华西口腔医学杂志 ›› 2021, Vol. 39 ›› Issue (2): 188-194.doi: 10.7518/hxkq.2021.02.010

• 临床研究 • 上一篇    下一篇

抗菌肽GH12对龋相关三菌种生物膜形貌及菌种构成的影响

李欣蔚(), 王雨霏, 姜文韬, 张凌琳()   

  1. 口腔疾病研究国家重点实验室 国家口腔疾病临床医学研究中心 四川大学华西口腔医院牙体牙髓病科,成都 610041
  • 收稿日期:2020-05-11 修回日期:2021-01-09 出版日期:2021-04-01 发布日期:2021-04-09
  • 通讯作者: 张凌琳 E-mail:lixinwei12956@163.com;zhll_sc@163.com
  • 作者简介:李欣蔚,学士,E-mail:lixinwei12956@163.com
  • 基金资助:
    国家自然科学基金面上项目(81771062);四川省青年科技创新研究团队项目(2020JDTD0014)

Effects of antimicrobial peptide GH12 on the morphology and composition of cariogenic three-species biofilm

Li Xinwei(), Wang Yufei, Jiang Wentao, Zhang Linglin()   

  1. State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Cariology and Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2020-05-11 Revised:2021-01-09 Online:2021-04-01 Published:2021-04-09
  • Contact: Zhang Linglin E-mail:lixinwei12956@163.com;zhll_sc@163.com
  • Supported by:
    The National Natural Science Foundation(81771062);Sichuan Province Youth Science and Technology Innovation Research Team Project(2020JDTD0014)

摘要: 目的

研究抗菌肽GH12对龋相关三菌种生物膜结构、形貌以及菌种构成的影响。

方法

龋相关三菌种生物膜含有致龋菌变异链球菌(S. mutans)以及共生菌血链球菌(S. sanguinis)和格式链球菌(S. gordonii)。以未处理组作为对照组,GH12(2、4、8 mg·L-1)作为实验组。通过结晶紫染色、扫描电镜和荧光原位杂交观察生物膜形貌变化,定性分析菌种构成,继而使用选择培养计数法定量分析生物膜内的S. mutans占比。

结果

与对照组相比,GH12处理后的龋相关三菌种生物膜生物量减少,密度降低,结构更松散;S. mutans占比逐渐降低甚至无法检测,而S. gordoniiS. sanguinis占比逐渐增加,占据主导地位。

结论

GH12可以降低龋相关三菌种生物膜中致龋菌占比,破坏其完整性,使其向更易清除的方向转换。

关键词: 抗菌肽, 牙菌斑, 变异链球菌, 血链球菌, 格氏链球菌

Abstract: Objective

To evaluate the effects of antimicrobial peptide GH12 designed de novo on the structure, morphology, and composition of a cariogenic three-species biofilm.

Methods

The cariogenic three-species biofilm consis-ted of the cariogenic Streptococcus mutans (S. mutans) and commensal bacteria Streptococcus sanguinis (S. sanguinis) and Streptococcus gordonii (S. gordonii). The biofilm was treated using GH12 (2, 4, and 8 mg·L-1), and untreated biofilm was used as the control. Changes in the morphology and structure of the three-species biofilm were evaluated through crystal violet staining, scanning electron microscopy (SEM), and fluorescent in situ hybridization (FISH). Moreover, S. mutans in the biofilm was selectively cultured, and its colony-forming units were counted.

Results

The biomass and density of the cariogenic three-species biofilm treated with GH12 decreased compared with those of the control. The number of S. mutans decreased gradually and eventually became undetectable, whereas the number of S. gordonii and S. sanguinis increased and became predominant in the biofilm.

Conclusion

GH12 can reduce the number of S. mutans within the cariogenic three-species biofilm, destroys its integrity, and consequently makes the biofilm easy to remove.

Key words: antimicrobial peptide, dental plaque, Streptococcus mutans, Streptococcus sanguinis, Streptococcus gordonii

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