New Adapted In Vitro Technology to Evaluate Biofilm Formation and Antibiotic Activity Using Live Imaging under Flow Conditions.
New Adapted In Vitro Technology to Evaluate Biofilm Formation and Antibiotic Activity Using Live Imaging under Flow Conditions.
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DOI：
10.3390/diagnostics11101746
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发表时间：
2021-09-23
期刊：
影响因子：
--
通讯作者：
Lavigne JP
中科院分区：
文献类型：
--
作者：
Pouget C;Dunyach-Remy C;Pantel A;Schuldiner S;Sotto A;Lavigne JP
生物膜的多微生物性质和慢性伤口内的细菌相互作用是理解细菌合作的关键。本研究的目的是开发一种在流动条件下研究和可视化实时成像中的生物膜的技术（Bioflux™ 200，Fluxion Biosciences）。 BiofluxTM 系统采用体外慢性伤口样介质 (CWM) 进行改造，模拟溃疡中遇到的环境。将金黄色葡萄球菌 (Newman) 和铜绿假单胞菌 (PAO1) 的两种参考菌株在 CWM 与对照培养基 (BHI) 中进行单一培养和共培养（比例 1:1，同时添加细菌）24 小时至 72 小时期间注射到 BiofluxTM 中。通过倒置显微镜评估每次生物膜形成的量化。 72小时后，在模拟伤口表面清创的流量自动增加后，将1x MIC、10x MIC和100x MIC的不同抗生素（头孢他啶、亚胺培南、利奈唑胺、苯唑西林和万古霉素）施用到系统中。生物膜研究强调，与 BHI 培养基相比，这两个物种（单独或联合）在 CWM 中构成了更快、更厚的生物膜。抗生素对成熟或“清创”生物膜的影响表明，一些临床上最常用的抗生素（如万古霉素或亚胺培南）无法破坏和减少生物膜生物量。使用体外 CWM 进行生命细胞成像是研究细菌生物膜和研究慢性伤口环境中微生物合作的一种有前景的工具。
The polymicrobial nature of biofilms and bacterial interactions inside chronic wounds are keys for the understanding of bacterial cooperation. The aim of this present study was to develop a technique to study and visualize biofilm in live imaging under flow conditions (Bioflux™ 200, Fluxion Biosciences). The BiofluxTM system was adapted using an in vitro chronic wound-like medium (CWM) that mimics the environment encountered in ulcers. Two reference strains of Staphylococcus aureus (Newman) and Pseudomonas aeruginosa (PAO1) were injected in the BiofluxTM during 24 h to 72 h in mono and coculture (ratio 1:1, bacteria added simultaneously) in the CWM vs. a control medium (BHI). The quantification of biofilm formation at each time was evaluated by inverted microscopy. After 72 h, different antibiotics (ceftazidime, imipenem, linezolid, oxacillin and vancomycin) at 1x MIC, 10x MIC and 100x MIC were administrated to the system after an automatic increase of the flow that mimicked a debridement of the wound surface. Biofilm studies highlighted that the two species, alone or associated, constituted a faster and thicker biofilm in the CWM compared to the BHI medium. The effect of antibiotics on mature or “debrided” biofilm indicated that some of the most clinically used antibiotic such as vancomycin or imipenem were not able to disrupt and reduce the biofilm biomass. The use of a life cell imaging with an in vitro CWM represents a promising tool to study bacterial biofilm and investigate microbial cooperation in a chronic wound context.
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影响因子：
4.9
作者：
Bowler, Laura L.;Zhanel, George G.;Saward, Laura L.
通讯作者：
Saward, Laura L.
DOI：
10.1099/00221287-6-1-2-95
发表时间：
1952-01-01
期刊：
JOURNAL OF GENERAL MICROBIOLOGY
影响因子：
--
作者：
DUTHIE, ES;LORENZ, LL
通讯作者：
LORENZ, LL
影响因子：
5.2
作者：
Gander, S;Hayward, K;Finch, R
通讯作者：
Finch, R
DOI：
10.1007/978-1-4939-9118-1_16
发表时间：
2019-01-01
期刊：
ACINETOBACTER BAUMANNII
影响因子：
--
作者：
Naudin, Bertrand;Heins, Anja;Nicol, Marion
通讯作者：
Nicol, Marion
影响因子：
3.8
作者：
Mirani, Zulfigar Ali;Aziz, Mubashir;Khan, Seema Ismat
通讯作者：
Khan, Seema Ismat