Neural Mechanisms Contributing to Dysphagia in Mouse Models.

Investigative research into curative treatments for dysphagia is hindered by our incomplete understanding of the neural mechanisms of swallowing in health and disease. Development of translational research models is essential to bridge this knowledge gap by fostering innovative methodology. Toward this goal, our laboratory has developed a translational research assessment tool to investigate the neural mechanistic control of swallowing in unrestrained, self-feeding mice. Here we describe our initial development of synchronous brainstem neural recordings with a videofluoroscopic swallow study assay in healthy mice across the life span. Refinement of this combined methodology is currently underway. Ultimately, we envision that this assessment tool will permit systematic analysis of therapeutic interventions for dysphagia in preclinical trials with numerous mouse models of human conditions that cause dysphagia, such as amyotrophic lateral sclerosis, Parkinson's disease, stroke, and advanced aging.

Decellularized extracellular matrix microparticles as a vehicle for cellular delivery in a model of anastomosis healing.

Extracellular matrix (ECM) materials from animal and human sources have become important materials for soft tissue repair. Microparticles of ECM materials have increased surface area and exposed binding sites compared to sheet materials. Decellularized porcine peritoneum was mechanically dissociated into 200 µm microparticles, seeded with fibroblasts and cultured in a low gravity rotating bioreactor. The cells avidly attached and maintained excellent viability on the microparticles. When the seeded microparticles were placed in a collagen gel, the cells quickly migrated off the microparticles and through the gel. Cells from seeded microparticles migrated to and across an in vitro anastomosis model, increasing the tensile strength of the model. Cell seeded microparticles of ECM material have potential for paracrine and cellular delivery therapies when delivered in a gel carrier. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1728-1735, 2016.

Adapting human videofluoroscopic swallow study methods to detect and characterize dysphagia in murine disease models.

This study adapted human videofluoroscopic swallowing study (VFSS) methods for use with murine disease models for the purpose of facilitating translational dysphagia research. Successful outcomes are dependent upon three critical components: test chambers that permit self-feeding while standing unrestrained in a confined space, recipes that mask the aversive taste/odor of commercially-available oral contrast agents, and a step-by-step test protocol that permits quantification of swallow physiology. Elimination of one or more of these components will have a detrimental impact on the study results. Moreover, the energy level capability of the fluoroscopy system will determine which swallow parameters can be investigated. Most research centers have high energy fluoroscopes designed for use with people and larger animals, which results in exceptionally poor image quality when testing mice and other small rodents. Despite this limitation, we have identified seven VFSS parameters that are consistently quantifiable in mice when using a high energy fluoroscope in combination with the new murine VFSS protocol. We recently obtained a low energy fluoroscopy system with exceptionally high imaging resolution and magnification capabilities that was designed for use with mice and other small rodents. Preliminary work using this new system, in combination with the new murine VFSS protocol, has identified 13 swallow parameters that are consistently quantifiable in mice, which is nearly double the number obtained using conventional (i.e., high energy) fluoroscopes. Identification of additional swallow parameters is expected as we optimize the capabilities of this new system. Results thus far demonstrate the utility of using a low energy fluoroscopy system to detect and quantify subtle changes in swallow physiology that may otherwise be overlooked when using high energy fluoroscopes to investigate murine disease models.

Improving the Utility of Laryngeal Adductor Reflex Testing: A Translational Tale of Mice and Men.

OBJECTIVES: Evaluation of the laryngeal adductor reflex (LAR) entails delivering air through an endoscope positioned 1 to 2 mm from the arytenoid mucosa to elicit bilateral vocal fold (VF) closure. This short working distance limits visualization to only the ipsilateral arytenoid and results in quantification of a single LAR metric: threshold pressure that evokes the LAR. Our goal was to evolve the LAR procedure to optimize its utility in clinical practice and translational research. STUDY DESIGN: Prospective translational experiment. SETTING: Academic institution. SUBJECTS: Young healthy human adults (n = 13) and 3 groups of mice: healthy, primary aging mice (n = 5), a transgenic mouse model of amyotrophic lateral sclerosis (ALS; n = 4), and young healthy controls (n = 10). METHODS: The VFs were visualized bilaterally during supramaximal air stimulation through an endoscope. Responses were analyzed to quantify 4 novel metrics: VF adduction phase duration, complete glottic closure duration, VF abduction phase duration, and total LAR duration. RESULTS: The 4 LAR metrics are remarkably similar between healthy young humans and mice. Compared to control mice, aging mice have shorter glottic closure durations, whereas ALS-affected mice have shorter VF abduction phase durations. CONCLUSIONS: We have established a new LAR protocol that permits quantification of novel LAR metrics that are translatable between mice and humans. Using this protocol, we showed that VF adduction is impaired in primary aging mice, whereas VF abduction is impaired in ALS-affected mice. These preliminary findings highlight the enhanced diagnostic potential of LAR testing.

Validation of computational fluid dynamics-based analysis to evaluate hemodynamic significance of access stenosis.

PURPOSE: Stenosis in a vascular access circuit is the predominant cause of access dysfunction. Hemodynamic significance of a stenosis identified by angiography in an access circuit is uncertain. This study utilizes computational fluid dynamics (CFD) to model flow through arteriovenous fistula to predict the functional significance of stenosis in vascular access circuits. METHODS: Three-dimensional models of fistulas were created with a range of clinically relevant stenoses using SolidWorks. Stenoses diameters ranged from 1.0 to 3.0 mm and lengths from 5 to 60 mm within a fistula diameter of 7 mm. CFD analyses were performed using a blood model over a range of blood pressures. Eight patient-specific stenoses were also modeled and analyzed with CFD and the resulting blood flow calculations were validated by comparison with brachial artery flow measured by duplex ultrasound. RESULTS: Predicted flow rates were derived from CFD analysis of a range of stenoses. These stenoses were modeled by CFD and correlated with the ultrasound measured flow rate through the fistula of eight patients. The calculated flow rate using CFD correlated within 20% of ultrasound measured flow for five of eight patients. The mean difference was 17.2% (ranged from 1.3% to 30.1%). CONCLUSIONS: CFD analysis-generated flow rate tables provide valuable information to assess the functional significance of stenosis detected during imaging studies. The CFD study can help in determining the clinical relevance of a stenosis in access dysfunction and guide the need for intervention.