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Carbohydrate biofunctionalized nanomaterials as antiadhesion and detection agents for bacterial pathogens.

机译：碳水化合物生物功能化的纳米材料作为细菌病原体的抗粘附和检测剂。

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摘要

The increasing incidences of multiple-drug-resistant bacteria have made the search for alternatives to conventional antibiotic treatment a high priority. Both glycotechnology and nanotechnology hold promise for antimicrobial treatment and pathogen detection. Many pathogenic microbes initiate diseases through carbohydrate protein interactions. This knowledge, along with the advances in chemical glycobiology, provides a scientific basis for new approaches using carbohydrates for antiadhesion therapy and pathogen detection. Nanosized materials also have the potential to be employed in antimicrobial applications and pathogen detection using their inherent physical and chemical properties. In addition, they serve as excellent platforms to link biomolecules.;This dissertation researches the antiadhesion and pathogen detection capabilities of carbohydrate-biofunctionalized nanomaterials (nanoparticles and nanotubes) as determined both qualitatively and quantitatively in vitro. First, nanoparticles were tested with three strains of Escherichia coli (ORN178, 208 and O157:H7) expressing different carbohydrate binding specificities. Microscopic studies showed the agglutination of bacteria mediated by nanoparticles of specific carbohydrate biofunctionalities. Cross reactions were observed perhaps suggesting that bacteria can express both a primary and a minor adhesin. Colony forming unit (CFU) assay, utilized to quantify the magnitude of bacterial agglutination, showed a 99% CFU reduction after E. coli ORN178 was incubated with mannose biofunctionalized nanoparticles (Man-NPs). Second, Man-NPs were demonstrated as inhibitors in preventing the adhesion of Campylobacter jejuni to Caco-2, a human colon cancer cell line. Third, single-walled nanotubes functionalized with two configurations of galactose (Gal-SWNT and Ga12-SWNT) were explored for their antiadhesion and detection capabilities against enterohaemorrhagic E. coli (EHEC) O157:H7. The divalent Gal2-SWNT was proved to be a stronger ligand in binding both at the molecular level (with a lectin through a turbidimetric assay) and at the cellular level (with bacteria through a precipitation assay, a fluorescence microscopy and a CFU assay). The fact that a small amount (25 μL) of galactose SWNT could capture ∼10 4 bacterial cells supports its use in pathogen detection. Fourth, a carbohydrate array was fabricated using various probes including different carbohydrate nanoparticles to capture/detect pathogens. In conclusion, the results of this research indicate that carbohydrate biofunctionalized nanomaterials have significant potential as agents for both antiadhesion therapy and pathogen detection.
机译：多重耐药细菌的发病率不断上升，因此寻找传统抗生素治疗替代品成为当务之急。糖技术和纳米技术都有望用于抗菌治疗和病原体检测。许多病原微生物通过碳水化合物蛋白质相互作用引发疾病。这些知识以及化学糖生物学的进步为使用碳水化合物进行抗粘连治疗和病原体检测的新方法提供了科学依据。纳米材料还具有利用其固有的物理和化学特性在抗菌应用和病原体检测中应用的潜力。此外，它们还为连接生物分子提供了极好的平台。本论文研究了在体外定性和定量测定的碳水化合物生物功能化纳米材料（纳米颗粒和纳米管）的抗粘附和病原体检测能力。首先，用三种表达不同碳水化合物结合特异性的大肠杆菌菌株（ORN178、208和O157：H7）测试纳米颗粒。显微镜研究显示由特定碳水化合物生物功能的纳米粒子介导的细菌凝集。观察到交叉反应，可能表明细菌可以表达初级和次级粘附素。用于量化细菌凝集程度的菌落形成单位（CFU）分析显示，在将大肠杆菌ORN178与甘露糖生物功能化纳米颗粒（Man-NPs）孵育后，CFU降低了99％。其次，Man-NPs被证明是防止空肠弯曲杆菌粘附至人结肠癌细胞系Caco-2的抑制剂。第三，研究了用两种构型的半乳糖（Gal-SWNT和Ga12-SWNT）功能化的单壁纳米管对肠出血性大肠杆菌（EHEC）O157：H7的抗粘附和检测能力。在分子水平（通过比浊法测定与凝集素）和细胞水平（通过沉淀法，荧光显微镜和CFU测定与细菌结合），二价Gal2-SWNT被证明是更强的配体结合力。少量（25μL）的半乳糖SWNT可以捕获约10 4个细菌细胞这一事实支持了其在病原体检测中的用途。第四，使用包括不同碳水化合物纳米颗粒的各种探针制造碳水化合物阵列以捕获/检测病原体。总之，这项研究的结果表明，碳水化合物生物功能化的纳米材料具有作为抗粘连治疗和病原体检测的重要潜力。

著录项

  • 作者

    Luo, Pengju (George).;

  • 作者单位

    Clemson University.;

  • 授予单位 Clemson University.;
  • 学科 Biology Microbiology.
  • 学位 Ph.D.
  • 年度 2006
  • 页码 122 p.
  • 总页数 122
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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