Abstract

The zebrafish Weberian apparatus is an emerging model for human conductive hearing system. Their Weberian apparatus comprises minute bones and ligamentary links, and conducts sound pressure transmission from the gas bladder to inner ear through four pairs of Weberian ossicles along the vertebral column. We herein present a methodological study using MicroCT to image the Weberian apparatus for biomechanical and morphological analysis. The aim of this work is to evaluate computational models generated from multiple MicroCT scans with different parameters, to identify the most feasible scan combination for practical (minimized scan time) yet accurate (relative to highest resolution) biomechanical simulations. We segmented and created 3D models from CT scan image stacks at 4.64 μm, 5.05 μm, 9.30 μm and 13.08 μm voxel resolutions, respectively. Then, we used geometric morphometrics analysis to quantify inter-model shape differences, as well as a series of finite element modal and harmonic analyses to simulate auditory signal vibrations. Relative to the highest resolution and most accurate model, the Model 9.30 is closest in overall geometry and biomechanical behavior of all lower resolution models. The differences in resolution and quality of the CT substantially affect the segmentation and reconstruction process of the three-dimensional model of the ossicles, and the subsequent analyses. We conclude that scan voxel resolution is a key factor influencing outcomes of biomechanical simulations of delicate and minute structures, especially when studying the harmonic response of minute ossicles connected by ligaments using finite element modeling. Furthermore, contrast variations in CT images as determined by x-ray power and scan speed, also affect fidelity in 3D models and simulation outcomes.

Highlights

  • Zebrafish (Danio rerio) is a versatile model organism in biomedical and organismal biology research

  • Developmental biology evidence of zebrafish hearing suggests that the Weberian ossicles gradually increase the range of frequencies available to the inner ear during zebrafish growth, just as middle ear development increases frequency range in mammal ontogeny (Higgs et al, 2003)

  • We present a study using MicroCT to image the Weberian apparatus for biomechanical analysis. The aims of this project are two-fold: 1) evaluate models generated from multiple MicroCT scans with different parameters to find the most feasible scan combination for practical yet accurate images for biomechanical simulations; 2) hypothesize practical scan parameter combinations for similar-sized structures in zebrafish, or the same structure in other small animal models

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Summary

Introduction

Zebrafish (Danio rerio) is a versatile model organism in biomedical and organismal biology research. The Weberian apparatus in zebrafish conducts sound pressure transmission from gas bladder to inner ear through Weberian ossicles, and is analogous to the human middle ear, which conducts sound vibration and transmit sound pressure from the temporal membrane to the inner ear through three middle ear ossicles (Evans, 1925; Weber, 1820). Developmental biology evidence of zebrafish hearing suggests that the Weberian ossicles gradually increase the range of frequencies available to the inner ear during zebrafish growth, just as middle ear development increases frequency range in mammal ontogeny (Higgs et al, 2003). Weberian ossicles conduct sound pressure from gas filled bladder to the fluid filled sinus impar of

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