Periodontal disease and dental caries, which are two major oral diseases in the oral cavity, are major causes of tooth loss [1]. Two major oral diseases are caused by a combination of various causative factors, among which the bacteria in the plaque play the largest role [2]. It is important to remove plaque, the main cause of dental caries and periodontal disease, in order to prevent both oral diseases0 [1,2]. A plaque is a membrane that covers a part of the tooth surface by bacterial masses in the oral cavity stuck to the pellicle of glycoprotein attached to the tooth surface. Also called plaque, and biofilm. The most effective way to prevent plaque is brushing, and the dentifrice used is also important [1,3].

The dentifrice is an auxiliary material used in the brushing process to clean the surface of the tooth, and corresponds to a quasi-drug product such as soap or detergent [2]. dentifrice is a drug for cleaning and polishing teeth consisting of inorganic abrasives, detergents, wetting agents, binders and flavoring agents [3]. And it is a detergent used to effectively clean the tooth surface. The effects of dentifrice include dental caries, prevention of periodontal disease, whitening effect, slowing perception, and suppressing halitosis [3-5].

Recently, dentifrices have been developed and used in various ways according to their use and target, and they are trying to research to improve the efficacy of dentifrice by developing technologies by remixing commonly used components differently from the existing ones [3]. Through this paper, we studied dentifrice containing resveratrol, catechin, and Baikalin ingredients. Resveratrol, a dentifrice component, is a type of polyphenol known as a natural extract and is called grape extract [5]. Resveratrol has strong antioxidant power, inhibits active acidity, prevents cell oxidation, and has anti-inflammatory effects, which are abundant in peanuts and wine [5,6]. Resveratrol is a pyltoalexin that is produced when a plant becomes a fungus-like environment, and is a polyylphenol-based material. This substance is known to be helpful in the treatment of various diseases, and it has been reported that it has anti-cancer, anti-inflammatory, and life-span effects. It is not soluble in water but soluble in alcohol [6].

In addition, Baicalin, called golden extract (root extract), is contained in the root of the family Lamiaceae and is one of the flavonoids [7]. Baikallin has been used in various living bodies for its efficacy such as antibacterial, diuretic, anti-inflammatory, metamorphosis and spasm prevention, and anticancer effects [7]. Catechin of green tea is a polyphenolic compound. The content of green tea is 120.3~153.7 mg/g, and the content varies depending on the type of tea, the presence or absence of shading, the amount of sunlight, and the period of growth. Catechin is an astringent ingredient in tea that prevents tooth decay and prevents dry mouth, inhibits carcinogenesis, has antibacterial, detoxifying, and anti-inflammatory properties [8].

This paper studied the formulation dentifrice in the form of effervescent tablets containing resveratrol, bicalin, and catechin. The main ingredient of the effervescent tablet is sodium hydrogen carbonate, and a large amount of sodium is added therein. Effervescent tablets are tablets that dissolve in water to release carbon dioxide and form bubbles. The released carbon dioxide increases the solubility of tablets and neutralizes the phase to protect the acid-sensitive drugs. Therefore, the purpose of this paper was to compare the effect of plaque removal by using the general dentifrice and the foamed dentifrice. Using dentifrice containing resveratrol, catechin, and bicalin, we compared gingival and plaque measurements and compared changes to determine whether it is effective in preventing gingivitis, plaque reduction, and periodontal disease.

1. Subject

Subjects in this paper regularly brushed at least twice a day among men and women between 20 and 50 years old who agreed to participate in the study, and 120 subjects were selected as subjects who had more than 24 residual teeth without cervical caries or restorations.

The subject dentifrice used in the study was an experimental dentifrice, a comparative dentifrice, and a standard dentifrice and a control gargle. Experimental dentifrice is sodium bicarbonate (sodium bicarbonte), calcium carbonate, xylitol, hydroxypropyl methyl cellulose, fragrance powder as a basic component, catechin (green tea extract), resveratrol (grape extract), bikalin (golden extract) as the main components was formulation. Comparative dentifrice is a dentifrice composed of sodium bicarbonate, calcium carbonate, xylitol, hydroxypropylmethylcellulose and fragrance powder. The main components of the experiment dentifrice were catechin, resveratrol, and baicalin.

The standard dentifrice is a dentifrice composed mainly of dental type silica. The control gargal is a solution in which only basic ingredients such as foaming agent, wetting agent and fragrance are added.

2. Method

In this paper, for 4 weeks, the subjects who signed the informed consent by parallel comparison and randomization were judged to be suitable for the selection criteria and exclusion criteria after registration. Thereafter, suitable subjects were randomly assigned to one of the experimental group, the comparative group, the negative control group, and the positive control group according to the order of registration after a standardization period of 1 week, and 4 groups were selected by distributing 30 subjects per group.

This experiment was conducted with the approval of Dankook University Clinical Trial Committee (approval number DKU2017-09-004-004).

How to use dentifrice and period of use

The subject must use dentifrice distributed for 4 weeks to create an oral environment with the same dentifrice, and if the same dentifrice was not used twice in the morning and evening (1 day), it was excluded from the study subjects. In all four groups, the standard dentifrice was used for brushing using the rotation method for 3 minutes after breakfast and dinner every day. And the experimental group was to wash the mouth using the experiment dentifrice after lunch. In the comparison group, after lunch, oral cleaning was performed using a comparative gargle. The negative control group had no additional mouthwash except after breakfast and dinner. The positive control group was used to wash the mouth after lunch using a control gargle. The control gargle was made by spitting 10 cc of the proper amount within 1 minute after lunch.

① PMA index

The PMA index was measured on the gingiva of the maxillary and mandibular anterior teeth.

② Talbott, Mandel and Chiltond’s gingival index

The gingivitis index was divided into mesial, centrifugal, and central gingivitis according to the rating criteria, and 6 parts were measured. And the individual gingivitis index was obtained by dividing the sum of the measurements by each part by the number of teeth to be examined. The teeth to be measured were 16, 11, 26, 46, 31, and 36 teeth, totaling 6 teeth.

① Plaque management ability index (patient hygiene performance index, PHP index)

It was measured using plaque index, and the target tooth was measured with teeth 16, 11, 26, 46, 31, and 36.

② Turesky's plaque index evaluation criteria

Plaque index was measured on teeth 15, 13, 26, 44, 32 and 36.

The spss 24.0 program was used for the collected data in this study, and the significance level was set to 0.05 for statistical significance. Comparison between groups was analyzed by one-way ANOVA, and intragroup comparison was analyzed by paired t-test.

Table 1 . Gender and age distribution of subjects

Gender	Age group
	All	20s	30s	40s
Total	120	71	29	20
Male	33	14	14	5
Female	87	57	15	15

Table 2 . Main ingredient content of target toothpaste

Toothpaste	Resveratrol	Baicalin	Catechin
Toothpaste1	0.05%	0.1%	0.3%
Toothpaste2	0%	0%	0%
Toothpaste3	0%	0%	0%
Toothpaste4	0%	0%	0%

Toothpaste1: Experiment toothpaste, Toothpaste2: Comparative toothpaste, Toothpaste3: Standard toothpaste, Toothpaste4: Control toothpaste solution.

1. Gingivitis test results

Table 3 and Table 4 show the gingivitis test results. In the PMA index, no significant differences were observed in all four groups even in the pre-experiment, one-week, and two-week investigations. After 4 weeks, the results were shown in the experimental group 10.20, the comparative group 11.07, the negative control 11.90, and the positive control 10.20. It was confirmed that the experimental group and the positive control group had a lower PMA index than the negative control group (p<0.01). Talbott's gingivitis index was not significantly different before and after 1 week. Significant results were investigated after 2 weeks, and after 4 weeks, the experimental group was 1.19, the comparative group 1.26, the negative control group was 1.44, and the positive control group was 1.12. After 2 and 4 weeks, it was confirmed that the gingivitis index of the experimental group and the positive control group were significantly lower than before the experiment (p<0.01).

Table 3 . Changes in PMA index by group and measurement period

Group	N	Base			1 week			2 weeks			4 weeks
		Mean	SD		Mean	SD		Mean	SD		Mean	SD
E1	30	11.27	2.95		11.03	2.89		10.93	2.42		10.20	2.30*a
E2	30	11.30	2.59		11.07	2.86		10.80	2.01		11.07	2.21ab
NC	30	11.33	2.88		11.67	2.95		11.63	2.39		11.90	2.26b
PC	30	11.30	2.63		11.10	2.81		10.80	1.95		10.20	2.27*a
p-value+		1.000			.805			.401			.011

SD: standard deviation, E1: experimental group, E2: comparison group, NC: negative control group, PC: positive control group.

⁺p-value by one way ANOVA, ^abSame letter means no statistical difference by Scheffe test.

*p<0.05 by paired t-test between base and after.

Table 4 . Changes in Talbott’s gingival index by group and measurement period

Group	N	Base			1 week			2 weeks			4 weeks
		Mean	SD		Mean	SD		Mean	SD		Mean	SD
E1	30	1.33	0.19		1.31	0.16		1.24	0.13*a		1.19	0.13*ab
E2	30	1.33	0.17		1.35	0.15		1.33	0.11b		1.26	0.12b
NC	30	1.36	0.15		1.37	0.10		1.41	0.09c		1.44	0.10*c
PC	30	1.35	0.16		1.31	0.16		1.22	0.13*a		1.12	0.11*a
p-value+		.814			.334			.000			.000

SD: standard deviation, E1: experimental group, E2: comparison group, NC: negative control group, PC: positive control group.

⁺p-value by one way ANOVA, ^abcSame letter means no statistical difference by Scheffe test.

*p<0.05 by paired t-test between base and after.

Figure 1. Changes in PMA index by group and measurement period.

Figure 2. Changes in Talbott’s gingival index by group and measurement period.

2. Plaque test result

As a result of plaque test, the amount of change is shown in Table 5 and Table 6. The PHP index did not show any significant difference in all four groups before, after 1 week, and after 2 weeks. After 4 weeks, the experimental group was 2.04, the comparative group 2.09, the negative control group 2.35, and the positive control group 2.08, indicating that the experimental group had significantly lower PHP than the negative control group. Turesky's plaque index was not significantly different between 4 groups before, 1 and 2 weeks after the experiment. After 4 weeks, the experimental group was 2.44, the comparative group 2.47, the negative control group 2.67, and the positive control group 2.45, indicating that the experimental group and the positive control group had significantly lower plaque indices than the negative control group (p<0.01).

Table 5 . Changes in PHP index by group and measurement period

Group	N	Base			1 week			2 weeks			4 weeks
		Mean	SD		Mean	SD		Mean	SD		Mean	SD
E1	30	2.23	0.39		2.18	0.36		2.07	0.32		2.04	0.37a
E2	30	2.16	0.49		2.07	0.45		2.15	0.35		2.09	0.43ab
NC	30	2.16	0.37		2.21	0.30		2.21	0.36		2.35	0.35b
PC	30	2.21	0.41		2.06	0.44		2.09	0.34		2.08	0.41ab
p-value+		.833			.376			.424			.011

SD: standard deviation, E1: experimental group, E2: comparison group, NC: negative control group, PC: positive control group.

⁺p value by one way ANOVA, ^abSame letter means no statistical difference by Scheffe test.

Table 6 . Changes in Turesky’s plaque index by group and measurement period

Group	N	Base			1 week			2 weeks			4 weeks
		Mean	SD		Mean	SD		Mean	SD		Mean	SD
E1	30	2.58	0.38		2.53	0.32		2.55	0.31		2.44	0.27a
E2	30	2.60	0.36		2.53	0.29		2.47	0.29		2.47	0.30ab
NC	30	2.55	0.35		2.56	0.28		2.57	0.22		2.67	0.26b
PC	30	2.58	0.38		2.51	0.29		2.49	0.30		2.45	0.30a
p-value+		.959			.911			.503			.004

SD: standard deviation, E1: experimental group, E2: comparison group, NC: negative control group, PC: positive control group.

⁺p-value by one way ANOVA, ^abSame letter means no statistical difference by Scheffe test.

As people's living standards increase, interest in oral health increases, and with these changes, consumption of oral hygiene care products such as oral gargle is also increasing [9]. In this trend, oral products such as gargle and toothpaste have been developed a lot, and many studies and products have been released for the purpose of preventing dental caries and preventing periodontal disease [10]. As described above, in the test of this paper, dentifrice in the form of foam containing resveratrol, baicalin, and catechin is used. We wanted to see the inhibitory effect of plaque (dental plaque) formation.

First, plaque formed on the tooth surface along with food intake is a kind of bacterial membrane composed of various bacteria such as dental caries and causative agent of periodontal tissue disease [2,10]. Theoretically, as long as the plaque can be completely removed, it is possible to prevent dental caries, prevent periodontal tissue disease, and prevent oral diseases caused by bacteria present in the plaque [11]. However, it is impossible to completely remove the bacterial film on the tooth surface. Therefore, the most common and effective method of oral environment management is brushing [10,11].

Figure 3. Changes in PHP index by group and measurement period.

Figure 4. Changes in Turesky’s plaque index by group and measurement period.

In addition, gingivitis is the initial stage of periodontal tissue disease and is an inflammation of gingivitis [12]. The typical symptoms of gingivitis are redness and bleeding, and may progress to periodontitis if progressed by neglecting gingivitis [13]. Therefore, thorough plaque management is needed to prevent and treat periodontal tissue disease in which the majority of the people are affected. In order to manage plaques known to cause periodontal tissue disease, several methods of tooth brushing have been devised and educated. Dentifrice also, by mixing various substances in dentifrice, we are trying to prevent gingivitis and dental caries [12,13]. So, as a component cultured in this test, it was targeted to the grape branch extract of Campbell Early, which is the most cultivated in Korea, and after verifying its antioxidant effect, antioxidative and anti-inflammatory effects were verified in activated rodent-derived macrophages, RAW 264.7 cells and human-derived mast cells, HMC-1 cells [5].

In addition, Baikalin is contained in the roots of Koganevana Scutellaria baicalensis Georgi (Lamiaceae) (12.5% yield from dry roots). It is said that the golden hydrothermal extract containing Baikalin can exert anti-inflammatory effects through immunomodulation [7,14]. It has been shown that the golden hydrothermal extract maintains the initial inflammatory cytokine, which promotes inflammation, at a constant level to prevent the progression of excessive inflammation and progression to chronic inflammation, and does not exhibit excessive inhibitory effect, thereby maintaining an appropriate immune level in the body [7].

Green tea extract (catakin) does not have sufficient antibacterial activity when used alone. Methyl xanthine showed no intrinsic inhibitory activity against the bacterial strain tested. When green tea extract and catechin were used in combination with gentamicin, the MIC value of gentamicin against standard strains and clinical isolates was reduced, and synergistic effects were observed [8,14].

Therefore, in this study, 120 adult men and women were tested for 4 weeks using resveratrol, catechin, and baicalin combination dentifrice to see the effect of reducing gingivitis and plaque. Before the experiment, all subjects were trained in the rotation method, and the experiment was conducted by using the corresponding dentifrice for 3 minutes 3 times a day. In order to see gingivitis and plaque through the experiment dentifrice of this experiment, the gingivitis index (PMA index) and Talbott's gingival index were examined in gingivitis. And in the plaque examination, plaque management ability index (PHP index) and Turesky's plaque index were investigated. The most effective way to prevent gingivitis is to remove the bacterial film on the tooth surface at an early stage by using a silver salt prevention detergent. Ingredients or teeth that prevent oral diseases [12,15].

If you brush your teeth using a toothbrush containing ingredients that help in healing, the prevention and treatment of oral diseases can be increased [16,17]. The use of resveratrol, catechin and baicalin combination dentifrice reduced gingivitis and the plaque. Also, according to another study, esveratrol, catechin and baicalin toothpaste are effective not only in reducing dental plaque but also in reducing bad breath [18,19].

Therefore, patients with gingivitis and plaque are considered to be effective to use dentifrice with green tea extract, resveratrol, and bicalin. And It should be possible to use dentifrice according to each individual's oral disease and characteristics, and it is considered that multiple development of dentifrice is necessary.

No potential conflict of interest relevant to this article was reported.