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

项目摘要 功能呼吸组织的体外工程仍然具有挑战性。然而，新的 生物工程技术发展迅速。气管和大呼吸道的3D生物打印， 结合了细胞和仿生材料，越来越多地被应用于治疗新生儿和 儿科呼吸系统疾病。理想情况下，在医学成像的引导下，患者自己的呼吸道和/或肺泡 干细胞/祖细胞可以用来产生一种新的针对患者的3D打印构建体。然而， 目前用于3D生物打印的仿生材料无法概括出 天然肺并不适合支持肺干/祖细胞的生长、分化和 功能。此外，所使用的常见仿生材料不会随着患者年龄的增长而增长。的目标是 我们的建议是通过3D打印技术生物工程一种功能良好的呼吸道，模仿天然介质 以及机械和组成复杂的大型呼吸道。由于细胞外基质(ECM)已被 被证明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是呼吸道支架3D生物打印的最佳底物，并将促进 典型分化的呼吸道上皮干/祖细胞的生长、分化和功能。 为了验证我们的假设，我们开发了以下目标：目标1，人类的机械优化 DECM 3D生物打印的呼吸道结构；目标2，3D生物打印的细胞活性和表型的优化 DECM呼吸道结构。这项提议的意义将在于证明decm是一种最优的 基质不仅用于支持和分化有功能的呼吸道上皮细胞，而且还用于3D 仿生呼吸道组织的打印。这项建议的创新之处将是对材料的优化 用于生产功能性植入性3D打印新生儿和儿童呼吸道结构以治疗 呼吸道疾病，如囊性纤维化、支气管肺发育不良和肺发育不良 先天缺陷。

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.