WO2022054629A1

WO2022054629A1 - Agent for improving and maintaining oral flora, and agent for removing dental plaque/teeth plaque and preventing tartar formation

Info

Publication number: WO2022054629A1
Application number: PCT/JP2021/031820
Authority: WO
Inventors: 武司片岡; 壮介小川; 知樹小林; 邦義清水
Original assignee: 株式会社アカシアの樹; 国立大学法人九州大学
Priority date: 2020-09-10
Filing date: 2021-08-31
Publication date: 2022-03-17
Also published as: JP7448122B2; JP2022046082A

Abstract

[Problem] The state of an oral flora not only influences the oral health status but also affects an intestinal flora. Deterioration of the intestinal flora leads to various diseases, autoimmune disorders, depression, etc. The balance of the oral flora is disrupted by the formation of an oral biofilm as well as the prolonged presence thereof. Therefore, oral biofilms become a pressing issue. [Solution] An agent for improving and maintaining an oral flora, said agent comprising an acacia bark extract as an active ingredient. This agent lowers the oral biofilm formation rate and releases biofilm-forming bacteria from the biofilm. As a result, the agent inhibits the growth of gram-negative bacteria and removes the gram-negative bacteria together with other bacteria to thereby enable the oral flora to keep a good balance.

Description

Oral flora improving / maintaining agent, plaque / plaque removal and tartar formation preventive agent

The present invention relates to an oral flora improving / maintaining agent for improving the disorder of the oral flora (bacterial flora) or preventing and maintaining the disorder.

Periodontal disease, which is one of the oral diseases, is a disease that occurs in the periodontal tissue consisting of four tissues: soft gingiva, root membrane and hard tissue cement, and alveolar bone. Announced by the Ministry of Health, Labor and Welfare (FY2017) ), The total number of patients with "gingival inflammation and periodontal disease" is said to be 3,983,000. There are 300 to 400 types of bacteria in the oral cavity, but when the periodontal disease bacteria contained in them grow abnormally in the gingival sulcus (the boundary between the teeth and gums), a periodontal pocket is formed and the attachment part is the tooth. It peels off the surface, followed by gingival swelling, causing destruction of the alveolar bone.

In the past, oral diseases such as periodontal disease were thought to be caused by specific pathogens, so antibacterial agents targeting specific pathogens are mainly studied in research aimed at preventing or improving oral diseases. Was there.

For example, Patent Document 1 describes an oral composition containing a polyphenol-containing natural product extract and lysine as active ingredients and exhibiting an antibacterial effect against pathogenic bacteria of periodontal disease (particularly Porphyromonas gingivalis). Has been done.

Japanese Unexamined Patent Publication No. 2019-21252

Periodontal disease has been considered to be a mixed infection caused by several types of periodontal disease bacteria. However, in recent years, periodontal disease is not caused by a single pathogen, but is a disease in which the pathogenicity of biofilm increases as a result of the interaction of various bacterial species including indigenous bacteria, that is, the oral cavity. It has come to be regarded as a disease caused by the disorder of flora (bacterial flora) (Dysbiosis).

Oral flora refers to a community of 300 to 400 types of bacteria existing in the oral cavity, and its condition (quality and balance of bacteria, etc.) affects not only the oral cavity but also the health of the whole body. Bacteria that make up the oral flora are roughly composed of three types, good bacteria, bad bacteria, and opportunistic bacteria, and they are fighting for power with each other. The higher the proportion of good bacteria, the healthier it is, and the more bad bacteria it has, the more likely it is that health problems will occur. Optimum bacteria are the bacteria with the highest proportion, but they are harmless when the flora is balanced, but when the number of bad bacteria increases, they become active in the bad bacteria.

In a healthy oral cavity, about 75% of biofilms are indigenous bacteria (gram-positive aerobic bacilli) and are not pathogenic to periodontal disease. However, long-term presence of biofilm in the oral cavity causes gram-negative bacteria such as Fusobacterium, Porphyromonas, Treponema, Prevotella, and Aggregatibacter to grow in the biofilm, with gram-negative anaerobic bacilli occupying about 75% of the oral cavity. As a result, the balance of the oral flora is lost, and diseases and troubles such as periodontal disease and bad breath occur.

In addition, the condition of the oral flora does not merely affect the health condition of the oral cavity, but also affects the intestinal flora. The intestine is a place that absorbs nutrients and is an important organ that controls 70% of immunity, and intestinal bacteria are greatly involved in these functions. Not only that, it is becoming clear that the products produced by intestinal bacteria affect various organs of the body and even the brain. Deterioration of intestinal flora can be caused by various diseases such as colon cancer, breast cancer and diabetes, obesity, dementia, allergic diseases such as hay fever and atopic dermatitis, autoimmune diseases, and depression. Involved in mental illness.

And even though the oral cavity and intestines are separated, they are connected by a tube and affect each other. That is, when the oral flora is predominantly bad bacteria and P. gingivalis increases and flows in large quantities from the mouth to the gastrointestinal tract, most of it is sterilized in the stomach, but some P. gingivalis survive and intestine. It has been reported that the inner flora is out of balance. On the contrary, when the environment of the intestinal flora deteriorates and the number of bad bacteria in the intestine increases, the mucosal barrier function of the intestine that prevents the invasion of harmful substances decreases, and the intestinal bacteria invade the intestinal cells and become inflamed. At the same time, it weakens the immune system of the whole body, causes systemic inflammation, and becomes susceptible to infections. Therefore, it is possible that the environment is such that periodontal disease bacteria are likely to increase, and the condition of the oral flora also deteriorates.

In view of the above circumstances, the present invention particularly suppresses the growth of gram-negative bacteria by reducing the formation rate of the biofilm and releasing the fungi constituting the biofilm from the biofilm. By removing Gram-negative bacteria in combination with other bacteria, the oral flora can be kept in a good balance.

Therefore, in order to solve the above-mentioned problems, as a first invention, an oral flora improving / maintaining agent containing an acacia bark extract as an active ingredient is provided.

The second invention is described in the first invention, which promotes the withdrawal of Gram-negative bacteria and / and Gram-positive bacteria containing acacia bark extract as an active ingredient from the oral biofilm (plaque, plaque). Provides an oral flora improving / maintaining agent.

As the third invention, the oral cavity according to the first invention, which suppresses the formation of an oral biofilm (plaque, plaque) of Gram-negative bacteria and / and Gram-positive bacteria containing acacia bark extract as an active ingredient. Provides an inner flora improving / maintaining agent.

As a fourth invention, the gram-negative bacterium and / and the gram-positive bacterium is a second invention or a second invention containing at least two or more of Streptococcus gordonii, Streptococcus mitis, Fusobacterium nucleatum, P. gingivalis, Treponema spp, and Tannerella forsythia. Provided is the oral flora improving / maintaining agent according to the third invention.

As the fifth invention, the food and drink containing the oral flora improving / maintaining agent according to any one of the first to fourth inventions is provided.

As the sixth invention, an oral cleansing agent containing the oral flora improving / maintaining agent according to any one of the first invention to the second invention is provided.

As the seventh invention, a dentifrice containing the oral flora improving / maintaining agent according to any one of the first invention to the second invention is provided.

The eighth invention provides a plaque / plaque removal agent and a tartar formation preventive agent containing an acacia bark extract as an active ingredient.

The ninth invention is the removal of plaque / tartar according to the eighth invention, which promotes the withdrawal of Gram-negative bacteria and / and Gram-positive bacteria containing acacia bark extract as an active ingredient from the oral biofilm. And an agent for preventing tartar formation.

The tenth invention includes the removal of plaque / tartar according to the eighth invention, which suppresses the formation of an oral biofilm of Gram-negative bacteria and / and Gram-positive bacteria containing an acacia bark extract as an active ingredient. To provide a tartar formation preventive agent.

In the eleventh invention, the gram-negative bacteria and / and the gram-positive bacteria include at least two or more of S. gordonii, S. mitis, F. nucleatum, P. gingivalis, Treponema spp, and T. forsythia. Provided are the plaque / plaque removal and anti-stone formation preventive agent according to the ninth invention or the tenth invention.

As the twelfth invention, the food and drink containing the plaque / plaque removal and tartar formation preventive agent according to any one of the eighth to ninth inventions is provided.

As the thirteenth invention, the oral cleansing agent containing the plaque / plaque removal and tartar formation preventive agent according to any one of the eighth to ninth inventions is provided.

As the fourteenth invention, the dentifrice containing the plaque / plaque removal and tartar formation preventive agent according to any one of the eighth to eleventh inventions is provided.

As the fifteenth invention, the plaque / plaque removal and tartar formation preventive agent according to any one of the eighth to eleventh inventions is used to remove plaque / plaque and prevent tartar formation. Provided are methods for removing plaque / plaque and preventing tartar formation.

As the sixteenth invention, an oral flora improving / maintaining method for improving / maintaining an oral flora by using the oral flora improving / maintaining agent according to any one of the first to fourth inventions. offer.

INDUSTRIAL APPLICABILITY The present invention provides an oral flora improving / maintaining agent for improving the disorder of the oral flora or preventing and maintaining the disorder, and also provides a plaque / plaque removal and tartar formation preventive agent. be able to.

The figure which shows the formation model of the intraoral biofilm Conceptual diagram showing the method of biofilm formation inhibition test Conceptual diagram showing the method of biofilm removal test The figure which shows the result of the biofilm formation inhibition test The figure which shows the result of the biofilm removal test

Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. The present invention should not be limited to these embodiments, and may be implemented in various embodiments without departing from the gist thereof.
<Embodiment 1>
<Overview>

This embodiment is an oral flora improving / maintaining agent containing an acacia bark extract as an active ingredient. Further, it is a food or drink, an oral cleanser and a dentifrice containing the oral flora improving / maintaining agent. In addition, the oral flora improving / maintaining agent is intended to suppress or decompose the formation of an oral biofilm.
<Structure>

"Acacia" means a tree belonging to the genus Acacia of the leguminous family. Examples of acacia include Acacia mernsii De Wild., Acacia mangium Wild., Acacia dealbata Link, Acacia decurrens Wild., Acacia pycnantha, and the like. Although not particularly limited, the acacia is preferably Acacia mearnsii De Wild. Or Acacia mangium Wild., And more preferably Acacia mearnsii De Wild.

This embodiment uses an acacia bark extract. The tree trunk structure is "outer bark-cork forming layer-inner bark-vascular cambium-tree part" from the outside, and generally "outer bark-cork forming layer-inner bark" is used as the bark. I'm calling. Similarly, the bark in this embodiment is also referred to as "outer bark-cork cambium-inner bark".

Bark extract refers to a substance eluted from the bark with water or an organic solvent. The water is preferably hot water. The organic solvent is preferably an alcohol, more preferably an alcohol having 1 to 4 carbon atoms, and particularly preferably ethanol. As the extraction solvent, one kind of solvent may be used alone, or two or more kinds of solvents may be used in combination. Further, the bark may be crushed and then extracted, or the bark may be crushed and dried before extraction.

It is known that about three-quarters of acacia bark extract consists of proanthocyanidins and about one-fourth consists of sugars such as sucrose. (Reference 1. Rie Kusano, et al., Α-Amylase and Lipase Inhibitory Activity and Structural characterization of Acacia Bark Proanthocyanidins, J Nat Prod. 2011 Feb 25; 74 (2): 119-28.) Matsuo, et al, characterization of the α-Amylase Inhibitory Activity of Oligomeric Proanthocyanidins from Acacia learnsii Bark Extract, Natural product Communications, Vol.11 No.12 1851-1854, 2016).

Further, proanthocyanidins in the bark extract of Acacia mearnsii are obtained by polymerizing 5-deoxyflavan-3-ols such as robinetinidol and fisetinidol and flavan-3-ols such as catechin and galocatechin (see Reference 1.).

The oral flora improving / maintaining agent of the present embodiment has an action of promoting the withdrawal of gram-negative bacteria and / and gram-positive bacteria from the oral biofilm. Further, the oral flora improving / maintaining agent of the present embodiment has an action of suppressing the formation of an oral biofilm of Gram-negative bacteria and / and Gram-positive bacteria.

In addition to Streptococcus mutans and Streptococcus sobrinus described in detail below, periodontal disease-related fungi, Pseudomonas aeruginosa, etc. are known to synthesize large amounts of glucan from sugar in order to actively form biofilms. (Refer to 3. Hanada, Control of Bacterial Biofilm in Oral Geriatric Dentistry, Vol. 16, No. 3, 2002) Furthermore, because sucrose has a high-energy bond, bacteria become sticky due to hydrolysis. -It is known to make insoluble glucan and form biofilms. (Refer to Reference 4. Hanada, Clinical Biology of Biofilm, J Health Care Dent. 2003; 5: 4-30.) As mentioned above, sugars such as sucrose promote the formation of biofilm and inhibit the formation. It does not promote withdrawal. Therefore, it is considered that proanthocyanidins in the acacia bark extract are involved components.
<Gram-negative bacteria>

Gram-negative bacteria are bacteria that do not stain purple by the Gram stain method. Examples of Gram-negative bacteria include P. gingivalis, Treponema spp, and T. forsythia, and these Gram-negative bacteria are classified into the category with the highest pathogenicity of periodontal disease. It has been reported that T. forsythia aggregates F. nucleatum and T. forsythia, suggesting that preventing F. nucleatum from adhering to the tooth surface makes it difficult for T. forsythia to adhere. In addition, Actinobacillus actinomycetemcomitans, Prevotella intermediate, Prevotella denticola, and Prevotella loescheii are also causative bacteria of periodontal disease. In addition, F. nucleatum is the causative agent of periodontal disease and plays a central role in the formation of oral biofilm. Prevotella nigrescens is the causative agent of periodontal disease, and it has been pointed out that it is particularly involved in adolescent gingival inflammation and gestational gingival inflammation. It is also known as a cause of chronic persistent inflammation in periodontal disease. It has also been reported that it co-aggregates with F. nucleatum. In addition, as early colony-forming bacteria of oral biofilm, Capnocytophaga gingivalis, Capnocytophaga ochracea, Capnocytophaga sputigena, Eikenella corrodens, Veillonella atypica, Haemophilus parainfluenzae, Campylobacter rectus, Peptostreptococcus micros, Campylobacter There are parvula and so on.

Gram-negative bacteria have LPS (lipopolysaccharide) and are not easily released from the cell wall, but are released by thawing and destroying cells when the bacteria die, which acts on animal cells and the like. It is toxic by doing so.
<Gram-positive bacteria>

Gram-positive bacteria are bacteria that stain purple by the Gram stain method. For example, Eubacterium spp is the causative bacterium of periodontal disease, and Eubacterium nodatum is said to be the causative bacterium. In addition, S. gordonii and S. mitis are classified as early adherent bacteria of biofilm, and it is said that 60 to 90% of the initial colonization is in the genus Streptococcus. In addition, as early colony-forming bacteria of oral biofilm, there are Actinomyces israelii, Actinomyces naeslundii, Propionibacterium acnes, Selenomonas flueggei, Streptococcus oralis, Streptococcus sanguis, Streptococcus constellatus, Streptococcus intermedius, Actinomyces odontolytius and the like.
<Intraoral biofilm>

The oral biofilm is a white and sticky film in which bacteria grow on the surface of the teeth and the residue of food is sometimes called plaque or plaque. An oral biofilm is a film in which various types of bacteria gather and begin to have a relationship with each other over time, compensating for each other's weaknesses and forming a film. When this oral biofilm is formed, various bacteria that have propagated on the teeth are protected by the cover of the oral biofilm, and the bactericidal power of saliva becomes less effective, leading to the progression of tooth decay and periodontal disease. I will go.

Since the present invention promotes the decomposition of the biofilm in the oral cavity, it inhibits the biofilm from continuing to grow or exist for a long time in the oral cavity, and as a result, the development of the relationship between the bacteria in the biofilm. Inhibits. Inhibition of the development of this relationship results in suppression of the growth of bad bacteria in the biofilm and contributes to maintaining the health of the oral flora.

Tartar is a plaque that adheres to the teeth and reacts with calcium and phosphoric acid contained in saliva to calcify, becoming hard like stones and sticking to the surface of the teeth. Tartar is a collection of dead bacteria and does not itself cause periodontal disease like the oral biofilm. The surface of the tartar is uneven, so the oral biofilm tends to adhere. Therefore, it results in further inflammation of the gums.

Biofilms formed by bacteria that propagate in the oral cavity are found in halitosis (Reference 5. Koji Shibuya, Study on the components and origin of physiological halitosis, Oral Hygiene Journal, J. Dent. Hlth, 51: 778? 792, 2001, Reference 6). . J. Washio, et al. (2005) Hydrogen sulfide-producinng bacteria in tongue biofilm and their relationship with oral malodour. J. Med. Microbiol., 54, 889-895.), Oral flora imbalance, oral cavity Unsanitary conditions in the mouth, periodontal disease (Refer to 7. Tomoko Kadowaki et al., Periodic disease and gingipine, Nikkei Journal (Folia Pharmacol. Jpn.) 122, 37-44 2003.), and related to periodontal disease. It is known to be the cause of systemic diseases (see 8. Naoko Hirohata et al., Halitosis and systemic diseases, Nihon University Medical Journal, 73 (5): 211-218 (2014).). Oral biofilms are not composed of one type of fungus, but are formed by the complex propagation of various fungi. More specifically, the mechanism of establishment of pathogenic biofilms is divided into three stages. (Reference 9. Masae Kuboniwa, et al., Subgingival biofilm formation. Periodontology, 2000 52, 38-52. 2010., Reference 10. Paul E. Kolenbrander, et al., Oral multispecies biofilm development and the key cell distance. Nat. Rev. Microbiol. 8, 471-480. 2010., Reference 11. Paul E. Kolenbrander, et al., Communication amon Oral Bacteria, MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, Sept. .reference).

FIG. 1 (see Reference 11.) is a diagram showing a formation model of an intraoral biofilm. The process of forming the intraoral biofilm is as follows. First, proteins in saliva adhere to the surface of teeth and form a film called pellicle. Subsequently, initial adhering bacteria such as S. gordonii and S. mitis adhere to the surface of the solid layer to form colonies on the pellicle. Next, the mediator F. nucleatum adheres to the flora, and then P. gingivalis, T. denticola, P. intermedia, A. actinomycetemcomitans, which are pathogens that are the main cause of periodontal disease as late adherent bacteria, are bacteria. The biofilm is formed in three stages: attachment to the flora.

From this, it is considered that if the formation of biofilm formed by the early adherent bacteria S. gordonii, S. mitis and the mediator F. nucleatum can be inhibited or eliminated, the late adherent bacteria cannot be established. .. As a result, imbalance of oral flora caused by oral biofilm, prevention of halitosis (see References 5 and 6), oral unsanitization, prevention of periodontal disease (see Reference 7), teeth It is considered that systemic diseases caused by peri-illness (see Reference 8.) can be prevented.
<Characteristics of initial adhering bacteria and mediators>

It is known that Gram-positive bacteria S. gordonii, Gram-positive bacteria S. mitis, etc., which are early adherent bacteria, and Gram-negative bacteria F. nucleatum, which are mediators, form biofilms and cause various oral problems. ing. For example, S. gordonii is not only involved in the formation of biofilm in the oral cavity and the subsequent onset of dental caries and gingival inflammation, but is also attracting attention in the fields of dentistry and medicine as the causative agent of infective endocarditis. (Refer to Reference 12. Yukihiro Takahashi et al., Adhesin of Oral Streptococcus pyogenes, Journal of Japanese Bacteriology, 62 (2), 283-293, 2013.).

S. gordonii is said to play an important role as an adhesion factor when P. gingivalis becomes a constituent bacterium of dental plaque. Furthermore, it directly destroys periodontal tissue and stimulates immune cells such as macrophages in the periodontal tissue to produce mediators such as inflammatory cytokines, causing damage to the tissue and periodontitis-related cells. It has been shown that it may be involved in periodontitis by indirect mechanisms such as colonization of the lesion and influence of proliferation (Reference 13. Shihoko Tajika et al., Oral cavity of healthy adults and patients with periodontitis. For the distribution of Lenza bulbs, see Iwa Medical University Dental Journal, 21, 66-77, 1996.). S. mitis produces a substance that induces the production of inflammatory cytokines outside the cells, suggesting its potential as a causative agent of various inflammatory diseases.

In F. nucleatum, it is known as a high-producing bacterium of methyl mercaptan (CH ₃ SH), which is an odorous substance that causes halitosis, and it is the main cause of halitosis (Reference 5., Reference 14. Miki Matsui et al.) , The condition of gingiva and the involvement of bacteria in halitosis and tongue moss on halitosis in young people without periodontitis.
<Halitosis>

The odorous substances that cause halitosis are hydrogen sulfide (H ₂ S) and methyl mercaptan (CH ₃ SH) produced by bacteria in the oral cavity using amino acids based on proteins present in the oral cavity as substrates. Among volatile sulfur compounds (VSC: Volatile Sulfur Compounds), methyl mercaptan is always present at a high concentration relative to the cognitive threshold, and the sensory evaluation value (discomfort) is high or low in exhaled breath and mouth with an unpleasant odor. It has been confirmed that this is the main component of the unpleasant odor. Methyl mercaptan has been found to have high production capacity in periodontal disease-related bacteria Porphyromonas and Fusobacterium (see References 5 and 6).

F. nucleatum is a long linear gram-negative anaerobic bacterium that is present in large volume ratios in oral biofilms and the like. It is resident in the human oral cavity and is also called a spindle bacterium because both ends of the bacterium are sharp and the central part is slightly thick. F. nucleatum is one of the periodontopathogenic bacteria that plays a central role in the formation of oral biofilms and forms an oral biofilm by co-aggregating with other bacteria. (See Ref. 9, Ref. 10, and Ref. 11.). In addition, it produces butyric acid, which has no sugar resolution and causes bad odor (halitosis). (See Reference 5.).
<Unbalanced oral flora>

In a healthy oral cavity, about 75% of biofilms are indigenous bacteria (gram-positive aerobic bacilli) and are not pathogenic for periodontal disease. On the other hand, in the periodontal pocket of adult periodontitis, gram-negative bacteria such as Fusobacterium, Porphyromonas, Treponema, Prevotella, and Aggregatibacter grow in the biofilm, and gram-negative anaerobic bacilli occupy about 75% of the oral cavity. And the balance of the oral flora is lost.
<Candidiasis>

It is a disease that causes pain, discomfort, and dysgeusia due to the increase of fungal bacteria in the mouth. It is especially likely to occur in the elderly with weakened immunity. In addition, candidiasis of the pharynx and esophagus may also occur.
<Aspiration pneumonia>

Pneumonia can occur when oral bacteria accidentally enter the lungs through the trachea and spread inflammation. It begins with symptoms such as frequent eating, taking longer than before, and difficulty breathing. Particular attention should be paid to elderly people whose swallowing ability is weakened and patients whose swallowing ability does not function well due to cerebrovascular disease or muscle disease. In severe cases, it can be life-threatening.
<Infective endocarditis>

Bacteria in the oral cavity may get on the blood and spread throughout the body, infecting the heart and causing heart disease. In addition, it may be fatal due to seriousness.
<Other diseases involving oral flora>

For example, oral flora such as sepsis, myocarditis, arteriosclerosis, diabetes, dermatitis, nephritis, rheumatoid arthritis, and osteoporosis is involved.
<Unsanitary in the oral cavity>

F. nucleatum is a long linear gram-negative anaerobic bacterium. It occupies a large volume in the oral biofilm. It is resident in the human oral cavity and has a spindle shape. This bacterium is the central bacterium for the formation of oral biofilm, and forms an oral biofilm by aggregating with other bacteria.
<Periodontal disease>

P. gingivalis, a gram-negative obligate anaerobic bacterium that is a periodontal disease bacterium, is a pathogenic bacterium that is regarded as important in the onset and progression of periodontitis, and produces a strong protease on the surface of the bacterium and outside the bacterium. It is thought to produce various conditions related to periodontal disease (see Reference 7.). In addition, P. gingivalis is known to adhere to the oral biofilm, and it is considered important to control the oral biofilm to which P. gingivalis adheres in order to prevent periodontal disease.

P. gingivalis produces pathogenic factors such as internal toxins (LPS), proteases, fibrous hair, and butyric acid, which penetrates into the resorption of gingiva and alveolar bone and enters the tissue gap, and is involved in periodontal tissue destruction. It is thought to be closely related to disease-induced systemic diseases. In addition, there are pathogenic bacteria that are regarded as important in the onset and progression of periodontal disease, and A. actinomycetemcomitans, which is said to be closely related to invasive periodontitis, is extrabacterial. It has vesicles, and toxins such as internal toxins and leukotoxins are present, which induces cell apoptosis, and P. intermedi is associated with invasive periodontitis for adolescent gingitis and gestational gingitis. It has been pointed out that they are involved.
<Mechanism of inflammation caused by P. gingivalis>

Toxins such as LPS, gingipain, and leukotoxin produced by the bacteria forming the biofilm cause an inflammatory reaction locally in the periodontal tissue. Furthermore, various cytokines and enzymes such as IL-1β and TNF-α destroy periodontal tissue. If the periodontal tissue is continuously stimulated chronically, the host cells cause a continuous or excessive immune reaction in the periodontal tissue, and the periodontal tissue is repeatedly destroyed.
<Beyond periodontal disease>

In recent years, attention has also been paid to the relationship with systemic diseases such as aspiration pneumonia, diabetes, arteriosclerosis, and pregnancy complications. This is because periodontal disease is not limited to local inflammation in the oral cavity, but is caused by inflammatory factors such as inflammatory cytokines (IL-1β, 6, 8, TNF-α, etc.) produced by the bacteria themselves and immune reactions. By affecting the whole body through the bloodstream, it often causes deterioration of the general condition. Therefore, the spread of inflammation to the whole body through periodontal pathogens and their immune reactions has been attracting attention as a link between periodontal disease and systemic disease (see Reference 8.).
<Supplementary information on oral biofilm formation>

On the tooth surface, what is called a pericle to which proteins and organic substances are bound is formed. Next, it is a specific oral indigenous bacterium called an early colonizers that attaches to the protein receptor of the pellicle and begins to form a colony. Specifically, S. mitis, S. gordonii, etc. are established as an early colonization group among indigenous oral bacteria. When colonies formed by the early colonization bacteria are formed on the tooth surface, the pellicle is hidden and disappears. Next, the bacterial group having adhesin against the surface protein on the bacterial surface of the early colonization group grows as a receptor. Then, the bacteria settle and proliferate in a coherent manner due to the relationship between the receptor and adhesin. In this way, colonies of late colonization bacteria are completed. At this time, the spindle fungus F. nucleatum plays a central role in the oral cavity.

These late colonization groups continue to metabolize by utilizing the metabolites of the early colonization bacteria group, and at the same time, they produce their own metabolites, and other bacteria use these metabolites in order. It can be seen that A. actinomycetemcomitans, P. intermedia, P. gingivalis, and T. denticola are typical periodontal disease bacteria, and periodontal disease bacteria are bacteria that multiply and appear unless the teeth are brushed.
<Embodiment 1 test>
<Test method>
<Preculture of microorganisms>

Brainheart infusion bouillon medium (Nissui Pharmaceutical Co., Ltd.) with S. gordonii (initial adherent bacteria), S. mitis (initial adherent bacteria), F. nucleatum subsp. Nucleatum (JCM No. 8532) 6 to 7 respectively. Precultured for days at 37 ° C. under anaerobic conditions (Aneropack Kenki, Mitsubishi Gas Chemicals Co., Ltd.).
<Biofilm formation inhibition test>

FIG. 2 is a conceptual diagram showing a method of a biofilm formation inhibition test. First, a 96-well plate was immersed in an acacia bark extract sample for 5 minutes in a clean bench, then the acacia bark extract sample was removed and air-dried for 60 minutes. Acacia bark extraction is performed by inoculating pre-cultured S. gordonii (initial adherent bacteria), S. mitis (initial adherent bacteria), and F. nucleatum subsp. Nucleatum (JCM No. 8532) in 1% each of the medium. 100 μL was added to each of the 96-well plates coated with the product sample. Then, it was statically cultured for 2 days at 37 ° C. under anaerobic conditions to form a biofilm. Two days later, the supernatant was removed, the biofilm was stripped by pipetting, and the turbidity of the liberated bacteria was measured at a wavelength of 570 nm. Berberine, cetylpyridinium chloride (CPC) and mouthwash GUM Night Care (non-alcoholic type) (Sunstar Co., Ltd.) were used as positive controls.
<Biofilm removal test>

FIG. 3 is a conceptual diagram showing a method of a biofilm removal test. First, 96 inoculated 1% each of pre-cultured S. gordoni (initial adherent bacteria), S. mitis (initial adherent bacteria), and F. nucleatum subsp. Nucleatum (JCM No. 8532) into the medium. 100 μL each was added to the well plate. Then, it was statically cultured for 6 days at 37 ° C. under anaerobic conditions to form a biofilm. After removing the supernatant, the biofilm formed on a 96-well plate was immersed in an acacia bark extract sample for 3 minutes. The turbidity of the liberated bacteria was measured at a wavelength of 570 nm. Berberine, cetylpyridinium chloride (CPC) and mouthwash GUM Night Care (non-alcoholic type) (Sunstar Co., Ltd.) were used as positive controls.
<Test results Inhibition of biofilm formation>

FIG. 4 shows the results of the biofilm formation inhibition test, showing the proportion of biofilm formed on 96-well plates pretreated with acacia bark extract samples. As shown in the figure, a decrease in the biofilm formation rate was confirmed in the test plots pretreated with the acacia bark extract sample as compared with the control. In particular, in the acacia bark extract sample 3 mg / mL concentration treatment group, it was found that the formation of biofilm was significantly inhibited as compared with the control (water).
<Test result Biofilm removal>

FIG. 5 is a diagram showing the results of the biofilm removal test, showing the turbidity of the bacteria released by the biofilm formed on the 96-well plate by the treatment of the acacia bark extract sample. As a result, it was confirmed that the bacteria were significantly released from the biofilm in all the acacia bark extract sample treatment groups tested in this study as compared with the control (water). It was also suggested that the biofilm removing ability of the acacia bark extract sample was high even when compared with the positive control.
<Food and drink>

There are various forms of the oral flora improving / maintaining agent of the present embodiment, and they can be provided in the form of solid, granular, liquid, gel, powder, paste and the like. Further, by using a known method, it can be provided as a food or drink, an oral cleanser, a dentifrice or the like containing the oral flora improving / maintaining agent.

For example, when it comes to food and drink, various beverages such as tea, soft drinks, lactic acid bacteria beverages, vegetable beverages, sports beverages, coffee, cocoa, and liquor, canned foods, frozen foods, retort foods, and instant soups (miso juice), Improved oral flora for various foods and drinks such as seasonings such as sauce, dressing and sauce, noodles, dairy products, jams, candy, caramel, gum, chocolate, biscuits, cakes, Japanese sweets, ice cream, cereals, processed fish meat products, etc. -By appropriately containing a maintenance agent, it can be provided as a food and drink for improving and maintaining the oral flora.
<Oral cleanser>

When used as an oral cleansing agent (also called mouthwash, mouthwash, dental rinse, mouthwash, etc.), chlorhexidine gluconate, cetylpyridium chloride, benzethonium chloride, popidone iodine, essential oil, triclosan, ethanol and an appropriate amount By appropriately containing the oral flora improving / maintaining agent together with the water, it can be provided as an oral cleaning agent for improving / maintaining the oral flora.
<Toothpaste>

When used as a dentifrice, calcium carbonate, hydroxyapatite, calcium hydrogenphosphate, aluminum hydroxide, alumina, silica, lauroyl sarcosin soda, sodium lauryl sulfate, sucrose fatty acid ester, dextranase base, alkyl glycoside, Toothpaste for improving and maintaining oral flora by appropriately containing an oral flora improving / maintaining agent together with sorbitol, glycerin, propylene glycol, sodium alginate, carboxymethyl cellulose, sodium fluoride, dextranase, chlorhexidine, sodium chloride, etc. It can be provided as an agent.

Claims

An oral flora improving / maintaining agent containing acacia bark extract as an active ingredient.
The oral flora improving / maintaining agent according to claim 1, which promotes the withdrawal of gram-negative bacteria and / and gram-positive bacteria containing acacia bark extract as an active ingredient from the oral biofilm (plaque, plaque).
The oral flora improving / maintaining agent according to claim 1, which suppresses the formation of oral biofilms (plaque, plaque) of gram-negative bacteria and / and gram-positive bacteria containing acacia bark extract as an active ingredient.
The oral flora according to claim 2 or 3, wherein the gram-negative bacterium and / and the gram-positive bacterium includes at least two or more of Streptococcus gordonii, Streptococcus mitis, Fusobacterium nucleatum, Porphyromonas gingivalis, Treponema spp, and Tannerella forsythia. Improvement / maintenance agent.
A food or drink containing the oral flora improving / maintaining agent according to any one of claims 1 to 4.
An oral cleansing agent containing the oral flora improving / maintaining agent according to any one of claims 1 to 4.
A dentifrice containing the oral flora improving / maintaining agent according to any one of claims 1 to 4.
A tartar formation preventive agent containing acacia bark extract as an active ingredient.
The tartar formation preventive agent according to claim 8, which promotes the withdrawal of gram-negative bacteria and / and gram-positive bacteria containing acacia bark extract as an active ingredient from the oral biofilm.
The tartar formation preventive agent according to claim 8, which suppresses the formation of an oral biofilm of Gram-negative bacteria and / and Gram-positive bacteria containing acacia bark extract as an active ingredient.
The prevention of dentin formation according to claim 9 or claim 10, wherein the gram-negative bacterium and / and the gram-positive bacterium includes at least two or more of Streptococcus gordonii, Streptococcus mitis, Fusobacterium nucleatum, Porphyromonas gingivalis, Treponema spp, and Tannerella forsythia. Agent.
A food or drink containing the tartar formation preventive agent according to any one of claims 8 to 11.
An oral cleanser containing the tartar formation preventive agent according to any one of claims 8 to 11.
A dentifrice containing the tartar formation preventive agent according to any one of claims 8 to 11.
A method for preventing tartar formation using the tartar formation preventive agent according to any one of claims 8 to 11.
An oral flora improvement / maintenance method for improving / maintaining an oral flora using the oral flora improving / maintaining agent according to any one of claims 1 to 4.