Galla chinensis extracts and calcium induce remineralization and antibacterial effects of enamel in a Streptococcus mutans biofilm model

Eun-Jeong Kim; Bo-Hyoung Jin

doi:10.11149/jkaoh.2018.42.3.90

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Kim and Jin: Galla chinensis extracts and calcium induce remineralization and antibacterial effects of enamel in a Streptococcus mutans biofilm model

Original Article

J Korean Acad Oral Health 2018;42(3):90-96.

Published online: 15 September 2018

DOI: https://doi.org/10.11149/jkaoh.2018.42.3.90

Galla chinensis extracts and calcium induce remineralization and antibacterial effects of enamel in a Streptococcus mutans biofilm model

Eun-Jeong Kim, Bo-Hyoung Jin

Department of Preventive and Social Dentistry, School of Dentistry, Seoul National University, Seoul, Korea

Corresponding Author: Bo-Hyoung Jin Department of Preventive and Social Dentistry, School of Dentistry, Seoul National University, 28 Yeongun-dong, Jongro-gu, Seoul 03080, Korea Tel: +82-2-740-8783 Fax: +82-2-766-8781 E-mail: jjbh@snu.ac.kr

Received 11 July 2018 Revised 12 September 2018 Accepted 12 September 2018

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

The purpose of this study was to investigate the combined effects of Galla chinensis extract (GCE) and calcium (CA) on enamel remineralization. The antibacterial effect of G. chinensis on Streptococcus mutans biofilm was also evaluated by examining the bacterial growth, acidogenesis, and morphology of the biofilm in vitro.

Methods

S. mutans biofilm was formed on bovine enamel specimens over a 72-h period and treated for 10 min with 1.0 mol CA, 4,000 ppm aqueous solution of GCE, or a combination of the two (GCE+CA). The enamel specimens were analyzed for enamel surface microhardness after remineralization. We tested the anti-cariogenic effects of GCE based on the inhibition of acid production, antibacterial activity, and morphological changes in S. mutans. The differences between the groups and antibacterial effects were analyzed using one-way analysis of variance.

Results

GCE+CA group showed the highest efficacy in enhancing remineralization. The GCE group showed the highest antibacterial activity against S. mutans biofilm. Although the GCE+CA group showed significant antibacterial activity, it was less than that of the GCE group (P<0.05). Both GCE and GCE+CA groups maintained a pH of approximately 7.0 for 1 h whereas the pH of the control group decreased rapidly from pH 7.3 to pH 6.1. SEM imaging revealed that S. mutans treated with GCE and GCE+CA showed irregular cell wall structure and showed fewer cells in the chain than the typical long chains observed in the control group.

Conclusions

This study found that natural G. chinensis significantly enhances enamel remineralization, and exerts synergistic effects with calcium. It also exerts strong bactericidal activity and inhibits acid production and changes in the microstructure of S. mutans biofilm.

Keywords: Anti-bacterial agents, Combined effect, Galla chinensis, Remineralization

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Fig. 1.

Flowchart of the in vitro experimental study design.

Fig. 2.

Acidogenicity of S. mutans biofilms was determined by measuring the pH of media.

Fig. 3.

SEM images of the S. mutans biofilm after 10 min treatment. (A) GCE, (B) GCE+CA, (C) CA, and (D) Control.

Table 1.

Comparison of the surface microhardness of different groups after remineralization

Condition	N	VHN	VHN	DVHN	P
Condition	N	Baseline	After artificial caries formation	After remineralization	P
GCE	24	308.8±5.1	48.70±5.00	41.8±3.9^a	0.009*
GCE+ CA	24	308.9±6.3	46.85±4.95	44.2±3.9^a
CA	24	303.2±6.3	42.72±5.19	27.1±5.4^b
Control	12	307.3±7.3	44.85±4.85	0.5±3.9^c

Values are mean±SD. DVHN = After remineralization VHN - Before treatment VHN (baseline). CA, immersed in 1.0 mol CaCl₂ for 10 min; GCE, immersed in 4,000 ppm GCE for 10 min; CA+GCE, immersed in 1.0 mol CaCl₂ and 4,000 ppm GCE for 10 min; Control, no treatment. *Statistically significant by repeated measured ANOVA at the a=0.05 level.

^a -cThe different lower case letters indicate statistically significant differences between same groups by Tukey’s HSD post-hoc test at P<0.05.

Table 2.

Antibacterial effects of the GCE against S. mutans biofilms

Condition	Exposure time
	CFU (×10⁸)
	0 min	5 min	10 min
GCE	48.8±10.6	5.2±1.4^a	3.1±0.9^a
GCE+CA	49.5±6.7	7.8±0.2^a	6.5±2.2^a
CA	51.2±8.8	34.1±7.8^b	30.1±8.4^b
Control	50.5±9.7	56.5±9.7^c	60.5±10.8^c

The data shown are the Mean±SD. The different superscripts in the same column indicate statistically significant difference from each group (P<0.05).

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