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）：定殖于牙齿表面的生物膜和衬于牙周袋/龈沟（龈下牙菌斑）的上皮细胞是自然界中存在的最复杂的生物膜之一，并导致牙周疾病。我们的长期目标是开发一种临床上合适的方法来增强光敏化合物（或光敏剂）在人类牙菌斑中的渗透和有效性，以预防，控制和/或治疗牙周炎。待检验的假设是：a）装载亚甲基蓝（MB）的聚合物纳米颗粒可以完全穿透体外由天然人牙菌斑形成的口腔多物种生物膜，和B）在用装载MB的纳米颗粒致敏之后，随后暴露于665 nm的红光之后，这些微生物生物膜在体外完全光破坏是可能的。这一R21提案的具体目标是：1。制备并表征MB-生物相容性和可生物降解的聚合物纳米粒。我们将（i）制备MB-封装的聚（ii）通过ZetaPALS仪器和扫描电子显微镜评估它们的尺寸、表面电荷和形态，（iii）通过光谱学验证它们的MB包封能力和效率，和（iv）研究MB在模拟环境中的体外释放，以确保足够的试剂可用于有效的光动力疗法。2.研究MB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作用。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负载纳米颗粒的内化，（iii）通过共聚焦扫描激光显微镜评估它们对生物膜的渗透，以及（iv）通过总活菌计数、使用DNA探针和PCR进行物种识别来评估它们对生物膜物种的光动力学效应。 公共卫生关系：该项目是一种尝试，利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。