Sytems Analysis of Nanoparticle Biocompatibility

描述（由申请人提供） 我们提出了一种定量构效关系（QSAR）的方法来研究特定的物理和化学表面性质，影响纳米粒子的生物相容性。选择无定形二氧化硅作为实验颗粒是因为其在消费品中的广泛使用，并且因为其易于以广泛的限定尺寸和表面化学合成。我们最近的工作表明，无定形二氧化硅的生物活性在<50 nm的颗粒中大大增强。我们推测，这是由于在硅烷醇网站的表面化学的颗粒直径减小的变化。 我们的方法包括三个目标：1）一组纳米颗粒诱导的分泌蛋白将被确定使用先进的基于质谱的蛋白质组学分析的条件培养基从巨噬细胞暴露于无定形二氧化硅，加上我们现有的基因微阵列数据。将进行生物信息学途径分析，以选择约20种途径生物标志物，这些生物标志物将用于修饰基于抗体的蛋白质ELISA微阵列平台，以进行多重响应分析。2)将制备一系列硅基颗粒，其中尺寸和表面化学性质随官能团选择性改变，并表征其尺寸、电荷、聚集状态、溶解产物和硅烷醇类型。X射线吸收近边光谱将用于识别吸附到模拟细胞膜的胶束的颗粒中的表面硅异构体。将在使用巨噬细胞的多孔细胞试验中评估每个颗粒的生物学反应，使用ELISA微阵列平台提供约20种不同途径标志物的剂量反应的定量测量。将对测得的理化参数和生物反应数据进行QSAR分析，以确定最强相关性的关系。3)将在通过气管内滴注暴露的小鼠中进一步测试从QSAR分析中选择的颗粒，并且将通过组织病理学沿着支气管灌洗液的ELISA微阵列分析来确定生物反应。目标2和目标3中获得的QSAR结果的比较将确定体外试验的预测性，并强调对体内生物相容性最重要的颗粒特征。结果将确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，从这项工作中开发的方法和生物标志物将提供一个筛选平台，可以在未来部署到各种纳米材料。

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.