用于声带组织工程的三层水凝胶支架《Biomacromolecules》

Trilayered Hydrogel Scaffold for Vocal Fold Tissue Engineering

R. Kevin Tindell, Michael J. McPhail, Cheryl E. Myers, Juergen Neubauer, Justin M. Hintze, David G. Lott*, and Julianne L. Holloway*

Chemical Engineering; School for Engineering of Matter, Transport and Energy, Arizona State University, Tempe, Arizona 85287, United States

Biomacromolecules 2022, XXXX, XXX, XXX-XXX

Publication Date: October 26, 2022

https://doi.org/10.1021/acs.biomac.1c01149

Abstract

The lamina propria within the vocal fold (VF) is a complex multilayered tissue that increases in stiffness from the superficial to deep layer, where this characteristic is crucial for VF sound production. Tissue-engineered scaffolds designed for VF repair must mimic the biophysical nature of the native vocal fold and promote cell viability, cell spreading, and vibration with air flow. In this study, we present a unique trilayered, partially degradable hydrogel scaffold that mimics the multilayered structure of the VF lamina propria. Using thiol-norbornene photochemistry, trilayered hydrogel scaffolds were fabricated via layer-by-layer stacking with increasing polymer concentration from the top to middle to deep layer. Mechanical analysis confirmed that hydrogel modulus increased with increasing polymer concentration. Partially degradable hydrogels promoted high cell viability and cell spreading in three dimensions as assessed via live/dead and cytoskeleton staining, respectively. Importantly, partially degradable hydrogels maintained some degree of the three dimensional polymer network following protease exposure, while still enabling encapsulated cells to remodel their local environment via protease secretion. Finally, the trilayered hydrogel scaffold successfully vibrated and produced sound in proof-of-concept air flow studies. This work represents a critical first step toward the design of a multilayered, hydrogel scaffold for vocal fold tissue engineering.

SUBJECTS:

• Atmospheric chemistry,

• Cells,

• Hydrogels,

• Layers,

• Scaffolds

摘要

声带（VF）内的固有层是一种复杂的多层组织，从表层到深层硬度增加，这一特性对VF声音的产生至关重要。设计用于心室颤动修复的组织工程支架必须模仿天然声带的生物物理性质，并促进细胞活力、细胞扩散和气流振动。在这项研究中，我们提出了一种独特的三层、部分可降解的水凝胶支架，它模拟了心室颤动固有层的多层结构。使用硫醇降冰片烯光化学，通过逐层堆叠制备了三层水凝胶支架，聚合物浓度从顶层到中层再到深层增加。力学分析证实，水凝胶模量随着聚合物浓度的增加而增加。部分可降解水凝胶分别通过活/死和细胞骨架染色评估，促进了高细胞活力和三维细胞扩散。重要的是，部分可降解的水凝胶在蛋白酶暴露后保持了一定程度的三维聚合物网络，同时仍使被包裹的细胞能够通过蛋白酶分泌重塑其局部环境。最后，三层水凝胶支架在概念验证气流研究中成功振动并产生声音。这项工作是设计用于声带组织工程的多层水凝胶支架的关键第一步。