细菌外排泵抑制剂对光动力疗法防龋效果的影响  

Study of the Effective of Efflux Pump-Inhibitors in Photodynamic Therapy's Anticaries Effect

作　　者：陶亚东[1] 邹朝晖[1] 孙盼盼[1] 阴慧娟 

机构地区：[1]天津医科大学口腔医学院,天津300070 [2]中国医学科学院生物医学工程研究所,天津300070

出　　处：《应用激光》2013年第1期91-96,共6页Applied Laser

摘　　要：目的:探讨细菌外排泵抑制剂(microbial efflux pump Inhibitor,EPI)———维拉帕米对以甲苯胺蓝O(toluidine blue O,TBO)为光敏剂的光动力疗法(photodynamic therapy,PDT)抑制牙菌斑生物膜内主要致龋菌作用的影响。方法:以变形链球菌、血链球菌、嗜酸性乳杆菌和粘性放线菌为实验菌株,建立牙菌斑生物膜模型。以维拉帕米作为细菌外排泵抑制剂,根据在PDT过程中加入维拉帕米和光敏剂的先后顺序,实验分为5组。A组:生理盐水处理组。B组:单纯PDT处理组。C组:同时加入光敏剂和维拉帕米PDT处理组。D组:先加入维拉帕米后加入光敏剂PDT处理组。E组:先加入光敏剂后加入维拉帕米PDT处理组。平板菌落计数观察牙菌斑生物膜活性,并运用激光共聚焦显微镜(Confocal laser scanning microscopy,CLSM)对处理后的牙菌斑生物膜进行断层扫描、分析。结果:与生理盐水处理组相比,单纯PDT处理组牙菌斑内致龋菌存活的数量(CFU/mL)明显减少(P<0.05),其抑菌率为81.02%;与单纯PDT处理组相比,加入维拉帕米的PDT处理组内牙菌斑内致龋菌存活数量有下降趋势。其中先加入维拉帕米后加入光敏剂PDT处理组牙菌斑内致龋菌存活数量显著性下降(P<0.05),抑菌率高达93.60%;激光共聚焦显微镜观察发现菌斑生物膜发生明显的变化。结论:实验表明光动力疗法有明显的防龋效果,且细菌外排泵抑制剂对PDT防龋效果有促进作用。Objective： To Study the effective of efflux pump-inhibitors--- Verapamil in photodynamic therapy using toluidine blue O as photosensitizer on the cariogenic bacteria in dental plaque biofilms. Methods： According to the order of the verapamil and photosensitizer in the PDT process, Streptococcus mutans, Streptococcus sanguis, eosinophilic Lactobacillus and Actinomyces viscosus to establish the dental plaque biofilm model. The experiment is divided into five groups, Group A： saline-treated group, Group B： only PDT group, Group C：Cells were incubated with the photosensitizer and verapamil group, Group D：Cells were incubated with verapamil before incubated the photosensitizer group, Group E：Cells were incubated with the photosensitizer before incubated verapamil group, the influence of the dental plaque biofilms was observed according to plate counting of bacteria method. The changes in the structure of biofilms after PDT were analysed by means of confocal laser scanning microscopy. Results： Compared to saline- treated group, in artificial caries model of only PDT group, the number of living cariogenic bacteria （CFU/mL） in plaque biofilms reduced more significantly （P〈0. 05）, the bactericidal rate upward to 81.02%, comperesd to group B, Cells were incubated with group D, there are a significant Declineing trend in the number of cariogenic bacteria （P〈0.05）, the sterilization rate was 93.60%. And the structure of dental plaque biofilms changed evidently. Conclusion： PDT is an effective method in eliminate cariogenic bacteria of dental plaque biofilms. And bacterial efflux pump inhibitors can improve PDT inhibition cariogenic bacteria in dental plaque biofilm.

关 键 词：光动力疗法 龋齿 菌斑生物膜 细菌外排泵 激光共聚焦显微镜 

分 类 号：R73[医药卫生—肿瘤]