Investigating phenotypic changes in wound biofilms in response to antimicrobial treatment using Raman Spectroscopy.

使用拉曼光谱研究伤口生物膜对抗菌治疗的表型变化。

• 批准号: 2902109
• 金额: --
• 依托单位: University of Strathclyde
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2902109
• 关键词: Investigating; phenotypic; changes; wound; biofilms

Wound infections such as diabetic foot ulcers (DFUs) can be highly problematic in terms of clinical management. Such infections carry a significant socioeconomic burden, costing the NHS an estimated £8.3b in 2017/18. DFUs have significant mortality rates, with 5-year mortality rates comparable with cancer. However, worryingly this is not reflected in the level of funding for chronic wound compared to cancer research.The main difficulty that clinicians face during the treatment of chronic wounds is the risk of infection at the wound site, arising from the accumulation of microbial aggregates in the wound bed. Infected wounds arise from the formation of complex biofilms that can be largely resistant to antibiotic therapy: most of which are polymicrobial in nature . This further complicates treatment regimes to alleviate specific pathogenic microorganisms within the mixed community. Given the increased complexity of the wound microbiome as well as the increasing threat of antimicrobial resistance, clinicians are faced with the challenging prospect of targeted topical therapy or systemic antibiotics. This can be extremely difficult as point-of-care technologies in wound care are limited with regards to microbial detection, with clinicians relying on basic culture or molecular methodologies that can take days for results. Thus there remains a requirement for rapid, sensitive, yet non-invasive methodologies for microbial identification at the bedside, to aid clinical therapy choices during wound management.Raman is an information rich vibration technique that gives label-free biochemical information at the sub-cellular level, however high-resolution imaging at a single cell level is a relatively slow process making it challenging for live cell imaging. Stimulated Raman scattering (SRS) is a more sensitive and much faster technique which can provide high resolution information on larger cell populations compared with normal Raman. SRS provides detailed biochemical information with sub-cellular resolution including determination of protein, lipids and nucleic acid location. Due to its speed and ability to image larger areas in a label free manner, it has great potential for imaging bacterial biofilms giving both biochemical information as well as structural information on biofilm formation, as well as changes to biofilm in response to antimicrobial treatments. To date there is evidence that different bacterial species exhibit unique spectral profiles depending on their chemical structures at a cellular level. Existing research is largely limited to single cells, with few studies investigating biofilm kinetics using SRS.The overall aims of the proposal are to test the feasibility of using SRS to monitor biofilm formation and assess unique Raman spectral profiles or signatures of different bacterial and fungal species related to wound infections.1 Monitoring growth dynamics of mono-species wound biofilm models using SRS - comparing spectral profiles in different clinically-relevant microbial species (gram+, gram- bacterial and fungal organisms) during biofilm attachment, maturation and dispersal.2 Investigating mono-species biofilm response to antibiotic therapy using a variety of microbiological, molecular, microscopic and spectroscopic techniques - SRS to be used to assess chemical signatures in the biofilms in response to bacterio- (or fungi-) static or -cidal antibiotics and other novel compounds.3 Assessing changes in biofilm formation in a multi-species biofilm model using SRS. Visualizing phenotypic changes in biofilm chemical structures containing a consortia of different microbial species.4 SRS to be used to profile real-world biofilms (those generated from clinical samples from wound infections which will be generated through an existing EPSRC grant EP/V005839/1). Characteristic changes in chemical structures identified by Raman Spectroscopy using simple biofilm models with known composition.

糖尿病足溃疡 (DFU) 等伤口感染在临床治疗方面可能会带来严重的问题，这种感染会带来重大的社会经济负担，2017/2018 年 NHS 的损失估计为 8.3 英镑，并且 DFU 的死亡率很高。年死亡率与癌症相当。然而，令人担忧的是，与癌症研究相比，这并没有反映在慢性伤口的资助水平上。农民在治疗慢性伤口过程中面临的主要困难是伤口部位的感染风险，感染伤口是由于复杂的生物膜的形成而产生的，这些生物膜在很大程度上对抗生素治疗具有抵抗力：其中大多数本质上是多种微生物，这使得减轻伤口内特定病原微生物的治疗方案变得更加复杂。鉴于伤口微生物群的复杂性增加以及抗菌素耐药性的威胁日益增加，牧师面临着局部治疗或全身抗生素的挑战性前景，因为这可能非常困难。伤口护理中的护理点技术在微生物检测方面受到限制，顾客依赖于基本培养或分子方法，可能需要数天才能得到结果，因此仍然需要快速、灵敏且非侵入性的微生物方法。床边识别，以帮助伤口处理期间的临床治疗选择。拉曼是一种信息丰富的振动技术，可在亚细胞水平提供无标记的生化信息，但是单细胞水平的高分辨率成像是一个相对缓慢的过程这使得活细胞成像具有挑战性。受激拉曼散射 (SRS) 是一种更灵敏、更快速的技术，与普通拉曼相比，它可以提供更大细胞群的高分辨率信息，提供具有亚细胞分辨率的详细生化信息，包括蛋白质、脂质和核酸位置的测定。由于其速度快且能够以无标记的方式对较大区域进行成像，因此它在细菌生物膜成像方面具有巨大的潜力，可提供生物膜形成的生化信息和结构信息，以及生物膜因抗菌治疗而发生的变化。迄今为止，有证据表明，不同的细菌物种根据其在细胞水平上的化学结构表现出独特的光谱特征，现有的研究主要局限于单细胞，很少有研究使用 SRS 来研究生物膜动力学。该提案的总体目标是测试使用 SRS 监测生物膜形成并评估与伤口感染相关的不同细菌和真菌物种的独特拉曼光谱图或特征的可行性。1 使用 SRS 监测单一物种伤口生物膜模型的生长动态 - 比较不同物种的光谱图生物膜附着、成熟和分散过程中的临床相关微生物种类（革兰氏+、革兰氏-细菌和真菌生物体）。2 使用各种微生物、分子、显微和光谱技术研究单种生物膜对抗生素治疗的反应 - 使用 SRS评估生物膜中对细菌（或真菌）静态或杀灭抗生素和其他新型化合物的化学特征。3 评估生物膜形成的变化在使用 SRS 的多物种生物膜模型中，可视化包含不同微生物物种的生物膜化学结构的表型变化。4 SRS 用于分析真实世界的生物膜（通过伤口感染的临床样本生成的生物膜）。现有的 EPSRC 拨款 EP/V005839/1）通过拉曼光谱使用具有已知成分的简单生物膜模型识别化学结构的特征变化。