Antimicrobial mechanisms of action zinc oxide nanoparticles

氧化锌纳米粒子的抗菌作用机制

• 批准号: 9385809
• 负责人: J SCOTT VANEPPS
• 金额: $ 19.44万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-23 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9385809
• 关键词: Address; Adhesions; Affect; Anti-Bacterial Agents; Antibiotic Therapy; Antibiotics; Appointment; Bacteria; Behavior; Binding; Biocompatible Materials; Biomimetics; Cell Death; Cell Respiration; Cell Wall; Cell physiology; Cells; Cessation of life; Chemistry; Clinical; Clinical Medicine; Complex; Data; Development; Development Plans; Device Designs; Devices; Educational workshop; Engineering; Enzyme Inhibition; Enzyme Inhibitor Drugs; Enzymes; Evaluation; Funding; Generations; Genes; Genomics; Goals; Grant; Growth; Healthcare; Immune response; Implant; Infection; Infection prevention; Lead; Libraries; Life; Liquid Chromatography; Literature; Mammalian Cell; Mass Spectrum Analysis; Mediating; Medical Device; Medicine; Membrane; Mentored Clinical Scientist Development Award (K08); Mentors; Mentorship; Metabolic Pathway; Michigan; Microbial Biofilms; Microbiology; Modification; Molecular Biology; NP protein; Operating Rooms; Operative Surgical Procedures; Oxidative Stress; Pathogenicity; Patients; Predisposition; Preparation; Process; Property; Proteins; Proteomics; Publications; Reactive Oxygen Species; Research; Resources; Scientist; Secure; Sepsis; Shapes; Staphylococcus aureus; Surface; System; Techniques; Technology; Testing; Therapeutic; Time; Training; Translational Research; Universities; Work; Zinc Oxide; antimicrobial; antimicrobial peptide; biological adaptation to stress; career; career development; cohort; college; combat; commercialization; cost; didactic education; experience; extracellular; gel electrophoresis; graduate student; implant material; implantable device; in vivo; innovation; interest; materials science; medical implant; meetings; microbial; multidisciplinary; mutant; nanomaterials; nanoparticle; new technology; novel; pathogen; prevent; product development; programs; protein complex; research and development; symposium

PROJECT SUMMARY: Despite a decade of engineering advancements and clinical process improvements, 1 million healthcare- associated infections in the U.S. can be attributed to indwelling medical devices annually. Zinc oxide nanoparticles (ZnO-NPs) are one of the most promising emerging antimicrobials with potential to combat device related infection. ZnO-NPs are inexpensive, stable, and easy to prepare with broad antimicrobial spectrum and wide therapeutic window. However, the antimicrobial mechanism of action of ZnO-NPs remains elusive. This proposal is specifically motivated to better understand the mechanism of action of ZnO-NPs. Such understanding is necessary to guide the design of device coatings that preserve antibacterial function in vivo. Reactive oxygen species (ROS) generation or membrane disruption are hypothesized mechanisms of action. However the literature is inconsistent and our preliminary data suggests that these NP effects are not sufficient. We recently demonstrated that ZnO-NPs have shape-dependent, biomimetic, reversible, enzyme inhibition properties. The central research question for this career development grant is: To what extent does ZnO-NP behavior as an enzyme inhibitor contribute to antimicrobial activity? I have multidisciplinary training in medicine, engineering, and molecular biology that is well-suited to address this question. My ultimate career goal is to become a clinician-scientist. I plan to have a clinical interest in sepsis as it relates to indwelling medical devices and an independently funded research program focused on the development of novel biomaterials to resist microbial contamination and infection. This proposal was developed to solidify my expertise, formalize my research niche, and garner the resources for the next phase of career development. My specific career development objectives for the next four years are to: 1. Solidify my expertise in microbiology (including biofilm microbiology), microbial-surface interaction, nanoparticle technology, and translational research. 2. Master techniques in evaluating mechanisms of action of antimicrobial and anti-biofilm materials. 3. Generate sufficient preliminary data and publication record to obtain independent research funding. 4. Secure my niche as an expert in bacterial-nanomaterial interactions. 5. Obtain secondary appointment in the College of Engineering so that I can work with and mentor graduate students in their research and career development. I have assembled a mentorship team of experts co-localized at the University of Michigan North Campus Research Complex with experience in clinical medicine, microbiology, material science and engineering, and product development/commercialization. Together we have devised a highly-individualized, project-oriented training plan that includes regular mentorship meetings, formal didactic education, career development workshops, and presentation at local and national conferences. Partnered with this career development plan is an innovative research plan. By synthesizing ZnO-NPs that are identical in surface chemistry but differ only in shape we can control the potential for enzyme inhibition and address the central research question above. Using these novel preparations, we will test the hypothesis: Pyramidal ZnO-NPs inhibit a cohort of bacterial enzymes which are critical to survival. Our research specific aims are to: 1. Quantify aerobic metabolism, membrane integrity, and microbial death in a commonly isolated medical device pathogen (i.e., Staphylococcus aureus) as a function of exposure time to spherical vs pyramidal ZnO-NPs. 2. Identify genes involved in enzyme inhibition by ZnO-NPs using a mariner transposon mutant library of S. aureus. 3. Determine the subset of S. aureus proteins that specifically complex with ZnO-NPs in a shape- dependent manner by 2D-gel electrophoresis followed by liquid chromatography paired with tandem mass spectroscopy (LC-MS/MS).

项目摘要： 尽管有十年的工程进步和临床过程改进，但100万个医疗保健 美国相关的感染每年可归因于留置医疗设备。氧化锌 纳米颗粒（ZnO-NP）是最有前途的抗菌剂之一，具有对抗的潜力 设备相关感染。 Zno-NP廉价，稳定且易于使用宽阔的抗菌素 光谱和宽阔的治疗窗口。但是，Zno-NP的作用抗菌机理仍然存在 难以捉摸。该建议是特别动机，以更好地理解ZnO-NP的作用机理。 这种理解对于指导在保留抗菌功能的设备涂料的设计中是必要的 体内。活性氧（ROS）产生或膜破坏是假设的机制 行动。但是，文献不一致，我们的初步数据表明这些NP效应不是 充足的。我们最近证明ZnO-NP具有形状依赖性，仿生，可逆的酶 抑制特性。这项职业发展赠款的中心研究问题是： ZnO-NP作为酶抑制剂的行为有助于抗菌活性？ 我接受了医学，工程和分子生物学的多学科培训，非常适合解决 这个问题。我的最终职业目标是成为临床医生科学家。我计划对 败血症与留置医疗设备和一个独立资助的研究计划有关 新型生物材料的发展以抵抗微生物污染和感染。该提议是 开发旨在巩固我的专业知识，正式化我的研究利基市场，并获得下一阶段的资源 职业发展。我未来四年的特定职业发展目标是： 1。巩固我在微生物学方面的专业知识（包括生物膜微生物学），微生物表面相互作用， 纳米颗粒技术和转化研究。 2。评估抗菌和抗生物膜材料作用机理的主要技术。 3。生成足够的初步数据和出版记录，以获得独立的研究资金。 4。确保我作为细菌纳米材料相互作用专家的利基市场。 5。在工程学院获得次要任命，以便我可以与 研究生研究和职业发展。 我已经组建了一个在密歇根大学北校区共定位的专家的指导团队 研究综合体具有临床医学，微生物学，材料科学和工程的经验，以及 产品开发/商业化。我们一起设计了一个高度个性化的，面向项目的 培训计划，包括定期指导会议，正式教学教育，职业发展 讲习班，并在地方和民族会议上演讲。 与该职业发展计划合作是一项创新的研究计划。通过合成ZnO-NP 表面化学的相同，但仅在形状上有所不同，我们可以控制酶抑制的潜力 解决上面的中心研究问题。使用这些新颖的准备工作，我们将检验以下假设： 锥体ZnO-NP抑制了对生存至关重要的细菌酶。我们的研究 具体目的是： 1。定量有氧代谢，膜完整性和微生物死亡 装置病原体（即金黄色葡萄球菌）与球形与金字塔的暴露时间有关 Zno-NP。 2。使用水手Transposon突变库中的ZnO-NP抑制酶抑制酶的基因 S.金黄色葡萄酒。 3。确定金黄色葡萄球菌蛋白的子集，该蛋白质与ZnO-NP特别复杂 2D凝胶电泳的依赖方式，然后是液相色谱与串联配对 质谱法（LC-MS/MS）。