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蓝光激活姜黄素处理后变形链球菌活力的流式细胞术评估

Flow cytometric assessment of Streptococcus mutans viability after exposure to blue light-activated curcumin.

作者信息

Manoil Daniel, Filieri Anna, Gameiro Cécile, Lange Norbert, Schrenzel Jacques, Wataha John C, Bouillaguet Serge

机构信息

Endodontics Unit, Section of Dental Medicine, University of Geneva, 19 rue Barthelemy Menn, CH-1205 Geneva, Switzerland.

Flow Cytometry Facility, Faculty of Medecine, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva 4, Switzerland.

出版信息

Photodiagnosis Photodyn Ther. 2014 Sep;11(3):372-9. doi: 10.1016/j.pdpdt.2014.06.003. Epub 2014 Jun 25.

Abstract

BACKGROUND

Streptococcus mutans biofilms are considered as primary causative agents of dental caries. Photodynamic antimicrobial chemotherapy (PACT) has been recently proposed as a strategy for inactivating dental biofilms. This study aimed to investigate the effect of blue light-activated curcumin on S. mutans viability and to explore its potential as a new anti-caries therapeutic agent. The effect of different concentrations and incubation times of photo-activated curcumin on the survival of S. mutans in planktonic and biofilm models of growth was assessed by flow cytometry.

METHODS

Streptococcus mutans in planktonic suspensions or biofilms formed on hydroxyapatite disks were incubated for 5 or 10min with curcumin prior to blue light activation. Bacteria were labeled with SYTO 9 and propidium iodide before viability was assessed by flow cytometry. Results were statistically analyzed using one-way ANOVA and Tukey multiple comparison intervals (α=0.05).

RESULTS

For planktonic cultures, 0.2μM of light-activated curcumin significantly reduced S. mutans viability (p<0.05). For biofilm cultures, light-activated curcumin at concentration of 40-60μM only suppressed viability by 50% (p<0.05). Independently of the mode of growth, incubation time has no significant effect on PACT efficiency.

CONCLUSION

This study indicates that blue light-activated curcumin can efficiently inactivate planktonic cultures of S. mutans whereas biofilms were more resistant to treatment. Flow cytometry allowed the detection of bacteria with damaged membranes that were unable to replicate and grow after cell sorting. Further studies seem warranted to optimize the efficacy of light-activated curcumin against S. mutans biofilms.

摘要

背景

变形链球菌生物膜被认为是龋齿的主要致病因素。光动力抗菌化学疗法(PACT)最近被提议作为一种使牙齿生物膜失活的策略。本研究旨在调查蓝光激活的姜黄素对变形链球菌活力的影响,并探索其作为一种新型抗龋治疗剂的潜力。通过流式细胞术评估了光激活姜黄素的不同浓度和孵育时间对变形链球菌在浮游和生物膜生长模型中存活率的影响。

方法

在蓝光激活之前,将浮游悬浮液或羟基磷灰石圆盘上形成的生物膜中的变形链球菌与姜黄素孵育5或10分钟。在用流式细胞术评估活力之前,用SYTO 9和碘化丙啶对细菌进行标记。使用单因素方差分析和Tukey多重比较区间(α=0.05)对结果进行统计学分析。

结果

对于浮游培养物,0.2μM的光激活姜黄素显著降低了变形链球菌的活力(p<0.05)。对于生物膜培养物,浓度为40 - 60μM 的光激活姜黄素仅使活力抑制了50%(p<0.05)。无论生长模式如何,孵育时间对PACT效率均无显著影响。

结论

本研究表明,蓝光激活的姜黄素可有效使变形链球菌的浮游培养物失活,而生物膜对治疗更具抗性。流式细胞术能够检测到细胞膜受损且在细胞分选后无法复制和生长的细菌。似乎有必要进行进一步研究以优化光激活姜黄素对变形链球菌生物膜的疗效。

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