NIRT: Ligand Nanodisplay for Cellular Internalization and Super-Activation

0609000 MogheThis提案是响应纳米科学和工程倡议，NSF 05-610，类别NIRT。 本研究的目的是（1a）设计、制备和表征用基质蛋白片段功能化的纳米颗粒;（1b）阐明纳米颗粒尺寸、配体序列和配体负载对细胞运动和基质组装的作用;（2）鉴定介导纳米颗粒内化和增加细胞活化的关键分子信号通路;以及（3）开发成像模式以量化纳米颗粒运输和内化动力学。 该方法涉及用纤连蛋白（配体）的各种片段对白蛋白衍生的纳米颗粒进行功能化。 该研究的第一阶段将研究基于配体纳米颗粒的基底的最佳配置如何显著改变皮肤衍生细胞（角质形成细胞、成纤维细胞）的形态和动力学（运动性、基质组装）。 第二阶段将侧重于使用蛋白质和基因水平的信号转导分析来确定潜在的生物学机制。 第三阶段将使用高分辨率双光子显微镜和具有纳米级保真度的磁响应生物传感器平台来探测纳米颗粒与细胞相互作用的性质和动力学，这项研究可以帮助设计改进的纳米材料，用于伤口愈合，组织工程，药物输送和癌症治疗中的潜在细胞靶向应用。这里设计的纳米颗粒是可生物降解的，可以靶向细胞，并且可以通过改变它们的大小来定制细胞功能。 较小的纳米颗粒可以刺激表皮细胞中增加的运动性（与烧伤、溃疡中的皮肤愈合相关），而较大的纳米颗粒促进皮肤收缩性和基质组装（与伤口修复相关）。 该项目通过罗格斯大学及其他大学的广泛多样性网络（研究生，博士后，本科生）扩展其推广范围，并与罗格斯大学关于生物材料细胞生物界面工程的三个新的综合研究生课程，生物界面研究生培训计划和国际研究合作产生共鸣。

0609000MogheThis proposal was received in response to Nanoscale Science and Engineering initiative, NSF 05-610, category NIRT. The objectives of this research are (1a) to design, fabricate, and characterize nanoparticles functionalized with matrix protein fragments, and (1b) to elucidate the role of nanoparticle size, ligand sequence and ligand loading on cell motility and matrix assembly; (2) to identify the key molecular signaling pathways that mediate nanoparticle internalization and increased cell activation; and (3) to develop imaging modalities to quantify nanoparticle trafficking and internalization dynamics. The approach involves the functionalization of albumin-derived nanoparticles with various fragments of fibronectin (ligand). The first phase of the study will examine how optimal configurations of substrates based on the ligand-nanoparticles can significantly alter the morphology and dynamics (motility, matrix assembly) of skin derived cells (keratinocytes, fibroblasts). The second phase will focus on identifying the underlying biologic mechanisms using protein and gene level signaling assays. The third phase will probe the nature and kinetics of nanoparticle-cell interactions using high resolution, two-photon microscopy and a magnetically responsive biosensor platform with nanoscale fidelity.This research can help design improved nanomaterials for potential cell targeting applications in wound healing, tissue engineering, drug delivery, and cancer therapy. The nanoparticles designed here are biodegradable, can be targeted to cells, and can be customized to cell functions by altering their size. Smaller nanoparticles can stimulate increased motility in epidermal cells (relevant for skin healing in burns, ulcers) while larger nanoparticles promote skin contractility and matrix assembly (relevant to wound repair). This project extends its outreach through the wide diversity network (graduate, postdoc, undergraduate) at Rutgers and beyond, and resonates with three new integrative graduate courses at Rutgers on engineering of cellular biointerfaces with biomaterials, a graduate training program on biointerfaces, and an international research collaboration.