Construction of Conjunctival Equivalents Using Molecular Deposition Techniques

摘要A1-R01授权书：利用分子沉积技术构建结膜等效物 技术 具有杯状、层状鳞状和未分化细胞的结膜坚固、有功能，对于 透明的角膜和健康的前眼。眼表疾病，包括感染性(沙眼)、 自身免疫(眼瘢痕性类天疱疮)，遗传性疾病，以及烧伤和史蒂文斯-约翰逊 综合症，可通过结疤损伤眼表，尤其是结膜。虽然结膜 有很大的修复潜力，这些疾病是痛苦的炎症性疾病，患者 迫切希望得到更好的治疗。目前的治疗方法包括使用多余结膜或羊膜。 薄膜。新型生物相容材料的开发在以下领域提供了改进的治疗方法 再生医学。对于这项提议，我们将利用新开发的分子层 沉积(MLD)技术用于构建生物活性材料涂层。我们的总体目标是开发一种 由三种类型的人结膜组成的可移植的功能性结膜结构 上皮细胞(未分化、层状鳞状和杯状)生长在我们的新表面上。表面是 基于沉积在丝素基质上的MLD膜，易于操作用于外科移植。 对于SA1，我们将鉴定生长在MLD丝素构建物上的结膜上皮细胞的表型 将其与生长在羊膜上的细胞进行比较。对于SA2，我们将评估这三种类型的功能 在我们构建的MLD丝上生长的上皮细胞对细胞功能和生理刺激的反应 将它们与生长在羊膜上的细胞进行比较。对于SA3，我们将测量对 MLD丝构建物上细胞的机械损伤与羊膜的比较。细胞计数， 分子和免疫荧光显微镜技术将被用来评估数量和表型 细胞生长。将使用物理技术来确定结构强度。CA2成像，ELISA用于 MUC5AC分泌，以及其他生理技术将被用来确定细胞在 建造。将使用免疫荧光显微镜和多种细胞因子测量来评估伤口 治愈。在MLD丝上生长的三种结膜上皮细胞的最终产品将具有功能，以及 可移植，非常适合修复病变和受损的结膜。

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.