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.

项目摘要：静脉静脉症或“口干”是一种挑战临床状况，是由唾液细胞损害造成的腺。这可能是由于多种组织损伤引起的，包括头部放射治疗急性损害颈部癌症，Sjogren病中的进行性自动免疫反应或衰老的其他未知病因。当前的治疗只能暂时缓解症状，而相关口腔健康的分辨率差衰变。这种情况的成本是相当大的，无论是生活质量还是增加的财务燃烧牙科护理。组织工程和再生医学领域为潜力提供了许多工具在支持性水凝胶材料中重建健康的唾液衍生细胞，但其中很少提供足够的空间和临时分辨率来恢复复杂的分支结构和精确空间主要唾液网格的表型图。但是，基于激光的水凝胶降解中的新发现（LBHD）可用于通过完整的水凝胶平板“雕刻”途径，并具有精确的亚细胞分辨率 XYZ，并提供了一种指导在3个维度中成长的唾液上皮芽的方法。我们关于目前的建议是，我们可以使用基于多光子的LBHD来延长给定的多细胞簇方向，并重现本地腺的关键要素。为此，我们将利用实验室的专业知识从健康组织中分离原代人唾液衍生的茎/祖细胞（HS/PC），封装为响应式3D多细胞球体簇，可自定义，生物相容性透明质酸酸（HA）水凝胶。我们正在进行的工作表明，通过调整这些水凝胶的孔隙率及其生物活性表位的浓度，我们可以影响簇的大小，形态，并与周围细胞外基质。我们将通过以下特定目的检验我们的假设：目标1。建立通过HA水凝胶雕刻“隧道”的参数，并促进HS/PC群集插入。目标2。将系统调整到可以使成纤维细胞共培养的替代物质中交联，以便于制造更容易的制造。目标3。评估不断增长的簇的表型，在其尾随和领先边缘和分支末端，用于分化表型的迹象。如果成功，该系统可以用作研究人唾液细胞组织和分化机制的有用模型；系统可能也是生产组织工程腺更换的早期原型。 R03机制将为必要的飞行员和可行性研究提供支持，以证明这些证明了技术可以合并以产生一个新颖的平台。