Predictive Toxicological Paradigms to Establish Inhalation Toxicology Models

建立吸入毒理学模型的预测毒理学范式

• 批准号: 7942901
• 负责人: Andre Elias Nel
• 金额: $ 53.33万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7942901
• 关键词: Academia; Address; Adjuvant; Allergic; Allergic inflammation; Animal Model; Animals; Antibody Formation; Antigens; Back; Biochemical; Biological; Bone Marrow; Bone Marrow Cells; Breathing; California; Cell Count; Cellular Membrane; Characteristics; Charge; Chemical Engineering; Collaborations; Conflict (Psychology); DNA Damage; Dendritic Cells; Disease model; Dyes; Electronics; Eligibility Determination; Engineering; Enzyme-Linked Immunosorbent Assay; Epithelial; Extravasation; Generations; Germany; Goals; Government Agencies; Hazard Assessment; Histology; Housing; Human; Hypersensitivity; IgE; IgG1; Immune; Immune response; In Vitro; Industry; Inflammation; Inflammatory; Inhalation Toxicology; Injury; Institutes; International; Investigation; Ions; Iron; Lead; Libraries; Link; Liquid substance; Lung; Measurement; Measures; Methods; Microscope; Microscopy; Mitochondria; Modeling; Molecular; Mus; Myelogenous; Nose; Oxidants; Particle Size; Pathogenesis; Pathway interactions; Pharyngeal structure; Phase; Pneumonia; Production; Property; Protocols documentation; Reactive Oxygen Species; Research; Resources; Safety; Screening procedure; Signal Transduction; Silicon Dioxide; Surface; Testing; Time; Toxic effect; Toxicology; Universities; Variant; Work; airway inflammation; allergic airway inflammation; bronchial epithelium; cell injury; combinatorial; cost; cytokine; cytotoxic; cytotoxicity; data sharing; density; design; eosinophilic inflammation; experience; in vivo; inflammatory marker; mouse model; multidisciplinary; nano; nanomaterials; nanoparticle; nanosystems; neutrophil; novel; oxidant stress; particle; public health relevance; response; tissue/cell culture; toxic metal; zeta potential

DESCRIPTION (provided by applicant): A multidisciplinary group at UCLA (Andre Nel, Jeffrey Zink, Tian Xia, Ning Li) in collaboration with Dr. Vince Castranova at NIOSH and Dr. Lutz Mddler at Bremen University (Germany), aim to establish a mouse inhalation toxicology model to screen combinatorial nanomaterials libraries that are linked to mechanistic injury pathways in tissue culture cells. This research will address the lack of reproducible screening protocols for assessment of nanomaterial (NM) safety. We hypothesize that two combinatorial libraries, which have been designed to (i) adjust the toxic effects of ZnO nanoparticles by changing Zn++ release through iron doping, (ii) adjust the cytotoxicity of cationic mesoporous silica nanoparticles (MSNP) by scaling back the surface cationic density, will be useful to establish a link between in vitro toxicology in human bronchial epithelium (NHBE) and myeloid dendritic cells (DC) and non-allergic and allergic pulmonary inflammation in mice. We posit that the link between non-allergic airway inflammation and cytotoxic injury will be explicable by nanoparticle properties that lead to oxidant injury, shedding of toxic metal ions, and ability to trigger mitochondrial injury. In contrast, the linkage of the material properties to allergic airway inflammation is likely to impact the generation of "danger signals" to DC, which allow them to initiate an immunostimulatory pathway that promotes allergic inflammation. To achieve our long-term goal of developing a predictive inhalation toxicology model, we propose in Aim 1 to characterize the bio-physicochemical properties of NM libraries that are expected to induce differential cytotoxic and pro-inflammatory effects in NHBE and bone marrow-derived DC. In vitro toxicity screening will be carried out by the high content screening (HCS) facility in the California Nano Systems Institute at UCLA as well as cytokine measurements by ELISA. We will also assess ROS production. The NM properties that will be assessed include particle size, size distribution, dispersibility, zeta-potential, TEM, XRD and BET analysis. HCS is carried out with a cocktail of dyes that reveal cellular membrane leakage, DNA damage, mitochondrial depolarization and intracellular Ca2+ flux. The particle characterization and in vitro screening will be carried out at the beginning of year 1 and when new batches are synthesized. Aim 2 will use the libraries to establish standardized protocols for allergic and non-allergic pulmonary inflammation in mice by an inhalation-aspiration approach. The protocol for non-eosinophilic inflammation (Castranova) will measure markers of inflammation, damage, and oxidant stress in the BAL along with lung histology for period of 1 day up to 2 months post-exposure. The allergic sensitization model, which relies on particle and OVA aspiration from the nose of anesthesized animals, will assess OVA-specific IgE levels in parallel with BAL differential cell counts and lung histology. These studies will be carried out in phases through years 1-2. We expect to develop a novel and predictive inhalation toxicology paradigm as a component of the consortium activities to establish in vitro and in vivo study protocols for NM safety screening. PUBLIC HEALTH RELEVANCE: This project addresses the design of cellular and animal screening procedures that can be used to assess the potential danger of engineered nanomaterials that are currently being introduced into the marketplace. We delineate a unique approach in which we use in-house synthesized nanoparticles to develop a predictive toxicological model, meaning that we will develop test protocols in tissue culture cells to obtain basic information about the nanoparticle properties that may render them dangerous and then test those ideas out in a mouse model that may reflect what could happen in the lung when those particles are inhaled. The goal of this project is to develop reliable and reproducible protocols that can be used by academic centers, industry and government agencies as a screen for nanomaterial safety.

描述(由申请人提供)：加州大学洛杉矶分校的一个多学科小组(Andre Nel，Jeffrey Zink，Tian Xia，NIOSH的Vince Castranova博士和德国不来梅大学的Lutz Mddler博士)与NIOSH的Vince Castranova博士和德国不来梅大学的Lutz Mddler博士合作，旨在建立小鼠吸入毒理学模型，以筛选与组织培养细胞中的机械损伤途径有关的组合纳米材料库。这项研究将解决缺乏用于评估纳米材料(NM)安全性的可重复筛选方案的问题。我们推测，两个组合文库的设计将有助于建立人支气管上皮(NHBE)和髓系树突状细胞(DC)的体外毒理学与小鼠非过敏性和过敏性肺部炎症之间的联系。这两个组合文库旨在(I)通过铁掺杂改变锌++的释放来调节纳米氧化锌的毒性作用，(Ii)通过缩小表面阳离子密度来调节阳离子介孔二氧化硅纳米颗粒(MSNP)的细胞毒性。我们假设，非过敏性呼吸道炎症和细胞毒性损伤之间的联系将可以通过纳米颗粒的性质来解释，纳米颗粒导致氧化损伤，有毒金属离子的脱落，以及触发线粒体损伤的能力。相比之下，材料特性与过敏性气道炎症的联系可能会影响DC的“危险信号”的产生，这使它们能够启动一条促进过敏性炎症的免疫刺激途径。为了实现我们开发预测吸入毒理学模型的长期目标，我们在目标1中建议表征NM文库的生物物理化学性质，这些性质有望在NHBE和骨髓来源的DC中诱导不同的细胞毒性和促炎效应。体外毒性筛选将由加州大学洛杉矶分校加州纳米系统研究所的高含量筛选(HCS)设施进行，并通过ELISA法进行细胞因子测量。我们还将评估ROS的产量。将评估的纳米材料性能包括颗粒大小、粒度分布、分散性、Zeta电位、透射电子显微镜、X射线衍射仪和BET分析。HCS是用多种染料混合进行的，显示细胞膜泄漏、DNA损伤、线粒体去极化和细胞内钙离子流动。颗粒表征和体外筛选将在第一年年初和合成新批次时进行。目的2将利用这些文库通过吸入-吸入法建立小鼠过敏性和非过敏性肺部炎症的标准化方案。非嗜酸性炎症试验(Castranova)将在暴露后1天至2个月期间测量BAL中炎症、损伤和氧化应激的标志物以及肺组织学。过敏性致敏模型依赖于麻醉动物鼻子中的颗粒和OVA吸入，将评估OVA特异性IgE水平，同时评估BAL细胞分类和肺组织学。这些研究将在第一至第二年分阶段进行。我们希望开发一种新的和可预测的吸入毒理学范例作为联盟活动的一个组成部分，以建立NM安全性筛选的体外和体内研究方案。 与公共健康相关：该项目致力于细胞和动物筛选程序的设计，这些程序可用于评估目前引入市场的工程纳米材料的潜在危险。我们描述了一种独特的方法，在这种方法中，我们使用内部合成的纳米颗粒来开发一个可预测的毒理学模型，这意味着我们将在组织培养细胞中制定测试方案，以获得有关纳米颗粒特性的基本信息，这些特性可能会使它们变得危险，然后在小鼠模型中测试这些想法，这些想法可能会反映吸入这些颗粒时肺部可能发生的情况。该项目的目标是开发可靠和可重复使用的协议，这些协议可以被学术中心、工业和政府机构用作纳米材料安全的屏蔽物。