Vocal folds are soft laryngeal connective tissues with distinct layered structures and complex multicomponent matrix compositions that endow phonatory and respiratory functions. This delicate tissue is easily damaged by various environmental factors and pathological conditions, altering vocal biomechanics and causing debilitating vocal disorders that detrimentally affect the daily lives of suffering individuals. Modern techniques and advanced knowledge of regenerative medicine have led to a deeper understanding of the microstructure, microphysiology, and micropathophysiology of vocal fold tissues. State-of-the-art materials ranging from extracecullar-matrix (ECM)-derived biomaterials to synthetic polymer scaffolds have been proposed for the prevention and treatment of voice disorders including vocal fold scarring and fibrosis. This review intends to provide a thorough overview of current achievements in the field of vocal fold tissue engineering, including the fabrication of injectable biomaterials to mimic in vitro cell microenvironments, novel designs of bioreactors that capture in vivo tissue biomechanics, and establishment of various animal models to characterize the in vivo biocompatibility of these materials. The combination of polymeric scaffolds, cell transplantation, biomechanical stimulation, and delivery of antifibrotic growth factors will lead to successful restoration of functional vocal folds and improved vocal recovery in animal models, facilitating the application of these materials and related methodologies in clinical practice.

Vocal fold tissue engineering requires biomimetic scaffolds with an appropriate matrix stiffness closely matching that of the natural vocal folds to maintain function. Traditionally, poly(ε-caprolactone) (PCL) and thermoplastic polyurethane (TPU) have been employed as the primary matrix materials for vocal fold electrospun scaffolds. However, not all of the scaffolds fabricated thus far matched the human vocal fold tissues. Poly(glycerol sebacate) (PGS) is a non-cytotoxic and biodegradable soft elastomer that has shown promising results for soft tissue engineering applications. However, no work has been done to employ this biomaterial to construct vocal fold scaffolds. In this study, PGS has been synthesized and blended with thermoplastic polyurethane (TPU) to produce vocal fold scaffolds with improved hydrophilicity and compliance by electrospinning. The resulting scaffolds were found to have mechanical properties mimicking those of the vocal fold lamina propria extracellular matrix (ECM). An unusual leaf-like structure was obtained when using 1,1,1,3,3,3-hexafluoroisopropanol (HFIP) as the solvent. Other suitable fibrous scaffolds were also obtained when using acetic acid and 2,2,2-trifluoroethanol (TFE) as binary solvents. A biological evaluation of these TPU/PGS scaffolds showed better cell spreading and significantly improved cell proliferation as compared to TPU-only scaffolds (p < 0.01), thereby suggesting potential applications for vocal fold tissue engineering.

Vocal folds are connective tissues housed in the larynx, which can be subjected to various injuries and traumatic stimuli that lead to aberrant tissue structural alterations and fibrotic-induced biomechanical stiffening observed in patients with voice disorders. Much effort has been devoted to generate soft biomaterials that are injectable directly to sites of injury. To date, materials applied toward these applications have been largely focused on natural extracellular matrix-derived materials such as collagen, fibrin or hyaluronic acid; these approaches have suffered from the fact that materials are not sufficiently robust mechanically nor offer sufficient flexibility to modulate material properties for targeted injection. We have recently developed multiple resilin-inspired elastomeric hydrogels that possess similar mechanical properties as those reported for vocal fold tissues, and that also show promising in vitro cytocompatibility and in vivo biocompatibility. Here we report studies that test the delivery of resilin-based hydrogels through injection to the subcutaneous tissue in a wild-type mice model; histological and genetic expression outcomes were monitored. The rapid kinetics of crosslinking enabled facile injection and ensured the rapid transition of the viscous resilin precursor solution to a solid-like hydrogel in the subcutaneous space in vivo; the materials exhibited storage shear moduli in the range of 1000-2000 Pa when characterized through oscillatory rheology. Histological staining and gene expression profiles suggested minimal inflammatory profiles three weeks after injection, thereby demonstrating the potential suitability for site-specific in vivo injection of these elastomeric materials. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2229-2242, 2018.