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、Ning Li）与 NIOSH 的 Vince Castranova 博士和不来梅大学（德国）的 Lutz Mddler 博士合作，旨在建立一个小鼠吸入毒理学模型，用于筛选与组织培养细胞中机械损伤途径相关的组合纳米材料库。这项研究将解决评估纳米材料 (NM) 安全性时缺乏可重复筛选方案的问题。我们假设两个组合库被设计为（i）通过铁掺杂改变 Zn++ 释放来调整 ZnO 纳米颗粒的毒性作用，（ii）通过减少表面阳离子来调整阳离子介孔二氧化硅纳米颗粒（MSNP）的细胞毒性密度，将有助于建立人支气管上皮（NHBE）和髓样树突细胞（DC）的体外毒理学与非过敏性和过敏性肺部炎症之间的联系在小鼠中。我们认为，非过敏性气道炎症和细胞毒性损伤之间的联系可以通过导致氧化损伤、有毒金属离子脱落以及触发线粒体损伤的能力的纳米颗粒特性来解释。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“危险信号”的产生，从而使它们能够启动促进过敏性炎症的免疫刺激途径。为了实现我们开发预测性吸入毒理学模型的长期目标，我们在目标 1 中建议表征 NM 文库的生物理化特性​​，这些特性预计会在 NHBE 和骨髓来源的 DC 中诱导不同的细胞毒性和促炎作用。体外毒性筛查将由加州大学洛杉矶分校加州纳米系统研究所的高内涵筛查 (HCS) 设施进行，并通过 ELISA 进行细胞因子测量。我们还将评估 ROS 的产生。将评估的纳米材料特性包括粒径、尺寸分布、分散性、zeta 电位、TEM、XRD 和 BET 分析。 HCS 使用混合染料进行，可揭示细胞膜渗漏、DNA 损伤、线粒体去极化和细胞内 Ca2+ 通量。颗粒表征和体外筛选将在第一年年初以及合成新批次时进行。目标 2 将利用这些库通过吸入-抽吸方法建立针对小鼠过敏性和非过敏性肺部炎症的标准化方案。非嗜酸性炎症方案 (Castranova) 将在暴露后 1 天至 2 个月的时间内测量 BAL 中的炎症、损伤和氧化应激标志物以及肺组织学。过敏致敏模型依赖于麻醉动物鼻子中的颗粒和 OVA 吸入，将与 BAL 分类细胞计数和肺组织学并行评估 OVA 特异性 IgE 水平。这些研究将在第一年至第二年分阶段进行。我们期望开发一种新颖的预测性吸入毒理学范例，作为联盟活动的一部分，以建立 NM 安全筛选的体外和体内研究方案。 公共健康相关性：该项目涉及细胞和动物筛选程序的设计，可用于评估目前引入市场的工程纳米材料的潜在危险。我们描绘了一种独特的方法，使用内部合成的纳米粒子来开发预测毒理学模型，这意味着我们将在组织培养细胞中开发测试方案，以获得有关可能使其危险的纳米粒子特性的基本信息，然后测试这些想法在小鼠模型中得出的结果可能反映了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且可重复的方案，可供学术中心、行业和政府机构用作纳米材料安全性的筛选。