Investigation of the differential cytotoxicity of oxide nanoparticles using zebrafish model

这项由纳米技术环境健康与安全项目授予的国家科学基金会奖，支持博伊西州立大学的Alex Punnoose教授与俄勒冈州立大学的Jeffrey a . Greenwood教授和Robert Tanguay教授合作，以斑马鱼为模型系统研究氧化物半导体纳米颗粒的细胞毒性。市场上已经有成千上万种纳米技术驱动的消费产品，这些产品的年收入接近数万亿美元。许多半导体氧化物，如氧化锌，在所有尺寸范围内（包括纳米级），目前被食品和药物管理局标记为“普遍认可的安全”物质。然而，Punnoose教授的小组最近利用人类免疫细胞进行的体外实验表明，氧化锌纳米颗粒表现出显著的细胞毒性，这引起了人们对氧化锌纳米颗粒对环境安全和健康影响的关注。所观察到的细胞毒性随细胞类型的不同而不同，其中对癌细胞的作用最强。基于这些结果，本项目利用斑马鱼对一组常用的半导体氧化物（如锡（IV）氧化物、铈（IV）氧化物和铜（II）氧化物）的细胞毒性进行了详细的体内研究，这是一种更相关的细胞毒性评估模型系统。此外，迄今为止在体外研究中观察到的氧化锌纳米颗粒对癌细胞的细胞特异性细胞毒性反应将在体内通过将荧光染料标记的人类癌细胞注射到48小时的转基因斑马鱼中，并通过无创监测肿瘤细胞的侵袭、转移和病理血管生成来研究。最后，将研究氧化纳米颗粒的各种物理化学性质在其细胞毒性行为中的作用。基于这些信息，将采用谨慎的材料工程方法来开发新的设计规则(i)去除氧化纳米颗粒对正常健康斑马鱼的有害毒性，使其成为更安全的纳米材料；（ii）提高选择性杀死癌细胞的任何氧化纳米颗粒的选择性和细胞毒性，以便它们可以被开发为潜在的抗癌剂。该项目将为研究生、本科生和高中生提供研究机会。拟议研究的结果将整合到几个现有的课程中，并且将引入纳米生物研究领域的研究生水平研讨会，向学生和当地社区介绍相关的新兴领域，如纳米技术、纳米毒理学和纳米医学。

1134468 PunnooseThis NSF award by the Environmental Health and Safety of Nanotechnology program supports work by Professor Alex Punnoose at Boise State University along with collaborators Professors Jeffrey A. Greenwood and Robert Tanguay at Oregon State University to investigate the cytotoxicity of oxide semiconductor nanoparticles using zebrafish as a model system. Thousands of nanotechnology-enabled consumer products are already available on the market and the annual revenue from such products is approaching several trillion dollars. Many semiconductor oxides, such as zinc oxide, at all size ranges (including nanoscale) are currently labeled as "generally recognized as safe" substances by the Food and Drug Administration. However, in vitro experiments by Professor Punnoose's group employing human immune cells have recently demonstrated that zinc oxide nanoparticles display significant cytotoxicity, raising concerns about the environmental safety and health impact of oxide nanoparticles in general. The observed cytotoxicity varies with cell type, and cancer cells showed the strongest effect. Based on these results, this project undertakes detailed in vivo investigations of the cytotoxicity of a panel of commonly used semiconducting oxides such as tin(IV) oxide, cerium(IV) oxide and copper(II) oxide using zebrafish, a more relevant model system for cytotoxicity evaluation. Furthermore, the cell specific cytotoxic response of zinc oxide nanoparticles to cancerous cells observed so far using in vitro studies will be investigated in vivo by injecting fluorescent dye labeled human cancer cells into 48 hour-old transgenic zebrafish, and by noninvasively monitoring the tumor cell invasion, metastasis and pathological angiogenesis. Finally, roles of the various physicochemical properties of oxide nanoparticles in their cytotoxic behavior will be investigated. Based on this information, careful materials engineering approaches will be employed to develop new design rules (i) to remove unwanted toxicity of oxide nanoparticles to normal healthy zebrafish making them safer nanomaterials and (ii) to improve the selectivity and cytotoxicity of any oxide nanoparticles that display selective cancer cell killing so that they can be developed as potential anti-cancer agents. This project will provide research opportunities for several graduate, undergraduate and high school students. Results from the proposed research will be integrated into several existing courses, and a graduate level seminar in the area of nano-bio research will be introduced to educate students and local community about related emerging fields such as nanotechnology, nanotoxicology and nanomedicine.