Biofunctional Materials Containing Protein Mimetics

描述（由申请人提供）：巨噬细胞在指导宿主-生物材料相互作用和许多疾病病因中起着核心作用。我们假设可以从巨噬细胞活性蛋白（纤维连接蛋白和白细胞介素-1）的功能结构设计仿生寡肽，并用于探测配体-受体相互作用和连接后事件，作为寻求更深入了解巨噬细胞行为的分子控制机制的基础。此外，这些肽被用作开发新型生物材料的平台技术的组成部分，在体外和体内部分调节巨噬细胞的功能。在这篇竞争性更新申请的再提交中，我们假设观察到的肽介导的巨噬细胞行为是由表面固定时的优先配体构象和调节巨噬细胞功能的多种细胞内事件的存在所贡献的。为了实现这些目标，制定了三个相互关联的目标：(1)确定固定在化学上不同底物上的肽的构象（TCPS，单甲氧基- peg -单丙烯酸-聚丙烯酸-共四甲基丙基丙烯酸酯共聚物，以及含有化学修饰的明胶和聚乙二醇-二丙烯酸酯的互穿网络），并将这种结构与配体-整合素识别和结合联系起来；(2)确定细胞内酪氨酸磷酸化蛋白，连接后的细胞内替代途径；使用单培养和共培养系统，在肽介导的粘附巨噬细胞中，结联后基因表达和细胞功能(3)量化和模拟巨噬细胞活性寡肽与整合素的结合动力学，并在肽-整合素络合时与鉴定的磷酸化整合素相关蛋白相关联。我们在探索材料-巨噬细胞相互作用的分子机制方面的持续努力，揭示了吞噬细胞功能的配体-受体关联和调节途径，这是理解生物医学设备宿主-材料相互关系的基础。

DESCRIPTION (provided by applicant): Macrophages are central in directing host-biomaterial interaction and many disease etiologies. We hypothesized that biomimetic oligopeptides can be designed from the functional structure of macrophage-active proteins (fibronectin and intedeukin-1) and employed to probe the ligand-receptor interaction and post-ligation events as a basis for seeking a greater understanding in the molecular control mechanism involved in macrophage behavior. Furthermore, these peptides were utilized as a component of a platform technology in the development of novel biomaterials to partly modulate macrophage function in vitro and in vivo. In this resubmission of the competing renewal application, we hypothesize that the observed peptide-mediated macrophage behavior is contributed by the preferential ligand conformation upon surface immobilization and the presence of multiple intracellular events in regulating macrophage function. To address these goals, three interrelated aims are formulated: (1) to determine the peptide conformation upon immobilization onto chemically distinct substrates (TCPS, copolymer of monomethoxy-PEG-monoacrylate-colacrylic acid-co-tetramethylolpropylacrylate, and interpenetrating networks containing chemically modified gelatin and polyethyleneglycol-diacrylate) and to correlate this structure with ligand-integrin recognition and binding, (2) to identify intracellular tyrosine phosphorylated proteins, alternative post-ligation intracellular pathways, post-ligation gene expression and cellular function in peptide-mediated adherent macrophages using mono- and co-culture systems, (3) to quantify and model the binding kinetics of macrophage-active oligopeptides with integrins, and to correlate with the identified phosphorylated integrin-associated proteins upon peptide-integrin complexation. Our continuing effort in probing the molecular mechanism of material-macrophage interaction has shed insights on ligand-receptor association and regulation pathways for phagocytic cell function, that are fundamental in understanding the host-material interrelationship for biomedical devices.