Directing Collective Epithelial Morphology in Space and Time Using a Light-Based Carving Tool

使用基于光的雕刻工具指导空间和时间上的集体上皮形态

基本信息

批准号：
9809041
负责人：
Daniel A Harrington
金额：
$ 15.4万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9809041
关键词：
3-Dimensional Ablation Acinus organ component Acute Affect Aging Architecture Autoimmune Responses Basement membrane Biological Biomanufacturing Caliber Cell Culture Techniques Cell Proliferation Cell Size Cell Survival Cells Cellular Morphology Cellular Spheroids Cessation of life Clinical Coculture Techniques Complex Connective Tissue Crosslinker Cues Dental Care Dental caries Deposition Development Disease Duct (organ) structure Ductal Elements Encapsulated Engineering Epithelial Epitopes Etiology Extracellular Matrix Face Feasibility Studies Fibroblasts Financial Hardship Future Gel Gland Goals Grant Growth Growth Factor Harvest Head and Neck Cancer Human Hyaluronic Acid Hydrogels In Vitro Integrins Laboratories Lasers Length Ligands Light Major salivary gland structure Maps Matrix Metalloproteinases Membrane Proteins Mesenchymal Methods Microscopy Morphology Myoepithelial Neurons Neurotransmitter Receptor Neurotransmitters Oral health Organ Pain Palliative Care Pathway interactions Patients Pattern Peptide Signal Sequences Phenotype Physics Pilot Projects Polymers Porosity Positioning Attribute Printing Proliferating Quality of life Radial Radiation induced damage Radiation therapy Regenerative Medicine Resolution Salivary Salivary Gland Tissue Salivary Glands Shapes Signal Transduction Sjogren&apos s Syndrome Source Stem cells Structure Study models Syringes System Technology Testing Therapeutic Time Tissue Engineering Tissues Travel Variant Work Xerostomia absorption base biomaterial compatibility bioprinting cell growth cell motility cell type cost crosslink implantable device implantation improved lithography migration monolayer new technology novel progenitor prototype reconstitution reduce symptoms regenerative therapy response salivary acinar cell salivary assay salivary cell standard of care stem temporal measurement three dimensional cell culture time use tool two-photon

项目摘要

Project Summary: Xerostomia, or “dry mouth”, is a challenging clinical condition, caused by damage to the cells of the salivary gland. It may result from a variety of tissue insults, including acute damage from radiation therapy for head and neck cancers, progressive auto-immune response in Sjogren’s disease, or other unknown etiology from aging. Current treatments offer only temporary relief of symptoms, and poor resolution of associated oral health decay. The cost of this condition is considerable, both in quality of life and the financial burden of increased dental care. The fields of tissue engineering and regenerative medicine offer many tools for the potential reconstitution of healthy salivary-derived cells within supportive hydrogel matrices, but few of these options provide sufficient spatial and temporal resolution to restore the complex branched structure and precise spatial phenotype map of the major salivary glands. However, new discoveries in laser-based hydrogel degradation (LBHD) can be used to “carve” pathways through intact hydrogel slabs, with pinpoint, subcellular resolution in xyz, and offer a method to guide a growing salivary epithelial bud in 3 dimensions. Our hypothesis for the present proposal is that we can use multiphoton-based LBHD to elongate a multicellular cluster in a given direction, and recreate key elements of the native gland. To do this, we will employ our laboratory’s expertise in isolation of primary human salivary-derived stem/progenitor cells (hS/PCs) from healthy tissues, and encapsulation as responsive 3D multicellular spheroid clusters within customizable, biocompatible hyaluronic acid (HA) hydrogels. Our ongoing work has shown that, by tailoring the porosity of these hydrogels and their concentration of bioactive epitopes, we can impact cluster size, morphology, and interaction with the surrounding extracellular matrix. We will test our hypothesis through the following Specific Aims: Aim 1. Establish parameters to carve “tunnels” through HA hydrogels and promote HS/PC cluster ingrowth. Aim 2. Adapt the system to alternate matrices that enable fibroblast co-culture, or incorporate photolabile crosslinkers for easier fabrication. Aim 3. Assess phenotype of the growing cluster, at its trailing and leading edges and branched termini, for signs of differentiated phenotype. If successful, this system could serve as a useful model for studying mechanisms of human salivary cell organization and differentiation; the system might also be an early prototype for manufacturing tissue engineered gland replacements. The R03 mechanism will provide support for the necessary pilot and feasibility studies, to demonstrate that these proven technologies can be combined to produce a novel platform.

项目概要：口干症或“口干”是一种具有挑战性的临床病症，由唾液细胞损伤引起它可能是由多种组织损伤引起的，包括头部和颈部放射治疗造成的急性损伤。颈部癌症、干燥病的进行性自身免疫反应或其他未知的衰老病因。目前的治疗只能暂时缓解症状，并且无法解决相关的口腔健康问题这种情况的代价是相当大的，无论是生活质量还是经济负担的增加。牙科护理领域为发挥潜力提供了许多工具。在支持性水凝胶基质中重建健康的唾液来源的细胞，但这些选择很少提供足够的空间和时间分辨率来恢复复杂的分支结构和精确的空间然而，基于激光的水凝胶降解的新发现。（LBHD）可用于通过完整的水凝胶板“雕刻”路径，并具有精确的亚细胞分辨率 xyz，并提供了一种在 3 个维度上引导唾液上皮芽生长的方法。目前的建议是，我们可以使用基于多光子的 LBHD 来延长给定的多细胞簇为此，我们将利用我们实验室的专业知识。从健康组织中分离原代人唾液干细胞/祖细胞 (hS/PC)，以及封装为可定制、生物相容性透明质酸中的响应性 3D 多细胞球体簇我们正在进行的工作表明，通过调整这些水凝胶及其孔隙率。生物活性表位的浓度，我们可以影响簇的大小、形态以及与我们将通过以下具体目标来检验我们的假设：目标 1. 建立参数以通过 HA 水凝胶雕刻“隧道”并促进 HS/PC 簇向内生长。目标 2. 使系统适应替代基质，以实现成纤维细胞共培养，或纳入不稳定性光目标 3. 评估生长簇的表型（尾随和前导）。边缘和分支末端，用于区分表型的迹象如果成功，该系统可以作为一个。该系统可能是研究人类唾液细胞组织和分化机制的有用模型； R03 机制也将成为制造组织工程腺体替代品的早期原型。为必要的试点和可行性研究提供支持，以证明这些经过验证的技术可以组合起来产生一个新颖的平台。