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中华口腔医学研究杂志(电子版) ›› 2018, Vol. 12 ›› Issue (02) : 83 -88. doi: 10.3877/cma.j.issn.1674-1366.2018.02.003

所属专题： 文献；

基础研究

载银氧化石墨烯对变异链球菌生长及生物膜形成的影响
吴瑞雪1, 杨鑫1, 陈依静1, 符远翔2, 余东升1, 赵玮1,†()   
  1. 1. 510055　广州，中山大学光华口腔医学院·附属口腔医院，广东省口腔医学重点实验室
    2. 510275　广州，中山大学化学工程与技术学院
  • 收稿日期:2017-10-25 出版日期:2018-04-01
  • 通信作者: 赵玮
  • 基金资助:
    国家自然科学基金(81472526); 广东省科技计划(2016A020215094); 广东省自然科学基金(2014A030313126)

Effect of graphene oxide-silver nanocomposites on Streptococcus mutans proliferation and biofilm formation

Ruixue Wu1, Xin Yang1, Yijing Chen1, Yuanxiang Fu2, Dongsheng Yu1, Wei Zhao1,†()   

  1. 1. Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
    2. School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou 510275, China
  • Received:2017-10-25 Published:2018-04-01
  • Corresponding author: Wei Zhao
  • About author:
    Corresponding author:Zhao Wei,Email:
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引用本文：

吴瑞雪, 杨鑫, 陈依静, 符远翔, 余东升, 赵玮. 载银氧化石墨烯对变异链球菌生长及生物膜形成的影响[J]. 中华口腔医学研究杂志(电子版), 2018, 12(02): 83-88.

Ruixue Wu, Xin Yang, Yijing Chen, Yuanxiang Fu, Dongsheng Yu, Wei Zhao. Effect of graphene oxide-silver nanocomposites on Streptococcus mutans proliferation and biofilm formation[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2018, 12(02): 83-88.

目的

探讨载银氧化石墨烯（GO/Ag）纳米复合物对变异链球菌（S.mutans）生长及生物膜形成的抑制作用。

方法

应用梯度稀释法检测GO/Ag、氧化石墨烯（GO）、纳米银（AgNPs）对变异链球菌的最小抑菌浓度（MIC）；菌落形成单位计数法比较GO/Ag、GO、AgNPs对变异链球菌的抑制作用；采用结晶紫染色法和XTT法测定变异链球菌24 h生物膜的生成量和活性；激光共聚焦扫描显微镜（CLSM）观察生物膜形态和计算24 h生物膜的活菌比例。数据以单因素方差分析（One-Way ANOVA）和LSD-t检验进行统计分析。

结果

GO/Ag对变异链球菌的MIC为0.16 mg/mL。0.64 mg/mL GO/Ag对变异链球菌的抑菌率为（82.90 ± 4.87）%，与对照组间的差异有统计学意义（t= 30.804，P<0.001）。而GO和AgNPs材料浓度提升至2.56 mg/mL时，肉眼观察细菌仍未见明显减少。在0.16 mg/mL GO/Ag浓度下，变异链球菌生物膜生成量和活性的减少率分别为（25.12 ± 0.01）%和（38.90 ± 3.42）%，活菌比例为（42.76 ± 19.48）%，与空白对照组间的差异均有统计学意义（t生成量= 7.274，P生成量<0.001；t活性= 7.765，P活性<0.001；t活菌比例= 10.412，P活菌比例<0.001）。

结论

与AgNPs和GO相比，GO/Ag新型纳米复合物对变异链球菌的生长具有较好的抑制作用，并能抑制其生物膜形成。

关键词: 变异链球菌, 纳米结构，银, 氧化物，石墨烯, 抗菌性
Objective

To investigate the effect of graphene oxide-silver nanocomposites (GO/Ag) on the proliferation and biofilm formation of Streptococcus mutans (S.mutans) .

Methods

Microdilution method was used to determine the minimum inhibitory concentration (MIC) of graphene oxide (GO) , silver-nanoparticles (AgNPs) , and GO/Ag nanocomposites against the S.mutans. For comparison of antibacterial efficiency of GO, AgNPs and GO/Ag, a colony-forming units counting method was applied. The biomass and the metabolic activities of biofilms with incubation of 24 h were tested by Crystal violet biofilm assay and XTT assay. Confocal laser scanning microscopy (CLSM) was performed to observe and calculate the percentage of live bacteria of S.mutans biofilm. The data were analyzed by One-Way ANOVA and LSD-t test.

Results

The MIC of GO/Ag nanocomposites against S.mutans was found to be 0.16 mg/mL. The bacteriostatic rate of 0.64 mg/mL GO/Ag was (82.90 ± 4.87) % and had significantly difference with the control group (t= 30.804, P<0.001) . In the GO and AgNPs groups, no apparent decrease of S.mutans could be observed, even after raising the concentration up to 2.56 mg/mL. In the 0.16 mg/mL GO/Ag group, reduction rate of biofilm biomass and metabolic activity were (25.12 ± 0.01) % and (38.90 ± 3.42) %, the percentage of live bacteria was (42.76 ± 19.48) %. All of these had significant difference with the blank control group (reduction rate of biofilm biomass: t= 7.274, P<0.001; reduction rate of metabolic activity: t= 7.765, P<0.001; percentage of live bacteria: t= 10.412, P<0.001) .

Conclusions

Comparing with AgNPs and GO, GO/Ag nanocomposites exhibited enhanced inhibition effect on the proliferation and biofilm formation of S.mutans.

Key words: Streptococcus mutans, Nanocomposites, silver, Oxides, graphene, Anti-bacterial property
图1 不同浓度的氧化石墨烯（GO）、纳米银（AgNPs）、载银氧化石墨烯（GO/Ag）对变异链球菌的抑菌率
表1 不同实验组对变异链球菌24 h生物膜生成量、活性的影响（%， ± s）
组别 生成量的减少率 活性的减少率
0.08 mg/mL GO/Ag组 2.46 ± 0.54a 8.95 ± 4.38
0.12 mg/mL GO/Ag组 17.34 ± 5.76a 30.23 ± 4.55a
0.16 mg/mL GO/Ag组 25.12 ± 0.01a 38.90 ± 3.42a
0.16 mg/mL GO组 4.58 ± 4.96 4.82 ± 1.16
0.16 mg/mL AgNPs组 0.80 ± 0.42 4.67 ± 0.28
表2 不同实验组对变异链球菌24 h生物膜活菌比例的影响（%， ± s）
组别 生物膜活菌比例 t值 P值
0.08 mg/mL GO/Ag组 52.22 ± 2.14 6.775 0.000
0.12 mg/mL GO/Ag组 44.52 ± 1.47 9.735 0.000
0.16 mg/mL GO/Ag组 42.76 ± 19.48 10.412 0.000
0.16 mg/mL GO组 57.56 ± 2.88 4.718 0.000
0.16 mg/mL AgNPs组 64.85 ± 0.59 1.915 0.062
图2 变异链球菌24 h生物膜的三维激光共聚焦扫描显微镜图像（荧光染色中倍放大）
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