Synchrotron imaging of crystalline biofilms in urinary catheters

导尿管中结晶生物膜的同步加速器成像

• 批准号: 7849922
• 负责人: AARON I PACKMAN
• 金额: $ 18.48万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7849922
• 关键词: Acute; Address; Affect; Antibiotic Therapy; Antibiotics; Antimicrobial Effect; Biocide; Biological Models; Bladder; Calcium; Calculi; Catheters; Cells; Chemical Structure; Chemicals; Chronic; Complex; Confocal Microscopy; Coupled; Data; Deposition; Development; Devices; Dimensions; Disease; Drainage procedure; Effectiveness; Electron Microscopy; Electrons; Environment; Evaluation; Evolution; Failure; Foundations; Funding; Growth; Hydrolysis; Image; Image Analysis; In Situ; In Vitro; Infection; Infection Control; Infection prevention; Investigation; Kidney; Laboratories; Long-Term Care; Magnesium; Mechanics; Methodology; Methods; Microbial Biofilms; Minerals; Mirabilis; Modeling; Monitor; Morbidity - disease rate; Nutrient; Optical Methods; Organism; Patients; Pattern; Performance; Phase; Phenotype; Precipitation; Process; Property; Proteus; Proteus Infections; Proteus mirabilis; Research; Research Project Grants; Resolution; Role; Sampling; Scanning Electron Microscopy; Science; Solid; Spectrometry; Spectrum Analysis; Stents; Structure; Surface; Symptoms; Synchrotrons; System; Testing; Time; Treatment Efficacy; Ultrasonics; Uncertainty; United States National Institutes of Health; Universities; Urea; Urinary Calculi; Urinary Catheterization; Urinary tract; Urinary tract infection; Urine; Virulence; Virulence Factors; Work; analytical method; base; catheter associated UTI; cell motility; fluid flow; improved; in vivo; innovation; inorganic phosphate; method development; microbial; mortality; mucoid; novel strategies; pathogen; pharmacokinetic model; prevent; prophylactic; public health relevance; simulation; solute; therapy development; tomography; urinary; Acute; Address; Affect; Antibiotic Therapy; Antibiotics; Antimicrobial Effect; Biocide; Biological Models; Bladder; Calcium; Calculi; Catheters; Cells; Chemical Structure; Chemicals; Chronic; Complex; Confocal Microscopy; Coupled; Data; Deposition; Development; Devices; Dimensions; Disease; Drainage procedure; Effectiveness; Electron Microscopy; Electrons; Environment; Evaluation; Evolution; Failure; Foundations; Funding; Growth; Hydrolysis; Image; Image Analysis; In Situ; In Vitro; Infection; Infection Control; Infection prevention; Investigation; Kidney; Laboratories; Long-Term Care; Magnesium; Mechanics; Methodology; Methods; Microbial Biofilms; Minerals; Mirabilis; Modeling; Monitor; Morbidity - disease rate; Nutrient; Optical Methods; Organism; Patients; Pattern; Performance; Phase; Phenotype; Precipitation; Process; Property; Proteus; Proteus Infections; Proteus mirabilis; Research; Research Project Grants; Resolution; Role; Sampling; Scanning Electron Microscopy; Science; Solid; Spectrometry; Spectrum Analysis; Stents; Structure; Surface; Symptoms; Synchrotrons; System; Testing; Time; Treatment Efficacy; Ultrasonics; Uncertainty; United States National Institutes of Health; Universities; Urea; Urinary Calculi; Urinary Catheterization; Urinary tract; Urinary tract infection; Urine; Virulence; Virulence Factors; Work; analytical method; base; catheter associated UTI; cell motility; fluid flow; improved; in vivo; innovation; inorganic phosphate; method development; microbial; mortality; mucoid; novel strategies; pathogen; pharmacokinetic model; prevent; prophylactic; public health relevance; simulation; solute; therapy development; tomography; urinary

DESCRIPTION (provided by applicant): A suite of synchrotron-based x-ray imaging methodologies will be used to assess the development of crystalline biofilms formed by Proteus mirabilis in urinary catheters. Proteus is an extremely common pathogen in urinary tract infections, and is particularly problematic in patients subject to long-term urinary catheterization. P. mirabilis hydrolyzes urea, leading to an increase in the pH of urine and the formation of mineral deposits within the urinary tract. These deposits form stones and encrustations in the bladder and kidneys, as well as encrustations on inserted devices such as catheters and ureteral stents. Approximately half of patients with long-term urinary catheters develop catheter blockage as a result of these infections. Blockage incidents can cause serious acute symptoms and are a contributing factor to morbidity and mortality in catheterized patients. The need to monitor and replace urinary catheters also presents a substantial burden for long-term management of these patients. One major challenge in understanding the development of crystalline biofilms and the associated catheter blockage is that the mineral deposits prevent use of optical methods such as confocal microscopy. To overcome this limitation, Proteus biofilms within urinary catheters will be imaged using sophisticated synchrotron-based methods that we have recently developed for non-invasive investigations of mineral samples. Synchrotron x-ray computed micro-tomography will be used to image the pore structure of Proteus biofilms in three dimensions with near-micron-scale resolution, and synchrotron x- ray diffraction will also be used to assess the distribution of different mineral phases within the biofilms. In addition, environmental scanning electron microscopy will be used to assess cell distributions within biofilms, and analytical electron microscopy will be performed using energy-dispersive spectrometry and electron diffraction to determine chemical distributions within the crystalline deposits. Analyses will be performed on intact catheters containing crystalline biofilms grown in vitro using a model bladder system, as well as on blocked catheters retrieved from patients. Quantitative descriptions of the biofilm pore structure will be developed using image analysis methods, and the internal transport environment within each biofilm will be evaluated by using the tomographic data in conjunction with lattice Boltzmann simulations of pore fluid flow. Results obtained for different Proteus strains and different growth durations will be compared in order to assess the interplay of pore structure, internal transport conditions, and crystalline biofilm development, as well as the overall effects of biofilm growth on catheter blockage. PUBLIC HEALTH RELEVANCE: Catheter blockage from Proteus infections still represents a major, ongoing, and serious problem for patients and for long-term care facilities. A lack of understanding of the structure and composition of crystalline biofilms has hindered the development of new treatments to prevent catheter infections and control catheter blockage. The project results will indicate how the crystalline structure influences the development of Proteus biofilms on urinary catheters and suggest improved strategies for treating these infections and for preventing catheter blockage.

描述（由申请人提供）：将使用一套基于同步加速器的X射线成像方法来评估尿液导管中Proteus mirabilis形成的晶体生物膜的发展。 Proteus是尿路感染中极为常见的病原体，在长期尿管插入术的患者中尤其有问题。 Mirabilis水解尿素，导致尿液的pH升高和尿路内矿物沉积物的形成。这些沉积物在膀胱和肾脏中形成石头和掩盖，以及在插入的设备（例如导管和输尿管支架）上的包裹。由于这些感染，大约一半的长期尿导管患者会出现导管阻塞。堵塞事件可能导致严重的急性症状，是导管患者发病率和死亡率的一个因素。监测和替换尿导管的需求还为这些患者的长期管理带来了巨大的负担。了解晶体生物膜的发展和相关导管阻塞的一个主要挑战是，矿物沉积物阻止使用光学方法，例如共聚焦显微镜。为了克服这一限制，将使用基于复杂的同步加速器的方法对尿导管中的Proteus生物膜进行成像，我们最近开发了用于矿物样品的非侵入性研究。同步加速器X射线计算的微型摄影将用于成像三个维度的Proteus Biofilms的孔结构，并以近米尺度的分辨率成像，并且同步X-射线X-射线衍射还将用于评估生物膜内不同矿物相的分布。此外，环境扫描电子显微镜将用于评估生物膜内的细胞分布，并将使用能量分散性光谱法和电子衍射来确定晶体沉积物中的化学分布。分析将对使用模型膀胱系统以及从患者检索到的封闭的导管中在体外生长的结晶生物膜的完整导管进行分析。将使用图像分析方法开发生物膜孔结构的定量描述，并将每个生物膜内的内部传输环境与孔流的晶格Boltzmann模拟一起评估每个生物膜内的内部传输环境。为了评估孔结构，内部运输条件和晶体生物膜发育的相互作用，将比较不同蛋白质菌株和不同生长持续时间的结果，以及生物膜生长对导管阻滞的总体影响。公共卫生相关性：从Proteus感染中的导管阻塞仍然代表了患者和长期护理设施的重大，持续且严重的问题。缺乏对晶体生物膜的结构和组成的了解，阻碍了新处理的发展，以防止导管感染和控制导管阻塞。该项目的结果将表明晶体结构如何影响Proteus生物膜在尿导管上的发展，并提出改进的治疗这些感染和预防导管阻塞的策略。