北京大学学报(医学版) ›› 2014, Vol. 46 ›› Issue (1): 30-34.

• 论著 • 上一篇    下一篇

过氧化物美白剂对釉质表面变形链球菌生物膜的影响

郑春艳1,潘洁2,王祖华1△,汪洋1   

  1. (北京大学口腔医学院·口腔医院 1. 牙体牙髓科,2.综合科,北京100081)
  • 出版日期:2014-02-18 发布日期:2014-02-18

Effects of hydrogen peroxide-containing bleaching on the growth of Streptococcus mutans biofilm on enamel disc surface

ZHENG Chun-yan1,PAN Jie2 , WANG Zu-hua1△ , WANG Yang1   

  1. (1.Department of Cariology and Endodontology; 2.Department of General Dentistry, Peking University School and Hospital of Stomatology, Beijing 100081, China)
  • Online:2014-02-18 Published:2014-02-18

摘要: 目的:观察高浓度过氧化物美白剂对于釉质表面变形链球菌生物膜的影响。方法: 选择拔除的离体牙,制备直径5 mm的带有天然釉质表面的圆盘20个;分为两组,其中10个圆盘用含有35%(质量分数)过氧化氢凝胶(Beyond公司,美国)进行美白,另外10个为对照组。20个釉质圆盘于灭菌人唾液中浸泡3.5 h形成获得性膜,加入脑心浸液肉汤(brain heart infusion broth, BHI)和变形链球菌(UA 159菌株)混悬液,放入5% (体积分数)CO2孵箱进行孵育。在孵育3、7、14、21、28 d后,每组各取出2个圆盘,其中一个圆盘刮去釉质表面的生物膜,系列稀释,接种于脑心浸液琼脂(BHI agar supplemented with blood, BHIS))血平皿菌落计数;另一个圆盘以荧光染色后,用激光共聚焦显微镜 (confocal laser scanning microscope, CLSM)分层扫描观察生物膜。结果: 培养3、7、14 d后美白牙面的生物膜内活菌数均显著少于对照组。在培养3 d后,美白釉质表面生物膜内活菌面积[(3 595 ± 2 903) μm2 vs. (89 155±65 963) μm2,t= 8.71,P=0.00]和活菌比例[(26.0% ±16.4%) vs.(92.2% ±10.9%),t= 19.93,P=0.00)]显著低于对照组;但培养21 d后出现逆转:美白釉质表面生物膜内活菌面积[(66 262±23 772)μm2 vs.(51 184±20 502)μm2,t=2.59,P=0.012]显著超过对照组。传统培养法得出的活菌数与CLSM结果一致。结论:美白后釉质表面短期内可抑制变形链球菌生物膜的生长,但在21 d后,美白组生物膜生长高于对照组。美白治疗在短期内有一定抗龋作用,但长期作用有待进一步研究。

关键词: 牙漂白, 牙釉质, 生物膜, 链球菌, 变异, 显微镜检查, 共焦

Abstract: Objective:To evaluate the effects of a commercial bleaching agent containing 35% (mass fraction) hydrogen peroxide on the growth of Streptococcus mutans biofilm on enamel disc surface. Methods:A total of 20 enamel disks were made from human extracted teeth and the enamel surfaces were kept intact. The discs were autocalved and randomly divided into two groups: bleaching group and control group. Each group contained 10 discs. For bleaching group, the enamel discs were whitened by commercial 35% hydrogen peroxide according to the instruction (BeyondTM Professional Dental Whitening Kit, Beyond Technology, TX,USA ); no treatment for control group. All the discs were kept in sterile human saliva for 3.5 hours, and then the mixture of brain heart infusion broth (BHI) medium and Streptococcus mutans were added. The discs and Streptococcus mutans were incubated together in BHI medium with 5% CO2 (volume fraction), at 37 ℃. After 3, 7, 14, 21 and 28 d’s incubation, two discs of each group were taken out and the biofilms on the enamel surfaces were evaluated by using conventional bacteria counts and confocal laser scanning microscope (CLSM). The bacteria in the biofilm on one disc enamel surface were analyzed by plating on BHIS agar and the colony-forming units were counted. The biofilm on the other disc surface was stained using a twocolour fluorescent dye kit (Bacerial Viability Kit L-7012) for CLSM. Results:The vital bacteria counts of vital cells in the 3, 7, and 14 d’s biofilms of the bleaching group were significantly fewer than those of the control group. Especially in the 3 days’ biofilm on the whitened surface, the vital bacteria counts [(3 595±2 903) μm2 vs. (89 155±65 963) μm2,t=8.71,P=0.00] and proportion of vital bacteria [(26.0%±16.4%) vs.(92.2%±10.9%),t=19.93,P=0.00] were significantly fewer than those of the control. While, for the 21d’s biofilm, the vital bacteria counts and the percentage of the vital cells of the bleaching group were more than those of the control group significantly [(66 262±23 772) μm2 vs. (51 184±20 502) μm2,t=2.59,P=0.012].Conclusion:The hydrogen peroxide-containing bleaching agent may inhibit the growth of Streptococcus mutans biofilm for about 3 weeks; but after 3 weeks, it seems that the bleached surface will increase the growth of biofilm. Whether the whitening therapy will increase caries susceptibility of the bleached surface needs further research.

Key words: Tooth bleaching, Dental enamel, Biofilms, Streptococcus mutans, Microscopy, confocal

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