Sytems Analysis of Nanoparticle Biocompatibility

描述(由申请人提供) 我们提出了一种定量结构活性关系(QSAR)方法来研究影响纳米颗粒生物相容性的特定物理和化学表面性质。无定形二氧化硅之所以被选为实验颗粒，是因为它在消费产品中的广泛应用，而且它很容易在各种定义的尺寸和表面化学成分中合成。我们最近的工作表明，无定形二氧化硅的生物活性在50 nm的颗粒中得到了极大的增强。我们假设这是由于随着颗粒直径的减小，硅醇表面化学发生了变化。 我们的方法涉及三个目标：1)通过对暴露于无定形二氧化硅的巨噬细胞的条件培养液进行先进的基于质谱学的蛋白质组分析，结合我们现有的基因芯片数据，将鉴定一组纳米颗粒诱导的分泌蛋白质。将进行生物信息途径分析，选择~20个途径生物标志物，用于改进基于抗体夹心法的蛋白质ELISA微阵列平台，用于多重反应分析。2)制备了一系列尺寸和表面化学随官能团选择性改变的硅基粒子，并对其大小、电荷、聚集状态、溶解产物和硅醇类型进行了表征。X射线吸收近边光谱学将被用来识别吸附在胶束上的颗粒中的表面硅异构体，以模拟细胞膜。每个颗粒的生物反应将在巨噬细胞的多孔细胞试验中进行评估，使用ELISA微阵列平台为大约20个不同的途径标记物提供剂量-反应的定量测量。QSAR分析将与测量的物理化学参数和生物反应数据一起进行，以确定相关性最强的关系。3)从QSAR分析中筛选出的颗粒将进一步在气管内滴注暴露的小鼠身上进行测试，并通过组织病理学和支气管灌洗液的ELISA微阵列分析来确定生物学反应。在AIMS 2和AIMS 3中获得的QSAR结果的比较将确定体外试验的预测性，以及突出对决定体内生物相容性最重要的颗粒特征。这些结果将决定纳米级无定形二氧化硅的性质，决定其生物相容性，并揭示与其他类型的纳米材料相关的一般原理。此外，通过这项工作开发的方法和生物标记物将提供一个筛选平台，可以在未来部署到各种纳米材料上。

DESCRIPTION (provided by applicant) We propose a quantitative structure activity relationship (QSAR) approach to investigate the specific physical and chemical surface properties that influence nanoparticle biocompatibility. Amorphous silica is chosen as an experimental particle due to its widespread use in consumer products, and because it is readily synthesized in a wide range of defined sizes and surface chemistries. Our recent work shows that the bioactivity of amorphous silica is greatly enhanced in particles <50 nm. We hypothesize that this is due to changes in the silanol site surface chemistry as particle diameter is decreased. Our approach involves three aims: 1) A panel of nanoparticle-induced secreted proteins will be identified using advanced mass spectrometry-based proteomic analysis of conditioned medium from macrophages exposed to amorphous silica, coupled with our existing gene microarray data. Bioinformatic pathway analysis will be performed to select ~20 pathway biomarkers, which will be used to modify an antibody sandwich-based protein ELISA microarray platform for multiplexed response analyses. 2) A series of silicabased particles where size and surface chemistry is selectively altered with functional groups will be prepared and characterized for size, charge, aggregation state, dissolution products and silanol types. X-ray absorption near-edge spectroscopy will be used to identify surface silicon isoforms in particles adsorbed to micelles mimicking cell membranes. The biological responses of each particle will be assessed in multiwell cellular assays with macrophages, using the ELISA microarray platform to provide quantitative measures of dose-response for ~20 different pathway markers. QSAR analyses will be performed with the measured physicochemical parameters and biological response data to identify relationships that correlate most strongly. 3) Particles selected from QSAR analysis will be further tested in mice exposed by intratracheal instillation, and biological responses will be determined by histopathology along with ELISA microarray analysis of bronchial lavage fluid. Comparison of QSAR results obtained in aims 2 and 3 will determine how predictive the in vitro assay is, as well as highlight particle characteristics that are most important for dictating biocompatibility in vivo. The results will determine properties of nano-scale amorphous silica that determine its biocompatibility, and reveal general principals relevant to other types of nanomaterial. In addition, the approach and biomarkers developed from this work will provide a screening platform that can be deployed to a variety of nanomaterials in the future.