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过氧化氢光解产生的羟基自由基对变形链球菌生物膜的抗菌活性。

Antimicrobial activity of hydroxyl radicals generated by hydrogen peroxide photolysis against Streptococcus mutans biofilm.

机构信息

Laboratory for Redox Regulation, Tohoku University Graduate School of Dentistry, 4-1 Seiryo, Aoba-ku, Sendai 980-8575, Japan.

Laboratory for Redox Regulation, Tohoku University Graduate School of Dentistry, 4-1 Seiryo, Aoba-ku, Sendai 980-8575, Japan.

出版信息

Int J Antimicrob Agents. 2016 Oct;48(4):373-80. doi: 10.1016/j.ijantimicag.2016.06.007. Epub 2016 Jul 15.

DOI:10.1016/j.ijantimicag.2016.06.007
PMID:27449541
Abstract

Prevention of dental caries with maximum conservation of intact tooth substance remains a challenge in dentistry. The present study aimed to evaluate the antimicrobial effect of H2O2 photolysis on Streptococcus mutans biofilm, which may be a novel antimicrobial chemotherapy for treating caries. S. mutans biofilm was grown on disk-shaped hydroxyapatite specimens. After 1-24 h of incubation, growth was assessed by confocal laser scanning microscopy and viable bacterial counting. Resistance to antibiotics (amoxicillin and erythromycin) was evaluated by comparing bactericidal effects on the biofilm with those on planktonic bacteria. To evaluate the effect of the antimicrobial technique, the biofilm was immersed in 3% H2O2 and was irradiated with an LED at 365 nm for 1 min. Viable bacterial counts in the biofilm were determined by colony counting. The thickness and surface coverage of S. mutans biofilm increased with time, whereas viable bacterial counts plateaued after 6 h. When 12- and 24-h-old biofilms were treated with the minimum concentration of antibiotics that killed viable planktonic bacteria with 3 log reduction, their viable counts were not significantly decreased, suggesting the biofilm acquired antibiotic resistance by increasing its thickness. By contrast, hydroxyl radicals generated by photolysis of 3% H2O2 effectively killed S. mutans in 24-h-old biofilm, with greater than 5 log reduction. The technique based on H2O2 photolysis is a potentially powerful adjunctive antimicrobial chemotherapy for caries treatment.

摘要

最大限度地保留完整牙体组织预防龋齿仍然是口腔医学的一个挑战。本研究旨在评估 H2O2 光解对变形链球菌生物膜的抗菌效果,这可能是一种治疗龋齿的新型抗菌化疗方法。将变形链球菌生物膜种植在圆盘状羟磷灰石标本上。孵育 1-24 小时后,通过共聚焦激光扫描显微镜和活菌计数评估生长情况。通过比较生物膜和浮游菌的杀菌效果来评估抗生素(阿莫西林和红霉素)的耐药性。为了评估抗菌技术的效果,将生物膜浸泡在 3%的 H2O2 中,并在 365nm 处用 LED 照射 1 分钟。通过平板计数法确定生物膜中的活菌数。生物膜的厚度和表面覆盖率随时间增加,而活菌计数在 6 小时后达到平台期。当用能杀死浮游菌并使活菌减少 3 个对数级的最低抗生素浓度处理 12 小时和 24 小时的生物膜时,其活菌数没有明显减少,表明生物膜通过增加厚度获得了抗生素耐药性。相比之下,3%H2O2 光解产生的羟基自由基能有效杀灭 24 小时龄生物膜中的变形链球菌,减少超过 5 个对数级。基于 H2O2 光解的技术可能是一种治疗龋齿的强大辅助性抗菌化疗方法。

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