Biocompatible PEG-Functionalized Methacrylates

描述：（申请人的摘要）本项目调查的潜力， 用聚乙二醇官能化聚甲基丙烯酸甲酯 (PEG)抑制上皮细胞的生长和炎症反应， 前瞻性眼内透镜。PMMA显示出较强的后部附着，但 一些增长。用PEG的痕量官能化将进一步抑制不期望的 生物学反应，而不干扰强大的后部附着， PMMA。 合适的体外实验模型将以低成本测量这些反应 以确定体内研究是否值得。该模型使用人类 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、传代并置于不同的生物材料上。贴壁 细胞将使用倒置显微镜和计算机进行可视化和计数。 图像分析。粘附将基于铺展面积/细胞和细胞的分数。 粘附细胞占据的生物材料表面。将孵育HCE-1细胞 裸和蛋白质预吸附PEG功能化的PMMA上，以揭示 促进细胞粘附的蛋白质。选择的蛋白质是人类的 纤连蛋白，粘附性RGD蛋白，和血清白蛋白，非粘附性蛋白。 人纤连蛋白将从柠檬酸化的人全血中分离， 包括其生物活性。蛋白质吸附将是 使用氯胺-T方法，用125定量放射性标记蛋白质。 本项目将合成和表征聚甲基丙烯酸甲酯悬垂功能化 与聚（乙二醇），包括ESCA和接触角测角法。 蛋白质预吸附聚合物的测角法将是未来的研究方向。巨噬 将对粘连进行量化，以评价可能的炎症反应， 这些生物材料会引发什么将从人全血中分离出巨噬细胞 通过在Ficoll-Hypaque梯度上离心。巨噬细胞趋化性和 将评估吞噬作用。粘附的巨噬细胞和HCE- 还将用扫描电子显微镜检查I细胞。

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.