Nanoparticle-based Antimicrobial Photochemotherapy in Biofilms

基于纳米颗粒的生物膜抗菌光化学疗法

• 批准号: 7588512
• 负责人: NIKOLAOS SOUKOS
• 金额: $ 25.28万
• 依托单位: ADA FORSYTH INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7588512
• 关键词: Adhesions; Antibiotics; Bacteria; Biocompatible; Caliber; Cell Line; Cells; Charge; Clinical; Complex; DNA Probes; Dental Plaque; Effectiveness; Encapsulated; Environment; Epithelial Cells; Excision; Exposure to; FDA approved; Failure; Free Radicals; Glycolates; Goals; Human; In Vitro; Laboratories; Laser Scanning Confocal Microscopy; Lead; Light; Local Anti-Infective Agents; Mechanics; Medical Research; Methods; Methylene blue; Microbial Biofilms; Microscopic; Mono-S; Morphology; Nanotechnology; Nature; Oral; Organism; Particle Size; Penetration; Periodontal Diseases; Periodontal Pocket; Periodontitis; Pharmaceutical Preparations; Phase; Photochemotherapy; Photosensitizing Agents; Phototoxicity; Predisposition; Prevention; Reporting; Research; Resistance development; Saliva; Scanning Electron Microscopy; Simulate; Singlet Oxygen; Spectrum Analysis; Structure of gingival sulcus; Surface; System; Techniques; Testing; Time; Tissues; Transmission Electron Microscopy; Visible Radiation; antimicrobial; antimicrobial drug; base; cytotoxic; instrument; microorganism; nanoparticle; oral bacteria; oral biofilm; particle; pathogen; periodontopathogen; public health relevance; solute; subgingival biofilm; tooth surface; water channel

DESCRIPTION (provided by applicant): Biofilms that colonize tooth surfaces and epithelial cells lining the periodontal pocket/gingival sulcus (subgingival dental plaques) are among the most complex biofilms that exist in nature and lead to periodontal diseases. Our long-term goal is to develop a clinically appropriate way to enhance the penetration and effectiveness of photoactive compounds (or photosensitizers) in human dental plaque for prevention, control and/or treatment of periodontitis. The hypotheses to be tested are: a) Methylene blue (MB)-loaded polymeric nanoparticles can fully penetrate oral multispecies biofilms evolved from natural human dental plaque in vitro, and b) Complete photodestruction of these microbial biofilms in vitro may be possible after their sensitization with MB-loaded nanoparticles followed by exposure to red light of 665 nm. The specific aims of this R21 proposal are: 1. To prepare and characterize MB-loaded biocompatible and biodegradable polymeric nanoparticles. We will (i) prepare MB-encapsulated nanoparticles of poly(lactic-co-glycolic acid) with anionic and positive surface charge, (ii) evaluate their size, surface charge, and morphology by ZetaPALS instrument and scanning electron microscopy, (iii) verify their capacity and efficiency of MB encapsulation by spectroscopy, and (iv) investigate the in vitro release of MB in simulated environment to insure that sufficient agent is available for effective photodynamic therapy. 2. To investigate the distribution of MB-loaded nanoparticles in biofilms and their phototoxicity on biofilm microorganisms in vitro. We (i) develop multi-species microcosm laboratory biofilms in vitro using saliva and human dental plaque, (ii) investigate the internalization of MB-loaded nanoparticles by bacteria using transmission electron microscopy, (iii) assess their penetration into the biofilms by confocal scanning laser microscopy, and (iv) evaluate their photodynamic effects on biofilm species by total viable counts, an species identification using DNA probes and PCR. PUBLIC HEALTH RELEVANCE: This project is an attempt to target dental plaque bacteria using visible light and microscopic particles loaded with a photoactive drug. Our research may help develop photodynamic therapy as a non-invasive technique for prevention, control and/or treatment of periodontal disease.

描述（由申请人提供）：定植于牙齿表面和牙周袋/牙龈沟内壁上皮细胞（龈下牙菌斑）的生物膜是自然界中存在的最复杂的生物膜之一，会导致牙周病。我们的长期目标是开发一种临床上合适的方法来增强光活性化合物（或光敏剂）在人体牙菌斑中的渗透性和有效性，以预防、控制和/或治疗牙周炎。要测试的假设是：a）负载亚甲基蓝（MB）的聚合物纳米颗粒可以在体外完全穿透由天然人类牙菌斑演变而来的口腔多物种生物膜，以及b）这些微生物生物膜在其致敏后可能在体外完全光破坏载有 MB 的纳米颗粒，然后暴露于 665 nm 的红光下。该 R21 提案的具体目标是： 1. 制备并表征负载 MB 的生物相容性和可生物降解的聚合物纳米粒子。我们将 (i) 制备具有阴离子和正表面电荷的 MB 封装的聚乳酸-乙醇酸共聚物纳米粒子，(ii) 通过 ZetaPALS 仪器和扫描电子显微镜评估其尺寸、表面电荷和形态，(iii)通过光谱验证其 MB 封装的能力和效率，以及 (iv) 研究模拟环境中 MB 的体外释放，以确保有足够的试剂可用于有效的光动力治疗。 2. 体外研究MB负载纳米粒子在生物膜中的分布及其对生物膜微生物的光毒性。我们（i）使用唾液和人类牙菌斑在体外开发多物种微观实验室生物膜，（ii）使用透射电子显微镜研究细菌对MB负载纳米粒子的内化，（iii）通过共焦扫描评估它们对生物膜的渗透(iv) 通过总活菌数（使用 DNA 探针和 PCR 进行物种鉴定）评估其对生物膜物种的光动力效应。 公共健康相关性：该项目尝试使用可见光和载有光敏药物的微观颗粒来靶向牙菌斑细菌。我们的研究可能有助于开发光动力疗法作为预防、控制和/或治疗牙周病的非侵入性技术。