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.
Highlights
Human vocal folds have a distinctive tri-layered mucosal structure with rich extracellular matrix substances such as collagen, elastin and hyaluronan [1]
While the rabbit model has been used to evaluate different potential vocal fold regeneration therapies [36], the data obtained in these studies was limited to traditional histology such as Hematoxylin and Eosin (H&E), which cannot provide the entire three-dimensional tissue landscape when it comes to a spatial understanding of wound healing
The entire wound area could be observed and compared across animals over time. Such capability is important to standardize the location and size of the injury and to evaluate the wound healing outcome in a more consistent manner. Before such studies can be conducted, it is important to determine the ultrastructure of normal rabbit vocal folds and verify that the proposed virtual histology methods provide adequate resolution and contrast
Summary
Human vocal folds have a distinctive tri-layered mucosal structure with rich extracellular matrix substances such as collagen, elastin and hyaluronan [1]. The delicate structure of the VFs makes them prone to scarring as a consequence of injury, disease or surgery [2]. The thicker scar tissue inhibits proper phonation and leads to voice disorders or even the loss of voice, which significantly hampers the patient’s communicative ability and quality of life [3]. Surgical attempts to remove the scar often result in additional, iatrogenic scarring [4]. While studies have been conducted on potential therapies, effective treatment for the prevention or removal of VF scarring has not yet been achieved [5,6,7,8]. Proper understanding of the cellular mechanisms of the scarring process is required for the development of better treatment strategies [9]
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