Surgical Infection Research Group, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, NSW, Australia.
Central Military Laboratories and Blood Bank, Prince Sultan Military Medical City, Riyadh, Saudi Arabia.
Int Wound J. 2017 Dec;14(6):1160-1169. doi: 10.1111/iwj.12777. Epub 2017 Jun 23.
Increasing evidence within the literature has identified the presence of biofilms in chronic wounds and proposed that they contribute to delayed wound healing. This research aimed to investigate the presence of biofilm in diabetic foot ulcers (DFUs) using microscopy and molecular approaches and define if these are predominantly mono- or multi-species. Secondary objectives were to correlate wound observations against microscopy results in ascertaining if clinical cues are useful in detecting wound biofilm. DFU tissue specimens were obtained from 65 subjects. Scanning electron microscopy (SEM) and peptide nucleic acid fluorescent in situ hybridisation (PNA-FISH) techniques with confocal laser scanning microscopy (CLSM) were used to visualise biofilm structures. Next-generation DNA sequencing was performed to explore the microbial diversity. Clinical cues that included the presence of slough, excessive exudate, a gel material on the wound bed that reforms quickly following debridement, poor granulation and pyocyanin were correlated to microscopy results. Of the 65 DFU specimens evaluated by microscopy, all were characterised as containing biofilm (100%, P < 0·001). The presence of both mono-species and multi-species biofilms within the same tissue sections were detected, even when DNA sequencing analysis of DFU specimens revealed diverse polymicrobial communities. No clinical correlations were identified to aid clinicians in identifying wound biofilm. Microscopy visualisation, when combined with molecular approaches, confirms biofilms are ubiquitous in DFUs and form either mono- or multi-species biofilms. Clinical cues to aid clinicians in detecting wound biofilm are not accurate for use in DFUs. A paradigm shift of managing DFUs needs to consider anti-biofilm strategies.
越来越多的文献证据表明，生物膜存在于慢性伤口中，并提出它们会导致伤口愈合延迟。本研究旨在通过显微镜和分子方法来研究糖尿病足溃疡（DFU）中生物膜的存在，并确定它们主要是单种还是多种生物膜。次要目标是将伤口观察结果与显微镜结果相关联，以确定临床线索是否有助于检测伤口生物膜。从 65 名患者中获得 DFU 组织标本。使用扫描电子显微镜（SEM）和肽核酸荧光原位杂交（PNA-FISH）技术结合共聚焦激光扫描显微镜（CLSM）来观察生物膜结构。进行下一代 DNA 测序以探索微生物多样性。临床线索包括有坏死组织、过多渗出物、在清创后迅速重新形成的伤口床上凝胶状物质、肉芽组织不良和绿脓菌素，与显微镜结果相关联。在通过显微镜评估的 65 个 DFU 标本中，所有标本均被描述为含有生物膜（100%，P < 0·001）。即使对 DFU 标本的 DNA 测序分析显示出多种多微生物群落，也检测到同一组织切片中存在单种和多种生物膜。未发现有助于临床医生识别伤口生物膜的临床相关性。显微镜可视化，结合分子方法，证实生物膜在 DFU 中无处不在，并形成单种或多种生物膜。有助于临床医生检测伤口生物膜的临床线索在 DFU 中并不准确。管理 DFU 需要考虑抗生物膜策略，这需要范式转变。
