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Journal of Peking University (Health Sciences) ›› 2023, Vol. 55 ›› Issue (1): 38-43. doi: 10.19723/j.issn.1671-167X.2023.01.006

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Antibacterial effect of low-temperature plasma on Enterococcus faecalis in dentinal tubules in vitro

Ruo-qing ZHONG1,Meng-qian ZHU1,Ying-long LI2,*(),Jie PAN1,*()   

  1. 1. Department of General Dentistry, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
    2. Department of Stomatology, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China
  • Received:2022-10-10 Online:2023-02-18 Published:2023-01-31
  • Contact: Ying-long LI,Jie PAN E-mail:liyinglongpku@163.com;panjie72@sina.com
  • Supported by:
    the National Natural Science Foundation of China(81901064);Beijing Hospitals Authority Youth Program(QML20210304)

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Abstract:

Objective: To construct a model of Enterococcus faecalis (E. faecalis) infection in dentinal tubules by gradient centrifugation and to evaluate the antibacterial effect of low-temperature plasma on E. faecalis in dentinal tubules. Methods: Standard dentin blocks of 4 mm×4 mm×2 mm size were prepared from single root canal isolated teeth without caries, placed in the E. faecalis bacterial solution, centrifuged in gradient and incubated for 24 h to establish the model of dentinal tubule infection with E. faecalis. The twenty dentin blocks of were divided into five groups, low-temperature plasma jet treatment for 0, 5 and 10 min, calcium hydroxide paste sealing for 7 d and 2% chlorhexidine gel sealing for 7 d. Scanning electron microscopy and confocal laser scanning microscope were used to assess the infection in the dentinal tubules and the antibacterial effect of low-temperature plasma. Results: The results of scanning electron microscopy and confocal laser scanning microscopy showed that after 24 h of incubation by gradient centrifugation, E. faecalis could fully enter the dentinal tubules to a depth of more than 600μm indicating that this method was time-saving and efficient and could successfully construct a model of E. faecalis infection in dentinal tubules. Low-temperature plasma could enter the dentinal tubules and play a role, the structure of E. faecalis was still intact after 5 min of low-temperature plasma treatment, with no obvious damage, and after 10 min of low-temperature plasma treatment, the surface morphology of E. faecalis was crumpled and deformed, the cell wall was seriously collapsed, and the normal physiological morphology was damaged indicating that the majority of E. faecalis was killed in the dentinal tubules. The antibacterial effect of low-temperature plasma treatment for 10 min exceeded that of the calcium hydroxide paste sealing for 7 d and the 2% chlorhexidine gel sealing for 7 d. These two chemicals had difficulty entering deep into the dentinal tubules, and therefore only had a few of antibacterial effect on the bacterial biofilm on the root canal wall, and there was also no significant damage to the E. faecalis bacterial structure. Conclusion: Gradient centrifugation could establish the model of E. faecalis dentin infection successfully. Low-temperature plasma treatment for 10 min could kill E. faecalis in dentinal tubules effectively, which is superior to the calcium hydroxide paste sealing for 7 d and the 2% chlorhexidine gel sealing for 7 d.

Key words: Low-temperature plasma, Gradient centrifugation, Enterococcus faecalis, Dentinal tubules

CLC Number: 

  • R781.3

Figure 1

A low-temperature plasma device and schematic diagram A, schematic diagram of low-temperature plasma device; B, the low-temperature plasma device, with the jet 1 cm from the inner side of the root canal of the dentin block."

Figure 2

Results of Enterococcus faecalis dentinal tubule infection A, scanning electron microscope image of dentinal tubules; B, C, Enterococcus faecalis entering the dentinal tubules after 24 hours of incubation, the arrows show the growth of Enterococcus faecalis in the dentinal tubules; D, Enterococcus faecalis were basically alive (green fluorescence) after 24 hours of incubation, the width in the middle of the arrows is the depth of Enterococcus faecalis into the dentinal tubules. P', pulpal side; D', dentin."

Figure 3

Scanning electron microscopy results of control and treated groups A1, A2, low-temperature plasma treatment for 0 min; B1, B2, low-temperature plasma treatment for 5 min; C1, C2, low-temperature plasma treatment for 10 min; D1, D2, calcium hydroxide paste sealing for 7 days; E1, E2, 2% chlorhexidine gel sealing for 7 days. Yellow arrows point to Enterococcus faecalis."

Figure 4

Confocal laser scanning microscopy results of control and treated groups, the left side of the image shows the root canal wall side and the right side shows the cementum side A, low-temperature plasma treatment for 0 minute; B, low-temperature plasma treatment for 5 minutes; C, low-temperature plasma treatment for 10 minutes; D, calcium hydroxide paste sealing for 7 days; E, 2% chlorhexidine gel sealing for 7 days. Red fluorescence represents dead bacteria, green represents alive bacteria."

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