Nanoneurotoxicity of Metal Oxide Nanomaterials and Neurodegeneration

项目摘要/摘要 越来越多的证据表明，由于二氧化锰、氧化铜、二氧化钛等纳米颗粒的暴露， 越来越多地使用这些工程纳米材料(ENM)，会导致纳米神经毒性，可能会对 患有阿尔茨海默病(AD)、帕金森氏病(PD)等神经退行性疾病。 然而，体内暴露的机制及其对神经退行性变的潜在贡献并不是很好。 为人所知。因此，我们的长期目标是研究ENM的暴露，如金属氧化物纳米颗粒和 它们的纳米神经毒性及其与AD等人类疾病的潜在相关性 疾病模型和成像方式。这一提议的具体假设是CuO的暴露 纳米颗粒增加血脑屏障(BBB)通透性，加剧神经变性，加剧 大脑A-β淀粉样变性病理和相关的神经炎症和氧化还原应激，改变脑细胞因子， 生物金属和能量代谢图谱。我们基于之前的观察和当前的关键 试点数据表明：(一)长期暴露在严重空气污染中(极有可能接触金属 氧化物纳米颗粒)与神经炎症、血脑屏障破坏和Aβ1-42蓄积( 阿尔茨海默病的显著神经病理特征)；(Ii)暴露于CuO纳米颗粒可通过 吸入；(Iii)暴露于CuO纳米颗粒会在体外诱导神经毒性、炎症和氧化应激。 为了验证我们目前的假设，我们将(I)确定吸入纳米CuO对体内血液的影响。 脑屏障完整性和增强的神经变性；(Ii)评估吸入CuO纳米颗粒的效果 对神经炎症、脑氧化应激和β淀粉样蛋白病理的影响；(Iii)评估CuO的作用 纳米颗粒吸入对β淀粉样斑块的记忆功能、生物特征和脑代谢的影响 活动。我们将使用体内MicroSPECT(微单光子发射计算机断层扫描)成像 组织学检测方法、细胞因子微流控生物芯片检测、氧化应激(4-HNE)和Aβ-EL ISA 分析，高能X射线荧光显微镜(µ-XRM)，通过测量X射线吸收光谱 (µ-XAS)和X射线近边吸收光谱(µ-XANES)与激光捕获显微切割(LCM)联用 组织获取技术、MRI/MRS、Morris水迷宫记忆测试和PS1/APP AD转基因 小鼠模型来实现这些实验目的。使用我们的综合实验方法，我们相信 我们将获得有助于制定监管纳米粒子的公共政策的知识 暴露及其对中枢神经系统(CNS)的神经毒性效应 关于金属氧化物纳米颗粒暴露于神经变性的潜在风险。更重要的是，它将 建立一种实验范式，这将对研究纳米神经毒性及其 对阿尔茨海默病等神经退行性疾病病因的潜在贡献。

Project Summary/Abstract Growing evidence has indicated that exposure of nanoparticles such as MnO2, CuO, TiO2, etc., due to increasing use of these engineered nanomaterials (ENMs), induces nanoneurotoxicity that may pose risks for having neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), etc. However, the mechanisms of in vivo exposure and its potential contribution to neurodegeneration are not well known. Hence, our long-range objective is to study exposure of ENMs such as metal oxide nanoparticles and their nanoneurotoxicity and potential correlation to human diseases such as AD using both in vitro and in vivo disease models and imaging modalities. The specific hypothesis for this proposal is that exposure of CuO nanoparticles increases blood-brain barrier (BBB) permeability, enhances neurodegeneration, exacerbates cerebral Aβ amyloid pathology and associated neuroinflammation and redox stress, alter brain cytokine, biometal, and energy metabolic profiles. We base the hypothesis on previous observations and current key pilot data which suggest that: (i) long-term exposure to severe air pollution (highly possible exposure of metal oxide nanoparticles) is associated with neuroinflammation, BBB disruption, and Aβ1-42 accumulation (the salient neuropathological features of AD); (ii) CuO nanoparticle exposure increases BBB permeability via inhalation; (iii) exposure of CuO nanoparticles induce in vitro neurotoxicity, inflammation, and oxidative stress. To test our current hypothesis, we will (i) determine the effects of CuO nanoparticle inhalation on in vivo blood- brain barrier integrity and enhanced neurodegeneration; (ii) assess the effects of CuO nanoparticle inhalation on neuroinflammation, cerebral oxidative stress and Aβ amyloid pathology; (iii) evaluate the effects of CuO nanoparticle inhalation on memory function, biometal profiles in Aβ amyloid plaques and brain metabolic activities. We will use in vivo microSPECT (micro Single Photon Emission Computed Tomography) imaging and histology detection methods, cytokine microfluidic biochip assays, oxidative stress (4-HNE) and Aβ ELISA assays, high-energy X-ray fluorescence microscopy (µ-XRM) via the measurement of x-ray absorption spectra (µ-XAS) and x-ray absorption near edge spectra (µ-XANES) coupled with laser capture microdissection (LCM) tissue procuring technique, MRI/MRS, Morris Water Maze (MWM) memory test, and PS1/APP AD transgenic mouse model to achieve these experimental aims. Using our integrated experimental approaches, we believe that we will gain knowledge that can contribute to the making of public policy on regulating nanoparticle exposure and its neurotoxic effects upon the Central Nervous System (CNS), and further our understanding about potential risk of metal oxide nanoparticle exposure for neurodegeneration. More importantly, it will establish an experimental paradigm that will be very useful for investigating the nanoneurotoxicity and its potential contribution to etiopathogenesis of neurodegenerative diseases such as AD.