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: 8697035
• 负责人: James Castracane
• 金额: $ 48.25万
• 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 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8697035
• 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（由申请人提供）：数百万人患有口干症，或因缺乏唾液而导致的“口干症”，导致多种症状导致生活质量下降，包括龋齿增加、口咽感染、吞咽困难（吞咽困难）消化（粘膜炎）、味觉丧失和疼痛。由于目前对这些问题的治疗方法还不够充分 (1)，因此人们对制造人工唾液腺产生了很大的兴趣。体外维持唾液腺泡细胞分化和功能对于此类构建体的成功工程至关重要；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础科学知识， 这仍然是设计功能性人工唾液腺能力的重大限制。需要体外测定系统来识别所需的细胞外信号。我们假设通过 3D 支架中的微米级图案呈现给细胞的化学修饰纳米纤维组合将支持唾液腺泡细胞功能。这个应用程序 提出了一种创新的跨学科策略，以创建高通量、非侵入性检测系统，用于检测活细胞生长中的唾液腺泡细胞分泌功能 在支架材料上，这在以前是不可能的。将生产基于 MEMS 的唾液传感器，以准确测量在独特支架组合上生长的细胞分泌的唾液分泌蛋白的浓度。一种新型多重免疫细胞化学方法将用于评估生物传感器测定后固定细胞的分化程度。使用创新的方法组合，纳米形貌、微图案和化学信号将被独立调节，从而识别出支持腺泡细胞功能的特定参数组合。将评估支架使用原代细胞和细胞系的组合维持腺泡分化或促进再分化的能力。我们将通过四个具体目标来解决这一假设： 目标 1：开发高通量 MEMS 探针阵列测定系统来评估体外腺泡细胞功能。目标 2：确定支持唾液腺泡细胞分化的理想纳米纤维结构。目标 3：优化纳米纤维表面特性以增强唾液腺腺泡细胞功能。目标 4：确定促进腺泡细胞功能的最佳微地形。本申请中产生的体外唾液腺构建体将有助于识别调节腺泡功能的途径，并且可以 应用于药物筛选。由于该应用而开发的原理将在未来应用于旨在复制唾液腺和其他复杂分支器官的人工腺体工程。