KR101914491B1 - Oral antibacterial composition containing chinese quince as effective component - Google Patents

Abstract

The present invention relates to an antibacterial composition for oral cavity containing a quince extract as an active ingredient. Specifically, the quince extract of the present invention has excellent physiological activities and thermal stability, and shows outstanding antibacterial effects against oral bacteria. Therefore, the quince extract can be used as an antibacterial agent for inhibiting oral bacteria, while increasing applicability of quince by using abandoned quince as a resource.

Description

[0001] The present invention relates to an oral antibacterial composition containing chinese quince as effective component,

The present invention relates to an oral antimicrobial composition containing a quince as an active ingredient, and more particularly, to an antimicrobial composition for inhibiting oral bacteria containing an extract of quinine which can be used as a disinfectant for oral care agents and oral hygiene tools .

As the dietary culture of busy modern people becomes more diverse, the consumption of pungent products and saccharides continues to increase, and the immune function is continuously weakened due to drinking and stress, so that oral microorganisms are increasing. Oral diseases are caused by the mouth, such as tooth decay, periodontitis and stomatitis. Dental caries and periodontal disease are major diseases that threaten the oral health of the people of Korea. Because the oral cavity is also connected to the bronchial tree, the bronchial damage caused by the external environment is related to the oral health, and the oral hygiene state also has a great influence on the bronchial tree. A healthy person's oral flora is stable, but if it is not healthy, the bacteria in the mouth may form a bacterial membrane, causing bad breath, dental caries, gingivitis, and periodontitis. Therefore, there is a need for oral hygiene management to prevent diseases associated with oral bacteria such as inflammation, dental caries, periodontitis and gingivitis. Nowadays, oral hygiene products that can be easily obtained on the market are one of the oral hygiene management auxiliary products that prevent tooth decay and gum disease by giving a refreshing effect to the mouth through chemical action rather than brushing. This is mainly used when brushing is not possible or when you want a fresh mouth. Oral cleansing agents can not replace toothbrushes that wipe the surface of teeth, but they contain antibacterial and germicidal ingredients, such as fluoride, to help prevent tooth decay and gum disease and help to remove bad breath. However, when a person without oral disease is used for a long period of time, various side effects such as dry mouth due to evaporation of alcohol, high mucosal damage due to alcohol concentration, tooth discoloration and the like may occur. Therefore, it may not be beneficial to the human body. In addition, when oral cleaning agent is used for a long time to remove harmful bacteria in the oral cavity, normal bacteria may be removed and the immunity may be lowered. Therefore, in order to maintain healthy oral hygiene, it is required to develop oral hygiene product which can be used safely for a long time without side effects. Researches on the use of natural materials for this purpose have been actively carried out.

On the other hand, the Chinese quince is a fruit of a deciduous tree belonging to the rose family. It is strong, strong, and strong. It is known that moths have cough, cold and bronchitis relieving effects, and they are known to contain effective ingredients such as malic acid, which acts as a protein coagulant. However, the quince has a lower moisture content and a stronger flavor than the ordinary fruit, and it is used as a herbal medicine, a fragrance and an edible car because it has a stout cell and woody texture and is rough and has a difficulty in various processing. It is a fact that corn is discarded. Therefore, it is necessary to evaluate the new functional value of corncorner and to apply it to various industries, and it is expected that it will be possible to recycle the waste material to be used as a new income source of the farmhouse while increasing the utilization.

On the other hand, Korean Patent No. 1295242 discloses an antimicrobial composition for inhibiting oral bacteria containing an extract of Toadstool as an active ingredient. Korean Patent No. 1447121 discloses a composition for inhibiting xanthine oxidase Or a health functional food has been disclosed, there has been no disclosure of a composition for oral antimicrobial containing a quince of the present invention as an active ingredient.

The present invention provides an oral antimicrobial composition comprising an extract of a quince extract as an active ingredient, which is obtained by the above-mentioned demand, and is characterized in that the quince extract has thermal stability and has excellent antimicrobial effect against physiological activity and oral bacteria Thereby completing the present invention.

In order to solve the above-described problems, the present invention provides a composition for inhibiting oral bacteria, which comprises a quinone extract as an active ingredient.

The present invention also provides a quasi-drug for preventing or ameliorating oral diseases containing an extract of a quinceae as an active ingredient.

The present invention relates to a composition for oral antimicrobial use containing a quince as an active ingredient, and the quince extract of the present invention has thermal stability, and has excellent antimicrobial activity against bacteria and bacteria in the oral cavity. Therefore, The composition of the present invention can be used as a composition for oral antimicrobial inhibition, so that the utility of the quince can be increased.

Fig. 1 shows the total polyphenol content of Tobacco mushroom, Doroji, Morus alba, and Gold Ears. a, b, c, d and e show significant differences between the experimental groups, p <0.05.
Fig. 2 shows the total flavonoid contents of Tobacco, Macromuscular, Platycodon, Quercus and Galium. a, b, c, d and e show significant differences between the experimental groups, p <0.05.
Fig. 3 shows the DPPH radical scavenging activity of Tobacco mushroom, Bombyx mori, Platycodon grandiflorum, Quercus acutissima, and Quercetin. a, b, c, d and e show significant differences between the experimental groups, p <0.05.
Fig. 4 shows the ABTS radical scavenging ability of Tobacco mushroom, Platycodon sp. a, b, c, d and e show significant differences between the experimental groups, p <0.05.
Fig. 5 shows the nitrite scavenging activity of Tobacco mushroom, Macromyx, Platycodon grandiflorum, Quercus mongolica and Goldsmith. a, b, and c represent a significant difference between the experimental groups, p <0.05.
Fig. 6 shows the results of inhibition of bacteria in the oral cavity by the single or combined treatment of the quinceae and the gingkojum extract. QE is a group treated with a mossy ethanol extract, LJE is treated with a germanium ethanol extract, and QE + LJE is a combination of a mossy ethanol extract and a ginseng ethanol extract.
FIG. 7 shows the result of confirming the antibacterial effect of quinine extract on oral bacteria contaminated with a toothbrush. SM is Streptococcus mutans, SA is Streptococcus sanguinis, An is Streptococcus angiosus, Sa is Staphylococcus aureus, and Se is Staphylococcus epidermidis.

The present invention provides a composition for inhibiting oral microbes comprising a quinone extract as an active ingredient. Preferably, the extract of the quinck extract is prepared by using C ₁ to C ₄ lower alcohols as a solvent, more preferably using ethanol as a solvent, but the present invention is not limited thereto.

The oral bacteria which are inhibited by the above-mentioned extracts of Staphylococcus aureus include Streptococcus mutans, Streptococcus sanguinis, Streptococcus anginosus, Staphylococcus aureus, But is not limited to, one or more selected from the group consisting of Staphylococcus epidermidis.

In the composition for inhibiting oral bacteria according to an embodiment of the present invention, the quinone extract is a horsetail ethanol extract, the oral bacterium is Streptococcus mutans mutans ) or Streptococcus sanguinis , but are not limited thereto.

The composition for inhibiting oral antibiotics may be formulated into an oral or parenteral dosage form in the form of solid, semi-solid, liquid or temperature-sensitive sol-gel phase transition by adding an inorganic or organic carrier. In one embodiment of the present invention, the composition may be formulated into, but not limited to, oral fresheners, oral cleansers, toothpastes, oral sprays, toothbrush cleaners, or denture cleansers.

The present invention also provides a quasi-drug for preventing or ameliorating oral diseases containing an extract of a quinceae as an active ingredient.

The oral diseases include, but are not limited to, cavities, periodontitis, or stomatitis.

The term "quasi-drug" in the present invention means a fiber, a rubber product or the like used for the purpose of treating, alleviating, treating or preventing a disease of a human or an animal, a weak action on the human body, Or products similar to those which are not machinery, preparations used for sterilization, insecticides and similar uses for the prevention of infections, for the purpose of diagnosis, treatment, alleviation, treatment or prevention of diseases of human beings or animals Machinery, or apparatus, and that is not an apparatus, machine, or apparatus of an article used for the purpose of giving pharmacological effects to the structure or function of a person or animal, It also includes supplies.

When the composition of the present invention is contained in quasi-drugs for the purpose of preventing or improving periodontal disease and tooth decay, the composition may be used as it is or may be used together with other quasi-drugs, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use.

The quasi-drug of the present invention may contain various bases and additives necessary for formulation according to the formulation, and the kind and amount of these components can be easily selected by those skilled in the art. For example, when the oral composition is a dentifrice, it may be prepared by adding an abrasive, a wetting agent, a foaming agent, a binder, a sweetener, a pH adjuster, a preservative, a medicinal ingredient, a flavoring agent, a brightener, a pigment and a solvent.

Hereinafter, the present invention will be described in more detail with reference to Preparation Examples and Examples. It is to be understood by those skilled in the art that these preparations and examples are merely intended to explain the present invention more specifically and that the scope of the present invention is not limited thereto.

Manufacturing example  1. Preparation of Natural Oriental Herb Extract

In the present invention, dry matter from Jirisan herbal bone was purchased from IKA All Basic (IKA-Werke GmbH, Staufen, Gemany), and dried at 40 It was powdered by mesh and stored in a cow. Then, the powder was extracted with 70% (v / v) ethanol. Specifically, each of the powders and ethanol was mixed at a volume ratio of 1: 9, and then the mixture was shaken using a shaker (HANBAK Scientific Co.) The mixture was shaken at 150 rpm for 24 hours at room temperature and repeatedly extracted three times.

The powder was mixed with water in a volume ratio of 1: 9, and the mixture was refluxed twice at 80 ° C for 3 hours using a reflux condenser (CA-1112, Eyela Co, Tokyo, Japan). After extraction, the extract was recovered and concentrated at 60 ° C with a vacuum concentrator (WB 2000, Heidolph, Germany). The concentrated extract was stored frozen and used as a test sample.

Example  1. Search for antimicrobial activity of pathogenic bacteria and oral bacteria in extracts

The antimicrobial activity of the ethanol extract and hot water extract of Preparation Example 1 was confirmed. (Listeria monocytogenes, ATCC 19115), Pseudomonas fluorescens (ATCC 21541), Staphylococcus aureus (ATCC 29737), Salmonella typhimurium (ATCC 11806), enteritis Vibrio sp. (Tryptic soy broth, TSB, Difco, USA) treated with tryptone and tryptone-treated soybean agar (Vibrio parahaemolyticus, ATCC 17802) were purchased from the Korean Microorganism Preservation Center (tryptic soy agar, TSA, Difco, USA). Oral bacteria include Streptococcus mutans (ATCC 25175), Streptococcus sanguins (KCTC 3299), Staphylococcus aureus (ATCC 29737), Staphylococcus epidermidis Staphylococcus epidermidis, ATCC 12228) and Streptococcus anginosus (ATCC 25175) were purchased from the Korean Microorganism Conservation Center. Brain heart infusion (BHI, Difco, USA) broth and agar were used for culture and antimicrobial activity.

The antimicrobial activity of the natural herbal extracts of Preparation Example 1 (corn, germ, bronze, dolomite, gold, silver) extracts was measured by a paper disc method. That is, 0.05 ml of a pathogenic strain and an oral cavity bacterium which had been activated twice were inoculated onto a dried medium and uniformly plated with a spreader. 40 쨉 l of a 0.1% corn ethanol extract was dispensed into a circular paper disc at 4 째 C For 24 hours, and then incubated with the culture medium at 37 ° C for 24 hours to confirm the generation of a clear zone.

The antimicrobial activity of Gram-positive bacteria (Listeria motocytosenius, Staphylococcus aureus, Bacillus cereus) and Gram-negative bacteria (enteritis bacteria, Escherichia coli, and Enterobacteriaceae) of hot water and ethanol extracts of Tobacco, As shown in Table 1, the ethanol extracts of T. orientalis, P. japonicus, B. japonica and C. giltae showed antimicrobial activity against pathogenic bacteria, whereas hydrothermal extracts did not show antimicrobial activity due to the absence of zona pellucida.

As shown in Table 2, the hot-water extracts showed no antimicrobial activity, while the ethanol extracts showed antibacterial activity. In particular, the ethanol extracts of Staphylococcus aureus and Streptococcus Showed high antimicrobial activity against mutans.

Table 1 and Table 2 show that the hot water extract of natural oriental herb has no antimicrobial activity against pathogenic bacterium and oral bacteria but the ethanol extract has antimicrobial activity.

In Table 1 and 2, ++ means that the diameter of the transparent zone is 4 mm, + is 2 to 3 mm, and - means no antibacterial activity.

Measurement of antimicrobial activity against pathogenic bacteria Pathogenic microorganism Natural herbal extract Stone McMundong Bellflower Quince Gold silver ethanol Heat number ethanol Heat number ethanol Heat number ethanol Heat number ethanol Heat number Listeria motosaito zenith + - + - + - + - + - Staphylococcus aureus + - + - + - + - + - Bacillus cereus + - + - + - + - + - Enteritis + - + - + - + - + - Escherichia coli + - + - + - + - + - Vibrio parahaemolyticus + - + - + - + - + -

Measurement of antimicrobial activity against oral bacteria Oral bacteria Natural herbal extract Stone McMundong Bellflower Quince Gold silver ethanol Heat number ethanol Heat number ethanol Heat number ethanol Heat number ethanol Heat number Streptococcus mutans + - + - + - ++ - ++ - Streptococcus sanguineus + - + - + - ++ - ++ - Streptococcus angiosus + - + - + - + - + - Staphylococcus aureus + - + - + - + - + - Starfilacocus epidermidis + - + - + - + - + -

Example  2. Measurement of physiological activity of ethanol extract of natural oriental herb

The physiological activity of the ethanol extract of natural oriental herbal medicine having antibacterial activity in Example 1 was measured.

1) Total polyphenol content

The total polyphenol content was determined by the method of Singleton et al. (1958), and 1 ml of 0.2 N polynucleoty reagent was added to 1 ml of the extract, followed by reaction at room temperature for 3 minutes. Then, sodium carbonate (Na ₂ CO ₃ ) (75 g / l) was added, and the mixture was allowed to stand in a dark place for 1 hour, and absorbance was measured at 765 nm. Total polyphenol content was calculated by standard curve with gallic acid as a standard.

The total polyphenol contents of the ethanol extracts of 0.1% (v / v) corn steep liquor, corn steep liquor, bellflower, corn and gilt-silver were measured. As a result, the total polyphenol contents of the extracts of corn steep, Phenol contents were 295.78, 18.08, 21.74, 404.49 and 682.02 ㎍ / ㎖, respectively. Polyphenol content of Ganoderma lucidum extract was the highest, and polyphenol content of quince was the second.

2) Total flavonoid content measurement

According to the method of Saleh and Hameed (2008), 0.15 ml of 5% sodium nitrite was added to 0.1 ml of the sample, and the mixture was allowed to stand at 25 ° C for 6 minutes. Then, 0.3 ml of 10% aluminum chloride was added thereto, For 5 minutes. Then, 1 ml of 1N sodium hydroxide (NaOH) was added and stirred. Then, the absorbance at 510 nm was measured, and the content was calculated by the calibration curve of rutin hydrate (Sigma-Aldrich Co.).

As shown in FIG. 2, the content of total flavonoids in ethanol extracts of 0.1% (v / v), A. myrtaceae, bellflower, corn and gilt silver was 42.49, 40.67, 43.89, 59.91 and 53.79 g / ㎖, and the content of moth extract was the highest.

3) Measurement of DPPH radical scavenging ability

0.8 ml of 0.4 mM DPPH (α-α-diphenyl-β-picrylhydrazyl) ethanol solution was mixed with 0.4 ml of the extract by shaking, and the mixture was allowed to stand for 10 minutes. Then, a spectrophotometer (Pharmacia Biotech Ultrospec 1000, Cambridege, UK) at 525 nm and converted according to the following equation (1).

[Formula 1]

DPPH radical scavenging activity (%) = [1- (sample absorbance / control absorbance)] × 100

The DPPH radical scavenging activity of DPPH radical scavenging ability was 58.11% and the extract of McMundum ethanol was lower than that of DPPH radical scavenging ability as shown in FIG. 3 as a result of measuring the DPPH radical scavenging ability after treatment with 1 μg / 2.25% of total extract, 21.01% of bloody ethanol extract, 69.66% of morpholine ethanol extract and 71.71% of ginseng ethanol extract. Among them, ethanol extract of ginseng and giitta extract showed high DPPH radical scavenging ability.

4) Measurement of ABTS radical scavenging ability

ABTS [2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt; A9941, Sigma-Aldrich Co.] The radical scavenging activity was measured according to the method of Re et al. (1999). 7.4 mM ABTS and 2.6 mM potassium persulfate were mixed and allowed to stand in a dark room for 24 hours to form radicals. Then, the ABTS solution was added to phosphate buffer solution (PBS) so that the absorbance at the wavelength of 732 nm became 0.700 ± 0.030 (phosphate-buffered saline, PBS, pH 7.4). 950 μl of ABTS solution was added to 50 μl of the sample, reacted in a dark place for 10 minutes, absorbance was measured at a wavelength of 732 nm, and the activity was calculated by the following formula 2.

[Formula 2]

ABTS radical scavenging activity (%) = [1- (sample absorbance / control absorbance)] × 100

As a result, as shown in FIG. 4, the ABTS radical scavenging activity was 59.91% and 53.79% in the ABTS radical scavenging ability of 50% or more when treated with 1 μg / ml of the extracts of mulberry, mulberry, , Tobacco, McDonald, and Bellflower showed 42.49, 40.67 and 33.89%, respectively.

5) Nitrite scavenging ability measurement

To measure the nitrite scavenging effect, 1 ml of 1 mM sodium nitrite (NaNO ₂ ) solution was added according to the method of Kato et al. (1987), and 0.1 N hydrogen chloride (HCl) was added to make a total volume of 10 ml. This solution was reacted at 37 ° C for 1 hour and then 1 ml of each reaction solution was diluted with 4 ml of a 2% (v / v) acetic acid solution and 1 ml of a grease reagent (Griess reagent, 1% sulfanilic acid: 1% naphthylamine = 1: 1). The mixture was incubated at room temperature for 15 minutes, and the absorbance at 520 nm was measured.

As shown in FIG. 5, nitrite scavenging ability was high at 72.63, 72.26, 76.97, 77.31 and 78.95% when treated with 1 / / ml of the extract of T. orientalis, P. vaginalis, B. vaginalis and P. gingivalis.

Example  3. Inhibitory effect of ethanol extract of natural oriental herb on oral bacteria growth

To determine the growth inhibitory effects of Streptococcus mutans, Streptococcus mutansis, Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus anginosus on the extracts of corn, gilthead, bellflower, One loop was inoculated into BHI (Brain Heart Infusion), which was prepared so that the final concentration of the ethanol extract was 0.1%, and each bacterium in each oral cavity was inoculated at 37 ° C for 24 hours. Then, 0.1% peptone And then 1 ml was taken. BHI agar was inoculated and cultured at 37 ° C for 24 hours. The colonies were counted and compared with the control.

As a result, as shown in Table 3, ethanol extracts of natural oriental herb extracts against Streptococcus mutans and Streptococcus were inhibited by 6 to 8 log cycles compared with the control, Activity. Streptococcus anginosus, Staphylococcus aureus and Staphylococcus epidermidis were inhibited by 1 ~ 2 log cycle inhibition only on Ganoderma lucidum and Morus alba ethanol extracts. There was almost no inhibitory effect.

Measurement of growth inhibitory effect on oral bacteria (log No. CFU / ml) Strain Natural herbal extract Control Stone
ethanol McMundong
ethanol Bellflower
ethanol Quince
ethanol Gold silver
ethanol Streptococcus mutans 8.54 ± 0.06 ^f 1.30 ± 0.00 ^c 1.97 ± 0.02 ^e 1.71 + 0.04 ^d 0.78 ± 0.00 ^a 0.97 + 0.03 ^b Streptococcus sanguineus 8.84 ± 0.06 ^e 2.51 + - 0.01 ^d 2.38 ± 0.07 ^c 2.59 ± 0.01 ^d 1.47 ± 0.01 ^b 1.06 + 0.06 ^a Streptococcus angiosus 7.29 + - 0.01 ^c 6.08 ± 0.01 ^b 7.32 0.02 ^c 7.91 ± 0.48 ^d 5.31 ± 0.01 ^a 6.09 ± 0.09 ^b Staphylococcus aureus 8.34 ± 0.06 ^d 8.53 ± 0.00 ^e 8.02 0.03 ^c 8.28 ± 0.04 ^d 6.70 ± 0.00 ^a 6.84 ± 0.01 ^b Starfilacocus epidermidis 8.34 ± 0.06 ^d 8.59 ± 0.11 ^e 8.04 0.04 ^c 7.83 + 0.02 ^b 7.60 + - 0.11 ^b 7.28 ± 0.02 ^a

The colony forming unit (CFU) is a colony forming unit, and a, b, c, d, e and f in the row indicate significant differences from each other, which means that p <0.05.

Example  4. Minimum inhibitory concentration of ethanol extract

Liquid medium dilution was used to measure minimal inhibitory concentration (MIC). Each sample strain was inoculated into 10 ml of brain heart infusion (BHI) liquid medium and incubated at 37 ° C for 24 hours. Each of the cultured strains (1 × 10 ⁷ CFU / ml) was added to a 96-well plate containing BHI medium supplemented with morula ethanol and ghoulse ethanol extract (0, 0.05, 0.1, 0.3 and 0.5% Respectively. The plate was incubated at 37 ° C for 24 hours, and its absorbance was measured at an absorption wavelength of 490 nm to confirm the antimicrobial effect. The minimum concentration of the sample in which the growth of each strain did not occur was defined as the minimum inhibitory concentration (MIC).

As a result, as shown in Table 4, it was confirmed that the MIC of the extracts of Streptococcus mutans and Streptococcus japonica var. Japonica and Ganoderma lucidum had the lowest value of 0.05%, indicating a high antimicrobial activity in a small amount.

Minimum inhibitory concentration for oral bacteria Strain MIC (%) Mosaic Ethanol Extract Ethanol Extract Streptococcus mutans 0.05 0.05 Streptococcus sanguineus 0.05 0.05 Streptococcus angiosus 0.1 0.3 Staphylococcus aureus 0.3 0.3 Starfilacocus epidermidis 0.3 0.3

Example  5. Determination of Thermal Stability of Ethanol Extract of Quercus and Gum-Eul

The extracts were heat treated at 80 ℃ for 30 min, at 100 ℃ for 30 min, and at 121 ℃ for 15 min, respectively, to measure the thermal stability of the extracts of Culex gruel and Ganoderma lucidum. The clear zone was measured and the presence or absence of thermal stability was measured in comparison with that of the untreated extract.

As a result, as shown in Table 5, the antimicrobial activity of the guar gum and Ganoderma lucidum ethanol extracts showed the antimicrobial activity even after the heat treatment, and the antimicrobial activity tended to be slightly lowered in the heat treatment as compared with the control without heat treatment. The antimicrobial activity of the extracts of guar gum and Ganoderma lucidum was found to be relatively stable to heat, and it is expected that the industrial utilization will be very high.

In Table 5, + means antibacterial activity, - means no antibacterial activity.

Thermal stability of ethanol extracts extract Strain Control Heat treatment temperature (캜) 80 100 121 Quince
Ethanol extract Streptococcus mutans + + + + Streptococcus sanguineus + + + + Streptococcus angiosus + + + + Staphylococcus aureus + + + + Starfilacocus epidermidis + + + + Gold silver
Ethanol extract Streptococcus mutans + + + + Streptococcus sanguineus + + + + Streptococcus angiosus + + + + Staphylococcus aureus + + + + Starfilacocus epidermidis + + + +

Example  6. Effects of single or multiple extracts of guinea-pig and gilt-eu-ethanol on the viability of oral bacteria

To investigate the effect of single or complex ethanol extracts of ginseng and ginseng extracts on the viability of germs in oral cavity, extracts of ginseng and Ganoderma lucidum were added at a concentration of 0.1% (v / v) After inoculation, the viable cell counts were measured by incubating for 24 hours, and the control with water was also measured by the same method. As a result, as shown in Table 6, the number of viable cell counts of single or mixed ginseng and ginseng ethanol was reduced by 3 to 4 log cycles, compared with that of the control, and the viable cell counts of the single- It is confirmed that the reduction effect is better.

Effect of reducing viable cell counts on oral bacteria (log No. CFU / ml) Strain Sample Control (water) Mother and child Gold euphon alone Mosaic + Gold Eunhwa Streptococcus mutans 5.40 ± 0.08 ^c 0.94 + - 0.03 ^a 1.61 + 0.02 ^b 1.03 + 0.05 ^a Streptococcus sanguineus 5.70 + - 0.05 ^d 1.06 + - 0.10 ^a 1.50 0.04 ^c 1.29 + 0.01 ^b Streptococcus angiosus 5.50 + - 0.01 ^d 1.01 0.02 ^a 1.14 ± 0.01 ^b 1.31 + - 0.01 ^c Staphylococcus aureus 5.55 + 0.07 ^c 0.95 ± 0.00 ^a 1.50 0.04 ^b 1.01 0.02 ^a Starfilacocus epidermidis 5.73 ± 0.03 ^c 1.95 ± 0.00 ^a 2.18 ± 0.00 ^b 1.97 + 0.02 ^a

CFU (colony forming unit) is a colony forming unit, and a, b, c and d in the row indicate a significant difference from each other, which means that p <0.05.

Meanwhile, when the extracts were added to the sterilized distilled water at a concentration of 0.1% alone or in combination, the number of viable bacteria in the oral cavity was measured at 37 캜 for 30 minutes at intervals of 10 minutes. As shown in FIG. 6, And it was confirmed that the reduction effect of ethanol extract of mosses was the best in the single treatment.

Example  7. Oral bacterial planktonic cells of the extract of mosses and ethanol planktonic  Antibacterial activity against cell

The preparation of the plankton cells was carried out by cooling the culture medium of five kinds of oral bacteria (Streptococcus mutans, Streptococcus sanguineus, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus angiosus) cultured for 24 hours After centrifugation at 3,000 rpm for 10 minutes using a centrifuge (Centrifuge 5810 R, Germany), the supernatant was discarded and washed twice with PBS (pH 6.8). The OD value of the bacterial culture was adjusted to 0.1. To determine the viability of the extracts, the bacterial strains were inoculated into a solution of 0.1%, 0.2% and 0.3% (v / v) of ethanol extracts of corn and ethanol extracts at 10, 20 and 30 minutes intervals. And the number of viable cells was compared with that before treatment (0 min).

As a result, as shown in Table 7, the decrease of the viable cell count was confirmed in all the treatments. Streptococcus mutans and Streptococcus mutansis were completely killed upon treatment with 0.2% and 0.3% of corn ethanol extract for 30 minutes.

The antimicrobial activity of planktonic cells of the bacterium in the oral cavity of the extract of corn ethanol (log No. CFU / ml) density(%) Strain 0 minutes 10 minutes 20 minutes 30 minutes 0.1 Streptococcus mutans 6.04 ± 0.04 ^d 4.64 + 0.04 ^c 4.18 ± 0.16 ^b 3.07 ± 0.06 ^a Streptococcus sanguineus 5.99 ± 0.03 ^d 5.10 + 0.09 ^c 4.94 + 0.04 ^b 3.61 ± 0.01 ^a Streptococcus angiosus 6.08 ± 0.07 ^d 4.58 + 0.02 ^c 4.29 ± 0.01 ^b 3.07 ± 0.06 ^a Staphylococcus aureus 6.03 ± 0.03 ^cd 4.59 + 0.02 ^b 4.18 ± 0.16 ^a 4.07 ± 0.06 ^a Starfilacocus epidermidis 5.95 ± 0.05 ^cd 5.89 ± 0.05 ^b 5.84 ± 0.06 ^b 4.93 ± 0.02 ^a 0.2 Streptococcus mutans 6.04 ± 0.04 ^d 3.35 ± 0.05 ^c 2.05 + 0.04 ^b 0.00 ± 0.00 ^a Streptococcus sanguinea 5.99 ± 0.03 ^d 2.05 ± 0.06 ^c 1.83 0.05 ^b 0.00 ± 0.00 ^a Streptococcus angiosus 6.08 ± 0.07 ^d 3.35 ± 0.05 ^c 2.05 + 0.04 ^b 1.96 ± 0.00 ^a Staphylococcus aureus 6.03 ± 0.03 ^cd 3.53 0.05 ^b 2.17 ± 0.15 ^a 2.09 ± 0.08 ^a Starfilacocus epidermidis 5.95 ± 0.05 ^cd 5.72 + 0.02 ^c 5.02 0.02 ^b 4.69 ± 0.01 ^a 0.3 Streptococcus mutans 6.04 ± 0.04 ^d 2.96 ± 0.00 ^c 2.26 + 0.07 ^b 0.00 ± 0.00 ^a Streptococcus sanguinea 5.99 ± 0.03 ^d 1.90 + - 0.05 ^c 1.31 + 0.01 ^b 0.00 ± 0.00 ^a Streptococcus angiosus 6.08 ± 0.07 ^d 3.29 + - 0.01 ^c 1.96 + 0.01 ^b 1.72 + 0.02 ^a Staphylococcus aureus 6.03 ± 0.03 ^cd 3.47 ± 0.01 ^c 2.28 0.02 ^b 2.02 0.02 ^a Starfilacocus epidermidis 5.95 ± 0.05 ^cd 5.04 0.04 ^c 4.85 ± 0.01 ^b 4.58 ± 0.02 ^a

CFU (colony forming unit) is a colony forming unit, and a, b, c and d in the row indicate a significant difference from each other, which means that p <0.05.

Example  8. Adhesive cells of oral bacteria in the extract of corn ethanol Attached cell ) On the antimicrobial activity

In order to measure the antimicrobial activity of adherent cells in the oral cavity of ethanol extracts of mosses, the surface to which the adherent cells were adhered was immersed in a BHI medium supplemented with 2% (w / v) agar, which is considered to be most similar to that of human oral mucosa And used after manufacturing. After drying in a 37 ° C incubator for 24 hours, 1 ml of bacterial strain in the oral cavity was inoculated, allowed to stand at room temperature for 1 hour, and washed twice with PBS (pH 6.8) to be used as an attached cell. The antimicrobial activity of the cell extracts on adherent cells was determined by treating the cells with 1 ml of extract (0.2 and 0.3%) at a concentration of ethanol and 10, 20, and 30 minutes, (pH 6.8), and then samples were collected using a sterile cotton swab. The samples were diluted with 0.1% peptone solution and inoculated into BHI agar. After incubation at 37 ° C for 24 hours, the colonies were counted CFU / ㎠, and the number of viable cells was compared with the control. As a result, as shown in Table 8, it was confirmed that the number of viable cells decreased by 1 to 3 log cycles when treated with the extract of corn ethanol.

The antimicrobial activity of the extract of mosses and ethanol on the attached cells of the oral bacteria (log No. CFU / ㎠) density(%) Strain 0 minutes 10 minutes 20 minutes 30 minutes 0.2 Streptococcus mutans 7.16 ± 0.17 ^c 4.73 ± 0.11 ^b 4.39 ± 0.10 ^b 4.09 0.05 ^a Streptococcus sanguineus 7.07 ± 0.05 ^d 4.47 ± 0.09 ^c 4.22 + 0.02 ^b 4.12 ± 0.09 ^a Streptococcus angiosus 7.35 + 0.01 ^d 4.49 ± 0.00 ^c 4.33 ± 0.01 ^b 4.12 + 0.04 ^a Staphylococcus aureus 7.20 ± 0.22 ^d 5.92 ± 0.86 ^c 5.02 0.03 ^b 4.99 ± 0.05 ^a Starfilacocus epidermidis 7.67 ± 0.10 ^cd 6.45 ± 0.15 ^b 5.91 ± 0.72 ^b 5.23 ± 0.46 ^a 0.3 Streptococcus mutans 7.16 ± 0.17 ^c 4.28 ± 0.02 ^b 4.09 ± 0.02 ^b 3.93 ± 0.00 ^a Streptococcus sanguinea 7.07 ± 0.05 ^d 4.06 ± 0.01 ^c 3.19 ± 0.17 ^b 3.09 ± 0.02 ^a Streptococcus angiosus 7.35 + 0.01 ^d 4.46 ± 0.01 ^c 4.05 + 0.01 ^b 3.89 ± 0.03 ^a Staphylococcus aureus 7.20 ± 0.22 ^d 5.90 + 0.07 ^c 4.95 ± 0.01 ^b 4.90 ± 0.07 ^a Starfilacocus epidermidis 7.67 ± 0.10 ^cd 5.78 ± 0.11 ^c 5.46 ± 0.25 ^b 5.09 ± 0.25 ^a

CFU (colony forming unit) is a colony forming unit, and a, b, c and d in the row indicate a significant difference from each other, which means that p <0.05.

Example  9. Antimicrobial Effect of Morus and Ethanol Extract on Oral Bacteria Contaminated with Toothbrush

To investigate the possibility of using toothpaste as a toothbrush disinfectant for extracts of corn ethanol, toothbrushes were contaminated with oral bacteria causing tooth decay and periodontitis. In order to contaminate the oral bacteria with the toothbrush, 1 ml of oral bacteria cultured twice at 37 ° C for 24 hours was added to 20 ml of peptone water to prepare a bacterial suspension. The toothbrush was immersed in the suspension for 30 seconds, then immersed again in sterile distilled water for 30 seconds, and then dried for 20 minutes. The contaminated toothbrush was immersed in 50 ml of physiological saline containing 0.2% (v / v) corn ethanol extract for 10 to 40 minutes and then dried for 20 minutes to measure the number of oral bacteria remaining on the toothbrush. As a result, as shown in Fig. 7, there was a difference according to the species, but all of the oral bacteria of the toothbrushes not immersed in the manganese ethanol showed about 10 ⁶ CFU / ㎖, and the number of oral bacteria decreased when the manganese ethanol extract was immersed Respectively. In particular, since 20 minutes and 30 minutes treatment showed similar effects, it was confirmed that sufficient disinfection effect could be obtained even after 20 minutes treatment.

Through the above examples, it has been confirmed that the extract of corn ethanol can be used as a disinfectant for various oral hygiene tools such as a toothbrush and an oral cleanser for oral hygiene (e.g., gauze).

Claims (7)

A composition for inhibiting oral bacteria in a Streptococcus mutans containing an extract of corn ethanol as an active ingredient. delete delete delete The composition for inhibiting oral bacteria according to claim 1, wherein the composition is one selected from the group consisting of an oral cleanser, an oral cleanser, a toothpaste, an oral spray, a toothbrush cleaner, and a denture cleanser. Quasi-drugs for the prevention or improvement of oral diseases caused by Streptococcus mutans containing an extract of corn ethanol as an active ingredient. delete

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