KR20230173479A - Composition for preventing, improving or treating oral disease comprising ovomucoid and uses thereof - Google Patents

Abstract

The present invention relates to a composition containing ovomucoid derived from egg white that has an inhibitory effect on Streptococcus mutans , which causes dental caries, and its use.
The ovomucoid of the present invention effectively inhibited the adhesion ability, aggregation ability, and biofilm formation of the Streptococcus mutans strain, and also decomposed the already formed biofilm. In addition, ovomucoid affected the EPS production and metabolic activity of Streptococcus mutans and reduced the expression of gtfA, ftf, vicR, comDE, relA, and brpA mRNA expression of factors related to dental caries induction. Therefore, the ovomucoid of the present invention can be effectively used as a composition for preventing, improving, or treating oral diseases related to Streptococcus mutans.

Description

Composition for preventing, improving or treating oral disease comprising ovomucoid and uses thereof {Composition for preventing, improving or treating oral disease comprising ovomucoid and uses thereof}

The present invention relates to a composition containing ovomucoid derived from egg white that has an inhibitory effect on Streptococcus mutans , which causes dental caries, and its use.

Various types of bacteria form colonies in the human oral cavity, and some microorganisms cause diseases such as dental caries (cavities). The causative bacteria that play an important role in the formation of dental plaque include Streptococcus mutans ( S. mutans ), and plaque is a bio-organism composed of glucan and fructan produced by S. mutans . It is a film (biofilm). As this plaque accumulates on the surface of the teeth, it becomes a habitat for bacteria, and organic acids such as lactic acid produced by microorganisms using nutrients in the mouth to metabolize carbohydrates create an acidic environment, demineralizing the enamel of the teeth, causing dental caries. . Glucan and fructan, which play an important role in the formation of the biofilm, are produced by bacterial glucosyltransferase (GTF) and fructosyltransferase (FTF), which produce extracellular polysaccharides. Accordingly, in order to prevent dental caries, various studies are being actively conducted on safe natural substances that can effectively remove bacteria in the oral cavity or inhibit the formation of biofilm plaque or the activity of related enzymes such as GTF and FTF.

Eggs are a complete food rich in various nutrients such as proteins, lipids, and minerals, and are also widely known as a protein source with an excellent composition of essential amino acids. Proteins such as ovalbumin, ovotransferrin, ovomucoid, ovomucin, or lysozyme exist in egg white. Among them, ovomucoid is one of the major glycoproteins present in egg white. It accounts for approximately 11% of egg white proteins and has a molecular weight of approximately 28,000.

KR 10-2012-0134924 (2012-11-27)

The present inventors have made diligent efforts to provide natural substances that can prevent, improve, or treat dental caries (cavities), and as a result, ovomucoid, a protein derived from egg white, inhibits Streptococcus mutans , a cavity-causing bacteria. After confirming what could be done, the present invention was completed.

Therefore, the purpose of the present invention is to provide a composition containing ovomucoid derived from egg white that has an inhibitory effect on Streptococcus mutans , which causes dental caries (cavities), and a use thereof.

The present invention provides a composition for preventing or treating oral diseases containing ovomucoid.

According to a preferred embodiment of the present invention, the ovomucoid is derived from egg white.

According to a preferred embodiment of the present invention, the oral disease is an oral disease caused by Streptococcus mutans .

According to a preferred embodiment of the present invention, the oral disease is cavities.

The present invention also provides a pharmaceutical composition for preventing or treating oral diseases containing ovomucoid.

The present invention also provides a health functional food composition for preventing or improving oral diseases containing ovomucoid.

The present invention also provides a composition for inhibiting biofilm formation in the oral cavity containing ovomucoid.

The ovomucoid of the present invention effectively inhibited the adhesion ability, aggregation ability, and biofilm formation of the Streptococcus mutans strain, and was also able to decompose already formed biofilm. In addition, ovomucoid also affected the EPS production and metabolic activity of Streptococcus mutans and reduced the expression of gtfA , ftf , vicR , comDE , relA and brpA mRNA expression of factors related to dental caries induction. Therefore, the ovomucoid of the present invention can be effectively used as a composition for preventing, improving, or treating oral diseases related to Streptococcus mutans.

Figure 1 shows the effect of ovomucoids on biofilm production of Streptococcus mutans.
Figure 2 shows a biofilm of Streptococcus mutans treated with ovomucoid observed using a scanning electron microscope.
Figure 3 shows the effect of ovomucoid on the decomposition of already formed biofilm of Streptococcus mutans.
Figure 4 shows the effect of ovomucoid on the production of extracellular polysaccharides (EPS) in Streptococcus mutans.
Figure 5 shows the effect of ovomucoids on the adhesion ability of Streptococcus mutans.
Figure 6 shows the effect of ovomucoids on the metabolic activity of Streptococcus mutans.
Figure 7 shows the effect of ovomucoids on mRNA expression of Streptococcus mutans.

Hereinafter, the present invention will be described in more detail.

'Prevention' of the present invention refers to all actions that suppress or delay the onset of oral diseases or related diseases due to the ovomucoid of the present invention.

'Improvement' or 'treatment' of the present invention refers to all actions that improve or benefit parameters related to oral diseases or related diseases, such as the degree of symptoms, due to the ovomucoid of the present invention.

In the present invention, the inhibitory effect of the protein ovomucoid derived from egg white on the caries-inducing characteristics of the Streptococcus mutans strain was confirmed. Ovomucoid effectively inhibited the adhesion, aggregation, and biofilm formation of Streptococcus mutans strains, and also decomposed already formed biofilms. Additionally, ovomucoids also affected EPS production and metabolic activity of Streptococcus mutans. As a result of examining the effect of ovomucoid on the expression of mRNA in Streptococcus mutans strains, the mRNA expression of gtfA , ftf , vicR , comDE , relA and brpA mRNA expression of factors related to dental caries induction was decreased upon treatment with ovomucoid. Therefore, the egg white-derived ovomucoid of the present invention can be effectively used as a composition for preventing, improving, or treating oral diseases, including cavities, or as a composition for inhibiting biofilm formation in the oral cavity.

Therefore, it is possible to provide a composition for preventing or treating oral diseases containing ovomucoid.

According to a preferred embodiment of the present invention, the ovomucoid is derived from egg white.

According to a preferred embodiment of the present invention, the oral disease is an oral disease caused by Streptococcus mutans .

According to a preferred embodiment of the present invention, the oral disease is cavities.

According to a preferred embodiment of the present invention, the composition may be in the form of toothpaste, gargling product, oral spray, oral ointment, mouthwash, or mouthwash.

Additionally, the present invention can provide a pharmaceutical composition for preventing or treating oral diseases containing ovomucoid.

According to a preferred embodiment of the present invention, the ovomucoid is derived from egg white.

According to a preferred embodiment of the present invention, the oral disease is an oral disease caused by Streptococcus mutans .

According to a preferred embodiment of the present invention, the oral disease is cavities.

When formulating the pharmaceutical composition of the present invention, one or more buffering agents (e.g. saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g. EDTA or glutathione), fillers, bulking agents, binders, adjuvants ( For example, aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrants or surfactants, diluents or excipients.

Solid preparations include tablets, pills, powders, granules, capsules, etc. These solid preparations contain one or more compounds and at least one excipient, such as starch (including corn starch, wheat starch, rice starch, potato starch, etc.) , calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose, methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethyl-cellulose or gelatin. It is prepared by mixing etc. For example, tablets or dragees can be obtained by combining the active ingredient with solid excipients, grinding them, adding suitable auxiliaries, and processing them into a granule mixture.

Additionally, in addition to simple excipients, lubricants such as magnesium stearate, talc, etc. are also used. Liquid preparations include suspensions, oral solutions, emulsions, or syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients may be included, such as wetting agents, sweeteners, fragrances, or preservatives. In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate can be added as a disintegrant, and anti-coagulants, flavoring agents, emulsifiers, solubilizers, dispersants, flavoring agents, antioxidants, and packaging agents. , pigments and preservatives may be additionally included.

The composition of the present invention is administered in a pharmaceutically effective amount. A pharmaceutically effective amount refers to an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level refers to the type of patient's disease, severity, activity of the drug, sensitivity to the drug, and administration time. , route of administration and excretion rate, duration of treatment, factors including concurrently used drugs, and other factors well known in the field of medicine. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. That is, the total effective amount of the composition of the present invention can be administered to the patient in a single dose, or by a fractionated treatment protocol in which multiple doses are administered over a long period of time. . Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.

The composition of the present invention can be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, and methods using biological response modifiers.

In addition, the present invention can provide a health functional food composition for preventing or improving oral diseases containing ovomucoid.

According to a preferred embodiment of the present invention, the ovomucoid is derived from egg white.

According to a preferred embodiment of the present invention, the oral disease is an oral disease caused by Streptococcus mutans .

According to a preferred embodiment of the present invention, the oral disease is cavities.

The food composition according to the present invention can be manufactured in various forms according to conventional methods known in the art. General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (e.g. canned fruit, bottled foods, jam, mamalade, etc.), fish, meat and their processed foods (e.g. ham, sausages, etc.) corned beef, etc.), bread and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine , vegetable proteins, retort foods, frozen foods, and various seasonings (e.g., soybean paste, soy sauce, sauce, etc.) can be produced by adding the ovomucoid of the present invention. In addition, nutritional supplements are not limited to this, but can be prepared by adding the ovomucoid of the present invention to capsules, tablets, pills, etc. In addition, the health functional food is not limited to this, but for example, the ovomucoid itself of the present invention is manufactured in the form of tea, juice, and drinks and liquefied, granulated, encapsulated, and powdered so that it can be consumed (health beverage). It can be consumed. Additionally, in order to use the ovomucoid of the present invention in the form of a food additive, it can be prepared and used in the form of a powder or concentrate. In addition, the ovomucoid of the present invention can be prepared in the form of a composition by mixing it with a known active ingredient known to be effective in preventing or improving oral diseases.

When using the ovomucoid of the present invention as a health drink, the health drink composition may contain various flavoring agents or natural carbohydrates as additional ingredients like a normal drink. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; polysaccharides such as dextrins and cyclodextrins; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as thaumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the composition of the present invention.

In addition, the ovomucoid of the present invention may be contained as an active ingredient in a health functional food composition for preventing or improving oral diseases, and the amount is not particularly limited to an amount effective to achieve the effect of preventing or improving oral diseases. It is preferably 0.01 to 100% by weight based on the total weight of the composition. The health functional food composition of the present invention can be prepared by mixing ovomucoid with other active ingredients known to be effective in oral diseases.

In addition to the above, the health functional food of the present invention includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, and preservatives. , may contain glycerin, alcohol, or carbonating agent. In addition, the health functional food of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverage, or vegetable beverage. These ingredients can be used independently or in combination.

In addition, the present invention can provide a composition for inhibiting biofilm formation in the oral cavity containing ovomucoid.

According to a preferred embodiment of the present invention, the ovomucoid is derived from egg white.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it is obvious to those skilled in the art that the scope of the present invention should not be construed as limited by these examples.

Strain culture and sample preparation

The Streptococcus mutans strains used in the present invention were S. mutans KCTC 5124 (KCTC 5124), S. mutans KCTC 5316 (KCTC 5316), and S. mutans KCTC 5458 (KCTC 5458), which were purchased from the Korea Biological Resources Center. Streptococcus mutans strain was inoculated into BHI liquid medium containing 0.1% sucrose, and 1 pack of AnaeroPack-MicroAero (MGC) was added to an anaerobic jar and cultured at 37°C. The ovomucoid used in the present invention was isolated from egg white. In the control group, an equal amount of distilled water was added instead of the sample.

Effect of ovomucoids on biofilm formation of Streptococcus mutans

Dental caries (cavities) is a disease closely related to microbial biofilms. Biofilms are structured communities of microorganisms attached to surfaces and are composed of proteins, polysaccharides, or metabolites. Microorganisms within biofilms have the problem of showing resistance to antibiotics and the host's immune system because biofilms play a role in inhibiting the penetration of antibacterial agents.

To determine the effect of ovomucoids on the biofilm formation of Streptococcus mutans, Streptococcus mutans strains were cultured in BHI broth containing ovomucoids (0.25-1 mg/mL) in a 24 well plate for 24 hours. cultured for a while. After culturing for 24 hours, 0.1% crystal violet solution was added to the biofilm in each well and stained for 30 minutes, then washed with distilled water and dried. After decolorizing by adding acid-alcohol solution, the plate was transferred to a new 96 well plate and the absorbance was measured at 570 nm, and the biofilm formation rate was calculated using the following [Calculation 1].

[Calculation Formula 1]

Biofilm formation rate (%) = (absorbance of sample/absorbance of control) x 100

For scanning electron microscopy, a glass coverslip was placed on a 6-well plate, and the Streptococcus mutans strain was cultured in BHI broth containing ovomucoid (1 mg/mL) for 24 hours. The biofilm formed on the glass coverslip was fixed using 2.5% glutaraldehyde, sequentially dehydrated in ethanol, and then completely dried. The dried sample was observed at magnifications of 5,000 and 10,000 times using a scanning electron microscope (SU8010, Hitachi High-Technologies Co., Tokyo, Japan).

As a result, as shown in the graph of [Figure 1], when treated with ovomucoid at a concentration of 0.25-1 mg/mL, biofilm formation was significantly inhibited in a concentration-dependent manner. When treated with 1 mg/mL ovomucoid, the biofilm formation rates of KCTC 5124, KCTC 5316, and KCTC 5458 decreased by 43.41%, 59.51%, and 73.55%, respectively.

In addition, as shown in [Figure 2], the Streptococcus mutans biofilm observed through FESEM also showed an inhibitory effect of ovomucoid (1 mg/mL) treatment on the Streptococcus mutans biofilm formation. These results meant that ovomucoid can effectively inhibit biofilm formation of Streptococcus mutans.

Effect of ovomucoids on biofilm decomposition of Streptococcus mutans formed

As described above, microorganisms in biofilms exhibit stronger antibacterial resistance than bacteria in the suspended state, and biofilms that have matured for a long period of time show greater antibacterial resistance than biofilms that have had a shorter maturation period. Therefore, an effective antibiofilm agent must be able to reduce not only the formation of biofilms but also already formed mature biofilms.

In order to determine the effect of ovomucoids on the decomposition of biofilm of already formed Streptococcus mutans, the Streptococcus mutans strain was cultured in a 24 well plate for 24 hours to form a biofilm. After removing the supernatant, BHI broth containing ovomucoid (0.25-1 mg/mL) was added to the formed biofilm and cultured for an additional 24 hours. Then, crystal violet staining was performed in the same manner as above to identify the biofilm. The formation rate was calculated using the following [Calculation Formula 2].

[Calculation Formula 2]

Biofilm formation rate (%) = (absorbance of sample/absorbance of control) x 100

As a result, as shown in [Figure 3], ovomucoid decomposed the biofilm formed for 24 hours. When treated with ovomucoid at a concentration of 1 mg/mL, the biofilm of KCTC 5124, KCTC 5316, and KCTC 5458 Film decreased by 12.71%, 14.97%, and 35.56%, respectively. Therefore, this result, together with [Figure 1] and [Figure 2], showed that ovomucoid not only inhibited the formation of biofilm at the initial stage, but could also decompose already formed biofilm.

Effect of ovomucoids on the production of water-soluble EPS in Streptococcus mutans

Streptococcus mutans uses sucrose to synthesize glucan and fructan using enzymes such as GTF and FTF, which affects the accumulation of acid-producing acid-resistant microorganisms. The production of extracellular polysaccharides (EPS) mediates bacterial attachment and adhesion between cells and contributes to the formation of biofilm. EPS formed on the tooth surface, especially water-insoluble glucan, supports the attachment of various oral microorganisms. It becomes a place.

To determine the effect of ovomucoids on the production of water-insoluble EPS by Streptococcus mutans, Streptococcus mutans strains were cultured in BHI broth containing ovomucoids (0.25-1 mg/mL) in a 12 well plate for 24 hours. incubated for some time. After culturing for 24 hours, the medium in each well was removed, the resulting biofilm was scraped, suspended in phosphate buffered saline (PBS), and centrifuged at 4000 × g for 10 minutes. The precipitate was suspended in 0.5 M NaOH at 37°C for 2 hours. After 2 hours, centrifugation was performed at 4000 EPS production rate was calculated.

[Calculation Formula 3]

EPS production rate (%) = (absorbance of sample/absorbance of control) x 100

As a result, the Streptococcus mutans strain treated with ovomucoid produced less EPS than the control group (Figure 4). At a concentration of 1 mg/mL, ovomucoid reduced the EPS production of KCTC 5124, KCTC 5316, and KCTC 5458 by 14.40%, 19.80%, and 32.56%, respectively. These results show that ovomucoids had a significant effect on EPS production and contributed to the reduction of biofilm formation.

Effect of ovomucoid on the adhesion and aggregation ability of Streptococcus mutans

The biofilm formation process is a complex process consisting of several stages, including initial attachment to a surface, aggregation, and maturation. Attachment of microorganisms is an essential characteristic for biofilm formation that also affects biofilm maturation, and aggregation ability is one of the important characteristics for attachment to surfaces. Therefore, since reducing adhesion to the tooth surface can be used as a major method to reduce oral biofilm, the effect of ovomucoid on the adhesion or aggregation ability of Streptococcus mutans strains was confirmed.

Specifically, to measure adhesion ability, Streptococcus mutans strain was inoculated into BHI broth containing ovomucoid (0.25-1 mg/mL) in a glass tube, and the glass tube was placed on a slope inclined at about 30°, 37 Cultured for 24 hours at °C. After incubation, the supernatant was removed, and the Streptococcus mutans strain attached to the wall of the glass tube was suspended in PBS, the absorbance was measured at 600 nm, and the adhesion ability was calculated using the following [Equation 4].

[Calculation Equation 4]

Adhesion ability (%) = (absorbance of sample/absorbance of control) x 100

In addition, to measure agglutination ability, Streptococcus mutans strain was cultured in BHI broth for 24 hours, and washed twice to remove culture supernatant to obtain only bacterial cells. Afterwards, it was suspended in PBS containing ovomucoid (0.25-1 mg/mL) at OD600=0.5 ± 0.02, the absorbance was measured at 600 nm after 24 hours, and the aggregation ability was calculated using the following [Equation 5].

[Calculation Equation 5]

Coagulation ability (%) = (1-(absorbance after 24 hours/absorbance after 0 hours)) x 100

As a result, the adhesion ability of Streptococcus mutans decreased in a concentration-dependent manner at 0.25-1 mg/mL of ovomucoid (FIG. 5). In particular, when treated with ovomucoid at a concentration of 1 mg/mL, the adhesion ability of KCTC 5124, KCTC 5316, and KCTC 5458 decreased by 28.75~43.47%.

Strain Ovomucoid Control 0.25 mg/mL 0.5mg/mL 1mg/mL KCTC 5124 48.41 ± 6.84 41.34 ± 4.97 40.70 ± 4.60 39.87 ± 2.55 KCTC 5316 31.95 ± 5.83 24.48 ± 5.09 23.16 ± 6.22 21.03 ± 2.47 KCTC 5458 48.13 ± 5.70 39.26 ± 4.42 34.52 ± 3.77 31.74 ± 5.73

In addition, the aggregation ability of KCTC 5124, KCTC 5316, and KCTC 5458 cultured with 1 mg/mL ovomucoid was significantly reduced to 39.87%, 21.03%, and 31.74%, respectively, compared to the respective controls (Table 1). Therefore, ovomucoid reduced the attachment and aggregation ability of Streptococcus mutans, which showed that ovomucoid can reduce oral biofilm formation by inhibiting the attachment of microorganisms to the surface.

Effect of ovomucoids on the metabolic activity of Streptococcus mutans

The MTT method was used to measure the effect of ovomucoids on the metabolic activity of Streptococcus mutans in biofilm.

Specifically, the Streptococcus mutans strain was cultured in BHI broth containing ovomucoid (0.25-1 mg/mL) for 24 hours in a 24 well plate. After culturing for 24 hours, the supernatant from each well was removed, and thiazolyl blue tetrazolium bromide (MTT) solution (2 mg/mL) was added for reaction. After 4 hours, dimethyl sulfoxide was added, absorbance was measured at 570 nm, and metabolic activity was calculated using the following [Calculation Formula 6].

[Calculation Equation 6]

Metabolic activity (%) = (absorbance of sample/absorbance of control) x 100

As a result, as shown in [Figure 6], when the Streptococcus mutans strain was treated with ovomucoid, the metabolic activity of all three strains decreased in a concentration-dependent manner. When treated with 1 mg/mL ovomucoid, the metabolic activity of KCTC 5124, KCTC 5316, and KCTC 5458 decreased by 33.31%, 27.83%, and 49.91%, respectively, compared to the control group.

Effect of ovomucoids on mRNA expression of Streptococcus mutans

In previous experiments to confirm the relative expression of genes related to biofilm formation of Streptococcus mutans by qRT-PCR when treated with ovomucoid, the inhibition effect by ovomucoid was the most among the three Streptococcus mutans strains. The experiment was conducted on the KCTC 5458 strain, which was found to be highly prevalent.

Specifically, the Streptococcus mutans strain cultured for 24 hours was inoculated into BHI broth and cultured with ovomucoid (1 mg/mL), and then an RNA isolation kit was used to isolate total RNA. After conversion to cDNA, qRT-PCR was performed with primers gtfA, ftf, comDE, vicR, brpA, and relA . Amplification results were calculated using the delta-delta Ct method. 16S rRNA was used as a housekeeping gene, and the primers used for PCR are shown in Table 2 below.

target direction Primer (5'→3') sequence number gtfA sense 5'-AGGAAGTGAAGCGGCCAGT-3' One anti-sense 5'-TCAATACGGCCATCCAAATCA-3' 2 ftf sense 5'-AAATATGAAGGCGGCTACAACG-3' 3 anti-sense 5'-CTTCACCAGTCTTAGCATCCTGAA-3' 4 comDE sense 5'-ACAATTCCTTGAGTTCCATCCAAG-3' 5 anti-sense 5'-TGGTCTGCTGCCTGTTGC-3' 6 vicR sense 5'-TGACACGATTACAGCCTTTGATG-3' 7 anti-sense 5'-CGTCTAGTTCTGGTAACATTAAGTCCAATA-3' 8 brpA sense 5'-GGAGGAGCTGCATCAGGATTC-3' 9 anti-sense 5'-AACTCCAGCACATCCAGCAAG-3' 10 relA sense 5'-ACAAAAAGGGTATCGTCCGTACAT-3' 11 anti-sense 5'-AATCACGCTTGGTATTGCTAATTG-3' 12 16S rRNA sense 5'-CCTACGGGAGGCAGCAGTAG-3' 13 anti-sense 5'-CAACAGAGCTTTACGATCCGAAA-3' 14

As shown in [Figure 7], when treated with ovomucoid at a concentration of 1 mg/mL, the expression level of the gtfA gene was reduced to 0.53 times that of the control medium, and the expression level of the ftf gene was reduced to 0.81 times that of the control medium. GTF and FTF are important factors in oral plaque formation. gtfA and ftf are genes involved in the formation of extracellular polymer matrix. The gtfA gene encodes sucrose phosphorylase, and the ftf gene encodes fructosyltransferase. These results showed that the use of sucrose for extracellular glucan and fructan production was reduced by ovomucoid treatment.

In addition, the mRNA expression of relA and brpA , which play an important role in acid resistance and biofilm formation in Streptococcus mutans, was suppressed by ovomucoid by 0.51-fold for relA and 0.52-fold for brpA compared to the control. relA , encoding guanosine tetra (penta)-phosphatesynthetase, is related to bacterial physiology, and brpA is a gene related to biofilm formation.

Lastly, the expression levels of vicR and comDE genes related to TCSTS (two-component signal transduction systems) were significantly reduced by 0.75-fold for vicR and 0.68-fold for comDE compared to the control group, respectively, by ovomucoid at a concentration of 1 mg/mL. did. The VicRK system, together with the ComCDE system, regulates microbial adhesion and biofilm formation by affecting the expression of genes encoding glucan matrix synthesis. vicR encodes the VicR response regulator, which is part of the VicRK system. QS factors play a role in transmitting signals between microorganisms, which regulates their biological activities. The QS system, which is involved in the maturation of Streptococcus mutans biofilms, is regulated by regulators encoded by the comD and comE genes that mediate downstream gene expression. Therefore, the above results show that the suppression of the dental caries-inducing properties of Streptococcus mutans is due to the effect of ovomucoids in down-regulating this system.

[Statistical processing]

The experiment took the mean ± standard deviation values. The statistical significance of the experimental results was tested at the P<0.05 level using the Student's t-test method and one-way ANOVA method using the SPSS software version 18 (SPSS Inc., Chicago, IL, USA) program. .

SEQUENCE LISTING <110> KONKUK UNIVERSITY INDUSTRIAL COOPERATION CORP. WithBio Inc. <120> Composition for preventing, improving or treating oral disease comprising ovomucoid and uses thereof <130> 1070984 <160> 14 <170> PatentIn version 3.2 <210> 1 <211> 19 <212> DNA <213> Artificial <220> <223> gtfA_sense <400> 1 aggagtgaa gcggccagt 19 <210> 2 <211> 21 <212> DNA <213> Artificial <220> <223> gtfA_anti-sense <400> 2 tcaatacggc catccaaatc a 21 <210> 3 <211> 22 <212> DNA <213> Artificial <220> <223> ftf_sense <400> 3 aaatatgaag gcggctacaa cg 22 <210> 4 <211> 24 <212> DNA <213> Artificial <220> <223> ftf_anti-sense <400> 4 cttcaccagt cttagcatcc tgaa 24 <210> 5 <211> 24 <212> DNA <213> Artificial <220> <223> comDE_sense <400> 5 acaattcctt gagttccatc caag 24 <210> 6 <211> 18 <212> DNA <213> Artificial <220> <223> comDE_anti-sense <400> 6 tggtctgctg cctgttgc 18 <210> 7 <211> 23 <212> DNA <213> Artificial <220> <223> vicR_sense <400> 7 tgacacgatt acagcctttg atg 23 <210> 8 <211> 30 <212> DNA <213> Artificial <220> <223> vicR_anti-sense <400> 8 cgtctagttc tggtaacatt aagtccaata 30 <210> 9 <211> 21 <212> DNA <213> Artificial <220> <223> brpA_sense <400> 9 ggaggagctg catcaggatt c 21 <210> 10 <211> 21 <212> DNA <213> Artificial <220> <223> brpA_anti-sense <400> 10 aactccagca catccagcaa g 21 <210> 11 <211> 24 <212> DNA <213> Artificial <220> <223> relA_sense <400> 11 acaaaaaggg tatcgtccgt acat 24 <210> 12 <211> 24 <212> DNA <213> Artificial <220> <223> relA_anti-sense <400> 12 aatcacgctt ggtattgcta attg 24 <210> 13 <211> 20 <212> DNA <213> Artificial <220> <223> 16S rRNA_sense <400> 13 cctacggggg gcagcagtag 20 <210> 14 <211> 23 <212> DNA <213> Artificial <220> <223> 16S rRNA_anti-sense <400> 14 caacagagct ttacgatccg aaa 23

Claims (7)

A composition for preventing or treating oral diseases containing ovomucoid.
The composition according to claim 1, wherein the ovomucoid is derived from egg white.
The composition according to claim 1, wherein the oral disease is caused by Streptococcus mutans .
The composition according to claim 1, wherein the oral disease is cavities.
A pharmaceutical composition for preventing or treating oral diseases containing ovomucoid.
A health functional food composition for preventing or improving oral diseases containing ovomucoid.
A composition for inhibiting biofilm formation in the oral cavity containing ovomucoid.