中国实用口腔科杂志

中国实用口腔科杂志

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冷常压等离子体对牙龈卟啉单胞菌抗菌作用的实验研究

顾舒扬1，李邵杰2，谢艳婷1，葛鑫1，田思睿1，赵宁宁1，王丹宁1，蔺增2，赵宝红1   

  1. 1. 中国医科大学口腔医学院·附属口腔医院种植中心，辽宁省口腔医学研究所口腔种植研究室，辽宁省口腔疾病重点实验室，辽宁省口腔疾病转化医学研究中心，辽宁  沈阳110002；2. 东北大学机械工程与自动化学院，辽宁  沈阳  110004
  • 出版日期:2020-01-15 发布日期:2020-03-02
  • 基金资助:
    辽宁省自然科学基金（20170541059）；辽宁省科技计划项目（2013225049）；沈阳市科技计划项目（F11-264-1-25）

  • Online:2020-01-15 Published:2020-03-02

摘要： 目的　评价冷常压等离子体（cold atmospheric plasma，CAP）对牙龈卟啉单胞菌（Porphyromonas gingivalis，P.g）的抗菌作用并观察其对P.g超微结构的影响。方法　用CAP对接种在钛片表面的P.g进行处理，依据处理距离和处理时间将实验组分为5 mm 30 s组、5 mm 60 s组、9 mm 30 s组和9 mm 60 s组，对照组不进行CAP处理。通过平板菌落计数法观察各组P.g数量的变化，场发射扫描电子显微镜（SEM）观察各组钛片表面P.g的形态和分布，并通过透射电镜（TEM）观察处理前后P.g超微结构的变化。结果　CAP处理时间和处理距离对各组菌落数均有显著影响，且各实验组菌落数与对照组相比，差异均有统计学意义（均P < 0.05）。SEM观察可见，随着CAP处理时间和距离的变化，菌体受到不同程度的破坏；5 mm 60 s组和9 mm 60 s组细菌溶解、破碎，几乎观察不到完整的细菌形态；对照组仍为完整的短杆状菌体结构。TEM观察可见，通过CAP处理，P.g细胞膜断裂、模糊不清，细胞质外溢，可观察到扭曲和完全分离的膜结构；对照组P.g菌体超微结构未发生明显改变，细胞壁、细胞膜及细胞质结构清晰。结论　CAP对P.g有较好的杀灭作用，并可引起P.g超微结构的变化，在一定范围内增加处理时间、缩减处理距离有助于增强抗菌效果。

关键词: 冷常压等离子体, 牙龈卟啉单胞菌, 抗菌作用, 场发射扫描电子显微镜, 透射电镜

Abstract: Objective　To evaluate the antibacterial effect of cold atmospheric plasma（CAP）on Porphyromonas gingivalis（P.g） and observe its effect on the ultrastructure of P.g. Methods　P.g on the titanium sheet surface was treated with CAP. According to the treatment distance and treatment time，the experimental group was divided into 5 mm 30 s group，5 mm 60 s group，9 mm 30 s group and 9 mm 60 s group；the control group was not treated with CAP. After treatment，the quantity of P.g was observed by plate colony counting method，the morphology and distribution of P.g on the surface of each group were observed by scanning electron microscope （SEM），and the ultrastructure of P.g was observed by transmission electron microscope （TEM）. Results　The treatment time and treatment distance of CAP had significant influence on P.g colony number. Compared with the control group，there were statistically significant differences in the antibacterial effect of P.g in each CAP treatment group（P < 0.05）. SEM showed that the bacteria were destroyed in varying degrees with the change of time and distance of CAP treatment；the bacteria in 5 mm 60 s group and 9 mm 60 s group were dissolved and broken，and almost no complete bacterial morphology could be observed. The P.g in control group was still a complete short rod cell structure. TEM showed that through CAP treatment，the cell wall and the cell membrane ruptured and blurred and cytoplasm overflowed，and the distorted and completely separated membrane structure could be observed. In control group，the ultrastructure of P.g cells did not change significantly，and the structure of cell wall，cell membrane and cytoplasm was clear. Conclusion　The CAP can damage the ultrastructure of P.g and has a good antibacterial effect on P.g，which is increased with increasing the treatment time and reducing the treatment distance within a certain range.

Key words: cold atmospheric plasma, CAP；Porphyromonas gingivalis, P.g；antibacterial effect；scanning electron microscope, SEM；transmission electron microscope, TEM