A Hydrogel-Based Cellular Model of the Human Vocal Fold

基于水凝胶的人类声带细胞模型

• 批准号: 10209183
• 负责人: Xinqiao Jia
• 金额: $ 51.15万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10209183
• 关键词: Acoustics; Adopted; Affect; Aging; Air; Air Movements; American; Animal Model; Apoptosis; Architecture; Basement membrane; Biochemical; Biology; Biomechanics; Biophysics; Cell model; Cells; Cellular Morphology; Chemical Exposure; Chemicals; Chimeric Proteins; Cicatrix; Clinical; Communities; Connective Tissue; Consensus; Cues; Custom; Data; Decision Making; Deposition; Development; Disease; Engineering; Epithelial; Epithelial Cells; Extracellular Matrix; FGF2 gene; Fibrinogen; Fibroblasts; Fibrosis; Functional disorder; Future; Gene Expression; Growth Factor; Health; Human; Hydrogels; In Situ; In Vitro; Intervention; Lamina Propria; Larynx; Ligation; Lung; Mechanical Stress; Mechanics; Mesenchymal; Mesenchymal Stem Cells; Methodology; Methods; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Monitor; Motion; Multipotent Stem Cells; Muscle; Myofibroblast; Operative Surgical Procedures; Organ; Parents; Pathologic; Permeability; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Phenotype; Phonation; Physiological; Physiology; Pliability; Predisposition; Process; Property; Protein Kinase; Research; Side; Signal Transduction; Stratified Epithelium; Stratified Squamous Epithelium; Structure; TGFB1 gene; Testing; Therapeutic; Tissue Microarray; Tissue Model; Tissues; Trachea; Treatment Efficacy; Voice; base; cohesion; cytokine; design; efficacious treatment; epithelial injury; fasudil; fibrogenesis; fundamental research; graphene; healing; human tissue; improved; induced pluripotent stem cell; inhibitor/antagonist; interfacial; interstitial; mechanical properties; mimetics; miniaturize; prevent; real time monitoring; repaired; rho; sensor; sound; spatiotemporal; tissue injury; vocal cord; vocalis muscle; wound healing

Project Summary Voice is produced when the vocal folds are driven into a wave-like motion by the airstream from the trachea, converting aerodynamic energy and airflow into acoustic energy in the form of sound. Each vocal fold consists of a pliable vibratory layer of connective tissue, known as the lamina propria (LP), sandwiched between a muscle and a stratified squamous epithelium (EP). Numerous environmental, mechanical and pathological factors can damage this delicate tissue, resulting in vocal fold scarring that affects millions of Americans with limited treatment options. Although there is a general consensus on the pathophysiology of vocal fold scarring, the molecular and cellular mechanisms that control unremitting fibrosis remain poorly understood. Studies on other fibrotic diseases suggest that fibroblasts, epithelial cells and the interstitial matrix are active players in fibrogenesis. This project aims to engineer a reliable, physiologically relevant in vitro tissue model that can be used to investigate vocal fold development, health, and disease, and more importantly, to facilitate the development and testing of new treatment options. We propose to develop a microengineered organ chip that integrates the epithelial and mesenchymal cells in a tissue-mimetic configuration with built-in airflow to stimulate phonation. Using the microfluidic model, we will investigate how damage to the epithelium initiates fibrosis, how the fibrotic extracellular matrix (ECM) sustains fibrosis and how myofibroblast proliferation and matrix deposition continue unabated. Finally, we will calibrate our model with an antifibrotic growth factor that has shown efficacy in treating vocal fold scarring, and test a promising pharmacological inhibitor that has not been previously tested in the context of vocal fold scarring. Highly efficient bioorthogonal tetrazine ligation will be used to establish the initial LP matrix surrounding healthy fibroblasts and to introduce compositional and mechanical alterations that promote fibroblast activation. Pluripotent and multipotent stem cells will be guided to differentiate into vocal fold- like epithelial cells and fibroblasts by adopting a development paradigm and through systematic manipulation of the engineered microenvironment. Piezoresistive strain sensors embedded in the sidewalls of the microfluidic channels will be used to monitor tissue stiffness and EP permeability in situ. The microengineered tissue model will be characterized in terms of cell phenotype, microstructure, mechanical properties and physiological function. For comparison purposes, a stand-alone, human-sized vocal fold model will be developed and characterized employing methodologies established in the laryngology field. Data generated from this project should significantly impact fundamental research related to vocal fold scarring and provide critical information on therapeutic decision-making in the near future.

项目摘要 当声带被气流从气流中驱动到波浪状的运动中时，会产生声音， 以声音形式将空气动力和气流转化为声能。每个人声折叠都是 夹在肌肉之间 和分层的鳞状上皮（EP）。许多环境，机械和病理因素可以 损坏这种精致的组织，导致声带疤痕，影响数百万的美国人 治疗选择。尽管关于声带疤痕的病理生理学有一个普遍的共识，但 控制无纤维化的分子和细胞机制仍然了解不足。对他人的研究 纤维化疾病表明成纤维细胞，上皮细胞和间质基质是活跃的参与者 纤维发生。该项目旨在设计一个可靠的，生理上相关的体外组织模型 用于调查声带的发展，健康和疾病，更重要的是，以促进 开发和测试新治疗方案。我们建议开发一个微工具的器官芯片 与内置的气流中整合组织模拟构型中的上皮和间质细胞以刺激 发声。使用微流体模型，我们将研究上皮的损害如何启动纤维化，如何 纤维化细胞外基质（ECM）维持纤维化以及肌纤维细胞增殖和基质沉积 继续没有减弱。最后，我们将用抗纤维化生长因子校准模型，该模型已显示出功效 在治疗声带疤痕和测试有希望的药理学抑制剂时，该抑制剂以前尚未进行过测试 在人声折叠的背景下。高效的生物方向双嗪结扎将用于建立 围绕健康成纤维细胞的初始LP基质，并引入组成和机械改变， 促进成纤维细胞激活。多能和多能干细胞将被引导分化为声带 - 像上皮细胞和成纤维细胞一样，通过采用开发范式并通过系统地操纵 工程的微环境。嵌入微流体侧壁中的压电应变传感器 通道将用于监测组织刚度和原位EP渗透性。微工程组织模型 将以细胞表型，微结构，机械性能和生理功能来表征。 为了进行比较，将开发和表征一个独立的人尺寸的人声折叠模型 采用在喉科领域建立的方法。该项目产生的数据应该 显着影响与声带疤痕有关的基本研究，并提供有关的关键信息 在不久的将来的治疗决策。