Biocompatible PEG-Functionalized Methacrylates

描述：(申请者摘要)这个项目调查的是 聚乙二醇官能化聚甲基丙烯酸甲酯 (聚乙二醇酯)抑制上皮细胞的生长和炎症反应 前瞻人工晶状体。PMMA表现出很强的后部依附性，但 一些成长中的。使用聚乙二醇化的跟踪功能将进一步抑制不需要的 在不干扰强后部附着的情况下的生物反应 PMMA。 合适的体外实验模型将以较低的成本测量这些反应。 以确定是否值得在体内进行追踪。这个模型使用的是人类 角膜上皮细胞系HCE-I作为上皮细胞模型。HCE-1细胞 将被培养、传代并放置在不同的生物材料上。追随者 细胞将使用倒置显微镜和计算机化进行可视化和计数 图像分析。附着力将基于铺展面积/单元和 贴壁细胞占据的生物材料表面。HCE-1细胞将被培养 裸露和蛋白质预吸附的聚乙二醇化聚甲基丙烯酸甲酯的作用 促进细胞黏附的蛋白质。选择的蛋白质是人类的 纤维连接蛋白是一种粘附性RGD蛋白，而血清白蛋白是一种非粘附性蛋白。 将从柠檬化的人全血中分离出人纤维连接蛋白 特征，包括它的生物活性。蛋白质的吸附将是 用氯胺-T法，用125对放射性标记蛋白进行定量。 该项目将合成和表征挂起官能化的PMMA 聚乙二醇法，包括ESCA和接触角测角法。 蛋白质预吸附聚合物的测角将努力进行。巨噬细胞 将对粘连进行量化，以评估可能的炎症反应 这些生物材料可以引发。巨噬细胞将从整个人类血液中分离出来 通过Ficoll-Hypaque梯度离心法。巨噬细胞趋化性和 将对吞噬功能进行评估。黏附性巨噬细胞和HCE-1的形态 还将对I细胞进行扫描电子显微镜检查。

DESCRIPTION: (Applicant's Abstract) This project investigates the potential of functionalizing poly methyl methcrylate (PMMA) with poly (ethylene glycol) (PEG) to inhibit epithelial cell in growth and inflammatory reactions to the prospective intraocular lens. PMMA displays strong posterior attachment, but some in-growth. Trace functionalization with PEG will further inhibit undesired biological responses without disturbing the strong posterior attachment of PMMA. A suitable in vitro experimental model will measure these responses at low cost to determine if in vivo pursuit is worthwhile. This model uses the human corneal epithelial cell line HCE- I as an epithelial cell model. HCE-1 cells will be cultured, passaged and placed onto the different biomaterials. Adherent cells will be visualized and counted using inverted microscopy and computerized image analysis. Adhesion will be based on the spread area/cell and fraction of biomaterial surface occupied by adherent cells. HCE-1 cells will be incubated on bare and protein-preadsorbed PEG-functionalized PMMA to reveal the role of proteins in promoting cell adhesion. The proteins selected are human fibronectin, an adhesive RGD protein, and serum albumin, a nonadhesive protein. Human fibronectin will be isolated from citrated whole human blood and characterized, including its biological activity. Protein adsorption will be quantified radiolabelling proteins with 125, using the chloramine-T method. This project will synthesize and characterize PMMA pendantly functionalized with poly(ethylene glycol), including ESCA and contact angle goniometry. Goniometry of protein-preadsorbed polymers will be endeavored. Macrophage adhesion will be quantified to appraise possible inflammatory reactions that these biomaterials can elicit. Macrophages will isolated from whole human blood by centrifugation on a Ficoll-Hypaque gradient. Macrophage chemotaxis and phagocytosis will be assessed. The morphology of adherent macrophages and HCE- I cells will also be examined with scanning electron microscopy.