A human nasal explant model to study Staphylococcus aureus biofilm in vitro
A human nasal explant model to study Staphylococcus aureus biofilm in vitro
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DOI：
10.1002/alr.21146
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发表时间：
2013-07-01
影响因子：
6.4
通讯作者：
Vreugde, Sarah
中科院分区：
文献类型：
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作者：
Cantero, Daniel;Cooksley, Clare;Vreugde, Sarah
背景金黄色葡萄球菌（S.金黄色葡萄球菌）生物膜与严重和顽固的慢性鼻窦炎（CRS）病例相关。然而，它在这种情况下的病理生理学的作用并不完全清楚。本研究旨在建立一种鼻腔鼻窦组织移植模型，以分析S。方法取5例垂体手术患者的鼻腔鼻窦组织标本，分别进行含金黄色葡萄球菌和不含金黄色葡萄球菌的培养。金黄色葡萄球菌生物膜的体外培养。在生物膜激发24小时后，对组织外植体进行使用Live/Dead BacLight染色的共聚焦扫描激光显微镜（CSLM）和组织学检查。IL-6的测量，在信使RNA（mRNA）水平（使用定量逆转录聚合酶链反应[qRT-PCR]）和蛋白质水平（使用酶联免疫吸附试验[ELISA]），进行评估生物膜粘膜interaction.Results的外植体的活力后24小时，证实了CSLM和组织学。虽然光学显微镜未能鉴定S。在金黄色葡萄球菌生物膜中，通过CSLM证实其存在于生物膜挑战的样品中。与对照组相比，生物膜处理组织中IL-6 mRNA转录水平上调4.9倍（p = 0.0485）。结论鼻腔鼻窦组织块是一种可行的功能性模型，能够分析生物膜-粘膜的直接相互作用，有助于我们进一步了解S.金黄色葡萄球菌生物膜在鼻窦炎中的作用。我们的模型表明，S。生长初始阶段的金黄色葡萄球菌生物膜不是惰性旁观者，而是在鼻窦粘膜中引发免疫应答。(C)2013 ARS-AAOA，LLC.
Background Staphylococcus aureus (S. aureus) biofilm has been associated with severe and recalcitrant cases of chronic rhinosinusitis (CRS). However, its role in the pathophysiology of this condition is not completely understood. This study aims to develop a sinonasal tissue explant model to analyze the interaction of S. aureus biofilm with the mucosa in vitro.Methods Sinonasal tissue samples from 5 control patients undergoing pituitary surgery were cultured with and without S. aureus biofilm in vitro. Confocal scanning laser microscopy (CSLM) using the Live/Dead BacLight stain and histology were performed on the tissue explants after 24hours of biofilm challenge. Measurements of IL-6, at both the messenger RNA (mRNA) level (using quantitative reverse-transcriptase polymerase chain reaction [qRT-PCR]) and the protein level (using enzyme-linked immunosorbent assay [ELISA]), were undertaken to evaluate biofilm-mucosa interaction.Results Viability of the explants after 24hours was confirmed by CSLM and histology. Although light microscopy failed to identify S. aureus biofilms, its presence was confirmed in the biofilm-challenged samples by CSLM. IL-6 mRNA transcript levels were 4.9-fold upregulated in biofilm-treated tissue compared to controls (p = 0.0485). A similar trend was observed at the protein level (p = 0.0313).Conclusion The sinonasal tissue explant is a viable and functional model capable of analyzing direct biofilm-mucosal interactions and can advance our understanding of the role played by S. aureus biofilm in sinus inflammation. Our model suggests that S. aureus biofilms in the initial phase of growth are not inert bystanders but elicit an immune response in the sinonasal mucosa. (C) 2013 ARS-AAOA, LLC.