Engineering Functioning Salivary Glands Using Micropatterned Scaffolds

描述（由申请人提供）：本申请的目标是利用一种创新的策略，将成人唾液腺细胞与微图案人工支架结合起来，设计一个复杂的3D人工唾液腺。我的研究计划的长期目标是促进转化研究通过工程人工唾液腺用于人类患者患有唾液腺功能减退。头颈部放射治疗和干燥综合征在不可逆的唾液腺组织损伤后都会导致唾液分泌减少。在这些患者中，唾液分泌不足会导致口腔干燥，导致吞咽困难、龋齿、口咽感染、黏膜炎和味觉丧失。一种恢复唾液流动的新策略是用附着在支架上的自组织细胞组成的工程组织代替受损的唾液组织。在这个应用中测试的假设是，自组织的唾液腺功能单元连接到一个微图案的人造支架上，可以创建一个功能性的人造腺体。这一假设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Aim 1中，将建立生长成年小鼠初级唾液腺细胞的条件，将创建一个功能化的微图案支架，并将建立唾液腺细胞附着在支架上的条件。在第二个目标中，我们将首先促进成人唾液腺细胞自组织成功能单元，然后通过功能化的成核位点将这些单元连接到支架上，形成一个3D人工腺体结构。这种人工腺体的功能将在未来的R01应用中进行体内测试。该方法将促进人类人工唾液腺的工程，也可作为工程其他复杂分支器官（如胰腺、肾脏和肺）的原型。

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.