Synthetic, self-oscillating vocal fold models for voice production researcha).

Sound for the human voice is produced by vocal fold flow-induced vibration and involves a complex coupling between flow dynamics, tissue motion, and acoustics. Over the past three decades, synthetic, self-oscillating vocal fold models have played an increasingly important role in the study of these complex physical interactions. In particular, two types of models have been established: "membranous" vocal fold models, such as a water-filled latex tube, and "elastic solid" models, such as ultrasoft silicone formed into a vocal fold-like shape and in some cases with multiple layers of differing stiffness to mimic the human vocal fold tissue structure. In this review, the designs, capabilities, and limitations of these two types of models are presented. Considerations unique to the implementation of elastic solid models, including fabrication processes and materials, are discussed. Applications in which these models have been used to study the underlying mechanical principles that govern phonation are surveyed, and experimental techniques and configurations are reviewed. Finally, recommendations for continued development of these models for even more lifelike response and clinical relevance are summarized.

Three-Dimensional Printing of Ultrasoft Silicone with a Functional Stiffness Gradient.

A methodology for three-dimensionally printing ultrasoft silicone with a functional stiffness gradient is presented. Ultraviolet-cure silicone was deposited via two independently controlled extruders into a thixotropic, gel-like, silicone oil-based support matrix. Each extruder contained a different liquid silicone formulation. The extrusion rates were independently varied during printing such that the combined selectively deposited material contained different ratios of the two silicones, resulting in localized control of material stiffness. Tests to validate the process are reported, including tensile testing of homogeneous cubic specimens to quantify the range of material stiffness that could be printed, indentation testing of cuboid specimens to characterize printed stiffness gradients, and vibratory testing of synthetic multilayer vocal fold (VF) models to demonstrate that the method may be applied to the fabrication of biomechanical models for voice production research. The cubic specimens exhibited linear stress-strain data with tensile elasticity modulus values between 1.11 and 27.1 kPa, more than a factor of 20 in stiffness variation. The cuboid specimens exhibited material variations that were visually recognizable and quantifiable via indentation testing. The VF models withstood rigorous phonatory flow-induced vibration and exhibited vibratory characteristics comparable to those of previous models. Overall, while process refinements are needed, the results of these tests demonstrate the ability to print ultrasoft silicone with stiffness gradients.

Aerodynamic-induced Effects of Artificial Subglottic Stenosis on Vocal Fold Model Phonatory Response.

OBJECTIVE: Subglottic stenosis (SGS) is characterized by a narrowing of the trachea near the cricotracheal junction and impairs breathing. SGS may also adversely affect voice quality, but for reasons that are not fully understood. The purpose of this study is to provide experiment-based data concerning the effects on phonation of airway obstruction due to SGS. STUDY DESIGN: Basic science METHODS: A device simulating a SGS of adjustable severity ranging from 36% to 99.8% obstruction was created. Self-oscillating synthetic VF models were mounted downstream of the device and data were acquired to evaluate the effects of the obstruction on phonatory response. RESULTS: Onset pressures were relatively insensitive to obstructions of up to approximately 80% to 90% reductions in subglottic airway area and sharply increased thereafter. Flow rate (under conditions of constant pressure), flow resistance, and fundamental frequency all exhibited similar degrees of sensitivity to SGS obstruction as onset pressure. High-frequency noise became significant by 80% obstruction. Glottal area appeared to be less sensitive, not being affected until approximately 90% obstruction. CONCLUSIONS: Consistent with previous computational studies, this study found that aerodynamic, acoustic, and vibratory responses of self-oscillating VF models were largely unaffected by SGS until approximately 80% to 90% obstruction, and significantly affected at higher obstructions. This suggests that Grades I and II stenoses are unlikely to introduce subglottic airway aerodynamic disturbances that are sufficient in and of themselves to significantly alter phonatory output. The SGS model introduces a framework for future benchtop studies involving subglottic and supraglottic airway constrictions.

Optimization of Synthetic Vocal Fold Models for Glottal Closure.

Synthetic, self-oscillating models of the human vocal folds are used to study the complex and inter-related flow, structure, and acoustical aspects of voice production. The vocal folds typically collide during each cycle, thereby creating a brief period of glottal closure that has important implications for flow, acoustic, and motion-related outcomes. Many previous synthetic models, however, have been limited by incomplete glottal closure during vibration. In this study, a low-fidelity, two-dimensional, multilayer finite element model of vocal fold flow-induced vibration was coupled with a custom genetic algorithm optimization code to determine geometric and material characteristics that would be expected to yield physiologically-realistic frequency and closed quotient values. The optimization process yielded computational models that vibrated with favorable frequency and closed quotient characteristics. A tradeoff was observed between frequency and closed quotient. A synthetic, self-oscillating vocal fold model with geometric and material properties informed by the simulation outcomes was fabricated and tested for onset pressure, oscillation frequency, and closed quotient. The synthetic model successfully vibrated at a realistic frequency and exhibited a nonzero closed quotient. The methodology described in this study provides potential direction for fabricating synthetic models using isotropic silicone materials that can be designed to vibrate with physiologically-realistic frequencies and closed quotient values. The results also show the potential for a low-fidelity model optimization approach to be used to tune synthetic vocal fold model characteristics for specific vibratory outcomes.

Embedded 3D printing of multi-layer, self-oscillating vocal fold models.

The biomechanics of human voice production are commonly studied using benchtop silicone vocal fold models that mimic the vibration of their in vivo counterparts. These models often have multiple layers of differing stiffness that represent human vocal fold tissue layers and are fabricated using a multi-step casting process. The purpose of the present study is to introduce and demonstrate a process for fabricating functional multi-layer vocal fold models using an alternative approach, termed embedded 3D printing, that is a hybrid of casting and 3D printing. In this paper the fabrication process is described. Analysis of the resulting geometric and stiffness characteristics of the layers, including layer elastic modulus values ranging from less than 1 kPa to approximately 40 kPa, is presented. The results of tests demonstrating that the models are capable of sustained phonomimetic vibration are given. Capabilities and limitations of the embedded 3D printing process are discussed. It is concluded that the process has the potential to contribute to voice biomechanics research by facilitating prospective improvements in the fabrication, design, and functionality of multi-layer vocal fold models.

3D Printing Low-Stiffness Silicone Within a Curable Support Matrix.

Embedded 3D printing processes involve extruding ink within a support matrix that supports the ink throughout printing and curing. In once class of embedded 3D printing, which we refer to as "removable embedded 3D printing," curable inks are printed, cured, then removed from the uncured support matrix. Removable embedded 3D printing is advantageous because low-viscosity inks can be patterned in freeform geometries which may not be feasible to create via casting and other printing processes. When printing solid-infill geometries, however, uncured support matrix becomes trapped within the prints, which may be undesirable. This study builds on previous work by formulating a support matrix for removable embedded 3D printing that cures when mixed with the printed silicone ink to solve the problem of trapped, uncured support matrix within solid-infill prints. Printed specimens are shown to have a nearly isotropic elastic modulus in directions perpendicular and parallel to the printed layers, and a decreased modulus and increased elongation at break compared to specimens cast from the ink. The rheological properties of the support matrix are reported. The capabilities of the printer and support matrix are demonstrated by printing a variety of geometries from four UV and addition-cure silicone inks. Shapes printed with these inks range by nearly two orders of magnitude in stiffness and have failure strains between approximately 50 and 250%, suggesting a wide range of potential applications for this printing process.

3D-Printed Synthetic Vocal Fold Models.

OBJECTIVE: Synthetic vocal fold (VF) models used for studying the physics of voice production are comprised of silicone and fabricated using traditional casting processes. The purpose of this study was to develop and demonstrate a new method of creating synthetic VF models through 3D printing in order to reduce model fabrication time, increase yield, and lay the foundation for future models with more life-like geometric, material, and vibratory properties. STUDY DESIGN: Basic science. METHODS: A 3D printing technique based on embedding a UV-curable liquid silicone into a gel-like medium was selected and refined. Cubes were printed and subjected to tensile testing to characterize their material properties. Self-oscillating VF models were then printed, coated with a thin layer of silicone representing the epithelium, and used in phonation tests to gather onset pressure, frequency, and amplitude data. RESULTS: The cubes were found to be anisotropic, exhibiting different modulus values depending on the orientation of the printed layers. The VF models self-oscillated and withstood the strains induced by phonation. Print parameters were found to affect model vibration frequency and onset pressure. Primarily due to the design of the VF models, their onset pressures were higher than what is found in human VFs. However, their frequencies were within a comparable range. CONCLUSION: The results demonstrate the ability to 3D print synthetic, self-oscillating VF models. It is anticipated that this method will be further refined and used in future studies exploring flow-induced vibratory characteristics of phonation.

Magnetic resonance imaging-based measurement of internal deformation of vibrating vocal fold models.

A method is presented for tracking the internal deformation of self-oscillating vocal fold models using magnetic resonance imaging (MRI). Silicone models scaled to four times life-size to lower the flow-induced vibration frequency were embedded with fiducial markers in a coronal plane. Candidate marker materials were tested using static specimens, and two materials, cupric sulfate and glass, were chosen for testing in the vibrating vocal fold models. The vibrating models were imaged using a gated MRI protocol wherein MRI acquisition was triggered using the subglottal pressure signal. Two-dimensional image slices at different phases during self-oscillation were captured, and in each phase the fiducial markers were clearly visible. The process was also demonstrated using a three-dimensional scan at two phases. The benefit of averaging to increase signal-to-noise ratio was explored. The results demonstrate the ability to use MRI to acquire quantitative deformation data that could be used, for example, to validate computational models of flow-induced vocal fold vibration and quantify deformation fields encountered by cells in bioreactor studies.

The evolution of the syrinx: An acoustic theory.

The unique avian vocal organ, the syrinx, is located at the caudal end of the trachea. Although a larynx is also present at the opposite end, birds phonate only with the syrinx. Why only birds evolved a novel sound source at this location remains unknown, and hypotheses about its origin are largely untested. Here, we test the hypothesis that the syrinx constitutes a biomechanical advantage for sound production over the larynx with combined theoretical and experimental approaches. We investigated whether the position of a sound source within the respiratory tract affects acoustic features of the vocal output, including fundamental frequency and efficiency of conversion from aerodynamic energy to sound. Theoretical data and measurements in three bird species suggest that sound frequency is influenced by the interaction between sound source and vocal tract. A physical model and a computational simulation also indicate that a sound source in a syringeal position produces sound with greater efficiency. Interestingly, the interactions between sound source and vocal tract differed between species, suggesting that the syringeal sound source is optimized for its position in the respiratory tract. These results provide compelling evidence that strong selective pressures for high vocal efficiency may have been a major driving force in the evolution of the syrinx. The longer trachea of birds compared to other tetrapods made them likely predisposed for the evolution of a syrinx. A long vocal tract downstream from the sound source improves efficiency by facilitating the tuning between fundamental frequency and the first vocal tract resonance.

Identity and novelty in the avian syrinx.

In its most basic conception, a novelty is simply something new. However, when many previously proposed evolutionary novelties have been illuminated by genetic, developmental, and fossil data, they have refined and narrowed our concept of biological "newness." For example, they show that these novelties can occur at one or multiple levels of biological organization. Here, we review the identity of structures in the avian vocal organ, the syrinx, and bring together developmental data on airway patterning, structural data from across tetrapods, and mathematical modeling to assess what is novel. In contrast with laryngeal cartilages that support vocal folds in other vertebrates, we find no evidence that individual cartilage rings anchoring vocal folds in the syrinx have homology with any specific elements in outgroups. Further, unlike all other vertebrate vocal organs, the syrinx is not derived from a known valve precursor, and its origin involves a transition from an evolutionary "spandrel" in the respiratory tract, the site where the trachea meets the bronchi, to a target for novel selective regimes. We find that the syrinx falls into an unusual category of novel structures: those having significant functional overlap with the structures they replace. The syrinx, along with other evolutionary novelties in sensory and signaling modalities, may more commonly involve structural changes that contribute to or modify an existing function rather than those that enable new functions.

Electrically conductive synthetic vocal fold replicas for voice production research.

A method of fabricating electrically conductive synthetic vocal fold replicas and monitoring their vibration via resistance measurement is presented. Normally non-conductive silicone replicas were coated with conductive graphite and subjected to long-term vibration tests. Synchronized resistance and imaging data using hemilarynx and full larynx configurations showed an inverse correlation between replica contact area and resistance during vibration, similar to clinical electroglottography (EGG) used to estimate vocal fold contact area. This method has potential for long-term replica vibration monitoring and studying basic physical relationships between resistance and contact area in vocal folds and vocal fold replicas.

Cavitation onset caused by acceleration.

Striking the top of a liquid-filled bottle can shatter the bottom. An intuitive interpretation of this event might label an impulsive force as the culprit in this fracturing phenomenon. However, high-speed photography reveals the formation and collapse of tiny bubbles near the bottom before fracture. This observation indicates that the damaging phenomenon of cavitation is at fault. Cavitation is well known for causing damage in various applications including pipes and ship propellers, making accurate prediction of cavitation onset vital in several industries. However, the conventional cavitation number as a function of velocity incorrectly predicts the cavitation onset caused by acceleration. This unexplained discrepancy leads to the derivation of an alternative dimensionless term from the equation of motion, predicting cavitation as a function of acceleration and fluid depth rather than velocity. Two independent research groups in different countries have tested this theory; separate series of experiments confirm that an alternative cavitation number, presented in this paper, defines the universal criteria for the onset of acceleration-induced cavitation.

A Flow Perfusion Bioreactor System for Vocal Fold Tissue Engineering Applications.

The human vocal folds (VFs) undergo complex biomechanical stimulation during phonation. The aim of the present study was to develop and validate a phono-mimetic VF flow perfusion bioreactor, which mimics the mechanical microenvironment of the human VFs in vitro. The bioreactor uses airflow-induced self-oscillations, which have been shown to produce mechanical loading and contact forces that are representative of human phonation. The bioreactor consisted of two synthetic VF replicas within a silicone body. A cell-scaffold mixture (CSM) consisting of human VF fibroblasts, hyaluronic acid, gelatin, and a polyethylene glycol cross-linker was injected into cavities within the replicas. Cell culture medium (CCM) was perfused through the scaffold by using a customized secondary flow loop. After the injection, the bioreactor was operated with no stimulation over a 3-day period to allow for cell adaptation. Phonation was subsequently induced by using a variable speed centrifugal blower for 2 h each day over a period of 4 days. A similar bioreactor without biomechanical stimulation was used as the nonphonatory control. The CSM was harvested from both VF replicas 7 days after the injection. The results confirmed that the phono-mimetic bioreactor supports cell viability and extracellular matrix proteins synthesis, as expected. Many scaffold materials were found to degrade because of challenges from phonation-induced biomechanical stimulation as well as due to biochemical reactions with the CCM. The bioreactor concept enables future investigations of the effects of different phonatory characteristics, that is, voice regimes, on the behavior of the human VF cells. It will also help study the long-term functional outcomes of the VF-specific biomaterials before animal and clinical studies.

Quantification of Porcine Vocal Fold Geometry.

OBJECTIVE: The aim of this study was to quantify porcine vocal fold medial surface geometry and three-dimensional geometric distortion induced by freezing the larynx, especially in the region of the vocal folds. STUDY DESIGN: The medial surface geometries of five excised porcine larynges were quantified and reported. METHODS: Five porcine larynges were imaged in a micro-CT scanner, frozen, and rescanned. Segmentations and three-dimensional reconstructions were used to quantify and characterize geometric features. Comparisons were made with geometry data previously obtained using canine and human vocal folds as well as geometries of selected synthetic vocal fold models. RESULTS: Freezing induced an overall expansion of approximately 5% in the transverse plane and comparable levels of nonuniform distortion in sagittal and coronal planes. The medial surface of the porcine vocal folds was found to compare reasonably well with other geometries, although the compared geometries exhibited a notable discrepancy with one set of published human female vocal fold geometry. CONCLUSIONS: Porcine vocal folds are qualitatively geometrically similar to data available for canine and human vocal folds, as well as commonly used models. Freezing of tissue in the larynx causes distortion of around 5%. The data can provide direction in estimating uncertainty due to bulk distortion of tissue caused by freezing, as well as quantitative geometric data that can be directly used in developing vocal fold models.

A synthetic, self-oscillating vocal fold model platform for studying augmentation injection.

OBJECTIVE: To design and evaluate a platform for studying the mechanical effects of augmentation injections using synthetic, self-oscillating vocal fold models. STUDY DESIGN: Basic science. METHODS: Life-sized, synthetic, multilayer, self-oscillating vocal fold models were created that simulated bowing via volumetric reduction of the body layer relative to that of a normal, unbowed model. Material properties of the layers were unchanged. Models with varying degrees of bowing were created and paired with normal models. Following initial acquisition of data (onset pressure, vibration frequency, flow rate, and high-speed image sequences), bowed models were injected with silicone that had material properties similar to those used in augmentation procedures. Three different silicone injection quantities were tested: sufficient to close the glottal gap, insufficient to close the glottal gap, and excess silicone to create convex bowing of the bowed model. The above-mentioned metrics were again taken and compared. Pre- and post-injection high-speed image sequences were acquired using a hemilarynx setup, from which medial surface dynamics were quantified. RESULTS: The models vibrated with mucosal wave-like motion and at onset pressures and frequencies typical of human phonation. The models successfully exhibited various degrees of bowing which were then mitigated by injecting filler material. The models showed general pre- to post-injection decreases in onset pressure, flow rate, and open quotient and a corresponding increase in vibration frequency. CONCLUSION: The model may be useful in further explorations of the mechanical consequences of augmentation injections.

Influence of numerical model decisions on the flow-induced vibration of a computational vocal fold model.

Computational vocal fold models are often used to study the physics of voice production. In this paper the sensitivity of predicted vocal fold flow-induced vibration and resulting airflow patterns to several modeling selections is explored. The location of contact lines used to prevent mesh collapse and assumptions of symmetry were found to influence airflow patterns. However, these variables had relatively little effect on the vibratory response of the vocal fold model itself. Model motion was very sensitive to Poisson's ratio. The importance of these parameter sensitivities in the context of vocal fold modeling is discussed.

Acoustically-coupled flow-induced vibration of a computational vocal fold model.

The flow-induced vibration of synthetic vocal fold models has been previously observed to be acoustically-coupled with upstream flow supply tubes. This phenomenon was investigated using a finite element model that included flow-structure-acoustic interactions. The length of the upstream duct was varied to explore the coupling between model vibration and subglottal acoustics. Incompressible and slightly compressible flow models were tested. The slightly compressible model exhibited acoustic coupling between fluid and solid domains in a manner consistent with experimental observations, whereas the incompressible model did not, showing the slightly compressible approach to be suitable for simulating acoustically-coupled vocal fold model flow-induced vibration.

Influence of subglottic stenosis on the flow-induced vibration of a computational vocal fold model.

The effect of subglottic stenosis on vocal fold vibration is investigated. An idealized stenosis is defined, parameterized, and incorporated into a two-dimensional, fully-coupled finite element model of the vocal folds and laryngeal airway. Flow-induced responses of the vocal fold model to varying severities of stenosis are compared. The model vibration was not appreciably affected by stenosis severities of up to 60% occlusion. Model vibration was altered by stenosis severities of 90% or greater, evidenced by decreased superior model displacement, glottal width amplitude, and flow rate amplitude. Predictions of vibration frequency and maximum flow declination rate were also altered by high stenosis severities. The observed changes became more pronounced with increasing stenosis severity and inlet pressure, and the trends correlated well with flow resistance calculations. Flow visualization was used to characterize subglottal flow patterns in the space between the stenosis and the vocal folds. Underlying mechanisms for the observed changes, possible implications for human voice production, and suggestions for future work are discussed.

Determining 3D flow fields via multi-camera light field imaging.

In the field of fluid mechanics, the resolution of computational schemes has outpaced experimental methods and widened the gap between predicted and observed phenomena in fluid flows. Thus, a need exists for an accessible method capable of resolving three-dimensional (3D) data sets for a range of problems. We present a novel technique for performing quantitative 3D imaging of many types of flow fields. The 3D technique enables investigation of complicated velocity fields and bubbly flows. Measurements of these types present a variety of challenges to the instrument. For instance, optically dense bubbly multiphase flows cannot be readily imaged by traditional, non-invasive flow measurement techniques due to the bubbles occluding optical access to the interior regions of the volume of interest. By using Light Field Imaging we are able to reparameterize images captured by an array of cameras to reconstruct a 3D volumetric map for every time instance, despite partial occlusions in the volume. The technique makes use of an algorithm known as synthetic aperture (SA) refocusing, whereby a 3D focal stack is generated by combining images from several cameras post-capture (1). Light Field Imaging allows for the capture of angular as well as spatial information about the light rays, and hence enables 3D scene reconstruction. Quantitative information can then be extracted from the 3D reconstructions using a variety of processing algorithms. In particular, we have developed measurement methods based on Light Field Imaging for performing 3D particle image velocimetry (PIV), extracting bubbles in a 3D field and tracking the boundary of a flickering flame. We present the fundamentals of the Light Field Imaging methodology in the context of our setup for performing 3DPIV of the airflow passing over a set of synthetic vocal folds, and show representative results from application of the technique to a bubble-entraining plunging jet.

Vibratory responses of synthetic, self-oscillating vocal fold models.

The flow-induced responses of four self-oscillating synthetic vocal fold models are compared. All models were life-sized and fabricated using flexible silicone compounds with material properties comparable to those of human vocal fold tissue. Three of the models had two layers of different stiffness to represent the body-cover grouping of vocal fold tissue. Two of the two-layer models were based on the "M5" geometry [Scherer et al., J. Acoust. Soc. Am. 109, 1616-1630 (2001)], while the third was based on magnetic resonance imaging data. The fourth model included several layers, including a thin epithelial layer, an exceedingly flexible superficial lamina propria layer, a ligament layer that included an anteriorly-posteriorly oriented fiber to restrict vertical motion, and a body layer. Measurements were performed with these models in full larynx and hemilarynx configurations. Data included onset pressure, vibration frequency, glottal flow rate, maximum glottal width, and medial surface motion, the latter two of which were acquired using high-speed imaging techniques. The fourth, multi-layer model exhibited onset pressure, frequency, and medial surface motion traits that are comparable to published human vocal fold data. Importantly, the model featured an alternating convergent-divergent glottal profile and mucosal wave-like motion, characteristics which are important markers of human vocal fold vibration.