Stem Cell-Based Hair Cell Regeneration and Therapy in the Inner Ear.

Hearing loss has become increasingly prevalent and causes considerable disability, thus gravely burdening the global economy. Irreversible loss of hair cells is a main cause of sensorineural hearing loss, and currently, the only relatively effective clinical treatments are limited to digital hearing equipment like cochlear implants and hearing aids, but these are of limited benefit in patients. It is therefore urgent to understand the mechanisms of damage repair in order to develop new neuroprotective strategies. At present, how to promote the regeneration of functional hair cells is a key scientific question in the field of hearing research. Multiple signaling pathways and transcriptional factors trigger the activation of hair cell progenitors and ensure the maturation of newborn hair cells, and in this article, we first review the principal mechanisms underlying hair cell reproduction. We then further discuss therapeutic strategies involving the co-regulation of multiple signaling pathways in order to induce effective functional hair cell regeneration after degeneration, and we summarize current achievements in hair cell regeneration. Lastly, we discuss potential future approaches, such as small molecule drugs and gene therapy, which might be applied for regenerating functional hair cells in the clinic.

Measurements of bone-conducted sound in the chinchilla external ear.

We measure bone-conduction (BC) induced skull velocity, sound pressure at the tympanic membrane (TM) and inner-ear compound-action potentials (CAP) before and after manipulating the ear canal, ossicles, and the jaw to investigate the generation of BC induced ear-canal sound pressures and their contribution to inner-ear BC response in the ears of chinchillas. These measurements suggest that in chinchilla: i.) Vibrations of the bony ear canal walls contribute significantly to BC-induced ear canal sound pressures, as occluding the ear canal at the bone-cartilaginous border causes a 10 dB increase in sound pressure at the TM (PTM) at frequencies below 2 kHz. ii.) The contributions to PTM of ossicular and TM motions when driven in reverse by BC-induced inner-ear sound pressures are small. iii.) The contribution of relative motions of the jaw and ear canal to PTM is small. iv.) Comparison of the effect of canal occlusion on PTM and CAP thresholds point out that BC-induced ear canal sound pressures contribute significantly to bone-conduction stimulation of the inner ear when the ear canal is occluded.

Monitoring pressure changes in the inner ear induced by middle ear pressure therapy with an EFET01 device in guinea pigs.

BACKGROUND: As a low-cost, portable, handheld air pressure generation tool not requiring a ventilation tube, the EFET01 device has shown clinical effectiveness for intractable Ménière's disease (MD) patients in Japan. However, no animal studies have investigated changes in inner ear pressure (PI) when applying this device. OBJECTIVE: To determine the PI properties in response to middle ear pressure therapy (MEPT) induced by the EFET01 in guinea pigs. MATERIAL AND METHODS: In seven healthy guinea pigs, bi-phasic pressure pulses from -5 to 12 cm H2O were delivered to the external ear canal and transmitted to the middle and inner ear cavities with an intact tympanic membrane. Hydrostatic pressure change in the inner ear perilymphatic compartment was measured by a servo-controlled micropipette system. RESULTS: From eight successful ears, pressure changes in the middle ear slightly decreased and were instantly transferred to the inner ear. The EFET01 produces a bi-phasic positive/negative pressure pulse, which is approximately twice as large as the monophasic pressure pulse. CONCLUSION: Our study clarified the EFET01's ability to transmit pressure and verified its effectiveness in MD patients as observed in clinical studies. SIGNIFICANCE: The PI properties in guinea pig response to MEPT with the EFET01 device were investigated.

Early Steps towards Hearing: Placodes and Sensory Development.

Sensorineural hearing loss is the most prevalent sensory deficit in humans. Most cases of hearing loss are due to the degeneration of key structures of the sensory pathway in the cochlea, such as the sensory hair cells, the primary auditory neurons, and their synaptic connection to the hair cells. Different cell-based strategies to replace damaged inner ear neurosensory tissue aiming at the restoration of regeneration or functional recovery are currently the subject of intensive research. Most of these cell-based treatment approaches require experimental in vitro models that rely on a fine understanding of the earliest morphogenetic steps that underlie the in vivo development of the inner ear since its initial induction from a common otic-epibranchial territory. This knowledge will be applied to various proposed experimental cell replacement strategies to either address the feasibility or identify novel therapeutic options for sensorineural hearing loss. In this review, we describe how ear and epibranchial placode development can be recapitulated by focusing on the cellular transformations that occur as the inner ear is converted from a thickening of the surface ectoderm next to the hindbrain known as the otic placode to an otocyst embedded in the head mesenchyme. Finally, we will highlight otic and epibranchial placode development and morphogenetic events towards progenitors of the inner ear and their neurosensory cell derivatives.

Inner ear biomechanics reveals a Late Triassic origin for mammalian endothermy.

Endothermy underpins the ecological dominance of mammals and birds in diverse environmental settings1,2. However, it is unclear when this crucial feature emerged during mammalian evolutionary history, as most of the fossil evidence is ambiguous3-17. Here we show that this key evolutionary transition can be investigated using the morphology of the endolymph-filled semicircular ducts of the inner ear, which monitor head rotations and are essential for motor coordination, navigation and spatial awareness18-22. Increased body temperatures during the ectotherm-endotherm transition of mammal ancestors would decrease endolymph viscosity, negatively affecting semicircular duct biomechanics23,24, while simultaneously increasing behavioural activity25,26 probably required improved performance27. Morphological changes to the membranous ducts and enclosing bony canals would have been necessary to maintain optimal functionality during this transition. To track these morphofunctional changes in 56 extinct synapsid species, we developed the thermo-motility index, a proxy based on bony canal morphology. The results suggest that endothermy evolved abruptly during the Late Triassic period in Mammaliamorpha, correlated with a sharp increase in body temperature (5-9 °C) and an expansion of aerobic and anaerobic capacities. Contrary to previous suggestions3-14, all stem mammaliamorphs were most probably ectotherms. Endothermy, as a crucial physiological characteristic, joins other distinctive mammalian features that arose during this period of climatic instability28.

In vitro and in vivo models: What have we learnt about inner ear regeneration and treatment for hearing loss?

The sensory cells of the inner ear, called hair cells, do not regenerate spontaneously and therefore, hair cell loss and subsequent hearing loss are permanent in humans. Conversely, functional hair cell regeneration can be observed in non-mammalian vertebrate species like birds and fish. Also, during postnatal development in mice, limited regenerative capacity and the potential to isolate stem cells were reported. Together, these findings spurred the interest of current research aiming to investigate the endogenous regenerative potential in mammals. In this review, we summarize current in vitro based approaches and briefly introduce different in vivo model organisms utilized to study hair cell regeneration. Furthermore, we present an overview of the findings that were made synergistically using both, the in vitro and in vivo based tools.

Purinergic signaling in the peripheral vestibular system.

The inner ear comprises the cochlea and vestibular system, which detect sound and acceleration stimulation, respectively. The function of the inner ear is regulated by ion transport activity among sensory epithelial cells, neuronal cells, non-sensory epithelial cells, and luminal fluid with a unique ionic composition of high [K+] and low [Na+], which enables normal hearing and balance maintenance. One of the important mechanisms regulating ion transport in the inner ear is purinergic signaling. Various purinergic receptors are distributed throughout inner ear epithelial cells and neuronal cells. To date, most studies have focused on the role of purinergic receptors in the cochlea, and few studies have examined these receptors in the vestibular system. As purinergic receptors play an important role in the cochlea, they would likely do the same in the vestibular system, which is fairly similar to the cochlea in cellular structure and function. Based on available studies performed to date, purinergic signaling is postulated to be involved in the regulation of ion homeostasis, protection of hair cells, otoconia formation, and regulation of electrical signaling from the sensory epithelium to vestibular neurons. In this review, the distribution and roles of purinergic receptors in the peripheral vestibular system are summarized and discussed.

Evolution of inner ear neuroanatomy of bats and implications for echolocation.

Phylogenomics of bats suggests that their echolocation either evolved separately in the bat suborders Yinpterochiroptera and Yangochiroptera, or had a single origin in bat ancestors and was later lost in some yinpterochiropterans1-6. Hearing for echolocation behaviour depends on the inner ear, of which the spiral ganglion is an essential structure. Here we report the observation of highly derived structures of the spiral ganglion in yangochiropteran bats: a trans-otic ganglion with a wall-less Rosenthal's canal. This neuroanatomical arrangement permits a larger ganglion with more neurons, higher innervation density of neurons and denser clustering of cochlear nerve fascicles7-13. This differs from the plesiomorphic neuroanatomy of Yinpterochiroptera and non-chiropteran mammals. The osteological correlates of these derived ganglion features can now be traced into bat phylogeny, providing direct evidence of how Yangochiroptera differentiated from Yinpterochiroptera in spiral ganglion neuroanatomy. These features are highly variable across major clades and between species of Yangochiroptera, and in morphospace, exhibit much greater disparity in Yangochiroptera than Yinpterochiroptera. These highly variable ganglion features may be a neuroanatomical evolutionary driver for their diverse echolocating strategies4,14-17 and are associated with the explosive diversification of yangochiropterans, which include most bat families, genera and species.

Stimulation efficiency of an actuator driven piston at the biological interface to the inner ear.

Direct acoustic cochlear stimulation uses piston motion to substitute for stapes footplate (SFP) motion. The ratio of piston to stapes footplate motion amplitude, to generate the same loudness percept, is an indicator of stimulation efficiency. We determined the relationship between piston displacement to perceived loudness, the achieved maximum power output and investigated stapes fixation and obliteration as confounding factors. The electro-mechanical transfer function of the actuator was determined preoperatively on the bench and intraoperatively by laser Doppler vibrometry. Clinically, perceived loudness as a function of actuator input voltage was calculated from bone conduction thresholds and direct thresholds via the implant. The displacement of a 0.4 mm diameter piston required for a perception equivalent to 94 dB SPL at the tympanic membrane compared to normal SFP piston displacement was 27.6-35.9 dB larger, consistent with the hypothesis that the ratio between areas is responsible for stimulation efficiency. Actuator output was 110 ± 10 eq dB SPLFF @1Vrms ≤ 3 kHz and decreased to 100 eq dB SPLFF at 10 kHz. Output was significantly higher for mobile SFPs but independent from obliteration. Our findings from clinical data strongly support the assumption of a geometrical dependency on piston diameter at the biological interface to the cochlea.

Temporal synchrony effects of optic flow and vestibular inputs on multisensory heading perception.

Precise heading perception requires integration of optic flow and vestibular cues, yet the two cues often carry distinct temporal dynamics that may confound cue integration benefit. Here, we varied temporal offset between the two sensory inputs while macaques discriminated headings around straight ahead. We find the best heading performance does not occur under natural condition of synchronous inputs with zero offset but rather when visual stimuli are artificially adjusted to lead vestibular by a few hundreds of milliseconds. This amount exactly matches the lag between the vestibular acceleration and visual speed signals as measured from single-unit-activity in frontal and posterior parietal cortices. Manually aligning cues in these areas best facilitates integration with some nonlinear gain modulation effects. These findings are consistent with predictions from a model by which the brain integrates optic flow speed with a faster vestibular acceleration signal for sensing instantaneous heading direction during self-motion in the environment.

Evaluation of the whole auditory pathway using high-resolution and functional MRI at 7T parallel-transmit.

PURPOSE: The aim of the present study is to show a MR procedure for the evaluation of simultaneous left and right auditory functions with functional MRI, and high-resolution acquisition of anatomical auditory pathway using parallel-transmit (pTx) methods at 7T. METHODS: The time-efficient MR acquisition included two steps: RF weights were optimized for the regions-of-interest and high-resolution MR images of the inner-ear were acquired for the first 30 min (400 µm-iso resolution) followed by functional MRI acquisitions along the whole auditory pathway during the next 20 minutes. Data was processed with a linear cross-correlation analysis to define frequency preferences for each voxel in the auditory relays. RESULTS: Tonotopic maps revealed ordered bilateral frequency gradients in the auditory relays whereas at the level of the cochlear nuclei and superior olivary complexes the frequency gradients were less evident. A 21% increase in transmit-field efficiency was achieved over the left/right inner-ear regions and thus its main structures were clearly discernible using the pTx methods, compared to a single transmit RF coil. CONCLUSION: Using 7T pTx allows a fast (less than 60 min in total) and qualitative evaluation of the simultaneous left and right auditory response along the entire auditory pathway, together with high-resolution anatomical images of the inner-ear. This could be further used for patient examination at 7T.

Nerve-associated Schwann cell precursors contribute extracutaneous melanocytes to the heart, inner ear, supraorbital locations and brain meninges.

Melanocytes are pigmented cells residing mostly in the skin and hair follicles of vertebrates, where they contribute to colouration and protection against UV-B radiation. However, the spectrum of their functions reaches far beyond that. For instance, these pigment-producing cells are found inside the inner ear, where they contribute to the hearing function, and in the heart, where they are involved in the electrical conductivity and support the stiffness of cardiac valves. The embryonic origin of such extracutaneous melanocytes is not clear. We took advantage of lineage-tracing experiments combined with 3D visualizations and gene knockout strategies to address this long-standing question. We revealed that Schwann cell precursors are recruited from the local innervation during embryonic development and give rise to extracutaneous melanocytes in the heart, brain meninges, inner ear, and other locations. In embryos with a knockout of the EdnrB receptor, a condition imitating Waardenburg syndrome, we observed only nerve-associated melanoblasts, which failed to detach from the nerves and to enter the inner ear. Finally, we looked into the evolutionary aspects of extracutaneous melanocytes and found that pigment cells are associated mainly with nerves and blood vessels in amphibians and fish. This new knowledge of the nerve-dependent origin of extracutaneous pigment cells might be directly relevant to the formation of extracutaneous melanoma in humans.

The proteome of the human endolymphatic sac endolymph.

The endolymphatic sac (ES) is the third part of the inner ear, along with the cochlea and vestibular apparatus. A refined sampling technique was developed to analyse the proteomics of ES endolymph. With a tailored solid phase micro-extraction probe, five ES endolymph samples were collected, and six sac tissue biopsies were obtained in patients undergoing trans-labyrinthine surgery for sporadic vestibular schwannoma. The samples were analysed using nano-liquid chromatography-tandem mass spectrometry (nLC-MS/MS) to identify the total number of proteins. Pathway identification regarding molecular function and protein class was presented. A total of 1656 non-redundant proteins were identified, with 1211 proteins detected in the ES endolymph. A total of 110 proteins were unique to the ES endolymph. The results from the study both validate a strategy for in vivo and in situ human sampling during surgery and may also form a platform for further investigations to better understand the function of this intriguing part of the inner ear.

Preoperative Differences in Intracranial Facial Versus Vestibular Schwannomas: A Four Nerve Assessment.

OBJECTIVES: Assesses whether preoperative functional testing can distinguish vestibular schwannomas from facial nerve schwannomas medial to the labyrinthine segment. STUDY DESIGN: Retrospective cohort. METHODS: Retrospectively review surgically managed intracranial facial and vestibular schwannomas between January 2015 and December 2019 at two tertiary care centers. Patients with neurofibromatosis 2 and surgery for recurrence were excluded. Preoperative functional testing to include House-Brackmann scores, electroneuronography (ENoG), cervical vestibular evoked myogenic potentials (cVEMP), caloric testing, acoustic brainstem responses (ABRs), acoustic reflexes, and audiograms was compared between the two groups of schwannomas. RESULTS: Twelve facial and 128 vestibular schwannomas met inclusion criteria. In only one case was a facial schwannoma diagnosed preoperatively from imaging. No statistically significant difference was found in preoperative House-Brackmann scores, ENoG, cVEMP, caloric testing, ABRs, or acoustic reflexes. Pure tone average was worse in the vestibular schwannoma group (63 dB [95% CI: 58-68 dB] vs. 46 dB [95% CI: 34-58 dB], P = .01), and the difference was more apparent in the lower frequencies. Word recognition score was better in the facial schwannoma group (66% [95% CI: 45-86%] vs. 41% [95% CI: 34-47%], P = .02). CONCLUSION: Specialized preoperative functional evaluation of the nerves of the internal auditory canal cannot reliably predict the presence of an intracranial facial schwannoma. Hearing is better in facial schwannomas, particularly in the lower frequencies. This should raise the index of suspicion for an intracranial facial schwannoma, especially in candidates for hearing preservation vestibular schwannoma surgery. LEVEL OF EVIDENCE: 3 Laryngoscope, 131:2098-2105, 2021.

Alterations to vestibular morphology in highly bred domestic dogs may affect balance.

The modern domestic dog (Canis lupus familiaris) provides an excellent model to examine the effects of cranial modification. Extreme variation in skull length among dog breeds due to high levels of selective breeding is known to be linked to disorders of the head and neck. Such alteration may also influence sensory organs including those of the vestibular system (VS), one of the most fundamental sense organs, essential in maintaining balance. Studies in mammals have shown that orientation of ipsilateral semicircular canals (SCCs) of the VS at right angles (orthogonality) is related to angular acceleration sensitivity. Due to their considerable variation in craniofacial form while exhibiting similar locomotion, domestic dogs provide an excellent natural experiment to examine if cranial alteration influences VS functional morphology. Our methods examine how change in cranial base length across dog breeds relates to SCC orthogonality using linear modeling and analyses of variance. The sample studied (29 bony labyrinths of 17 dog breeds) was obtained from a previous study on canid inner ear metrics. Results support the hypothesis that orthogonality between the anterior and posterior SCC + ampulla significantly correlates with cranial base length. This suggests a close relationship between the orientations of SCCs with their ampullae and cranial structure among dog breeds. Specifically, highly derived breeds, such as the brachycephalic pug, have anterior and posterior SCCs and ampullae that deviate the most from orthogonality. Therefore, such highly bred domestic dogs may also have altered vestibular function due to compressed cranial form.

Molecular Aspects of the Development and Function of Auditory Neurons.

This review provides an up-to-date source of information on the primary auditory neurons or spiral ganglion neurons in the cochlea. These neurons transmit auditory information in the form of electric signals from sensory hair cells to the first auditory nuclei of the brain stem, the cochlear nuclei. Congenital and acquired neurosensory hearing loss affects millions of people worldwide. An increasing body of evidence suggest that the primary auditory neurons degenerate due to noise exposure and aging more readily than sensory cells, and thus, auditory neurons are a primary target for regenerative therapy. A better understanding of the development and function of these neurons is the ultimate goal for long-term maintenance, regeneration, and stem cell replacement therapy. In this review, we provide an overview of the key molecular factors responsible for the function and neurogenesis of the primary auditory neurons, as well as a brief introduction to stem cell research focused on the replacement and generation of auditory neurons.

Molecular Mechanisms and Biological Functions of Autophagy for Genetics of Hearing Impairment.

The etiology of hearing impairment following cochlear damage can be caused by many factors, including congenital or acquired onset, ototoxic drugs, noise exposure, and aging. Regardless of the many different etiologies, a common pathologic change is auditory cell death. It may be difficult to explain hearing impairment only from the aspect of cell death including apoptosis, necrosis, or necroptosis because the level of hearing loss varies widely. Therefore, we focused on autophagy as an intracellular phenomenon functionally competing with cell death. Autophagy is a dynamic lysosomal degradation and recycling system in the eukaryotic cell, mandatory for controlling the balance between cell survival and cell death induced by cellular stress, and maintaining homeostasis of postmitotic cells, including hair cells (HCs) and spiral ganglion neurons (SGNs) in the inner ear. Autophagy is considered a candidate for the auditory cell fate decision factor, whereas autophagy deficiency could be one of major causes of hearing impairment. In this paper, we review the molecular mechanisms and biologic functions of autophagy in the auditory system and discuss the latest research concerning autophagy-related genes and sensorineural hearing loss to gain insight into the role of autophagic mechanisms in inner-ear disorders.

10.5 T MRI static field effects on human cognitive, vestibular, and physiological function.

PURPOSE: To perform a pilot study to quantitatively assess cognitive, vestibular, and physiological function during and after exposure to a magnetic resonance imaging (MRI) system with a static field strength of 10.5 Tesla at multiple time scales. METHODS: A total of 29 subjects were exposed to a 10.5 T MRI field and underwent vestibular, cognitive, and physiological testing before, during, and after exposure; for 26 subjects, testing and exposure were repeated within 2-4 weeks of the first visit. Subjects also reported sensory perceptions after each exposure. Comparisons were made between short and long term time points in the study with respect to the parameters measured in the study; short term comparison included pre-vs-isocenter and pre-vs-post (1-24 h), while long term compared pre-exposures 2-4 weeks apart. RESULTS: Of the 79 comparisons, 73 parameters were unchanged or had small improvements after magnet exposure. The exceptions to this included lower scores on short term (i.e. same day) executive function testing, greater isocenter spontaneous eye movement during visit 1 (relative to pre-exposure), increased number of abnormalities on videonystagmography visit 2 versus visit 1 and a mix of small increases (short term visit 2) and decreases (short term visit 1) in blood pressure. In addition, more subjects reported metallic taste at 10.5 T in comparison to similar data obtained in previous studies at 7 T and 9.4 T. CONCLUSION: Initial results of 10.5 T static field exposure indicate that 1) cognitive performance is not compromised at isocenter, 2) subjects experience increased eye movement at isocenter, and 3) subjects experience small changes in vital signs but no field-induced increase in blood pressure. While small but significant differences were found in some comparisons, none were identified as compromising subject safety. A modified testing protocol informed by these results was devised with the goal of permitting increased enrollment while providing continued monitoring to evaluate field effects.

Utilidad diagnóstica de la densidad ósea de la cápsula laberíntica en el diagnóstico de la otosclerosis por tomografía de alta resolución / Diagnostic utility of labyrinth capsule bone density in the diagnosis of otosclerosis with high resolution tomography

OBJETIVO: El objetivo de este estudio es comparar la densidad ósea alrededor de la cápsula ótica en pacientes otosclerosos con un grupo control y encontrar el límite de densidad ósea a partir del cual podemos diagnosticar la enfermedad. MATERIAL Y MÉTODOS: Se realizó un estudio retrospectivo de casos y controles. La densidad ósea en unidades de Hounsfield (HU) de 28 oídos otosclerosos fue comparada con la densidad de 33 cápsulas no otoscleróticas. La densidad fue medida en 8 áreas de interés (ROI) donde normalmente se encuentran los focos otoscleróticos. Adicionalmente se realizó la densidad media de estas regiones (PROMED). Además, se calcularon las curvas ROC de cada ROI y la densidad media (PROMED). RESULTADOS: Todas las densidades radiológicas en HU de cada ROI y la densidad media en pacientes otosclerosos fueron menores en comparación con los oídos no otosclerosos. El área bajo la curva ROC de cada ROI y la densidad media mostraron que las áreas con mayor rendimiento diagnóstico fueron la densidad media, la fissula antefenestram y la región precoclear, con valores de corte de 1.980, 1.750 y 2.114 HU, respectivamente. CONCLUSIÓN: La densidad media de la cápsula ótica (PROMED), la densidad en fissula antefenestram (ROI 1) y en la región precoclear (ROI 3) parecen ser los parámetros más útiles para realizar el diagnóstico de otosclerosis

OBJECTIVE: The aim of this study is to compare the bone density around the otic capsule in otosclerotic patients with a control group, and find the cut-off values of bone density from which we can diagnose the disease. MATERIAL AND METHODS: A retrospective case-control study was performed. Bone densities in Hounsfield units (HU) from 28 otosclerotic ears were compared to the densities of 33 non otosclerotic capsules. These densities were measured in eight regions of interest (ROI) where the otosclerotic foci are usually found. The mean density of these regions (PROMED) was taken. Furthermore, the ROC curves of each ROI and the mean density (PROMED) were calculated. RESULTS: All radiological densities in HU of each ROI and the mean density in otosclerotic patients were lower compared to non otosclerotic ears. The area under the ROC curve of each ROI and the mean density showed that the areas with greater accuracy for the diagnosis of otosclerosis were mean density, the fissula ante fenestram, and precochlear region, with cut-off values of 1980 HU, 1750 HU and 2114 HU, respectively. CONCLUSION: The mean density of the otic capsule (PROMED), the density in the fissula ante fenestram (ROI 1) and in the precochlear region (ROI 3) seem to be the most useful parameters to make a diagnosis of otosclerosis

The endocranium and trophic ecology of Velociraptor mongoliensis.

Neuroanatomical reconstructions of extinct animals have long been recognized as powerful proxies for palaeoecology, yet our understanding of the endocranial anatomy of dromaeosaur theropod dinosaurs is still incomplete. Here, we used X-ray computed microtomography (µCT) to reconstruct and describe the endocranial anatomy, including the endosseous labyrinth of the inner ear, of the small-bodied dromaeosaur, Velociraptor mongoliensis. The anatomy of the cranial endocast and ear were compared with non-avian theropods, modern birds, and other extant archosaurs to establish trends in agility, balance, and hearing thresholds in order to reconstruct the trophic ecology of the taxon. Our results indicate that V. mongoliensis could detect a wide and high range of sound frequencies (2,368-3,965 Hz), was agile, and could likely track prey items with ease. When viewed in conjunction with fossils that suggest scavenging-like behaviours in V. mongoliensis, a complex trophic ecology that mirrors modern predators becomes apparent. These data suggest that V. mongoliensis was an active predator that would likely scavenge depending on the age and health of the individual or during prolonged climatic events such as droughts.