Engineering Functional Salivary Glands Using Micropatterned Scaffolds

使用微图案支架工程功能性唾液腺

• 批准号: 8874957
• 负责人: James Castracane
• 金额: $ 47.94万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8874957
• 关键词: Acinar Cell; Address; Adverse effects; Affect; American; Architecture; Autoimmune Diseases; Biological Assay; Biosensor; Caliber; Cell Differentiation process; Cell Line; Cell Polarity; Cell physiology; Cells; Charge; Chemicals; Chitosan; Clinical; Complex; Deglutition; Deglutition Disorders; Dental caries; Devices; Diagnosis; Differentiation and Growth; Digestion; Dimensions; Engineering; Environment; Extracellular Matrix; Fiber; Future; Gland; Heparin; Hyaluronic Acid; Image; In Vitro; Infection; Knowledge; Life; Maintenance; Measures; Methods; Monitor; Mucositis; Mus; Nanotopography; Organ; Oropharyngeal; Pain; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Phenotype; Preclinical Drug Evaluation; Production; Property; Quality of life; Radiation therapy; Saliva; Salivary; Salivary Glands; Signal Transduction; Sjogren' s Syndrome; Structure; Surface; Surface Properties; Symptoms; System; Taste Perception; Time; Tissue Engineering; Tissues; United States; Work; Xerostomia; base; cell fixing; combinatorial; design; experience; extracellular; head and neck cancer patient; imaging modality; immunocytochemistry; implantable device; in vitro Assay; in vivo; innovation; interest; macromolecule; mimetics; nanofiber; novel; response; salivary acinar cell; scaffold; secretory protein; sensor; small molecule; surface coating; three dimensional structure

DESCRIPTION (provided by applicant): Millions of people suffer from xerostomia, or "dry mouth" resulting from lack of saliva, causing a decreased quality of life resulting from multiple symptoms, including increased dental caries, oropharyngeal infections, difficulties with swallowing (dysphagia) and digestion (mucositis), loss of taste, and pain. As current treatments for these problems are inadequate (1), there is considerable interest in creating an artificial salivary gland. Maintenance of salivary acinar cell differentiation and function in vitro is criticl to the successful engineering of such constructs; however, this breakthrough has not yet been achieved. This is primarily due to the current lack of basic scientific knowledge regarding the specific extracellular signals that are required to maintain or induce acinar cell differentiation, which remains a substantial limitation in the ability to engineer a functional artificial salivary gland. An in vitro assay system is needed to identify required extracellular signals. We hypothesize that the combination of chemically-modified nanofibers presented to cells via microscale patterning in a 3D scaffold will support salivary acinar cell function. This application proposes an innovative, interdisciplinary strategy to create a high-throughput, non-invasive assay system that will be used to sense salivary acinar cell secretory function in live cells grown on scaffold materials, which has not been previously possible. A MEMS-based saliva sensor will be produced to accurately measure the concentration of a salivary secretory protein secreted by cells grown on unique scaffold combinations. A novel multiplexed immunocytochemistry method will be used to assess the extent of differentiation in fixed cells following the biosensor assay. Using an innovative combination of methods, nanotopography, micropatterning, and chemical signaling will be independently modulated, such that specific combinations of parameters will be identified that support acinar cell function. Scaffolds will be assessed for their ability to maintin acinar differentiation or promote re-differentiation using a combination of primary cells and cell lines. We will address this hypothesis in four specific aims: Aim 1: Develop a high-throughput MEMS probe array assay system to evaluate acinar cell function in vitro. Aim 2: Identify the ideal nanofiber configuration to support salivary acinar cell differentiation. Aim 3: Optimize nanofiber surface properties to enhance salivary gland acinar cell function. Aim 4: Identify an optimal microtopography to promote acinar cell function. The in vitro salivary gland construct produced in this application will be useful to identify pathways regulating acinar function and can be applied for drug screening. The principles developed as a result of this application will be applied in the future towards engineering of artificial glands designed to replicate salivary gland and other complex branching organs.

描述(申请人提供)：数以百万计的人患有口干症，或口干，由于缺乏唾液，导致生活质量下降，由多种症状引起，包括龋齿增加，口咽感染，吞咽困难(吞咽困难)和消化困难(粘膜炎)，失去味觉和疼痛。由于目前对这些问题的治疗方法还不够充分，人们对制造人工唾液腺有相当大的兴趣。在体外维持唾液腺泡细胞的分化和功能是成功构建此类结构的关键，然而，这一突破尚未实现。这主要是由于目前缺乏关于维持或诱导腺泡细胞分化所需的特定细胞外信号的基本科学知识， 这对设计功能性人工唾液腺的能力仍然是一个实质性的限制。需要一种体外检测系统来识别所需的细胞外信号。我们假设，通过3D支架中的微尺度图案呈现给细胞的化学修饰纳米纤维的组合将支持唾液腺泡细胞的功能。此应用程序 提出了一种创新的跨学科策略，以创建一种高通量、非侵入性的检测系统，该系统将用于检测活细胞中培养的唾液腺泡细胞的分泌功能 在脚手架材料上，这在以前是不可能的。将生产一种基于MEMS的唾液传感器，以准确测量生长在独特支架组合上的细胞分泌的唾液分泌蛋白的浓度。在生物传感器检测之后，一种新的多重免疫细胞化学方法将被用于评估固定细胞的分化程度。利用一种创新的方法组合，纳米拓扑术、微图案化和化学信号将被独立调制，从而将识别支持腺泡细胞功能的特定参数组合。将评估支架维持腺泡分化或促进再次分化的能力，使用原代细胞和细胞系的组合。我们将在四个具体目标中解决这一假设：目标1：建立高通量MEMS探针阵列检测系统，以在体外评估腺泡细胞的功能。目的2：确定支持唾液腺泡细胞分化的理想纳米纤维构型。目的3：优化纳米纤维表面性能，增强涎腺腺泡细胞功能。目的4：寻找促进腺泡细胞功能的最佳微结构。在此应用中产生的体外唾液腺结构将有助于识别调节腺泡功能的途径，并可以 可应用于药物筛选。作为这一应用的结果而开发的原理将在未来应用于设计复制唾液腺和其他复杂分支器官的人造腺体的工程中。