Antibacterial effect of different concentrations of Galla Chinensis extract on cariogenic bacteria in a biofilm model

Eun-Jeong Kim; Bo-Hyoung Jin

doi:10.11149/jkaoh.2020.44.1.13

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Kim and Jin: Antibacterial effect of different concentrations of Galla Chinensis extract on cariogenic bacteria in a biofilm model

Original Article

J Korean Acad Oral Health 2020;44(1):13-19.

Published online: 15 March 2020

DOI: https://doi.org/10.11149/jkaoh.2020.44.1.13

Antibacterial effect of different concentrations of Galla Chinensis extract on cariogenic bacteria in a biofilm model

Eun-Jeong Kim¹, Bo-Hyoung Jin^2,

¹Dental Research Institution, Korea

²Department of Preventive & Social Dentistry, School of Dentistry, Seoul National University, Seoul, Korea

Corresponding Author: Bo-Hyoung Jin Department of Preventive & Social Dentistry, School of Dentistry, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 08826, Korea Tel: +82-2-740-8783 Fax: +82-2-766-8781 E-mail: jjbh@snu.ac.kr https://orcid.org/0000-0003-3526-6805

Received 19 February 2020 Revised 7 March 2020 Accepted 7 March 2020

This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Objectives

Galla chinensis inhibited the adherence of planktonic oral bacteria and acid production by cariogenic bacteria. However, little is known about the relevant conditions of Galla Chinensis extract (GCE) exposure time and concentration and the effect of GCE on the structural and functional activity of cariogenic bacteria. The antibacterial effects of natural G. Chinensis extract on S. mutans, S. sanguinis, and S. oralis biofilms were evaluated in vitro.

Methods

Biofilms formed on glass surfaces were treated with different concentrations of GCE at different exposure times. The effects were assessed by examining the bactericidal activity, acido-genesis, minimum inhibitory concentration, and morphology.

Results

There was a statistically significant difference in the bacterial growth inhibition depending on the concentration of the GCE, with bacterial growth being inhibited as the concentration of GCE increased. A concentration of 1.0 mg/ml GCE had similar bactericidal effects against S. mutans and S. oralis biofilms to those produced by 2.0 mg/ml CHX. In the 1.0 mg/ml GCE group, incomplete septa were also observed in the outline of the cell wall, together with disruption of the cell membrane. In addition, there was also a slight exudation of the intracellular content from the bacteria in the 1.0 mg/ml GCE and 2 mg/ml CHX groups.

Conclusions

These results indicate that GCE inhibits the growth of S. mutans, S. sanguinis, and S. oralis with increasing concentrations. It alters the microstructure of S. mutans biofilms. These results suggest that GCE might be a useful anti-bacterial agent for preventing dental caries.

Keywords: Antibacterial effects, Biofilms, Galla Chinensis, Streptococcus mutans

References

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Fig. 1.

The bacterial growth curve of S. mutans, S. sanguinis and S. oralis biofilms by GCE concentration. After the biofilms had been exposed to the test solutions for 3, 6, 9, 12, and 24 hours, the number of colonies was counted to determine the CFU. There was a significant difference over time at all concentrations.

Fig. 2.

TEM images of the S. mutans biofilm after 1 hour of treatment. The black arrows indicate the wall of S. mutans, and the white arrows indicate the intracellular contents. The scale bar is 100 nm. (A) 1% DMSO, (B) 2 mg/ml CHX, and (C) 1.0 mg/ml GCE. After the S. mutans biofilms had been exposed to 1% DMSO for 1 h as the negative control, the TEM showed a clear outline of the S. mutans cell wall and a peptidoglycan layer. However, most of the peptidoglycan layers of S. mutans in the CHX group had disappeared. In the 1.0 mg/ml GCE group, incomplete septa were also observed in the outline of the cell wall. In addition, there was a slight exudation of the intracellular contents in the 1.0 mg/ml GCE and 2 mg/ml CHX groups.

Table 1.

MIC induced by GCE at different concentrations

Treatment group		S. mutans*	S. sanguis*	S. oralis*
GCE 0.1 mg/ml	M±SD	2.43±0.04^a	2.58±0.03^a	2.60±0.02^a
	Proportion (%)	84.04	80.51	87.18
GCE 0.2 mg/ml	M±SD	2.32±0.04^a	2.14±0.12^a,b	2.16±0.04^b
	Proportion (%)	75.75	52.05	56.5
GCE 0.4 mg/ml	M±SD	1.95±0.14^b	2.10±0.07^a,b	2.41±0.06^b
	Proportion (%)	47.83	50.21	48.68
GCE 0.8 mg/ml	M±SD	1.69±0.08^b	2.08±0.08^a,b	1.95±0.05^c
	Proportion (%)	36.96	41.51	46.18
GCE 1.0 mg/ml	M±SD	1.48±0.10^c	1.54±0.12^c	1.35±0.08^d
	Proportion (%)	30.13	28.92	25.68
CHX 2.0 mg/ml	M±SD	1.40±0.30^c	1.35±0.06^c	1.25±0.05^d
	Proportion (%)	28.81	24.1	23.23
1% DMSO	M±SD	2.69±0.05^a	2.69±0.03^a,b	2.74±0.01^a
	Proportion (%)	102.35	89.72	107.59

Data are presented as log₁₀ colony-forming units (CFU)/disc and percentages. M, Ln (Log₁₀ CFU); SD, standard deviation; Proportion, If the OD value is less than 90-95% of the control 100%, the bacteria is affected and has not grown.

^a,b,c,d The different superscripts in the same column indicate a statistically significant difference from each group (P<0.05). Post hoc Tukey’s HSD, *P<0.05.

Table 2.

General linear model including time and treatment groups

Source	df	Mean square	F	Sig.	Partial eta square
Treatment groups	6	15.12	5694.59	<.001	0.995
Time	4	10.11	3806.47	<.001	0.989
Treatment groups×Time	24	0.89	337.88	<.001	0.979

R Squared=.997 (Adjusted R Squared=.996).

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