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: 7817003
• 负责人: NIKOLAOS SOUKOS
• 金额: $ 27.86万
• 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/7817003
• 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）：将牙齿表面和上皮细胞固定在牙周口袋/牙龈沟中的生物膜（牙块牙菌斑）是本质上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适当的方法来增强人牙牙菌斑中光活性化合物（或光敏剂）的渗透和有效性，以预防，控制和/或治疗牙周炎。要测试的假设是：a）甲基蓝（MB）的聚合物纳米颗粒可以完全穿透从天然人类牙菌斑体外进化而成的口服多物种生物膜，b）在MB-laind lage and MB-flage proff ins placts plactm plactm andm proffro innan nananoptime之后，可能会完全消极地剥夺这些微生物生物膜。该R21建议的具体目的是：1。准备和表征MB负载的生物相容性和可生物降解的聚合物纳米颗粒。我们将（i）用阴离子和正面电荷准备MB封闭的纳米颗粒，（ii）通过Zetapals仪器评估其大小，表面电荷和形态，并扫描电子显微镜（III）（iii）验证其对MB逐渐释放的能力和效率（IIV）的能力和（IIV），并验证MB的环境（IIV），并验证MB的环境，IIV和IIV的范围内的量代理可用于有效的光动力疗法。 2。研究生物膜中载有MB的纳米颗粒的分布及其在体外生物膜微生物上的光毒性。我们（i）使用唾液和人牙斑制定多种物种实验室实验室生物膜，（ii）（ii）通过细菌使用传输电子显微镜（iii）评估其通过公共扫描激光显微镜对生物膜的渗透（III）通过（III）评估其在生物膜中的渗透（III）来研究MB负载的纳米颗粒的内在化，并评估其（IV计数，使用DNA探针和PCR的物种鉴定。 公共卫生相关性：该项目是使用带有光活性药物的可见光和微观颗粒靶向牙齿斑块细菌的尝试。我们的研究可能有助于开发光动力疗法作为一种用于预防，控制和/或治疗牙周疾病的非侵入性技术。

项目成果

Photodynamic effects of methylene blue-loaded polymeric nanoparticles on dental plaque bacteria.

• DOI: 10.1002/lsm.21069
• 发表时间: 2011-09
• 期刊: LASERS IN SURGERY AND MEDICINE
• 影响因子: 2.4
• 作者: Klepac-Ceraj, Vanja;Patel, Niraj;Song, Xiaoqing;Holewa, Colleen;Patel, Chitrang;Kent, Ralph;Amiji, Mansoor M.;Soukos, Nikolaos S.
• 通讯作者: Soukos, Nikolaos S.