Ex vivo bioengineering of functional biomimetic airways for treatment of neonatal and pediatric respiratory conditions

项目摘要 功能性呼吸组织的离体工程化仍然具有挑战性。但新 生物工程技术发展迅速，气管和大气道的3D生物打印， 结合细胞和仿生材料，越来越多地被应用于新生儿和 小儿呼吸系统疾病。理想地，通过医学成像引导，患者自身的气道和/或肺泡 干细胞/祖细胞可用于产生新的患者特异性3D打印构建体。然而，在这方面， 目前在3D生物打印中使用的仿生材料未能重现生物打印的复杂微环境。 并且对于支持肺干细胞/祖细胞生长、分化和 功能此外，所使用的普通仿生材料不会随着患者年龄增长而生长。的目标 我们的建议是通过3D打印技术，模拟天然介质， 和大气道的机械和组成复杂性。由于细胞外基质（ECM）已经被 显示在肺发育和细胞过程的调节中起着不可或缺的作用，我们的假设 脱细胞ECM是3D生物打印气道支架的最佳基质， 代表性分化的气道上皮干/祖细胞的生长、分化和功能。 已经开发了以下目标来测试我们的假设：目标1，人体的机械优化 dECM 3D生物打印气道结构;目标2，3D生物打印中细胞活力和表型的优化 dECM气道结构。该建议的意义在于证明dECM是一种最佳方法。 基质不仅用于支持和分化功能性气道上皮细胞，而且用于3D 打印仿生气道组织。本提案的创新之处将是对这种材料的优化 用于生产功能性可植入3D打印的新生儿和儿科气道结构， 呼吸系统疾病，如囊性纤维化、支气管肺发育不良和肺发育不全， 先天性缺陷

Project Summary Ex vivo engineering of functional respiratory tissues continues to be challenging. However, new bioengineering techniques are developing at a rapid pace. 3D bioprinting of trachea and large airways, incorporating both cells and biomimetic materials, is increasingly being applied for treatment of neonatal and pediatric respiratory conditions. Ideally, guided by medical imaging, a patient's own airway and/or alveolar stem/progenitor cells can be utilized to generate a novel patient-specific 3D-printed construct. However, biomimetic materials currently utilized in 3D bioprinting fail to recapitulate the complex microenvironment of the native lung and are not optimal for supporting lung stem/progenitor cell growth, differentiation, and function. Further, common biomimetic materials utilized do not grow as the patient ages. The objective of our proposal is to bioengineer a functional airway, through 3D printing technology, that mimics native medium and large airways in mechanical and compositional complexity. As extracellular matrix (ECM) has been shown to play an integral part in lung development and the regulation of cellular processes, our hypothesis is that decellularized ECM is an optimal substrate for 3D bioprinting of airway scaffolds and will promote growth, differentiation, and function of representative differentiated airway epithelial stem/progenitor cells. The following aims have been developed to test our hypothesis: Aim 1, mechanical optimization of human dECM 3D bioprinted airway structures; Aim 2, optimization of cell viability and phenotype in 3D bioprinted dECM airway structures. The significance of this proposal will be in demonstrating that dECM is an optimal substrate for not only the support and differentiation of functional airway epithelial cells, but also for the 3D printing of biomimetic airway tissues. The innovation of this proposal will be the optimization of this material for use in the production of functional implantable 3D printed neonatal and pediatric airway constructs to treat respiratory conditions such as Cystic Fibrosis, Bronchopulmonary Dysplasia and lung hypoplasia due to congenital defects.