Biomimetic Microsystem for High Throughput Evaluation of Engineered Nanomaterials

用于工程纳米材料高通量评估的仿生微系统

• 批准号: 7941830
• 负责人: ROBERT M WORDEN
• 金额: $ 44.04万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-28 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7941830
• 关键词: Adjuvant; Affect; Allergens; Allergic; Allergic Disease; Animals; Antigens; Asthma; Behavior; Binding; Biological; Biological Assay; Biological Response Modifiers; Biomimetics; Breathing; CD8B1 gene; Cell Culture Techniques; Cell membrane; Cells; Charge; Chemistry; Computer Simulation; Data; Dendritic Cells; Deposition; Detection; Disease; Electrodes; Endocytosis; Engineering; Evaluation; Exhibits; Exposure to; Flow Cytometry; Fluorescence; Goals; Health; Health Status; Immune; Immune response; Immune system; In Vitro; Incidence; Inhalation Exposure; Interdisciplinary Study; Ligands; Lipid Bilayers; Lipopolysaccharides; Measurement; Measures; Membrane; Methods; Microelectrodes; Modeling; Modification; Molecular; Molecular Models; Mus; Ovalbumin; Oxidation-Reduction; Performance; Production; Property; Proteins; Quinones; Reaction; Reactive Oxygen Species; Risk Assessment; Role; Safety; Screening procedure; Severities; Spectrum Analysis; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Theoretical model; Toxic effect; Toxicity Tests; allergic airway disease; allergic airway inflammation; base; chemical group; chemical property; commercial application; crosslink; design; electric impedance; engineering design; functional group; in vivo; insight; mathematical model; meetings; microsystems; molecular modeling; nanomaterials; nanoparticle; public health relevance; receptor; research study; response; uptake

DESCRIPTION (provided by applicant): Engineered nanomaterials (ENM) have unique properties that can cause adverse health effects. Due to their small size and potential for airborne dispersion, inhalation exposure to ENM might contribute to the increased incidence and/or exacerbation of allergic airway disease. The overall goal of this project is to develop a multi- level toxicity testing platform for ENM that includes in vivo measurement of allergic airway disease in mice, in vitro measurement of T cell activation, high-throughput measurement of ENM's interactions with bilayer lipid membranes (BLM) that mimic cell membranes, and in silico prediction of ENM's molecular properties. The overarching hypothesis that ENM possess adjuvant-like properties that promote allergic airway disease will be tested using five specific aims (SA). SA1 is to synthesize well-characterized ENM having controlled functional groups that catalyze redox reactions or activate membrane receptors. SA2 is to determine the adjuvant potential of ENM on allergic airway sensitization and asthma-like disease in mice. SA3 is to determine the effects of ENM on dendritic cell-induced activation and effector function of CD4+ and CD8+ T cells. SA4 is to measure the direct effects of ENM on synthetic bilayer lipid membranes. SA5 is to develop and validate mathematical models that can correlate ENM physicochemical properties with their biological and toxicological effects at the animal, cell, and membrane levels for health risk assessment. Biodegradable poly(propargyl glycolide) nanoparticles will be synthesized and coated with chemical groups (lipopolysaccharide (LPS) and quinone) to generate ENM likely to stimulate the immune response. LPS bind to receptor proteins on cell membranes and trigger cellular uptake by endocytosis, and quinones can trigger oxidation-reduction reactions, including production of reactive oxygen species by immune cells. A multi-tiered approach will be used to determine whether addition of LPS and quinone to ENM increases the ENM's ability to promote airway disease by (1) increasing the murine immune system's response to the antigen ovalbumin, (2) increasing T cell activation by dendritic cells in response to ovalbumin, and (3) modifying the ELM's molecular interactions with BLM. Mice will be exposed to the ENM by inhalation, and severity of allergic airway disease will be histopathologically, morphometrically and biochemically assessed. Dendritic cells will be exposed to the ENM, and their ability to activate T cells will be measured using fluorescence assisted flow cytometry. BLM will be deposited on electrodes and exposed to the ENM. The resulting interactions between the ENM and BLM will be measured in a high-throughput mode using cyclic voltammetry and electrical impedance spectroscopy. Theoretical models will be developed that describe the molecular properties of the ENM and their interactions with cellular components. These models will be used to analyze the experimental data and help elucidate mechanisms by which ENM induce toxic effects. PUBLIC HEALTH RELEVANCE: This project will provide fundamental insight into how a nanoparticle's physical and chemical properties determine its ability to enhance allergic airway disease like asthma. This insight will aid in setting health and safety standards for engineered nanomaterials, provide new high- throughput methods for nanoparticle detection and safety screening, and facilitate design of new nanomaterials that simultaneously meet safety standards and exhibit desirable performance properties needed for commercial applications.

描述（由申请人提供）：工程纳米材料（ENM）具有可能导致不良健康影响的独特特性。由于它们的大小和空气传播的潜力，吸入接触ENM可能有助于增加过敏气道疾病的发病率和/或加剧。该项目的总体目标是为ENM开发一个多层次的毒性测试平台，其中包括小鼠过敏性气道疾病的体内测量，体外测量T细胞激活，ENM与双层脂质膜（BLM）的高通量测量，对二孔和硅质的相互作用的相互作用。 ENM具有辅助性促进过敏性气道疾病的总体假设将使用五个特定目标（SA）进行测试。 SA1是合成具有催化氧化还原反应或激活膜受体的受控官能团的良好特征的ENM。 SA2是为了确定ENM在小鼠过敏性气道敏化和类似哮喘的疾病上的辅助潜力。 SA3是为了确定ENM对CD4+和CD8+ T细胞的树突状细胞诱导的激活和效应子功能的影响。 SA4是测量ENM对合成双层脂质膜的直接影响。 SA5将开发和验证数学模型，以将ENM物理化学特性与它们在动物，细胞和膜水平上进行健康风险评估的生物学和毒理学作用相关联。可生物降解的聚乙二醇纳米颗粒将与化学基（脂多糖（LPS）和奎因酮）合成并覆盖，以产生可能刺激免疫反应的ENM。 LPS与细胞膜上的受体蛋白结合，并通过内吞作用引发细胞摄取，奎因酮可以触发氧化还原反应，包括免疫细胞产生活性氧。 A multi-tiered approach will be used to determine whether addition of LPS and quinone to ENM increases the ENM's ability to promote airway disease by (1) increasing the murine immune system's response to the antigen ovalbumin, (2) increasing T cell activation by dendritic cells in response to ovalbumin, and (3) modifying the ELM's molecular interactions with BLM.小鼠将通过吸入暴露于ENM，过敏性气道疾病的严重程度将在组织病理学，形态学上和生化评估上进行。树突状细胞将暴露于ENM，并使用荧光辅助流式细胞仪测量其激活T细胞的能力。 BLM将沉积在电极上并暴露于ENM。 ENM和BLM之间所得的相互作用将在高通量模式下使用环状伏安法和电阻抗光谱进行测量。将开发理论模型，以描述ENM的分子特性及其与细胞成分的相互作用。这些模型将用于分析实验数据，并有助于阐明ENM诱导有毒作用的机制。 公共卫生相关性：该项目将为纳米颗粒的物理和化学特性如何确定其增强过敏性气道疾病（如哮喘）的能力提供基本见解。这种见解将有助于为工程纳米材料设定健康和安全标准，为纳米颗粒检测和安全筛查提供新的高吞吐量方法，并促进设计新的纳米材料，同时符合安全标准并展示商业应用所需的可取性能。