J Periodontal Implant Sci.  2019 Oct;49(5):319-329. 10.5051/jpis.2019.49.5.319.

The bactericidal effect of an atmospheric-pressure plasma jet on Porphyromonas gingivalis biofilms on sandblasted and acid-etched titanium discs

• Lee JY
• Kim KH
• Park SY
• Yoon SY
• Kim GH
• Lee YM
• Rhyu IC
• Seol YJ

• KMID: 2461022
• DOI: http://doi.org/10.5051/jpis.2019.49.5.319

Abstract

PURPOSE
Direct application of atmospheric-pressure plasma jets (APPJs) has been established as an effective method of microbial decontamination. This study aimed to investigate the bactericidal effect of direct application of an APPJ using helium gas (He-APPJ) on Porphyromonas gingivalis biofilms on sandblasted and acid-etched (SLA) titanium discs.
METHODS
On the SLA discs covered by P. gingivalis biofilms, an APPJ with helium (He) as a discharge gas was applied at 3 different time intervals (0, 3, and 5 minutes). To evaluate the effect of the plasma itself, the He gas-only group was used as the control group. The bactericidal effect of the He-APPJ was determined by the number of colony-forming units. Bacterial viability was observed by confocal laser scanning microscopy (CLSM), and bacterial morphology was examined by scanning electron microscopy (SEM).
RESULTS
As the plasma treatment time increased, the amount of P. gingivalis decreased, and the difference was statistically significant. In the SEM images, compared to the control group, the bacterial biofilm structure on SLA discs treated by the He-APPJ for more than 3 minutes was destroyed. In addition, the CLSM images showed consistent results. Even in sites distant from the area of direct He-APPJ exposure, decontamination effects were observed in both SEM and CLSM images.
CONCLUSIONS
He-APPJ application was effective in removing P. gingivalis biofilm on SLA titanium discs in an in vitro experiment.

Keyword

Bacterial Load; Confocal Microscopy; Plasma Gases; Porphyromonas gingivalis; Scanning Electron Microscopy; Titanium

MeSH Terms

Bacterial Load
Biofilms*
Decontamination
Helium
In Vitro Techniques
Methods
Microbial Viability
Microscopy, Confocal
Microscopy, Electron, Scanning
Plasma Gases
Plasma*
Porphyromonas gingivalis*
Porphyromonas*
Stem Cells
Titanium*
Helium
Plasma Gases
Titanium

Figure

• 

  Figure 1 Photograph of the apparatus for He-APPJ generation and He-APPJ application to titanium discs in this study. He-APPJ: helium atmospheric-pressure plasma jet.

• 

  Figure 2 Scanning electron microscopy images of the central and marginal areas after He-APPJ treatment. The control group (no treatment) showed heavy bacterial accumulation on the discs. In the He gas–only treatment group, remaining bacterial biofilm was observed in both the central and marginal areas in the 3-minute treatment group, while bacterial rods had been eliminated from the central area of the 5-minute treatment group, but not in the marginal area. However, the He-APPJ group showed a clear titanium surface without a bacterial biofilm in both the 3-minute and 5-minute treatment groups. He-APPJ: helium atmospheric-pressure plasma jet.

• 

  Figure 3 Confocal laser scanning microscopy images of the central and marginal areas after He-APPJ application. Heavy live bacterial accumulation on the discs was observed in the control group. In the He gas–only treatment group, the amount of bacteria was higher in the central area than in the marginal area in the 3-minute treatment group. In the 5-minute treatment group, the amount of bacteria decreased in both the central and marginal areas, although more live bacteria were observed in the marginal area. in the He-APPJ treatment group, the amount of bacteria was lower in the central area than in the marginal area in both the 3-minute and 5-minute treatment groups. Only a few viable bacteria were observed in the central area of the 3-minute treatment group. In the marginal area, a mixture of live and dead bacteria was found, whereas most bacteria were dead in the 5-minute treatment group. He: helium, He-APPJ: helium atmospheric-pressure plasma jet.

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