Multimodal virtual histology of rabbit vocal folds by nonlinear microscopy and nano computed tomography

Abstract

Human vocal folds (VFs) possess a unique anatomical structure and mechanical properties for human communication. However, VFs are prone to scarring as a consequence of overuse, injury, disease or surgery. Accumulation of scar tissue on VFs inhibits proper phonation and leads to partial or complete loss of voice, with significant consequences for the patient's quality of life. VF regeneration after scarring provides a significant challenge for tissue engineering therapies given the complexity of tissue microarchitecture. To establish an effective animal model for VF injury and scarring, new histological methods are required to visualize the wound repair process of the tissue in its three-dimensional native environment. In this work, we propose the use of a combination of nonlinear microscopy and nanotomography as contrast methods for virtual histology of rabbit VFs. We apply these methods to rabbit VF tissue to demonstrate their use as alternatives to conventional VF histology that may enable future clinical studies of this injury model.

Quantitative evaluation of the human vocal fold extracellular matrix using multiphoton microscopy and optical coherence tomography

“…Recently, synchrotron X-ray microtomography have been used successfully on human cadavers 16 , and nanotomography on rabbits 17 to identify the tri-dimensional structures of the VF ECM, including its muscle fibers morphology. However, these techniques are not collagen or elastin sensitive and suffer from low contrast since they instead rely on tissue density (X-ray attenuation coefficient) as a contrast mechanism.…”

A micro-mechanical model for the fibrous tissues of vocal folds

“…This is mainly ascribed to their challenging experimental multiscale characterization. Despite the considerable progress made in 3D microimaging [43,52,18,73,51,32,3,48], vocal folds, along with their fibrous architectures, are hardly observable in vivo [71,23]. Although a large biomechanical database has been collected on excised vocal folds over the last twenty years [17,87,49,74,16,20], the 3D microscale rearrangement of the loaded tissues is still to be explored.…”

The 13th International Conference on Advances in Quantitative Laryngology, Voice and Speech Research (June 2–4, 2019, Montreal, Quebec, Canada)

“…Larynges were harvested for histological studies (n = 1 for hematoxylin and eosin, and PAS staining; n = 1 for scanning electron microscope) and immunohistochemistry (n = 3). The antibodies against occludin, ZO-1 and cldn1, 3,4,5,6,7,8,10,11,12,17 were used to clarify the localization of each TJ-related protein in the laryngeal glands. Anti-cldn antibodies were selected among those whose gene expression was confirmed by previously performed RT-PCR analysis with rat larynx.…”