KR102540804B1 - Composition containing Torreya nucifera extracts for preventing the formation of biofilm of mmicroorgaism - Google Patents

Abstract

The present invention relates to a composition containing a nutmeg tree extract having an effect of inhibiting biofilm formation, and more particularly, plaque, calculus, gingivitis or periodontitis containing as an active ingredient a nutmeg nut extract having a biofilm formation inhibitory effect of oral bacteria. It relates to a composition for preventing and improving oral diseases such as.

Description

Composition containing Torreya nucifera extracts for preventing the formation of biofilm of mmicroorgaism}

The present invention relates to a composition containing a nutmeg tree extract having an effect of inhibiting biofilm formation, and more particularly, plaque, calculus, gingivitis or periodontitis containing as an active ingredient a nutmeg nut extract having a biofilm formation inhibitory effect of oral bacteria. It relates to a composition for preventing, alleviating and improving oral diseases such as.

A biofilm is a microbial complex surrounded by various polymer materials formed by microorganisms, and is also called a biofilm or biofilm. These biofilms serve as a protective film that helps microorganisms adapt to various environments, and cells in the biofilm share metabolism with each other and acquire properties that are advantageous to survival through gene transfer. . Therefore, cells that form biofilms have much stronger resistance to harsh environments and antibiotics than when they exist in a general floating bacteria state, and biofilms formed in organs cause various diseases. Examples of such diseases or locations include dental caries, gingivitis, periodontitis, otitis media, voice prostheses, hydrocephalus hunts, and cystic fibrosis. ), valvular endocarditis, prosthetic heart valves, central venous catheter, prosthetic hip joint, prosthetic knee joint, chronic bacterial prostatitis , intrauterine devices, and urinary catheters.

On the other hand, it is known that about 1,000 kinds of bacteria exist in the oral cavity, and many bacteria inhabiting it form oral biofilms. Among them, bacteria associated with early oral biofilm formation are known as Streptococcus spp . These Streptococcus species are early attachment bacteria on the tooth surface, and in particular, Streptococcus gordonii and Streptococcus oralis play an important role in starting the formation of oral biofilms that cause tartar. In addition, Porphyromonas gingivalis is known as a major cause of gum disease such as periodontitis and gingivitis. Therefore, in inhibiting tartar formation, efforts to control these bacteria are continuing.

When metal ions such as calcium or magnesium in saliva are combined with the biofilm formed on the tooth surface, calcified calculus is formed. If tartar is not properly removed, it causes gum diseases such as gingivitis and periodontitis, and seriously causes tooth loss. In an effort to prevent the occurrence of periodontal disease, antibacterial agents such as chlorhexidine gluconate, cetylpyridium chloride, sanguinarine, and triclosan have been developed and applied to oral products such as toothpaste and toothpaste, but they are still the cause of gingivitis. Its occurrence cannot be fundamentally prevented. In addition, when these synthetic antibiotics are used for a long time, they not only stimulate the oral mucosa, but also destroy normal bacteria, rather increase resistance to pathogens in the oral cavity, and have great side effects on the human body, such as tooth discoloration and taste loss.

Therefore, there is a need for an oral product for preventing periodontal disease containing a natural antibacterial agent that can overcome the problems of existing products using synthetic compounds.

Bija tree ( Torreya nucifera ) is an evergreen tree of the yew family and lives in the mountains of the southern coast of Korea. The fruit, root bark and flowers are used medicinally. Efficacy such as skin moisturizing and inflammation relief (Republic of Korea Patent No. 10-1648466) and anti-stress (Republic of Korea Patent No. 10-1434449) have also been reported, but there has been no report on the efficacy of bija tree for oral periodontal disease.

Accordingly, the present inventors have devised the present invention to provide an oral composition containing nutmeg tree extract as an active ingredient and effective in preventing or improving tartar deposition and periodontal disease.

The problem to be solved by the present invention is to improve the disadvantages and problems of the prior art as described above, and one aspect of the present invention is to provide a composition for inhibiting biofilm formation.

Another object of the present invention is to provide an oral composition that is effective in preventing and improving tartar deposition and periodontal disease containing a nutmeg tree extract.

However, the problem to be solved by the present invention is not limited to those mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

In order to achieve the above object, one aspect of the present invention is to use at least one nutmeg extract selected from the group consisting of nutmeg stem extract, nutmeg leaf extract, nutmeg flower extract, nutmeg fruit extract and nutmeg seed extract as an active ingredient It provides a composition for inhibiting biofilm formation of microorganisms, characterized in that it comprises.

In the present invention, the microorganisms are Streptococcus gordonii , Streptococcus oralis , Porphyromonas gingivalis , Fusobacterium nucleatum , Prevotella Intermediate ( Prevotella intermedia ), Parvimonas micra ( Parvimonasmicra; Peptostreptococcus micros ) and Aggregatibacter actinomycetemcomitans ( Aggregatibacter actinomycetemcomitans; Actinobacillus actinomycetemcomitans ), but is not limited thereto. .

In order to achieve the above object, the present invention provides a functional food composition for preventing and improving periodontal disease.

In order to achieve the above object, another aspect of the present invention provides an oral composition for preventing, alleviating and improving periodontal diseases such as calculus, gingivitis and periodontitis, because the oral composition is effective in inhibiting biofilm formation. The oral composition may have one or more formulations selected from toothpaste, chewing gum, mouth freshener, mouth wash, and mouth spray.

According to the present invention, the composition for inhibiting biofilm formation of microorganisms containing a nutmeg tree extract has an antibacterial activity against oral bacteria and an inhibitory effect on biofilm formation, thereby preventing and improving periodontal diseases such as tartar, gingivitis, and periodontitis can do.

In addition, it is possible to provide a composition that is made of natural materials and is safe for the human body without side effects or risks caused by synthetic materials.

In addition, since it is safe for the human body, it can be applied in various ways as a product that directly touches or even enters the human body, such as a functional food composition, toothpaste, and mouth gargle.

The technical effects of the present invention are not limited to those mentioned above, and other technical effects not mentioned will be clearly understood by those skilled in the art from the description below.

1 is a photograph showing the biofilm formation pattern of those not treated with the nutmeg leaf extract and those treated with the nutmeg leaf extract.
Fig. 2 is a graph showing biofilm formation patterns in hydroxyapatite disks of those not treated with the nutmeg leaf extract and those treated with the nutmeg leaf extract.
3 is a graph showing the synthesis of water-insoluble glucans between those not treated with the nutmeg leaf extract and those treated with the nutmeg leaf extract.
Figure 4 is a graph showing the results of the cytotoxicity test of the nutmeg leaf extract on oral mucosal cells.

Hereinafter, the present invention will be described in more detail. However, this is only specifically described to aid understanding of the present invention, and the scope of the present invention is interpreted based on the claims.

The present invention is a microorganism characterized in that it comprises at least one nutmeg extract selected from the group consisting of nutmeg tree stem extract, nutmeg leaf extract, nutmeg flower extract, nutmeg fruit extract and nutmeg seed extract as an active ingredient It provides a composition for inhibiting biofilm formation of.

The nutmeg tree extract can be divided into a nutmeg stem extract, a nutmeg leaf extract, a nutmeg flower extract, a nutmeg fruit extract and a nutmeg seed extract according to the part used.

In the present invention, the "separate tree stem extract" is any type of extract obtained by subjecting the stem of a nutmeg tree to extraction, for example, an extract in the form of a juice obtained by applying heat directly or indirectly to a plant stem, or the upper part of a living plant stem. It includes all extracts in the form of extracts that flow out from cuttings, extracts obtained by extracting plants with organic solvents or water, extracts obtained by cooling the gas when burning plant stems, and the like.

In the present invention, the "fermented nut extract" includes an extract obtained by using an organic solvent or water, an extract obtained by pressing the nutmeg fruit, and an oil extract obtained by reducing the solvent extract.

In the present invention, the "seed extract of wisteria" includes an extract obtained by using an organic solvent or water, an extract obtained by pressing wisteria seeds, or an oil extract obtained by depressurizing the solvent extract.

In the present invention, "Bija tree leaf extract" means an organic solvent or water extract.

In the present invention, "Virginia flower extract" means an organic solvent or water extract.

For example, the nutmeg tree extract is extracted with an organic solvent such as alcohol having 1 to 4 carbon atoms, ethyl acetate, hexane, chloroform, water, or a mixture thereof after grinding each stem, leaf, flower, fruit, and seed may be an extract.

As the most preferred extract, an ethanol extract of nutmeg tree leaves may be used.

Cold extraction method, warm needle extraction method, or heat extraction method may be used as the method for preparing the extract, and a conventional extraction device, ultrasonic crushing extractor, or fractionator may be used.

According to another example, the nutmeg tree extract may be made into oil by depressurizing the organic solvent extract of the fruits and/or seeds.

The prepared extract may be filtered, concentrated, or dried to remove the solvent. Specifically, the filtration may be performed using a filter paper or a vacuum filter, and the concentration may be concentrated under reduced pressure using a vacuum concentrator, for example, a rotary evaporator. The drying may be performed by, for example, a freeze-drying method.

The composition according to the present invention is effective in inhibiting microbial biofilm formation.

Here, "microorganism" is a concept including all bacteria, fungi, yeast and algae that are particularly effective in inhibiting biofilm formation of microorganisms by the composition of the present invention. Therefore, in addition to the microorganisms directly confirmed to inhibit biofilm formation in the following examples and experimental examples, all microorganisms that can be expected and confirmed to inhibit biofilm formation based on the disclosure of the present specification within the scope of ordinary ability by those skilled in the art. It should be understood that microorganisms are included.

According to an embodiment of the present invention, in particular, Streptococcus gordonii, Streptococcus oralis , Porphyromonas gingivalis , Fusobacterium nucleatum , Prevotella intermedia ( Prevotella intermedia ), Parvimonas micra ( Parvimonas micra; Peptostreptococcus micros ) and Aggregatibacter actinomycetemcomitans ( Aggregatibacter actinomycetemcomitans; Actinobacillus actinomycetemcomitans ) It is effective in inhibiting biofilm formation.

The nutmeg tree extract according to the present invention can be appropriately used depending on the product used as long as it can inhibit biofilm formation. Typically, the nutmeg tree extract according to the present invention is contained in an amount of 0.001 to 5.0% by weight, preferably 0.2 to 2% by weight, based on the total weight of the composition, with respect to a composition exhibiting an inhibitory effect on microbial biofilm formation, including this composition. can be included

The composition has an antibacterial activity against periodontal disease-causing bacteria, and inhibits the formation of biofilm on the tooth surface, thereby suppressing calculus production, thereby exhibiting an excellent effect of preventing and improving gingivitis or periodontitis disease.

In the present invention, gingivitis refers to a disease in which inflammation occurs in the gums, which is a part of the oral mucosa, and symptoms of swelling, redness, pain, or local heat appear, and in severe cases, ulcers or gangrene may occur.

In the present invention, periodontitis (periodontitis) is also referred to as periodontitis, and refers to a periodontal disease in which teeth are lost as a result of destruction of cell tissue when a chronic infection is a disease mainly inventing gums and bony tissues.

The composition for inhibiting microbial biofilm formation according to the present invention can be directly applied to animals including humans.

The composition for inhibiting biofilm formation of microorganisms according to the present invention may further include various components in addition to the nutmeg tree extract depending on the method and purpose of use.

According to one embodiment of the present invention, it is possible to provide a functional food composition containing a nutmeg tree extract, for example, food or beverage.

In this specification, food means a natural product or processed product containing one or more nutrients, preferably means a state that can be eaten directly through a processing process to some extent, and as a general meaning, food, food additives, and health functional food and beverages, and includes various foods, beverages, gum, candy, tea, vitamin complexes, and functional foods.

According to one embodiment of the present invention, a composition for preventing and treating periodontitis or gingivitis is provided. Examples of such compositions include toothpastes, mouthwashes, mouthwashes, toothpaste creams, and the like.

When the composition according to the present invention is a toothpaste, it may further include an abrasive, a binder, a moisturizer, a foaming agent, a sweetener, a whitening agent or a flavoring agent in addition to the bean extract.

When the composition according to the present invention is a paste composition, the binder is garagenin, various cellulose derivatives for thickening, gums such as xanthan gum and tragacanth gum, polyvinyl alcohol, sodium polyacrylate, polyacrylic acid/maleic acid copolymer , organic binders such as synthetic polymer derivatives such as carboxyvinyl polymers, inorganic binders such as silica and laponite, etc. may be used alone or in combination.

The humectant may be sorbitol, glycerin, ethylene glycol, propylene glycol, polyether glycol, polypropylene glycol, or the like.

As the sweetener, sucrose, lactose, maltose, xylitol, sodium cyclamate, sodium saccharin, stevioside, aspartame, and the like may be used.

The fragrance is peppermint, menthol, anethol, eugenol, limonene, citronenol, alpha terpineol, salicylmethyl, cineol, linalol, itilinaroll, vanillin, thymol, spearmint oil, sage oil, Rosemary oil, cinnamon oil, etc. can be used.

In the case where the composition according to the present invention is a mouthwash solution, it can be prepared by mixing nutmeg extract with a common solvent and formulating it as a mouthwash solution, and cleaning the mouth 2 to 10 times a day to treat periodontal diseases such as periodontitis or gingivitis. You can book and treat.

When the composition according to the present invention is a toothpaste cream, it may further include a liquid including water and a moisturizing agent, a gelling agent or a water-insoluble brightening agent, a sweetener, a flavoring agent, and the like.

When the composition according to the present invention is a mouthwash, it may further include a toothpaste carrier, specifically non-toxic alcohol.

In addition to the above components, the composition may contain nutrients, vitamins, electrolytes, flavors, colorants, fillers, alginic acid and its salts, organic acids, thickeners, pH adjusters, flavors, stabilizers, preservatives, glycerin as appropriate according to the intended purpose of use. , alcohol, a carbonating agent, and the like may be further included.

According to one embodiment of the present invention, the oral composition of the present invention containing the nutmeg tree extract can be prepared and used in formulations such as toothpaste, chewing gum, mouthwash, mouthwash and mouthspray, but is not limited thereto no.

The toothpaste of the present invention includes a composition selected from the group consisting of an abrasive, a binder, a wetting agent, an active ingredient, a foaming agent, a sweetener, and a flavoring agent.

Hereinafter, experimental examples and examples will be described in detail to explain the present invention in detail. However, the experimental examples according to the present invention can be modified in many different forms, and the scope of the present invention should not be construed as being limited to the experimental examples and examples described in detail below. Experimental examples and examples of the present invention are provided to more completely explain the present invention to those skilled in the art.

Preparation Example 1: Manufacture of nutmeg extract

Dried nutmeg leaves were pulverized and exposed to an ultrasonic device for 30 minutes while immersed in twice the amount of ethanol. After soaking for 24 hours, the mixture was filtered through a filter paper, and then the solvent was removed using a rotary vacuum concentrator to prepare an extract.

Experimental Example 1: Oral strains and culture conditions

As strains, Streptococcus gordonii , Streptococcus oralis , and Porphyromonas gingivalis were purchased from the KCTC Center for Biological Resources and used.

BHI ( Streptococcus gordonii ), Streptococcus oralis , Porphyromonas gingivalis bacteria containing 5 ug/mL hemin and 1 ug/ml vitamin K ( After inoculation in brain heart infusion) broth medium, it was politically cultured at 37 ° C. under anaerobic conditions and used for experiments.

Experimental Example 2: Confirmation of antibacterial activity

A minimal inhibitory concentration (MIC) test was conducted to measure the antibacterial activity of the nutmeg tree extract against oral bacteria.

In a 96 well-plate, the bacterial culture medium at a concentration of about 5x10^5 CFU/mL was treated with extracts according to each concentration so that the final mixed solution was 200 uL. In the case of the extract, the extract of nutmeg tree extract concentrated under reduced pressure was dissolved in DMSO at a concentration of 50 mg/mL, and diluted with a sterilized liquid medium to a concentration of 500, 250, 125, 62.5, and 31.25 ug/mL.

A 96-well plate treated with the mixture including the bacterial culture medium and the extract was incubated at 37 ° C. for 24 to 72 hours under anaerobic conditions.

Determination of antimicrobial activity was performed by measuring the absorbance at 600 nm using a microplate reader immediately after sample treatment and after culturing for 24 to 72 hours for the absorbance of the experimental group compared to the control group without extract treatment, and calculating and comparing the change value.

S. gordonii S. oralis P. gingivalis S. gordonii + S. oralis + P. gingivalis MIC
(ug/mL) 125 31.3 >500 >500

Table 1 above shows the MIC values for Streptococcus gordoni, Streptococcus oralis, and Porphyromonas gingivalis single strains and complex strains, and the nutmeg tree extract was used for Streptococcus gorgonii and Streptococcus oralis, respectively. It was confirmed that antibacterial activity was exhibited at concentrations of 125 and 31.3 ug/mL or more.

Experimental Example 3: Measurement of Biofilm Formation Inhibiting Ability-Microplate

In order to confirm the biofilm formation inhibitory ability of the nutmeg tree extract, an experiment using a microplate was conducted.

About 5x10^5 CFU/mL of Streptococcus gordoni, Streptococcus oralis, and Porphyromonas gingivalis complex bacteria in a 96-well-plate, 200, 100, 50, 25, and 10 ug/mL of bija nut extract, respectively, 1 200 uL of liquid medium containing % sucrose was treated. The 96 well-plate was incubated at 37 °C under anaerobic conditions for 72 hours.

After culturing for 72 hours, the medium was removed, and 200 uL of purified water was added to each well to remove floating bacteria and bacteria weakly bound to the biofilm, and the operation was repeated three times. Thereafter, 200 uL of 0.1% crystal violet staining reagent was added to each well and reacted at room temperature for 15 minutes. After 15 minutes, it was washed three times with purified water to remove residual dye. After the stained biofilm was dried at room temperature, the degree of biofilm formation of each well was confirmed.

As shown in Figure 1, it was confirmed that biofilm formation was inhibited at a concentration of 200 ppm or more in the experimental group treated with the nutmeg tree extract of the present invention compared to the control group.

Experimental Example 4: Measurement of Biofilm Formation Inhibiting Ability-Hydroxyapatite disc

A hydroxyapatite disk experiment was conducted to confirm the biofilm formation inhibitory ability of the nutmeg tree extract.

Streptococcus gordoni, Streptococcus oralis, and Porphyromonas gingivalis complex bacteria about 5x10^5 CFU/ml, 200, 100, 50, 25 ug/mL, 1% sucrose of nutmeg tree extract, respectively, in a 24-well-plate 1 mL of liquid medium containing , and immersed in a hydroxyapatite disk coated with artificial saliva. The 24 well-plate was cultured at 37 °C under anaerobic conditions for 3 days.

After culturing for 72 hours, the medium was removed, and 1 mL of purified water was added to each well to remove floating bacteria and bacteria weakly bound to the biofilm, and the operation was repeated three times. Thereafter, 1 mL of 0.1% crystal violet staining reagent was added to each well and reacted at room temperature for 15 minutes. After 15 minutes, it was washed three times using purified water to remove residual staining reagent. After drying the dyed biofilm at room temperature, 95% ethanol was added to allow sufficient reaction. 100 uL of ethanol from which the dye was dissolved was transferred to a 96-well plate and absorbance was measured at 550 nm using a microplate reader.

As shown in Figure 2, it was confirmed that the experimental group treated with the extract of the present invention inhibited biofilm formation at a concentration of 100 ug / mL or more compared to the control group.

Experimental Example 5: Measurement of Insoluble Glucan Synthesis Inhibition

After culturing Streptococcus gordoni in BHI liquid medium for 24 hours, centrifugation (6,000 g, 4 ° C., 15 minutes) and the supernatant was taken. The extract was treated for each concentration in the supernatant of Streptococcus gordoni, and the concentration of sucrose was set to 1%.

After culturing at 37℃ for 18 hours, the reaction was stopped by adding the same amount of methanol, centrifuged (6,000 g, 4℃, 15 minutes), washed twice with purified water, and washed once with 70% ethanol to obtain water-insoluble glucan. Got a pellet.

After drying the residual solvent, the pellet was treated with a reagent in which anthrone was dissolved at 2 mg/mL in 75% H ₂ SO ₄ and reacted at 100 °C for 15 minutes. Then, after waiting until the temperature of the reactant dropped to room temperature, the absorbance was measured at 620 nm using a microplate reader.

As a result, as shown in FIG. 3, the water-insoluble glucan synthesis inhibitory activity of the nutmeg tree extract showed a water-insoluble glucan production rate of 42.4% and 78.7% at 200 and 100 ug/mL, respectively, compared to the non-treated nutmeg tree extract.

Experimental Example 6: Cytotoxicity to oral mucosal cells

An MTT experiment was conducted to measure the toxicity of nutmeg tree extract to oral mucosal cells.

hTERT-OME (immortalized human oral mucosal epithelial cell) was treated with about 1x10^4 cells/well in a 96 well plate, and then in RPMI-1640, 10% FBS, 1% penicillin-streptomycin medium at 37°C, 5% CO ₂ condition. incubated for 24 hours. After 24 hours, the medium used in the previous culture was removed, and the medium was treated with a nutmeg tree extract diluted to 200, 100, 50, 25, or 12.5 ug/mL, followed by reaction at 37°C and 5% CO ₂ for 24 hours. After treatment, 20 uL of 5 mg/mL MTT solution was added to each well, followed by reaction at 37°C and 5% CO ₂ for 3 hours. After the reaction, all the supernatant was removed, 150 uL of DMSO was added, reacted at room temperature for 10 minutes, and the absorbance was measured at 540 nm using a microplate reader.

As a result, as shown in FIG. 4, the nutmeg tree extract did not show cytotoxicity to oral mucosal cells at 200 ug/mL or less.

Example 1: Preparation of dentifrice composition containing nutmeg tree extract

Example - A toothpaste composition was prepared with the ingredients and composition ratios shown in Table 2 below.

The manufacturing procedure of toothpaste composition among oral compositions is to disperse a small amount of powdered ingredients such as carboxymethylcellulose sodium and saccharin sodium in amorphous sorbitol solution as a wetting agent, dilute with purified water, mix first in a mixer, and then mix silica and calcium carbonate. An abrasive was added and mixed a second time. And finally, a toothpaste composition was prepared by adding a foaming agent, a stabilizer, a flavoring agent, etc. and mixing the third time.

last name name Examples (% by weight) silica 20.0 Agriglycerin (98%) 30.0 Amorphous sorbitol solution (70%) 30.0 sodium lauryl sulfate 1.0 Sodium Carboxymethyl Cellulose 0.5 xanthan gum 0.3 sodium fluoride 0.22 Spices 1.0 sodium saccharin 1.0 citric acid Appropriate amount sodium bicarbonate 2.0 wisteria extract 0.02 Purified water To 100

The dentifrice composition of Example was prepared by including a nutmeg tree extract of 200 ug/mL, which is the minimum concentration that inhibits biofilm production.

From the above experiments, it was found that the birch tree extract was used to treat Streptococcus gordonii and Streptococcus oralis , which play an important role in the early stage of biofilm formation, which is the root cause of periodontal disease, and the foreskin causing gum disease. It was confirmed that it is a natural material that prevents or improves gum diseases such as periodontitis and gingivitis by inhibiting the formation of a bacterial film of Porphyromonas gingivalis .

Claims (7)

A composition for inhibiting biofilm formation of microorganisms, comprising a nutmeg leaf extract as an active ingredient. The composition for inhibiting biofilm formation according to claim 1, wherein the wisteria leaf extract is an extract extracted with alcohol having 1 to 4 carbon atoms, ethyl acetate, hexane, chloroform, or a mixed solvent thereof, or extracted with water. . The method of claim 1, wherein the microorganism is Streptococcus gordonii, Streptococcus oralis , Porphyromonas gingivalis , Fusobacterium nucleatum , Prevotella intermedia ( Prevotella intermedia ), Parvimonas micra ( Parvimonas micra; Peptostreptococcus micros ) and Aggregatibacter actinomycetem comitans ( Aggregatibacter actinomycetemcomitans; Actinobacillus actinomycetemcomitans ) Selected from one or more bacteria selected from the group consisting of Composition for inhibiting biofilm formation of microorganisms, characterized in that. The composition for inhibiting biofilm formation of microorganisms according to claim 1, wherein the wisteria leaf extract is contained in an amount of 0.001 to 5.0% by weight based on the total content of the composition for inhibiting biofilm of microorganisms. The composition for inhibiting biofilm formation of microorganisms according to any one of claims 1 to 4, wherein the composition for inhibiting biofilm of microorganisms is a health functional food composition for preventing or improving periodontitis or gingivitis. The composition for inhibiting biofilm formation of microorganisms according to any one of claims 1 to 4, wherein the composition for inhibiting biofilm of microorganisms is an oral composition for preventing and improving periodontal disease. The composition for inhibiting biofilm formation of microorganisms according to claim 6, wherein the composition has a formulation of at least one selected from toothpaste, chewing gum, mouthwash, mouthwash, and oral spray.

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