Highly resilient, hydrophilic bioelastomers for engineering vocal fold tissue

描述（由申请人提供）：受损的声音产生对个人健康、社会和职业功能以及社会生产力有重大影响；据估计，在美国，仅教师的声音问题所造成的社会成本每年就高达25亿美元。然而，用于治疗声带疾病的材料的开发一直受到声带严格的机械要求的阻碍，这些要求包括在高达1000Hz的频率下维持变形的能力，以及在瞬态拉伸高达200%后完全后缩的能力。迄今为止，尽管在开发用于声带组织工程的材料支架方面做出了广泛的努力，但尚未发现具有所需机械性能的材料。我们提出了一个全面的生物工程方法来解决这个问题。我们将采用基于昆虫蛋白弹性蛋白的新型弹性支架，弹性蛋白是昆虫发声、跳跃和飞行器官中的主要能量储存，在高达4000Hz的频率下变形后显示出无与伦比的弹性（拉伸后恢复）。我们将采用模块化重组方法来生成类弹性蛋白多肽（rlp）， rlp可以被设计成携带生物活性结构域，而不影响弹性蛋白结构域的机械特性，并且可以独立调节这些基质的多种特性，包括机械特性、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），并采用一套振荡流变学、拉伸测试和高频扭转波方法来表征细胞封装结构的机械特性。组织学、免疫组织学、western blot和基因表达技术将被用来证实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细胞/材料结构注入兔声带以改善声带瘢痕提供信息。因此，我们的研究将有助于在高频下表征和培养材料的方法的发展，并产生一类新的材料，可以优化声带组织的再生。我们的方法最终将有助于为机械要求苛刻的再生医学应用设计材料的通用平台。

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.