一种用于评估低频超声辅助伤口清创的新型体外伤口生物膜模型。
A novel in vitro wound biofilm model used to evaluate low-frequency ultrasonic-assisted wound debridement.
作者信息
Crone S, Garde C, Bjarnsholt T, Alhede M
机构信息
Professor, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen N, Denmark.
出版信息
J Wound Care. 2015 Feb;24(2):64, 66-9, 72. doi: 10.12968/jowc.2015.24.2.64.
OBJECTIVE
Bacterial biofilms remain difficult to treat. The biofilm mode of growth enables bacteria to survive antibiotic treatment and the inflammatory reaction. Low-frequency ultrasound has recently been shown to improve healing in a variety of settings. It is hypothesised that ultrasound disrupts the biofilm leaving bacteria more vulnerable to antiseptic or antibiotic treatment. The objective of this study is to develop a realistic model to elucidate the effect of ultrasound on biofilms.
METHOD
A novel in vitro wound biofilm model was developed. Biofilms of Staphylococcus aureus were casted in a semi-solid agar gel composed of either tryptic soy broth (TSB) or a wound simulating media (WSM; composed of Bolton broth with blood and plasma), to resemble the non-surface attached aggregates. The model was used to evaluate the antibiofilm effect of an ultrasonic-assisted wound debridement device (UAW) in the presence of saline irrigation and treatment with a polyhexamethylene biguanide (PHMB)-containing antiseptic. Confocal microscopy was used to evaluate the effect of treatments on biofilm disruption and cell viability counting measured the antibacterial effects.
RESULTS
Confocal microscopy showed that application of 10 seconds of moderate-intensity UAW could effectively disrupt semi-solid biofilms grown on both media settings. This treatment only had a small effect on the cell viability. A 24-hour treatment with PHMB was able to reduce the number of bacteria but not eradicate the biofilm in both media settings. Interestingly, the efficacy of the PHMB antiseptic was significantly higher when applied on biofilms grown in the more complex WSM media. However, we found a significant improvement in reducing the number of viable bacteria grown on both media when applying UAW before administration of the PHMB solution. Applying UAW in the presence of PHMB further improved the efficacy.
CONCLUSION
Using a realistic in vitro biofilm wound model, we show combining UAW with a PHMB-containing antiseptic has potential as an antibiofilm strategy in wound care.
DECLARATION OF INTEREST
The manufacturer of the ultrasonic-assisted wound debridement device, Söring GmbH, Germany, has supported the ultrasound studies. The funding company had no role in the design, data collection, analysis, review, or approval of the manuscript.
目的
细菌生物膜仍然难以治疗。生物膜生长模式使细菌能够在抗生素治疗和炎症反应中存活。最近研究表明,低频超声在多种情况下可促进愈合。据推测,超声可破坏生物膜,使细菌更易受到防腐剂或抗生素治疗的影响。本研究的目的是建立一个逼真的模型,以阐明超声对生物膜的作用。
方法
建立了一种新型的体外伤口生物膜模型。金黄色葡萄球菌生物膜接种于由胰蛋白胨大豆肉汤(TSB)或伤口模拟培养基(WSM;由含血液和血浆的博尔顿肉汤组成)制成的半固体琼脂凝胶中,以模拟非表面附着的聚集体。该模型用于评估在生理盐水冲洗和含聚六亚甲基双胍(PHMB)防腐剂处理的情况下,超声辅助伤口清创装置(UAW)的抗生物膜效果。共聚焦显微镜用于评估处理对生物膜破坏的影响,细胞活力计数用于测量抗菌效果。
结果
共聚焦显微镜显示,应用10秒中等强度的UAW可有效破坏在两种培养基环境中生长的半固体生物膜。这种处理对细胞活力影响较小。在两种培养基环境中,用PHMB处理24小时能够减少细菌数量,但不能根除生物膜。有趣的是,当将PHMB防腐剂应用于在更复杂的WSM培养基中生长的生物膜时,其效果显著更高。然而,我们发现,在应用PHMB溶液之前应用UAW,可显著减少在两种培养基中生长的活菌数量。在PHMB存在的情况下应用UAW可进一步提高疗效。
结论
使用逼真的体外生物膜伤口模型,我们表明将UAW与含PHMB的防腐剂联合使用,在伤口护理中作为一种抗生物膜策略具有潜力。
利益声明
超声辅助伤口清创装置的制造商德国索林有限公司支持了超声研究。资助公司在稿件的设计、数据收集、分析、审查或批准过程中没有任何作用。