Construction of Conjunctival Equivalents Using Molecular Deposition Techniques

使用分子沉积技术构建结膜等效物

基本信息

批准号：
9884619
负责人：
Darlene A Dartt
金额：
$ 39.4万
依托单位：
SCHEPENS EYE RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9884619
关键词：
Adhesions Affect Agonist Amino Acids Anterior Arginine Aspartic Acid Autoimmune Diseases Autoimmune Process Biochemical Biocompatible Materials Burn injury Cell Count Cell Proliferation Cell Survival Cell physiology Cells Characteristics Chemicals Cholinergic Agonists Cicatrix Conjunctival Epithelium Conjunctival Pterygium Cornea Cultured Cells Deposition Development Disease EGF gene Electrolytes Enzyme-Linked Immunosorbent Assay Epithelial Epithelial Cells Epithelium Eye Eyelid structure Fibroins Film Genetic Diseases Glass Glues Glycine Goals Goblet Cells Grant Growth Growth Factor Human Image Immunofluorescence Microscopy Infection Inflammation Inflammatory Ions Location MUC5AC gene Measurement Measures Mechanics Mediator of activation protein Medicine Membrane Metals Molecular Mucins Natural regeneration Nerve Growth Factors Ocular cicatricial pemphigoid Operative Surgical Procedures Pain Patients Phenotype Physiological Production Proliferating Publishing Reproducibility Serotonin Silk Squamous Cell Stevens-Johnson Syndrome Stimulus Stress Structure of molecular layer of cerebellar cortex Surface Surgical sutures Techniques Tight Junctions Tissues Titanium Trachoma Transplantation Trauma Undifferentiated Visual impairment Water base biomaterial compatibility cell growth cell injury cell type conjunctiva cytokine healing human female hydrophilicity improved male novel ocular surface physical property prevent relating to nervous system repaired response scaffold sex standard of care titanium dioxide water channel wound wound closure wound healing

项目摘要

Abstract A1-R01 Grant Title: Construction of Conjunctival Equivalents Using Molecular Deposition Techniques A robust, functional conjunctiva with goblet, stratified squamous, and undifferentiated cells is critical for a transparent cornea and a healthy anterior eye. Ocular surface disorders, including, infectious (trachoma), autoimmune (ocular cicatricial pemphigoid), and genetic diseases, as well as burns and Stevens-Johnson syndrome, can damage the ocular surface, especially the conjunctiva, by scarring. Although the conjunctiva has substantial repair potential, these diseases are painful inflammatory diseases and the patients are desperate for better treatments. Current treatments include use of redundant conjunctiva or amniotic membrane. Development of novel biocompatible materials is providing improved treatments in the field of regeneration medicine. For this proposal we will take advantage of the newly developed molecular layer deposition (MLD) technique to build coatings of bioactive materials. Our overall goal is to develop a transplantable and functional conjunctival construct consisting of the three types of human conjunctival epithelial cells (undifferentiated, stratified squamous, and goblet) growing on our novel surface. The surface is based on an MLD film deposited on a silk fibroin matrix that is easily manipulated for surgical transplantation. For SA1 we will characterize the phenotype of conjunctival epithelial cells growing on MLD silk construct and compare it with cells growing on amniotic membrane. For SA2 we will evaluate the function of the three types of epithelial cells growing on our constructs of MLD silk in response to physiological stimuli of cell function and compare them with cells growing on amniotic membrane. For SA3 we will measure the repair response to a mechanical injury of the cells on constructs of MLD silk compared to amniotic membrane. Cell counting, molecular, and immunofluorescence microscopy techniques will be used to evaluate amount and phenotype of cell growth. Physical techniques will be used to determine construct strength. Ca2+ imaging, ELISAs for MUC5AC secretion, and other physiological techniques will be used to determine function of cells on the construct. Immunofluorescence microscopy and multi-cytokine measurements will be used to evaluate wound healing. The final product of three types of conjunctival epithelial cells grown on MLD silk will be functional, and transplantable and excellent for repair of the diseased and damaged conjunctiva.

摘要 A1-R01 资助名称：利用分子沉积构建结膜等效物技巧具有杯状、复层鳞状和未分化细胞的强健、功能性结膜对于结膜至关重要。透明的角膜和健康的前眼。眼表疾病，包括传染性（沙眼）、自身免疫性疾病（眼部瘢痕性类天疱疮）和遗传病，以及烧伤和史蒂文斯-约翰逊病综合征，可通过疤痕损害眼表，尤其是结膜。虽然结膜具有巨大的修复潜力，这些疾病是痛苦的炎症性疾病，患者迫切需要更好的治疗方法。目前的治疗方法包括使用多余的结膜或羊膜膜。新型生物相容性材料的开发正在改善该领域的治疗方法再生医学。对于这个提案，我们将利用新开发的分子层沉积（MLD）技术构建生物活性材料涂层。我们的总体目标是开发一个由三种类型的人类结膜组成的可移植和功能性结膜结构上皮细胞（未分化、复层鳞状和杯状）在我们的新型表面上生长。表面是基于沉积在丝素蛋白基质上的 MLD 薄膜，易于进行手术移植。对于 SA1，我们将表征在 MLD 丝结构上生长的结膜上皮细胞的表型，并将其与羊膜上生长的细胞进行比较。对于SA2我们将评估三种类型的功能上皮细胞在我们的 MLD 丝结构上生长，以响应细胞功能的生理刺激，并且将它们与羊膜上生长的细胞进行比较。对于 SA3，我们将测量修复响应与羊膜相比，MLD 丝结构上的细胞机械损伤。细胞计数，分子和免疫荧光显微镜技术将用于评估细胞生长。物理技术将用于确定结构强度。 Ca2+ 成像、ELISA MUC5AC 分泌和其他生理技术将用于确定细胞的功能构造。免疫荧光显微镜和多细胞因子测量将用于评估伤口康复。在 MLD 丝上生长的三种类型的结膜上皮细胞的最终产品将具有功能性，并且可移植并且非常适合修复患病和受损的结膜。