Roger W. Chan

Characterization of Vocal Fold Scarring in a Canine Model

Objective The objective was to assess the histological and viscoelastic shear tissue properties of the scarred vocal fold lamina propria at 2 and 6 months postoperatively in a canine model.

Relative contributions of collagen and elastin to elasticity of the vocal fold under tension.

This study examined the contributions of collagen and elastin to the tensile elastic properties of the vocal fold lamina propria. Uniaxial stress–strain responses of vocal fold cover and vocal ligament specimens from 20 human larynges (12 males, 8 females) were quantified with sinusoidal stretch-release deformation in vitro. Mid-coronal sections of 12 specimens were examined histologically with Masson’s trichrome and elastin van Gieson stain to quantify the relative densities of collagen and elastin fibers. Results showed that significantly higher levels of collagen were found in the male vocal fold than female, for both the cover and the ligament. For male there was a significantly higher level of elastin in the cover than in the ligament. On average, the elastic modulus of the male cover was about twice that of the female at high-tensile strain (35–40%), whereas the male ligament was 3–5 times stiffer than the female in the same range. The ligament was stiffer than the cover for male, but the opposite was observed for female. These findings suggested that collagen and elastin could contribute differentially to elasticity of the cover and the ligament. The data may provide guidance for surgical reconstruction and tissue engineering of different lamina propria layers.

Effect of Postmortem Changes and Freezing on the Viscoelastic Properties of Vocal Fold Tissues

AbstractIt is common practice in laryngeal research laboratories to store excised larynges and vocal fold tissue specimens frozen, presumably to preserve viability and structural integrity of the specimens for physiologic and biomechanical experiments. However, little is known about the biomechanical effects of postmortem changes and frozen storage in vocal fold tissues. This study attempted to quantify the effects of postmortem changes and freezing on the viscoelastic shear properties of the canine vocal fold mucosa. Sixteen larynges were excised from adult dogs immediately postmortem in a viable state. Using a torsional rheometer, the complex shear modulus (G*) of the mucosal tissue from one vocal fold of each larynx was measured as a function of frequency (0.01–15 Hz) by linear small-amplitude oscillation experiments. Measurement was repeated for ten mucosal specimens after 24 h of postmortem storage in saline solution at room temperature. Eleven of the 16 larynges were frozen and stored at −20°C, six of them at a slow rate of cooling (storage in a regular freezer) and five others by quick freezing (chilling by liquid nitrogen prior to frozen storage). The larynges were thawed slowly overnight after one month and the viscoelastic shear properties of the contralateral vocal fold mucosa were measured. Results showed that the elastic shear modulus (G′) and dynamic viscosity (η ′) of the vocal fold mucosa did not seem to change significantly after 24 h of storage in saline at room temperature, nor after one month of frozen storage following quick freezing, whereas both G′ and η′ decreased significantly for tissues that were slowly frozen. These findings supported the feasibility of using quick freezing to preserve laryngeal tissues for in vitro biomechanical testing, for excised larynx experiments, and for tissue engineering applications.

A constitutive model of the human vocal fold cover for fundamental frequency regulation

The elastic as well as time-dependent mechanical response of the vocal fold cover (epithelium and superficial layer of the lamina propria) under tension is one key variable in regulating the fundamental frequency. This study examines the hyperelastic and time-dependent tensile deformation behavior of a group of human vocal fold cover specimens (six male and five female). The primary goal is to formulate a constitutive model that could describe empirical trends in speaking fundamental frequency with reasonable confidence. The constitutive model for the tissue mechanical behavior consists of a hyperelastic equilibrium network in parallel with an inelastic, time-dependent network and is combined with the ideal string model for phonation. Results showed that hyperelastic and time-dependent parameters of the constitutive model can be related to observed age-related and gender-related differences in speaking fundamental frequency. The implications of these findings on fundamental frequency regulation are described. Limitations of the current constitutive model are discussed.

A bovine acellular scaffold for vocal fold reconstruction in a rat model

With a rat model of vocal fold injury, this study examined the in vivo host response to an acellular xenogeneic scaffold derived from the bovine vocal fold lamina propria, and the potential of the scaffold for constructive tissue remodeling. Bilateral wounds were created in the posterior vocal folds of 20 rats, and bovine acellular scaffolds were implanted into the wounds unilaterally, with the contralateral vocal folds as control. The rats were humanely sacrificed after 3 days, 7 days, 1 month, and 3 months, and the coronal sections of their larynges were examined histologically. Expressions of key matrix proteins including collagen I, collagen III, elastin, fibronectin, hyaluronic acid, and glycosaminoglycans (GAGs) were quantified with digital image analysis. Significant infiltration of host inflammatory cells and host fibroblasts in the scaffold implant was observed in the acute stage of wound repair (3 days and 7 days postsurgery). The mean relative densities of collagen I, collagen III, and GAGs in the implanted vocal folds were significantly higher than those in the control after 3 days, followed by gradual decreases over 3 months. Histological results showed that the scaffolds were apparently degraded by 3 months, with no fibrotic tissue formation or calcification. These preliminary findings suggested that the bovine acellular scaffold could be a potential xenograft for vocal fold regeneration.

Viscoelastic properties of phonosurgical biomaterials at phonatory frequencies

The purpose of this study was to examine the functional biomechanical properties of several injectable biomaterials currently or potentially used for vocal fold augmentation.

Injection of human mesenchymal stem cells improves healing of vocal folds after scar excision--a xenograft analysis.

Using a xenograft model the aim was to analyze if injection of human mesenchymal stem cells (hMSC) into the rabbit vocal fold (VF), after excision of an established scar, can improve the functional healing of the VF.

Modeling of the transient responses of the vocal fold lamina propria

The human voice is produced by flow-induced self-sustained oscillation of the vocal fold lamina propria. The mechanical properties of vocal fold tissues are important for understanding phonation, including the time-dependent and transient changes in fundamental frequency (F(0)). Cyclic uniaxial tensile tests were conducted on a group of specimens of the vocal fold lamina propria, including the superficial layer (vocal fold cover) (5 male, 5 female) and the deeper layers (vocal ligament) (6 male, 6 female). Results showed that the vocal fold lamina propria, like many other soft tissues, exhibits both elastic and viscous behavior. Specifically, the transient mechanical responses of cyclic stress relaxation and creep were observed. A three-network constitutive model composed of a hyperelastic equilibrium network in parallel with two viscoplastic time-dependent networks proves effective in characterizing the cyclic stress relaxation and creep behavior. For male vocal folds at a stretch of 1.4, significantly higher peak stress was found in the vocal ligament than in the vocal fold cover. Also, the male vocal ligament was significantly stiffer than the female vocal ligament. Our findings may help explain the mechanisms of some widely observed transient phenomena in F(0) regulation during phonation, such as the global declination in F(0) during the production of declarative sentences, and local F(0) changes such as overshoot and undershoot.

Arytenoid Adduction for Correcting Vocal Fold Asymmetry: High-Speed Imaging

Objectives We hypothesized that high-speed digital imaging provides a quantitative method to evaluate the effect of arytenoid adduction for the correction of asymmetric and irregular vocal fold vibration in unilateral vocal fold paralysis. Methods Six subjects with unilateral vocal fold paralysis participated in the study (4 male, 2 female; mean [±SD] age, 52.5 ± 21.3 years). Videokymographic and laryngotopographic methods for image analysis were performed for highspeed recordings of vocal fold vibration for visualizing the glottal vibratory patterns, and for quantifying the frequency of vibration of each vocal fold, respectively. Comparisons of the paralyzed and the normal vocal folds were made before and after arytenoid adduction. Results Analysis of the laryngotopographs revealed 2 distinct frequencies of vibration for the paralyzed and the contralateral vocal folds for all subjects before surgery. After arytenoid adduction, the vibration frequencies became identical or nearly identical in all subjects. Conclusions Asymmetric vibration in vocal fold paralysis was exemplified by differences in vibration frequency between the vocal folds. The present data showed that after arytenoid adduction the vibration frequencies and the vibratory patterns of the contralateral vocal folds approached symmetry. This surgical procedure could improve the functional symmetry of the larynx for phonation.

Elasticity of the human false vocal fold

Objectives: Very little is known about the elasticity of the human ventricular fold (false vocal fold). To better understand the potential role of the false fold in the fluid dynamics and aeroacoustics of phonation, we made some measurements on the elastic properties of human ventricular fold tissues in vitro. Methods: Uniaxial tensile stress-strain characteristics of 6 male and 6 female false fold specimens were quantified with sinusoidal stretch-release deformation. Midcoronal sections of 3 specimens were examined to quantify the relative densities of collagen, elastin, seromucous glandular tissue, and adipose tissue by digital image analysis. Results: Nonlinear stress-strain curves with hysteresis (viscous energy loss) were observed, with large interindividual differences. A hybrid linear-exponential model was used to determine the elastic modulus (tangent Youngs modulus) of the false fold. On average, the male false fold was twice as stiff as the female at a tensile strain of 20% to 30%. Conclusions: This preliminary gender-related difference in elasticity could be attributed to a higher proportion of glandular tissue in the female false fold, due to the lower elastic modulus of glands. The present data allow one to develop a more comprehensive biomechanical model of phonation, for optimizing postoperative voice production following laryngeal reconstruction procedures.