Desrosiers Martin, Bendouah Zohra, Barbeau Jean
Department of Otolaryngology-Head and Neck Surgery, University of Montreal, Montreal, Canada.
Am J Rhinol. 2007 Mar-Apr;21(2):149-53. doi: 10.2500/ajr.2007.21.3007.
In vitro biofilm-producing capacity in isolates of Staphylococcus aureus and Pseudomonas aeruginosa collected from the sinus cavities after endoscopic sinus surgery (ESS) are associated with a poor outcome in patients with chronic rhinosinusitis (CRS). However, conventional oral antibiotic therapy is frequently ineffective in eradicating bacteria in the biofilm form. Increasing the concentration of antibiotics may offer a means of countering this resistance. The aim of this study was to determine the in vitro activity of moxifloxacin (MOXI) against S. aureus in biofilm form (recovered from patients with CRS at least 1 year post-ESS).
This study was performed in a research microbiology laboratory, where five isolates of S. aureus with known biofilm-forming capacity were cultured in Tryptic Soy Broth 0.5% glucose in 96-well plates at 37 degrees C for 24 hours. After visual confirmation of biofilm formation, plates were incubated in phosphate-buffered saline (PBS) or with MOXI at concentrations of 0.1x, Ix, 100x, and 1000x minimal inhibitory concentration (MIC) for an additional 24 hours. Biofilm from 3 wells of each concentration were collected and sonicated and the number of viable bacteria was determined by serial dilution and plating.
After incubation, the number of viable bacteria was similar for nontreated and MOXI-treated biofilms at MIC and sub-MIC levels. However, MOXI at 1000x (0.1-0.2 mg/mL) gave a 2 to 2.5 log reduction in number of viable bacteria.
In vitro results show that increased concentrations of antibiotics, easily attainable in topical solutions, are effective in killing bacteria in bacterial biofilms. This suggests a role for topical antibiotic therapies in the treatment of biofilm infections.
从内镜鼻窦手术(ESS)后鼻窦腔收集的金黄色葡萄球菌和铜绿假单胞菌分离株的体外生物膜形成能力与慢性鼻窦炎(CRS)患者的不良预后相关。然而,传统的口服抗生素治疗在根除生物膜形式的细菌方面常常无效。增加抗生素浓度可能提供一种对抗这种耐药性的方法。本研究的目的是确定莫西沙星(MOXI)对生物膜形式的金黄色葡萄球菌(从ESS后至少1年的CRS患者中分离得到)的体外活性。
本研究在一个研究微生物实验室进行,将五株已知具有生物膜形成能力的金黄色葡萄球菌分离株在含有0.5%葡萄糖的胰蛋白胨大豆肉汤中于96孔板中在37℃培养24小时。在肉眼确认生物膜形成后,将平板在磷酸盐缓冲盐水(PBS)中或与浓度为0.1倍、1倍、100倍和1000倍最小抑菌浓度(MIC)的MOXI一起再孵育24小时。收集每个浓度3孔的生物膜并进行超声处理,通过连续稀释和平板接种确定活菌数量。
孵育后,在MIC和亚MIC水平,未处理和MOXI处理的生物膜中的活菌数量相似。然而,1000倍(0.1 - 0.)的MOXI使活菌数量减少了2至2.5个对数级。
体外结果表明,在局部溶液中容易达到的抗生素浓度增加对杀死细菌生物膜中的细菌有效。这表明局部抗生素疗法在生物膜感染治疗中具有作用。