirmed the findings. The spectrum contained added subharmonics compared to modal voice. In the inverse filtered signal the amplitude of every second airflow pulse was considerably lowered. The ventricular folds appeared to modulate the sound by reducing the glottal flow of every other vocal fold vibratory cycle.     

Towards the Expression of Sex Hormone Receptors in the Human Vocal Fold

BACKGROUND: The human larynx is assumed to be a steroid receptor target organ. There are only very limited data on the evidence of steroid receptors in the vocal folds, although voice alterations due to hormonal influence and treatment have been found. GOAL OF THE STUDY: To investigate the expression of estrogen alpha, progesterone, and androgen receptors in human vocal folds (vocalis muscle, glands, lamina propria, epithelium). METHODS: Immunohistochemically, vocal fold cadaver specimens of 15 autopsied patients (6 women, 9 men), which were taken approximately 4 to 8 hours postmortem were investigated. Furthermore, one (male) vocal fold biopsy obtained intraoperatively during a laryngectomy was tested. RESULTS: No specific immunohistochemical staining for the different types of steroid hormones investigated could be observed in either the postmortem taken biopsies nor the intraoperatively one. However, several unspecific staining patterns could be observed. CONCLUSION: The results of this study contradict recently published data and question the expression of sex hormone receptors in the vocal folds. Main causes of false interpretations of unspecific staining are discussed.     

Three-dimensional biomechanical properties of human vocal folds: parameter optimization of a numerical model to match in vitro dynamics

The human voice signal originates from the vibrations of the two vocal folds within the larynx. The interactions of several intrinsic laryngeal muscles adduct and shape the vocal folds to facilitate vibration in response to airflow. Three-dimensional vocal fold dynamics are extracted from in vitro hemilarynx experiments and fitted by a numerical three-dimensional-multi-mass-model (3DM) using an optimization procedure. In this work, the 3DM dynamics are optimized over 24 experimental data sets to estimate biomechanical vocal fold properties during phonation. Accuracy of the optimization is verified by low normalized error (0.13 ± 0.02), high correlation (83% ± 2%), and reproducible subglottal pressure values. The optimized, 3DM parameters yielded biomechanical variations in tissue properties along the vocal fold surface, including variations in both the local mass and stiffness of vocal folds. That is, both mass and stiffness increased along the superior-to-inferior direction. These variations were statistically analyzed under different experimental conditions (e.g., an increase in tension as a function of vocal fold elongation and an increase in stiffness and a decrease in mass as a function of glottal airflow). The study showed that physiologically relevant vocal fold tissue properties, which cannot be directly measured during in vivo human phonation, can be captured using this 3D-modeling technique.     

Botulinum toxin injection in the treatment of vocal fold paralysis associated with multiple sclerosis: A case report

Botulinum toxin has been demonstrated clinically to be an effective treatment for a variety of laryngeal problems, most notably spasmodic dysphonia. As in other movement disorders, the theory behind the injection of this substance in the larynx has been a weakening of the vocal fold musculature to relieve uncoordinated and spasmodic movement of the vocal folds, presumably rebalancing the forces within the intralaryngeal musculature. Recently, this concept was applied to help reposition the arytenoid cartilage in acute and longstanding anteromedial cricoarytenoid dislocations. This same concept may apply to the paralyzed vocal fold. In support of this idea, a number of investigators have shown that immobile, clinically paralyzed vocal folds may still have partial voluntary motor unit activity. This voluntary activation may not produce clinically evident movement but may be sufficient to produce tone within the fold. If the voluntary motor units in the abductor musculature of the paralyzed fold are weakened with botulinum toxin, the continued pull of the functioning adductor musculature may be sufficient to medialize the paralyzed fold. This idea has been supported by animal experiments, which have shown that botulinum toxin may affect the ability of the fold to rebalance itself. With this evidence in mind, a patient with fold immobility secondary to multiple sclerosis was treated in an attempt at laryngeal rebalancing, using botulinum toxin to medialize the fold. However, instead of simply having the fold return fixed to the midline, the patient regained normal laryngeal mobility and voice. While it is unclear whether the botulinum toxin alone was responsible, the coincidence of this occurrence certainly requires reporting. This paper is a report of the first successful treatment of vocal fold paralysis using botulinum toxin to treat vocal fold fixation in a patient with multiple sclerosis.     

The shear modulus of the human vocal fold in a transverse direction

The aim of this study was to measure the shear modulus of the vocal fold in a human hemilarynx, such that the data can be related to direction of applied stress and anatomical context. Dynamic spring rate data were collected using a modified linear skin rheometer using human hemilarynges, and converted to estimated shear modulus via application of a simple shear model. The measurement probe was attached to the epithelial layer of the vocal fold cover using suction. A sinusoidal force of 3g was applied to the epithelium, and the resultant displacement logged at a rate of 1kHz. Force measurement accuracy was 20microg and position measurement accuracy was 4microm. The force was applied in a transverse direction at the midmembranous point between the vocal process and the anterior commissure. The shear modulus of the three female vocal folds ranged from 814 to 1232Pa. The shear modulus of the three male vocal folds ranged from 1021 to 1796Pa. These data demonstrate that it is possible to obtain estimates for the shear modulus of the vocal fold while preserving anatomical context. The modulus values reported here are higher than those reported using parallel plate rheometry. This is to be expected as the tissue is attached to surrounding structures, and is under natural tension.     

Chaotic vibrations of a vocal fold model with a unilateral polyp

A nonlinear model was proposed to study chaotic vibrations of vocal folds with a unilateral vocal polyp. The model study found that the vocal polyp affected glottal closure and caused aperiodic vocal fold vibrations. Using nonlinear dynamic methods, aperiodic vibrations of the vocal fold model with a polyp were attributed to low-dimensional chaos. Bifurcation diagrams showed that vocal polyp size, stiffness, and damping had important effects on vocal fold vibrations. An increase in polyp size tended to induce subharmonic patterns and chaos. This study provides a theoretical basis to model aperiodic vibrations of vocal folds with a laryngeal mass.     

A theoretical model of the pressure field arising from asymmetric intraglottal flows applied to a two-mass model of the vocal folds

A theoretical flow solution is presented for predicting the pressure distribution along the vocal fold walls arising from asymmetric flow that forms during the closing phases of speech. The resultant wall jet was analyzed using boundary layer methods in a non-inertial reference frame attached to the moving wall. A solution for the near-wall velocity profiles on the flow wall was developed based on a Falkner-Skan similarity solution and it was demonstrated that the pressure distribution along the flow wall is imposed by the velocity in the inviscid core of the wall jet. The method was validated with experimental velocity data from 7.5 times life-size vocal fold models, acquired for varying flow rates and glottal divergence angles. The solution for the asymmetric pressures was incorporated into a widely used two-mass model of vocal fold oscillation with a coupled acoustical model of sound propagation. Asymmetric pressure loading was found to facilitate glottal closure, which yielded only slightly higher values of maximum flow declination rate and radiated sound, and a small decrease in the slope of the spectral tilt. While the impact on symmetrically tensioned vocal folds was small, results indicate the effect becomes more significant for asymmetrically tensioned vocal folds.     

Laser Doppler Measurements of the Vocal Fold Blood Micro-Circulation

The authors have developed a laser Doppler method, referred to as LDF+LS (laser Doppler flowmetry by laryngostereometry), for measuring the vocal fold micro-circulation. The vocal folds of 103 patients were examined during general anesthesia. A laser Doppler probe was placed on defined positions at the vocal fold edge: midmembranous position (MP), 2 mm and 4 mm behind, and 1 mm anterior to MP. Three parameters were derived, ie, the concentration (CMBC) and velocity (V) of moving blood cells, and the product of both resulting in perfusion: P = CMBCxV. The results of 53 subjects with normal vocal fold status showed that the V at MP was significantly lower than 2 mm behind MP (P < 0.05). Men had significantly higher velocity than women (P < 0.05). Subjects older than 65 years had higher perfusion and CMBC as compared with younger subjects (P < 0.01). Smokers with normal vocal folds had higher velocity than non-smokers (P < 0.05). Measurements of healthy vocal folds were compared with benign vocal fold pathology. Vocal fold polyps had significantly higher perfusion than normal vocal folds (P < 0.01). In conclusion, the laser Doppler measurement is a sensitive tool estimating superficial blood flow in both normal and pathological vocal folds. The blood flow in normals differs with respect to gender and age probably due to variations in micro-circulation of the mucosa and lamina propria. Vocal fold pathology and external factors such as smoking result in blood flow changes.     

Phonation Threshold Power in Ex Vivo Laryngeal Models

This study hypothesized that phonation threshold power is measureable and sensitive to changes in the biomechanical properties of the vocal folds. Phonation threshold power was measured in three sample populations of 10 excised canine larynges treated with variable posterior glottal gap, variable bilateral vocal fold elongation, and variable vocal fold lesioning. Posterior glottal gap varied from 0 to 4 mm in 0.5 mm intervals. Bilateral vocal fold elongation varied from 0% to 20% in 5% intervals. Vocal fold lesion treatments included unilateral and bilateral vocal fold lesion groups. Each treatment was investigated independently in a sample population of 10 excised canine larynges. Linear regression analysis indicated that phonation threshold power was sensitive to posterior glottal gap (R² = 0.298, P < 0.001) and weakly to vocal fold elongation (R² = 0.052, P = 0.003). A one-way repeated measures analysis of variance indicated that phonation threshold power was sensitive to the presence of lesions (P < 0.001). Theoretical and experimental evidence presented here suggests that phonation threshold power could be used as a broad screening parameter sensitive to certain changes in the biomechanical properties of the larynx. It has not yet been measured in humans, but because it has the potential to represent the airflow-tissue energy transfer more completely than the phonation threshold pressure or flow alone, it may be a more useful parameter than these and could be used to indicate that laryngeal health is likely abnormal.     

Influence of supraglottal structures on the glottal jet exiting a two-layer synthetic, self-oscillating vocal fold model

A synthetic two-layer, self-oscillating, life-size vocal fold model was used to study the influence of the vocal tract and false folds on the glottal jet. The model vibrated at frequencies, pressures, flow rates, and amplitudes consistent with human phonation, although some differences in behavior between the model and the human vocal folds are noted. High-speed images of model motion and flow visualization were acquired. Phase-locked ensemble-averaged glottal jet velocity measurements using particle image velocimetry (PIV) were acquired with and without an idealized vocal tract, with and without false folds. PIV data were obtained with varying degrees of lateral asymmetric model positioning. Glottal jet velocity magnitudes were consistent with those measured using excised larynges. A starting vortex was observed in all test cases. The false folds interfered with the starting vortex, and in some cases vortex shedding from the false folds was observed. In asymmetric cases without false folds, the glottal jet tended to skew toward the nearest wall; with the false folds, the opposite trend was observed. rms velocity calculations showed the jet shear layer and laminar core. The rms velocities were higher in the vocal tract cases compared to the open jet and false fold cases.     

Vibration in a self-oscillating vocal fold model with left-right asymmetry in body-layer stiffness

This study compares the phonatory behavior of an asymmetric vocal fold model to that of each individual vocal fold model in a hemi-configuration. Although phonation frequencies of the two folds in hemi-configurations had a ratio close to 1:3, a subharmonic synchronization between the two folds was not observed in the asymmetric model. Instead, the vibratory behavior was dominated by the dynamics of one fold only, and the other fold was enslaved to vibrate at the same frequency. Increasing subglottal pressure induced a shift in relative dominance between the two folds, leading to abrupt changes in both vibratory pattern and frequency.     

Dilated Minute Chambers in Laryngeal Vocal Fold Polyps: Histopathological and Ultrastructural Features

In Reinke's space of human vocal fold, type III collagen forms a three dimensional network and this contains numerous minute chambers in between these fibers. These compartments are occupied by glycosaminoglycans and glycoproteins. In laryngeal fold lesions, such as Reinke's edema and vocal fold polyps, proteoglycan (PG)/hyaluronic acid (HA) components of extracellular matrix increased. We investigated the size and quantity of the minute chambers within Reinke's space, filled with PG/HA with the aid of transmission electron microscopy. Eight vocal fold polyps and 10 mucosal biopsies (as control group) were all evaluated by light microscopy and electron microscopy. We detected that PG/HA in extracellular matrix had been increased in vocal fold lesions when compared with control group, by Alcian Blue-pH 2.5 stain. The mean volume of the chambers in Reinke's space of normal larynx was measured as 0.040233 µm² whereas the mean volume of these chambers in vocal fold polyps was measured as 6.420221 µm². The difference between the volumes of these chambers in vocal fold polyps and in control group was statistically significant (P = 0.001). Within these chambers PG/HA were found and PG/HA filling these chambers were increased in vocal fold polyps. We think proteoglycan and glycosaminoglycans, especially HA, play an important role in determining biochemical properties of vocal fold lesions.     

Vocal fold and ventricular fold vibration in period-doubling phonation: physiological description and aerodynamic modeling

Occurrences of period-doubling are found in human phonation, in particular for pathological and some singing phonations such as Sardinian A Tenore Bassu vocal performance. The combined vibration of the vocal folds and the ventricular folds has been observed during the production of such low pitch bass-type sound. The present study aims to characterize the physiological correlates of this acoustical production and to provide a better understanding of the physical interaction between ventricular fold vibration and vocal fold self-sustained oscillation. The vibratory properties of the vocal folds and the ventricular folds during phonation produced by a professional singer are analyzed by means of acoustical and electroglottographic signals and by synchronized glottal images obtained by high-speed cinematography. The periodic variation in glottal cycle duration and the effect of ventricular fold closing on glottal closing time are demonstrated. Using the detected glottal and ventricular areas, the aerodynamic behavior of the laryngeal system is simulated using a simplified physical modeling previously validated in vitro using a larynx replica. An estimate of the ventricular aperture extracted from the in vivo data allows a theoretical prediction of the glottal aperture. The in vivo measurements of the glottal aperture are then compared to the simulated estimations.