Engineering Functioning Salivary Glands Using Micropatterned Scaffolds

描述（由申请人提供）：本申请的目标是使用将成人唾液腺细胞与微图案化人工支架相结合的创新策略来设计复杂的3D人工唾液腺。我的研究计划的长期目标是通过人工唾液腺的工程化来促进转化研究，用于患有唾液腺功能减退症的人类患者。头颈部放射治疗和干燥综合征都会导致不可逆的唾液腺组织损伤后唾液分泌减少。在这些患者中，由于口干导致言语困难、龋齿、口咽感染、粘膜炎和味觉丧失，唾液产生的缺乏导致显著的发病率。一种恢复唾液流的新策略是用由附着在支架上的自组织细胞组成的工程组织替换受损的唾液组织。在本申请中测试的假设是，附着于微图案化人工支架的自组织唾液腺功能单元可以产生功能性人工腺。这一假说是基于我们先前的证明，即胚胎唾液腺细胞具有自组织成功能性唾液腺组织的内在能力。在目标1中，将建立用于生长原代成年小鼠唾液腺细胞的条件，将建立功能化的微图案化支架，并且将建立唾液腺细胞可附着于支架的条件。在第二个目标中，我们将通过首先促进成年唾液腺细胞自组织成功能单元，然后通过功能化成核位点将这些单元附着到支架上，并形成3D人工腺体结构来组装工程化腺体。该人工腺体的功能将在未来的R01应用中进行体内测试。用于这种方法的方法将促进人类人工唾液腺的工程化，并且还可以作为工程化其他复杂分支器官如胰腺、肾脏和肺的原型。 公共卫生相关性：从这项资助获得的数据将推进有关成人唾液上皮细胞的能力的基本科学知识，以自我组织成唾液分泌结构时，放置在由适当的细胞外基质成分和结构组成的局部环境。我们还制造了一种人造支架，我们可以将唾液腺细胞附着在上面，它将作为一种管道，将唾液输送到口腔中。附着在人工支架上的自组织细胞将共同创建一个人工唾液腺，该唾液腺将在未来的R01应用中进行体内功能测试。这项工作将导致人类人工唾液腺将改善由于干燥综合征，放射治疗或其他原因导致唾液腺分泌不足的患者的口腔，牙齿和整体健康。

DESCRIPTION (provided by applicant): The goal of this application is to engineer a complex 3D artificial salivary gland using an innovative strategy combining adult salivary gland cells with a micropatterned artificial scaffold. The long-range goal of my research program is to facilitate translational research by engineering of an artificial salivary gland for use in human patients suffering from salivary hypofunction. Head and neck radiation therapy and Sjogren's syndrome both lead to decreased saliva production following irreversible salivary gland tissue damage. In these patients, lack of saliva production causes significant morbidity due to dry mouth that results in dysphasia, dental caries, oropharyngeal infections, mucositis, and loss of taste. A novel strategy for restoring salivary flow is to replace damaged salivary tissue with engineered tissue that is composed of self-organized cells attached to a scaffold. The hypothesis tested in this application is that self-organized salivary gland functional units that are attached to a micropatterned artificial scaffold can create a functioning artificial gland. This hypothesis is based on our previous demonstration that embryonic salivary gland cells have an inherent capacity to self- organize into functional salivary gland tissue. In Aim 1, conditions will be established for growing primary adult mouse salivary gland cells, a functionalized micropatterned scaffold will be created, and conditions whereby the salivary gland cells can attach to the scaffold will be established. In the second Aim, we will assemble an engineered gland by first facilitating self-organization of the adult salivary gland cells into functional units and then attach these units to the scaffold through functionalized nucleation sites, and form a 3D artificial gland structure. The function of this artificial gland will be tested in vivo in a future R01 application. The methods used for this approach will facilitate engineering of a human artificial salivary gland and also serve as a prototype for engineering other complex branched organs such as pancreas, kidney, and lung. PUBLIC HEALTH RELEVANCE: The data obtained from this grant will advance basic scientific knowledge regarding the ability of adult salivary epithelial cells to self-organize into saliva-secreting structures when placed in a local environment consisting of the appropriate extracellular matrix composition and structure. We also create an artificial scaffold to which we can attach salivary gland cells and that will function as a conduit for delivery of saliva into the oral cavity. Together, the self-organized cells attached to the artificial scaffold will create an artificial salivary gland that will be tested for function in vivo in a future R01 application. This work will lead to a human artificial salivary gland will improve the oral, dental, and overall health of patients suffering from lack of saliva production by the salivary gland due to Sjogren's syndrome, radiation therapy, or other causes.