Highly resilient, hydrophilic bioelastomers for engineering vocal fold tissue

描述(由申请人提供)：发声障碍对个人健康、社会和职业功能以及社会生产力具有重大影响；在美国，仅教师发声问题的社会成本估计就高达25亿美元左右。然而，用于治疗声带疾病的材料的发展一直受到声带严格的机械要求的阻碍，其中包括在高达1000赫兹的频率下保持变形的能力，以及在瞬时拉伸高达200%后完全回缩的能力。到目前为止，尽管声带组织工程材料支架的开发得到了广泛的努力，但还没有鉴定出具有所需力学性能的材料。我们提出了一个全面的生物工程方法来解决这个问题。我们将使用基于昆虫蛋白质resilin的新型弹性支架，resilin是昆虫发声、跳跃和飞行器官中的主要能量储存，并在高达4000赫兹的频率下变形后显示出无与伦比的弹性(拉伸后恢复)。我们将使用模块化重组方法来生成Resilin样多肽(RLP)，这些RLP可以在不影响Resilin结构域的机械性能的情况下被设计为携带生物活性结构域，并且可以独立调节这些基质的多种属性，包括机械性能、细胞结合和降解。我们将在静态和动态条件下在这些基质中培养人间充质干细胞(HMSCs)，并使用一套振荡流变学、拉伸测试和高频扭转波方法来表征细胞包裹结构的机械性能。组织学、免疫组织学、免疫印迹和基因表达技术将用于证实hMSCs的分化和声带细胞外基质的产生。这些研究将为我们选择注射到兔声带皱折以改善声带瘢痕的细胞/材料结构提供信息。因此，我们的研究将有助于开发高频表征和培养材料的方法，以及产生一类可能优化声带组织再生的新材料。我们的方法最终将在机械要求苛刻的再生医学应用的材料设计中作为一个通用平台有用。

DESCRIPTION (provided by applicant): Impaired voice production holds significant implications for individual health and wellness, social and occupational function, and societal productivity; the societal costs of voice problems in teachers alone have been estimated to be of the order of $2.5 billion annually in the United States. The development of materials for the treatment of vocal fold disorders, however, has been hampered by the stringent mechanical requirements of the vocal fold, which include the ability to both sustain deformation at frequencies as high as 1000Hz, and also completely recoil after transient stretch up to 200%. To date, despite widespread efforts in the development of materials scaffolds for the tissue engineering of the vocal fold, no materials with the required mechanical properties have been identified. We propose a comprehensive bioengineering approach to this problem. We will employ new elastomeric scaffolds based on the insect protein, resilin, which is the primary energy store in the sound-producing, jumping, and flight organs of insects, and demonstrates unmatched resilience (recovery after stretch) after deformation at frequencies up to 4000Hz. We will employ modular recombinant methods to generate resilin- like polypeptides (RLPs) that can be engineered to carry biologically active domains without compromising the mechanical properties of the resilin domain, and in which independent tuning of multiple properties of these matrices, including mechanical properties, cell binding, and degradation, is possible. We will culture human mesenchymal stem cells (hMSCs) in these matrices under both static and dynamic conditions, and employ a suite of oscillatory rheology, tensile testing, and high-frequency torsional-wave methods to characterize the mechanical properties of cell-encapsulated constructs. Histological, immunohistological, western blot, and gene expression techniques will be employed to confirm the differentiation of hMSCs and the production of vocal fold extracellular matrix. These studies will inform our choices of cell/materials constructs for injection into the vocal folds of rabbits to ameliorate vocal fold scarring. Our investigations wil thus contribute to the development of methods to characterize and culture materials at high frequencies, as well as yield a new class of materials that may optimize the regeneration of vocal fold tissue. Our approaches ultimately will be useful as a general platform in the design of materials for mechanically demanding regenerative medicine applications.