Tissue Engineering of Vocal Fold Lamina Propria

描述（由申请人提供）：人类发声依赖于柔性声带固有层，当被来自气管的气流驱动时，声带固有层在聚集在一起时可以振动。声音过度使用或滥用会导致疤痕，破坏固有层的自然柔韧性，导致声音嘶哑和其他声音功能障碍的症状。声带瘢痕的减少仍然是一个重大的治疗挑战。我们建议开发两种平行的组织工程方法，将导致声带固有层的再生。第一种方法将应用可注射的水凝胶来防止瘢痕形成，改善受损组织的柔韧性，启动主动组织重塑，并最终提供功能性声带固有层的体内再生。第二种方法依赖于通过细胞、人工细胞外基质（ECM）、生物线索和机械刺激的适当组合的体外功能性组织形成。我们正在开发基于交联颗粒网络（XPN）的人工ECM，该交联颗粒网络由透明质酸（HA）水凝胶颗粒（HGP）和水溶性功能聚合物组成。所述水凝胶颗粒被设计为显示受控的尺寸、限定的表面功能性、改善的酶稳定性和生物活性分子（包括抗纤维化药物、生长因子形态发生素和细胞粘附肽）的空间/时间显示。另一方面，XPN将具有可调的粘弹性和受控的降解，其捕获天然固有层的机械和生物学特性。存在两个水平的交联（在个体HGP内和个体HGP之间）提供了从机械应力中快速恢复的潜力。交联化学被设计为允许声带成纤维细胞（VFF）的原位包封。为了模拟声带组织所经历的机械环境，我们建议构建一种生物反应器，该生物反应器能够向细胞包封的支架提供明确的振动和拉伸应力。声带成纤维细胞、弹性和生物活性人工细胞外基质以及动态生物反应器的结合为声带固有层的体外组织工程提供了令人兴奋的机会。

DESCRIPTION (provided by applicant): Human voice production is dependent on the flexible vocal fold lamina propria that can vibrate when brought together while being driven by the airstream from the trachea. Voice overuse or abuse can lead to scarring that disrupts the natural pliability of the lamina propria and results in hoarseness and other symptoms of vocal dysfunction. The reduction of vocal fold scarring remains a significant therapeutic challenge. We propose to develop two parallel tissue engineering approaches that will lead to the regeneration of vocal fold lamina propria. The first method will apply injectable hydrogels to prevent scar formation, improve the pliability of damaged tissue, initiate active tissue remodeling, and ultimately, afford in vivo regeneration of functional vocal fold lamina propria. The second approach relies on in vitro functional tissue formation by the appropriate combination of cells, artificial extracellular matrices (ECM), biological cues and mechanical stimuli. We are developing artificial ECM based on crosslinked particle networks (XPN) that consist of hyaluronic acid (HA) hydrogel particles (HGP) and water soluble functional polymers. The hydrogel particles are designed to exhbit controlled sizes, defined surface functionality, improved enzymatic stability, and spatial/temporal display of biologically active molecules including antifibrotic drugs, growth factor morphogens and cell adhesion peptides. The XPN, on the other hand, will have tunable viscoelasticty and controlled degradation that capture the mechanical and biological characteristics of the native lamina propria. The existence of two levels of crosslinking (within and between individual HGP) offers potential for rapid recovery from mechanical stress. The crosslinking chemistry is designed to allow for in situ encapsulation of vocal fold fibroblasts (VFF). To mimic the mechanical environment experienced by the vocal fold tissue, we propose to construct a bioreactor that is capable of delivering well-defined vibrational and tensile stresses to the cell-encapsulated scaffolds. The combination of vocal fold fibroblasts, elastic and bioactive artificial ECM, and a dynamtic bioreactor offers exciting opportunity for in vitro tissue engineering of vocal fold lamina propria.