Penetration and Translocation of Nanoparticles through Skin

目的：在合成和控制工程纳米颗粒（NP）性能方面的进展已经升级了它们在从医学到能源的许多不同应用中的应用。目前，NPs被用于400多种消费品中(例如：防晒霜、食品容器)。纳米技术繁荣的一个后果是，人体最大的器官皮肤意外暴露于NP的风险增加。这项研究的动机是越来越多地在日常化妆品中使用金属氧化物（ZnO和TiO2） NPs来防止紫外线辐射（UVR）暴露。我们这项研究的主要目的是量化NP皮肤渗透，并通过对比UVR诱导的正常皮肤和屏障受损皮肤的渗透，对易位机制产生分子水平的见解。实验方法：我们将利用体内、离体和体外模型来量化UVR对NP渗透和转运的影响。通过研究渗透作为NP表面化学和载体的功能，包括化学渗透增强剂（CPE）的结合，通过提取或流化皮肤脂质来修饰皮肤屏障，将收集机理见解。提出的研究的中心假设是NP表面化学（电荷/疏水性）和水动力学尺寸是影响皮肤穿透的最重要的材料性质。组织清除率和毒性也取决于成分，但这些不是本提案的重点。具体目标1：建立一个体内小鼠模型，研究NP皮肤渗透，并产生NP易位的机制，作为NP大小、表面化学、载体和皮肤状态（正常与紫外线暴露）的函数。具体目标2：使用离体皮肤模型将目标1中获得的知识转化为人体系统，以量化皮肤生理和UVR对NP渗透的影响。具体目标3：建立体外分化角质细胞模型，研究NP摄取和细胞毒性作为NP表面化学和UVR的功能，并阐明在目标2中发现的定位方面的知识。预期结果：这些研究将是第一批产生广泛NP类型（量子点，荧光聚合物，金属氧化物）作为皮肤屏障状态函数的体内数据的研究之一。我们的研究将增加对皮肤屏障功能抵抗NP渗透的机制和驱动力的理解。这项研究的结果将为如何设计NPs来限制生物利用度和防止皮肤暴露的潜在副作用提供知识。我们将发展理解，这将使未来的工作能够更具体地针对纳米颗粒来防止或提高生物利用度。这一知识可用于开发强大的药物递送剂或减少化妆品纳米颗粒的渗透。更广泛的影响：这项工作的一个主要成果将是在一个高度跨学科的研究环境中，为研究生、本科生和高中生提供教育和指导，涵盖材料科学、表面化学、皮肤生物学和生物医学工程等方面。量子点和纳米毒性的概念将被整合到由PI提供的细胞与组织工程（BME462）和纳米生物光学（ECE580）课程中。参与这项研究的研究生和本科生将接受培训，在工业、政府和学术界的高科技和生物技术部门从事各种具有挑战性的职业道路。补充关键词：纳米技术，纳米材料，纳米颗粒，聚合物，量子点，金属，金属氧化物，半导体纳米晶体，人体皮肤，皮肤渗透，紫外线辐射，生物利用度，细胞培养，人体健康影响，皮肤接触，人体暴露。

CBET-0837891DeLouiseObjectives: Advances in the synthesis and control of engineered nanoparticles (NP) properties has escalated their use in numerous diverse applications ranging from medicine to energy. Currently NPs are formulated in over 400 consumer products (eg. sunblocks, food containers). A consequence of this nanotechnology boom is an increased risk of unintended NP exposure with human skin, the largest organ in the body. The proposed research is motivated by the increasing use of metal oxide (ZnO and TiO2) NPs in daily-use cosmetic products to protect against ultraviolet radiation (UVR) exposure. Our main objective of this research is to quantify NP skin penetration and to generate molecular level insight into the mechanisms of translocation by contrasting permeation through normal and barrier compromised skin induced by UVR. Experimental Approach: We will utilize in vivo, ex vivo and in vitro models to quantify the effects of UVR on NP permeation and translocation. Mechanistic insight will be gleaned by investigating penetration as a function of NP surface chemistry and vehicle including incorporation of chemical penetration enhancers (CPE) to modify the skin barrier by extraction or fluidization of skin lipids. The central hypothesis of the proposed research is that the NP surface chemistry (charge/hydrophobicity) and hydrodynamic size are the most important material properties that effect skin penetration. Tissue clearance and toxicity also depend on composition but these are not a focus of this proposal. Specific Aim 1: Develop an in vivo mouse model to investigate NP skin penetration and generate mechanistic insight on NP translocation as a function of NP size, surface chemistry, vehicle and skin status (normal vs. UV exposed). Specific Aim 2: Translate knowledge gained in Aim 1 to a human system using an ex vivo skin model to quantify the affect of skin physiology and UVR on NP penetration. Specific Aim 3: Develop an in vitro differentiated keratinocyte model to investigate NP up-take and cytotoxicty as a function of NP surface chemistry and UVR and to elucidate knowledge on localization aspects discovered in Aim 2. Expected Results: These studies will be among the first to generate in vivo data on a wide range of NP types (quantum dots, fluorescent polymer, metal oxides) as a function of skin barrier status. Our study will increase understanding of the mechanisms and driving forces of skin barrier function against NP penetration. Results from this research will generate knowledge on how to design NPs to limit bioavailability and prevent potential side effects from skin exposure. We will develop understanding that will enable future work to be done in more specific targeting of nanoparticles to prevent or enhance bio availability. This knowledge could be used to develop powerful drug delivery agents or to decrease penetration of cosmetic nanoparticles. Broader Impact: A primary outcome of this work will be to provide education and mentoring to graduate, undergraduate and high school students in a highly interdisciplinary research environment spanning aspects of material science, surface chemistry, skin biology, and biomedical engineering. The concepts of quantum dots and nanotoxicity will be integrated into lectures presented in Cell and Tissue Engineering (BME462) and Nanobiophotoics (ECE580) courses given by the PI. Graduate and undergraduate students involved in this research will be trained to pursue a variety of challenging career paths in the high-tech and biotech sectors of industry, government, and academia. Supplemental Keywords: nanotechnology, nanomaterials, nanoparticles, polymer, quantum dots, metals, metal oxides, semiconductor nanocrystals, human skin, skin permeation, UV radiation, bioavailability, cell culture, human health effects, dermal contact, human exposure.