KR20190094518A - Antibacterial brush bristle composition using natural material having non-toxic oral health function, its bristles and method for producing the same - Google Patents

Abstract

The present invention relates to an antibacterial bristle composition using a natural material having a non-toxic oral health function, bristles thereof, and a manufacturing method thereof. More particularly, the present invention relates to an antibacterial bristle composition using a natural material having a non-toxic oral health function, improved to contribute to maintaining oral cleanliness and oral health by having anti-inflammatory and antibacterial activities against the periodontal disease-causing bacteria, reducing bad breath, and inhibiting aging of gums; bristles thereof; and a manufacturing method thereof.

Description

Antibacterial brush bristle composition using natural material having non-toxic oral health function, its bristles and method for producing the same}

The present invention relates to an antimicrobial bristle composition using a natural product having a non-toxic oral health function and its bristles, and to a method for producing the same. More specifically, the antimicrobial and anti-inflammatory activity, bad breath improvement ability, and gum aging inhibitory activity against periodontal disease-causing bacteria The present invention relates to an antimicrobial bristle composition using a natural product having an improved nontoxic oral health function so as to contribute to maintaining oral cleanliness and oral health, and a bristle and a manufacturing method thereof.

In general, a toothbrush consists of a bristles, a head portion having the bristles and a handle portion connected to the head portion, which should always be hygienic as an important tool for oral hygiene.

In particular, the bristles in the head of the toothbrush must have a certain strength and elasticity, as well as be easily moved along the tooth, the tooth being brushed.

However, since these bristles are always in a humid and poorly ventilated state, they are difficult to handle and store hygienically, and thus often cause disease and hygiene problems caused by various bacterial propagation.

Because of this, bacteria breed easily, and when brushing with these bristles, various gum diseases such as inflammation of the mouth, bad breath, and aging of the gums are caused.

Therefore, in order to solve this problem, a method of chemically disinfecting or disinfecting the toothbrush with ultraviolet rays has been developed, but these methods have a disadvantage in that the procedure is very complicated.

In addition, sterilizers have been developed to suppress bacterial growth of the bristles, and a sterilization box has been developed as an example of such a device. Such a device is very expensive and difficult to be easily purchased in a general household, but also for continuous operation. Since power must be supplied, power is consumed, which is economically inefficient, and is difficult to carry due to disadvantages that are difficult to carry.

Recently, as an alternative method for developing an antibacterial toothbrush, methods for coating and mixing materials having antiseptic and antimicrobial properties on the bristles have been developed, but the effective ones are toxic and cannot exclude human hazards, and the others are nontoxic. Antibacterial, anti-inflammatory activity, bad breath improvement, gum aging inhibitory ability, etc. have not been tested, such as the situation is high concern.

Republic of Korea Patent No. 084183 (2007.12.04), toothbrush having bristles containing charcoal and tourmaline powder Republic of Korea Patent No. 0796574 (2008.01.15), Toothbrush manufacturing method containing pine needle powder and antibacterial ingredients and the toothbrush Republic of Korea Patent No. 0969854 (Jul. 6, 2010), Toothbrush and Toothbrush with Bamboo Salt Republic of Korea Patent Publication No. 10-2013-0012535 (2013.02.04), bristle containing microcapsules containing bamboo salt component and toothbrush comprising the same Republic of Korea Patent No. 1476812 (2014.12.19.), Antimicrobial bristle composition containing organic nano clay and antimicrobial bristle using the same and a method of manufacturing the same

The present invention was created in view of the above-mentioned problems in the prior art, and has been created to solve this problem, and has antibacterial and anti-inflammatory activity, bad breath improvement ability, and gum aging inhibitory ability against periodontal disease-causing bacteria. The main purpose of the present invention is to provide an antimicrobial bristle composition using the natural product having an improved non-toxic oral health function so as to contribute to the bristles and a manufacturing method thereof.

The present invention is a means for achieving the above object, the sulfur extract having a concentration of 1-5% dissolved in alcohol alcohol; It provides an antimicrobial bristle composition utilizing a natural product having a non-toxic oral health function, characterized in that it comprises a; polyester filament loaded with the rhubarb extract.

At this time, the composition is characterized in that it further comprises a rosemarine (Rosmarinic Acid) having a concentration of 1-100mg / ㎖.

In addition, the composition is characterized in that it further comprises Epigallocatechin gallate (EGCG) having a concentration of 1-10mg / ㎖.

In addition, the composition is characterized in that it further comprises EC (Epicatechin) having a concentration of 1-10mg / ㎖.

In addition, the composition is characterized in that it further comprises Rosemarinic acid (Rosmarinic Acid) having a concentration of 1-100mg / ㎖, and EGCG (Epigallocatechin gallate) having a concentration of 1-10mg / ㎖.

In addition, the composition is characterized in that it further comprises Rosemarinic acid (Rosmarinic Acid) having a concentration of 1-100mg / ㎖, and EC (Epicatechin) having a concentration of 1-10mg / ㎖.

In addition, the composition is characterized in that it further comprises EGCG (Epigallocatechin gallate) having a concentration of 1-10 mg / ㎖, and EC (Epicatechin) having a concentration of 1-10 mg / ㎖.

In addition, the composition includes Rosemarinic acid having a concentration of 1-100 mg / ml, Epigallocatechin gallate (EGCG) having a concentration of 1-10 mg / ml, and EC having a concentration of 1-10 mg / ml. (Epicatechin) is also included in the feature.

In addition, the polyester filament in the composition is characterized in that the propolis having a concentration of 0.125-0.5% dissolved in alcohol alcohol is supported with the sulfur extract.

In addition, the present invention comprises the steps of making an antimicrobial bristle composition consisting of a yellow filament extract having a concentration of 1-5% dissolved in alcohol alcohol, and a polyester filament loaded with the sulfur extract; Making a master batch after adding a functional additive to the antimicrobial bristle composition, Rosemarinic Acid, Rosegalic Acid, or EGCG (Epigallocatechin gallate) or EC (Epicatechin) alone or in a mixture of two or more; It also provides a bristles manufacturing method using an antimicrobial bristle composition using a natural product having a non-toxic oral health function, characterized in that the step of preparing a bristle by melting, spinning, cooling the masterbatch.

In addition, the present invention also provides bristles prepared using the bristle composition described above.

According to the present invention, the antibacterial activity against the periodontal disease-causing bacteria, anti-inflammatory activity, bad breath improvement ability, and gum aging inhibitory ability can be obtained to contribute to oral cleanliness and oral health promotion.

1 is an exemplary view showing an example of obtaining a sulfur extract constituting the composition according to the present invention.
Figure 2 is an exemplary view showing an example of obtaining a propolis constituting the composition according to the present invention.

Hereinafter, a preferred embodiment according to the present invention will be described in more detail.

Prior to the description of the present invention, the following specific structures or functional descriptions are merely illustrated for the purpose of describing embodiments according to the inventive concept, and the embodiments according to the inventive concept may be implemented in various forms, It should not be construed as limited to the embodiments described herein.

In addition, the embodiment according to the concept of the present invention may be variously modified and may have various forms, it should be understood to include all modifications, equivalents, or substitutes included in the spirit and scope of the present invention. .

The present invention is a master batch made of a polyester filament loaded with natural ingredients that can contribute to maintaining oral cleanliness and oral health by providing antibacterial and anti-inflammatory activity, bad breath improvement ability, and gum aging ability against periodontal disease-causing bacteria. (Master Batch) is made by melting and spinning the master batch to cool and cut to produce fine hair, that is, bristles, and the prepared bristles are planted on the toothbrush rod to complete the toothbrush.

Here, the master batch refers to the production of pellets by mixing the additives imparting functionality to the resin as a main raw material by mixing and rotating the mixture by a blender and extruding the raw material by a twin screw extruder having a suitable temperature.

Recently, interest in oral health has increased as the standard of living of the people has increased, and the average daily number of teeth brushing is 2.35 times, and the consumption of oral health care products has increased as the attitude toward oral health care changes. Is in.

In this regard, LG H & H, Perio, Median, Oralby, Ecumax, CJ LION, Median, DIOTEK Korea, and Triple Korea, which are oral health care product manufacturers, have differentiated oral health from competitors in the rapidly changing market environment. For example, Oralby's Pro-Expert Clinical Pro-Flex has created a power tip brush head that cleans every corner of the mouth and makes it hard to reach, making it an excellent cleaning effect with moving brush heads and tooth gaps through comb-shaped brush heads. The premium-grade product has been introduced to the gum massage care function by penetrating deeply to remove plaque, and various examples such as Perio's slim touch black pearl toothbrush and Median's green tea antibacterial care toothbrush are disclosed.

Nevertheless, the average annual increase rate of patients receiving periodontal disease is 7.3%. Therefore, the development of technology with effective characteristics such as toothpaste, toothbrush, etc., which is necessary for the prevention of malignant oral disease, is required.

In addition, in recent years, a new type of flu or hepatitis A, the toothbrush having antibacterial has become a major issue of the toothbrush system, in view of these problems, the present invention improves the human safety while using anti-toxic natural antibacterial material to increase the antimicrobial properties. It has led to the development of enhanced toothbrush technology.

In addition, most of the current brush bristles antimicrobial agent is an organic antimicrobial agent because it has excellent antimicrobial, but due to the restrictions on the human body due to restrictions on the use of non-toxic antibacterial function of the bristles is urgently developed.

The antimicrobial bristle composition using a natural product having a nontoxic oral health function according to the present invention is used as a nontoxic natural component, but not used as an extract.

More specifically, the antimicrobial bristle composition is composed of a yellow filament extract having a concentration of 1-5% dissolved in alcohol alcohol, and a polyester filament on which the yellow flock extract is carried.

At this time, the rhubarb is a perennial herbaceous plant belonging to the buttercup family, and includes medicinal ingredients such as Berberine, Copticine, Warenine, Palmintine, and the like, and particularly Berberine It is known to have a high healing effect on stomatitis, sulphitis and stomatitis.

In addition, it has been reported that these yellow lotuses suppress dysentery, stop pulmonary tuberculosis and soil formation, have a strong bactericidal action, and have anti-inflammatory and antipyretic effects.

The method of obtaining a sulfur extract from the sulfur can be obtained by rinsing the dried yellow lotus root finely as shown in FIG. 1 and then repeatedly extracting a total of 5 times while immersing 1200 g in 70% methanol for 24 hours at room temperature. This was filtered through filter paper, concentrated under reduced pressure at 60 ° C., and the concentrated extract was lyophilized for 24 hours to be powdered, which was dissolved in alcohol alcohol to have a concentration of 1-5%.

Here, the reason for limiting the concentration of the sulfur extract is because it was confirmed that the antimicrobial inhibitory ability is high while maintaining the non-toxic (active substance berberine does not affect the survival of oral normal cells) at the concentration in the above range. It is also intended to be in conformity with No. 2006-32 of the Agency.

In the process of further studying the characteristics of the sulfur in the present invention, in particular, the bactericidal power against six harmful bacteria (ATCC 33384, ATCC 43037, ATCC 25586, ATCC 25611, ATCC 33277, ATCC 2517) causing various diseases in the oral cavity Since it was confirmed that it was excellent, the technology was developed to realize the most effective sterilization power based on this.

The present inventors analyzed through a minimum bactericidal concentration (MBC: Minimun Bactericidal Concentration, mg / ㎖) test to confirm the bactericidal power of the sulfur extracts for six harmful bacteria.

As a result, 0.25 mg / ml for ATCC 33384 (Aggregatibacter actinomycetemcomitans), 0.03 mg / ml for ATCC 43037 (Tannerella forsythensis), 0.0125 mg / ml for ATCC 25586 (Fusobacterium nucleatum subsp.nucleatum), ATCC 25611 (Prevotella intermedia) 0.06 MBC was identified as mg / ml, ATCC 33277 (Porphyromonas gingivalis) 0.06 mg / ml, ATCC 2517 (Streptococcus mutans) 0.25 mg / ml.

In addition, the present invention may further include rosmarinic acid in the antimicrobial bristle composition.

Rosemarinic acid (Rosmarinic Acid) is for maximizing antiviral activity, antimicrobial activity, anti-inflammatory activity, antioxidant activity, Streptococcus sobrinus, Candia albicans, Enterobacter spp., Listeria monocytogenes, Pityrosporium ovale, Staphylococcus epidermidis, Propionibacterium acnes, etc. It is a substance that has antiproliferative ability against microorganisms.

However, in order to maintain non-toxicity (does not affect the survival of oral normal cells), it should have a concentration of 1-100 µg / ml.

Here, the cell survival rate, that is, the nontoxicity test was performed through the analysis of cytotoxicity by MTT according to the Korea Food and Drug Administration's notice standard, which was cultured human normal oral keratinocyte (HNOK), test groups such as sulfur, rosemary, EGCG (Epigallocatechin gallate), EC (Epicatechin) and Propolis (Propolis), which are described below, are independently treated to quantitative cytotoxicity evaluation according to the MTT method, and the reaction time is divided into 24 hours, 48 hours, and 72 hours. Proceeded.

At this time, MTT Assay measured the survival rate of human-derived oral fibroblast cells by MTT assay based on MTT reduction, which is converted to a purple formazan product by mitochondrial dehydrogenase of dense cells, and the cells were stored in DMEM medium at 1 × 10⁴ cells / well. The drugs were treated by concentration by suspension at the density of. After incubation, MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenylterazoli-um bromide) solution was added at a concentration of 0.5 mg / ml and incubated for 4 hours, and the MTT-formazan product was DMSO. Dissolve by adding 200 μl.

In addition, biosafety was determined by measuring the amount absorbed at 540 nm using a spectrophotometer (Molecular devices, Palo Alto, USA). The survival rate of the cells was expressed as the percentage of control cells without any treatment.

That is, it was calculated as viability (%) = 100 × (absorbance of treated sample) / (absorbance of control).

In addition, the preparation of the sample was used by dissolving sulfur, rosemary acid, EGCG (Epigallocatechin gallate), EC (Epicatechin) and Propolis samples by concentration (1-100 ㎍ / ㎖) in DMEM medium, the results described above and below Got.

In addition, the cell line was harvested in primary culture by extracting the site without inflammation when extracting the third molar in order to use human-derived oral fibroblast cells, and the tissues were washed three times or more with PBS, and cut into 1-2 mm size 0.15% Collagenae-dispase was treated for 30 minutes. And, after separating the epithelium and connective tissue with tweezers under a stereoscopic microscope, the epithelium was centrifuged by treatment with 0.25% trysin EDTA solution for 15 minutes, the medium was exchanged every 3 days using DMEM medium, 70-80% rich If grown, it was used after three to four passages.

In addition, the present invention may further comprise EGCG (Epigallocatechin gallate) in the antimicrobial bristle composition.

The EGCG is the main component of the catechin, it is added for this because it has caries prevention and antibacterial activity, antiviral activity, anti-inflammatory action, antioxidant activity, carcinogenic activity.

In particular, as shown in [Figure 1], as a result of the paper disk method, when treated with oral harmful bacteria at a concentration of 10mg / mL showed a very significant antimicrobial activity, and above all C. albicans, the causative agent causing stomatitis It has been found to inhibit very effectively.

[Figure 1]

However, if the concentration exceeds 10mg / mL, the viability of normal cells in the oral cavity is lowered to 70% or less, so it is judged that nontoxicity cannot be maintained and should be added while maintaining the concentration range of 1-10mg / mL.

In addition, the present invention may further include EC (Epicatechin) in the antimicrobial bristle composition.

The EC is a substance belonging to the catechins, it is added for this because it has caries prevention, antibacterial activity, antiviral activity, anti-inflammatory action, antioxidant properties.

In particular, as shown in [Figure 2], as a result of the paper disk method, when treated with oral harmful bacteria at a concentration of 10mg / mL showed a significant antimicrobial activity, and above all, it was confirmed that it effectively inhibits S. sobrinus It became.

[Figure 2]

However, if the concentration exceeds 10mg / mL, the viability of normal cells in the oral cavity is lowered to 70% or less, so it is judged that nontoxicity cannot be maintained and should be added while maintaining the concentration range of 1-10mg / mL.

In addition, the present invention may be added to the antimicrobial bristle composition by mixing two or more of the above materials.

For example, Rosemarinic Acid and Epigallocatechin gallate (EGCG), Rosemarinic Acid and EC (Epicatechin), Epigallocatechin gallate and EGCG (Epicatechin), Rosemarinic Acid and EGCG (Epigallocatechin gallate) Epicatechin) may be mixed. However, the concentration should maintain the concentration of the above-mentioned single component as it is.

In particular, these components may be a functional additive, according to the method of manufacturing bristles according to the following.

That is, the bristle manufacturing method according to the present invention comprises the steps of making an antimicrobial bristle composition consisting of a yellow filament extract having a concentration of 1-5% dissolved in alcohol alcohol, and a polyester filament carrying the sulfur extract; Making a master batch after adding a functional additive to the antimicrobial bristle composition, Rosemarinic Acid, Rosegalic Acid, or EGCG (Epigallocatechin gallate) or EC (Epicatechin) alone or in a mixture of two or more; Melting and spinning the masterbatch to prepare the bristles by cutting.

In addition, the prepared bristles can be planted on the toothbrush bar to complete the toothbrush.

In addition, the present invention may further include Propolis as a component supported on the polyester filament.

The propolis may also be used with the functional additives.

In the present invention, it is necessary to support propolis having a concentration of 0.125-0.5% dissolved in alcohol alcohol, which also maintains non-toxicity (does not affect the survival of oral normal cells) when the concentration is within the above range. Because it was confirmed to be high.

These propolis have excellent antibacterial, anti-inflammatory, anti-oxidative and immune-enhancing effects in relation to oral health, and as a result of confirming the antimicrobial activity by the paper disk method against oral harmful bacteria, it is very significant within the concentration range as shown in [Figure 3]. In conclusion, we found that S. mutans, an early causative agent of dental caries, can be effectively inhibited.

[Figure 3]

However, it was confirmed through experiments according to the notification standard of the Ministry of Food and Drug Safety that the concentration in the above range should be maintained in order not to affect oral cell viability, that is, to maintain nontoxicity.

At this time, the propolis was used as a Brazilian product provided by Rappa Propolis Research Institute Co., Ltd., and powdered propolis in a solid lump state, from which impurities were removed, as shown in FIG. Was dissolved in 50 ℃, sonication conditions as a solvent.

In this case, when alcohol was used as a solvent, the fat-soluble substance in the propolis was well dissolved, but when distilled water was used as the solvent, only the water-soluble substance in the propolis was dissolved and the fat-soluble substance was formed as a precipitate under the beaker. Therefore, in the present invention, it is preferable to utilize the alcohol using the alcohol as a solvent.

Hereinafter, an Example is described.

In the embodiment according to the present invention was confirmed whether the growth inhibitory ability against oral harmful bacteria causing tooth decay and periodontal disease bacteria centered on non-toxic natural components of sulfur and propolis supported on the polyester filament.

For the antimicrobial activity analysis used in the experiment, the paper disc method was used, and a single colony of each strain isolated from pure strains was inoculated into 10 ml bacterial growth liquid medium, and cultured three times for 18 to 24 hours at the growth temperature of each strain. It was used as an antibacterial test strain, and the preparation of the antibacterial test plate medium was sterilized by growth medium containing 15% of agar of each strain, and then divided into 15 ml in a petri dish to coagulate the medium for the substrate. After the optical density (O, D) value is 0.4 (10 ⁶ CFU / mL) at 650nm, add it aseptically to the medium layer containing 0.7% agar and mix well. It was coagulated evenly to prepare a double inoculation medium medium.

In addition, after placing a sterile 8 mm paper disc on a sufficiently solid medium, 40 μl in the range of 0, 0.125, 0.25, 0.5, 1% propolis concentration, 0, 1,5, 10, 20% After absorbing into the discs, the cells were incubated at 37 ° C for 24 hours, and then the clear zone around the disc was observed.

증식 proliferation inhibitory activity against S. mutans causing dental caries

Occurs in many elderly oral S. mutans is dental caries (Dental caries: caries disease), the enamel of teeth damaged by acid (acid) produced as decomposition such as sugar, starch by S. mutans pathogens that cause the initial This leads to tooth decay.

According to the experimental results of the present invention, as shown in [Fig. 4] below, in the case of sulfur, 1 to 20% of all concentration groups showed a band of 1.5 mm or more, and in propolis of 0.125 to 1% of all concentration groups, a band of 2 mm or more. There was no difference in bacterial inhibition between 0.125% and 0.25% concentrations.

[Figure 4]

증식 proliferation inhibitory activity against S. sobrinus causing periodontitis and caries disease

S. sobrinus is a causative agent that aggravates tooth decay by acting secondarily when tooth enamel is damaged by S. mutans.

  According to the experimental results of the present invention, as shown in the following [Figure 5], 1 to 20% of all concentration groups in the case of the rhubarb showed more than 2mm transparent zone, 0.1mm to 1.5% in all concentration groups of propolis The above opacity was observed and there was no difference in bacterial inhibition between 0.25% and 0.5% concentrations.

[Figure 5]

증식 proliferation inhibitory activity against C. albicans

Candida albicans, the causative agent of oral candidiasis (Candidiasis, Thrush, thrush, stomatitis), develops off-white or milky white spots on the surface of the oral mucosa.

According to the experimental results of the present invention, as shown in the following [Figure 6], in the case of the yellow soybeans showed a transparent band of 1mm or more in 5 ~ 20% concentration group, in the case of propolis 0.125 ~ 1% in all concentration groups more than 1mm Indicated.

[Figure 6]

⑷ S. mutans, S. sobrinus, C . goldthread for albicans, compare the antimicrobial activity of propolis

As a result of comparing the antimicrobial activity of the three oral harmful bacteria, as shown in Table 1 and 2, the antimicrobial activity of sulfur and proplis were all concentration-dependent, and as the concentration of natural products increased, the inhibitory activity against bacteria was also increased. In particular, the two extracts showed very strong inhibitory activity against S. mutans and S. sobrinus .

Therefore, the use of two extracts may be effective in inhibiting S. mutans , S. sobrinus, and C. albicans , which cause stomatitis, as the main causes of dental caries disease.

Microbial strains Inhibition zone (Dia, mm) Sample concentration One% 5% 10% 20% Control 0.0 0.0 0.0 0.0 C. calbicans 0.0 1.0 2.0 3.0 S. mutans 1.5 4.0 5.0 6.0 S. sobrinus 2.0 4.0 5.0 6.0

(Comparison of Antimicrobial Activity of Concentrations of Strain against S. mutans, S. sobrinus, and C. albicans )

Microbial strains Inhibition zone (Dia, mm) Sample concentration 0.125% 0.25% 0.5% 1.0% Control 0.0 0.0 0.0 0.0 C. calbicans 1.0 2.0 2.5 3.0 S. mutans 2.0 2.0 3.0 3.5 S. sobrinus 1.5 2.0 2.0 2.5

(Comparison of Antimicrobial Activities of Propolis against S. mutans, S. sobrinus, and C. albicans )

In addition, the antioxidant activity analysis also verified the prevention of early aging gums.

분석 SOD (Superoxide dismutase) similar activity assay

SOD-like activity was measured by the method of Beauchamp and Fridovich (1971). A solution containing 50 mM carbonic buffer (pH 10.2), 0.1 mM EDTA, 0.1 mM Xanthine, 0.025 mM nitroblue tetrazolium (NBT), and extracts was reacted at 25 ° C for 10 minutes, followed by Xanthine oxidase The reaction was measured. SOD activity was measured for 5 minutes at 550nm for 30 seconds.

At this time, the SOD-like activity of the sulfur extract was analyzed by SOD-like activity by concentration (0 ~ 2.5 mg / mL), showed an effective activity as shown in Table 3 in the concentration range of 0.15 ~ 2.5 mg / mL.

SOD- like activities (%) Extract concentration (mg / mL) 0.156 0.312 0.625 1.25 2.5 17 ± 0.4 20 ± 0.8 22 ± 0.4 30 ± 0.6 41 ± 0.4

In addition, SOD-like activity of propolis was analyzed by SOD-like activity by concentration (0 ~ 2.5 mg / mL) Propolis, showed an effective activity as shown in Table 4 in the concentration range of 0.15 ~ 2.5 mg / mL.

SOD- like activities (%) Extract concentration (mg / mL) 0.156 0.312 0.625 1.25 2.5 10 ± 0.2 13 ± 0.6 17 ± 0.5 23 ± 0.2 35 ± 0.6

⑹ DPPH radical scavenging activity analysis

Blois' method for measuring radical scavenging effect of sample was slightly modified. 100 μl of 1 × 10 ⁻⁴ M DPPH and 100 μl of each extract were left in the dark for 30 minutes, and the residual radical concentration was measured at 517 nm using an ELISA Reader (Bio-RAD, USA).

At this time, the DPPH radical scavenging ability of the sulfur extract was analyzed by DPPH radical scavenging ability of the sulfur extract, showed a significant activity as shown in Table 5 at all concentrations (12.5 ~ 50 mg / mL).

DPPH radical scavenging activity (%) Extract concentration (ug / mL) 6.25 12.5 25 50 100 200 32 ± 0.4 41 ± 0.7 55 ± 0.3 67 ± 0.5 71 ± 0.4 68 ± 0.8

In addition, DPPH radical scavenging ability of propolis showed significant activity as shown in Table 6 at all concentrations (12.5 ~ 50 mg / mL).

DPPH radical scavenging activity (%) Extract concentration (ug / mL) 6.25 12.5 25 50 100 200 40 ± 0.7 55 ± 0.4 68 ± 0.5 80 ± 0.9 84 ± 0.3 83 ± 0.3

⑺ nitrite scavenging activity analysis

10 mL of the reaction solution was prepared by adding 1 mL of 1 mM NaNO 2 solution to 2 mL of the diluted 10-fold dilution sample and adjusting the pH of the reaction solution to 1.2 using 0.1 N HCl and 0.2 M citrate buffer (pH 6.0). After the solution was allowed to stand at 37 ° C for 1 hour, 5 mL of 2% acetic acid and 0.2 mL of Griess reagent (1% napthylamine: 1% sulfanilic acid = 1: 1 in 30% acetic acid) were added to 1 mL of the sample solution. After standing at room temperature for 15 minutes, absorbance was measured at 517 nm to calculate remaining nitrite.

At this time, the nitrite scavenging ability of the rhubarb extract was found to be higher depending on the concentration of the extract as shown in the graph of Figure 7 below, and showed excellent scavenging ability close to 100% at the concentration of 1.25mg / mL.

[Figure 7]

In addition, the nitrite scavenging ability of propolis was found to be higher depending on the concentration of the extract, as in the graph of Fig. 8, below. It showed much higher scavenging activity, and at a concentration of 0.015 mg / mL, it showed a very surprising scavenging activity of more than 200%.

[Figure 8]

In addition, anti-inflammatory activity was also analyzed.

⑻ Nitric Oxide / Nitrite Production Analysis

In nitric oxide production, nitrite levels in culture media and serum can be measured indirectly by colorimetric analysis of Griess reagent. For measurement of NO murine macrophage cell line RAW264.7 cells were used, Griess reagent was prepared with 1% sulfanilamide, 0.1% naphthyl ethylenediamine dihydrochloride, 5% phosphoric acid. After incubating raw264.7 with DMEM (10% FBS, 1% P / S) and dispensing 5x10 ⁵ cells / mL in 96-well plate, the next day sample was processed according to the concentration, and after 1 hour the main inflammatory reaction Treats LPS (Lipopolysaccharide) which acts as urea. After 24 hours, the NO level was mixed with the supernatant of the sample and the Griess reagent in a 1: 1 ratio by using Griess reagent, and then left at room temperature for 10 minutes after blocking light, and the absorbance was measured at 540 nm of ELISA reader.

At this time, the inhibitory activity of NO production of sulfur extract was inhibited in NO concentration compared to the control in all concentration groups (0.625 ~ 5 ug / mL) treated with LPS as shown in Figure 9 below. .

[Figure 9

In addition, as shown in Figure 10 below, the inhibition of NO production of propolis significantly inhibited NO production compared to the control group in both concentration groups treated with LPS (0.625 ~ 5 ug / mL). When it showed very high anti-inflammatory activity, there was no particular difference by concentration.

[Figure 10]

In addition, the oral health efficacy of the masterbatch according to the present invention and the bristles prepared using the same were also verified.

At this time, the medium is LB medium which most bacteria can survive and multiply as a basic microbial medium, and after the masterbatch is administered to the LB medium at 0, 1 week, 2 weeks, 3 weeks, 4 weeks interval of bacteria Whether or not colonies occurred was confirmed by agar medium and Spreading Culture method.

무 Aseptic test of masterbatch

In the case of the masterbatch (MB) loaded with hull, the sterile force was maintained until 3 weeks of culture as shown in Figure 11 below, but the bacterial colony appeared from 4 weeks, so the sterile holding power of the masterbatch was safely maintained for 3 weeks. And it was found.

[Figure 11]

In the case of the masterbatch (MB) loaded with propolis, aseptic cultures were maintained until 2 weeks of culture as shown in Figure 12 below, but bacterial colonies appeared from 3 weeks, so that the aseptic retention of the masterbatch was safely maintained to 2 weeks. It was found to be maintained.

[Figure 12]

Therefore, it was expected that the effect would be even greater when both sulfur and propolis were loaded together.

무 Aseptic force test of brush head

In the case of the bristles prepared using the masterbatch, which was loaded with the rhubarb and propolis, as shown in Figure 13 below, the sterile force was maintained until 3 weeks of culture, but a few bacterial colonies began to appear from 4 weeks. Therefore, it could be confirmed that the sterile force is safely maintained until three weeks.

[Figure 13]

세균 Analysis of bacterial membrane and plaque removal ability of tooth surface using brush head according to the present invention

This was confirmed by analyzing the tooth plaque index.

For the oral examination, the tooth coloring liquid Chrom-O-Red, Germiphene, Canada) was evenly applied to the subject's tooth surface using a cotton ball and lightly washed with water.

In addition, the staining status was examined by using a probe and alveolar. The adhesion degree of the dental plaque was applied to Quigley and Hein's dental plaque rating criteria modified by Turesky et al. The individual Bacterial Bacterial Index was calculated by dividing the sum of the measured values by each site by the number of teeth to be examined.

(Colored surface number / total surface number to be examined) * 100

In addition, the dental plaque index of Quigley and Hein, modified by Turesky et al., Was scored as follows.

-0: Non-dental bacterium

-1: dot attached to gingival edge

-2: Adhesion along gingival margin (width less than 1mm)

-3: If there is dental plaque on the cervical side 1/3

-4: If there are dental plaques up to 2/3 of the cervical side

-5: If there is a bacterial membrane beyond the cervical side 2/3

For sensory evaluation, ten people in their late twenties were selected and evaluated.

Plain brush head Barrel Brush Head propolis bristle Participant 1 25 30 25 Participant 2 32 32 30 Participant 3 24 26 22 Participant 4 12 20 18 Participant 5 23 15 32 Participant6 23 19 20 Participant7 35 30 25 Participant8 30 32 12 Participant9 38 13 23 Participant10 39 25 20 Dense bacterial film index
medium
12.5
11
10.3

As a result, it was confirmed that the reduction rate of plaque bristles and propolis bristles increased in the plaque removal ability.

확인 Confirmation of bad breath improvement effect (Sensory evaluation)

Seven men and women in their late twenties were selected and checked for the improvement of bad breath by measuring the content of hydrogen sulfide, methyl mercaptan, and methyl sulfide, using a bad breath analyzer.

At this time, the test method was to use a 1ml syringe in the mouth without using a tooth brush or any oral supplies, breathing only through the nose without opening the mouth, collecting bad breath in the mouth. After 3 minutes, the syringe was pumped 2 ~ 3 times, 1mL was collected and taken out of the mouth, and the 1mL scale was correctly aligned and inserted into the breath gas detector (Sensor Gas Chromatograph ODSA-FIS Inc.) for 4 minutes. The bad breath components measured were analyzed based on methyl sulfide [(CH₃) ₂S], hydrogen sulfide (H₂S), and methyl mercaptan (CH₃SH).

As a result, in the case of the bristles bristles, hydrogen sulfide increased 29.3125, methylmercaptan decreased 4.0125, and methyl disulfide 0.2 increased.

In the case of propolis bristles, hydrogen sulfide increased 48.775, methyl mercaptan decreased 2.275, and methyl disulfide 13.0125 after use.

 Overall, the total maximum bad breath reduction rate is summarized as follows: hydrogen sulfide decreased 99.98% in propolis, methyl mercaptan decreased 60.60% in sulfur, and methyl disulfide also decreased by 14.65% in sulfur.

As a result, the average rate of reduction of maximum bad breath was 2.68% in Huangshan.

확인 Confirmation of oral harmful bacteria inhibition and bad breath improvement through clinical trial (Clinical trial)

A total of six months were included for this study, including four weeks of study duration and study preparation and preparation.

The investigator selected oral subjects for 31 subjects as shown in Table 8 by judging whether the subjects were eligible for the study by oral examination at the first visit.

All subjects received oral hygiene training, and each individual was given the same toothpaste as two toothbrushes and instructed to brush their teeth four or more times a day during the study and as oral hygiene training.

Number of Research Subjects (persons)
31

gender(%)
man
19 (61.3)
Woman
12 (38.7)
Age (Mean ± SD)
20.24 ± 1.18

At this time, the clinical trial material was a toothbrush prototype using bristle with a bristle extract and propolis.

In each visit, the subjects received clinical evaluation in order of evaluation of oral harmful bacterium inhibition ability and bad breath improvement ability.

Here, the evaluation of oral harmful bacterium inhibition activity was measured three times after the baseline, 2 weeks and 4 weeks after the generation of irritating saliva chewing paraffin wax in the subject.

In addition, the evaluation of bad breath improvement was performed in three test groups after brushing with a control toothbrush and an antimicrobial toothbrush prototype, and collected a bad breath in the mouth while breathing only the nose without opening the mouth with a 1 ml dose of bad breath syringe. After 3 minutes, pump the syringe 2 ~ 3 times, collect 1mL, take it out of the mouth, align the 1mL scale accurately, insert it into the Sensor Gas Chromatograph ODSA-FIS Inc., and push the air containing bad breath. Measured by. At this time, the measurement time was based on 4 minutes, the results are shown in graphs and tables, the bad breath causing substances hydrogen sulfide (H ₂ S), methyl mercaptan (CH ₃ SH), methyl disulfide [(CH ₃ ) ₂ S ] Was compared before and after the reduction degree.

That is, to summarize the clinical trial progress, it is shown in Table 9 below.

Contents Primary Secondary 3rd term 0 days 2 weeks of use 4 weeks of use evaluation Base-line
Evaluation of oral harmful bacterium
Bad breath improvement evaluation
Evaluation of oral harmful bacterium
Bad breath improvement evaluation
Oral harmful bacterium inhibition activity evaluation
Bad breath improvement evaluation Participants 31 people 31 people 31 people

As a result of the evaluation, as shown in Figure 14-16, the clinical evaluation result of oral harmful bacteria showed more than 45% reduction compared to before using the toothbrush (day 0) and after 4 weeks. clinical evaluation results are hydrogen sulfide (H ₂ S) causative agent of the three, methyl mercaptan (CH ₃ SH), disulfide, methyl [(CH _{3) 2} S] hydrogen sulfide (H ₂ S) from, methyl mercaptan for ( CH ₃ SH) showed an excellent reduction effect (more than 50%).

[Figure 14]

Here, Figure 14 is a graph showing the inhibitory ability against oral harmful bacteria after 4 weeks of toothbrush use.

[Figure 15]

Here, Figure 15 is a graph showing the reduction effect of three types of bad breath causes after 4 weeks of toothbrush use.

[Figure 16]

Here, Figure 16 is a graph showing overall bad breath reduction after 4 weeks of toothbrush use.

As such, the antimicrobial bristle composition using the non-toxic oral health function according to the present invention, the bristle and its manufacturing method has been found to be beneficial to the oral health of the user.

Claims (15)

Rhubarb extract having a concentration of 1-5% dissolved in alcohol alcohol;
The antibacterial bristles composition using a natural product having a nontoxic oral health function, characterized in that it comprises a; polyester filament loaded with the rhubarb extract.
The method according to claim 1,
The composition is an antimicrobial bristle composition utilizing a natural product having a nontoxic oral health function, characterized in that it further comprises a rosemarinic acid (Rosmarinic Acid) having a concentration of 1-100mg / ㎖.
The method according to claim 1,
The composition is an antimicrobial bristle composition utilizing a natural product having a nontoxic oral health function, characterized in that it further comprises EGCG (Epigallocatechin gallate) having a concentration of 1-10mg / ㎖.
The method according to claim 1,
The composition is an antimicrobial bristle composition utilizing a natural product having a nontoxic oral health function, characterized in that it further comprises EC (Epicatechin) having a concentration of 1-10mg / ㎖.
The method according to claim 1,
The composition comprises a natural product having a non-toxic oral health function, characterized in that it further comprises Rosemarinic acid (Rosmarinic Acid) having a concentration of 1-100mg / ㎖, and EGCG (Epigallocatechin gallate) having a concentration of 1-10mg / ㎖ Antimicrobial bristle composition utilized.
The method according to claim 1,
The composition utilizes a natural product having a non-toxic oral health function, characterized in that it further comprises rosmarinic acid having a concentration of 1-100 mg / ml and EC (Epicatechin) having a concentration of 1-10 mg / ml. One antibacterial bristles composition.
The method according to claim 1,
The composition further comprises EGCG (Epigallocatechin gallate) having a concentration of 1-10 mg / ml, and EC (Epicatechin) having a concentration of 1-10 mg / ml. One antibacterial bristles composition.
The method according to claim 1,
The composition includes Rosemarinic Acid having a concentration of 1-100 mg / ml, Epigallocatechin gallate (EGCG) having a concentration of 1-10 mg / ml, and EC (Epicatechin having a concentration of 1-10 mg / ml). Antimicrobial bristle composition utilizing natural products having a nontoxic oral health function, characterized in that it further comprises).
The method according to claim 1,
The polyester filament in the composition, characterized in that the propolis having a concentration of 0.125-0.5% dissolved in alcohol alcohol is supported with a sulfur extract, antimicrobial bristle composition utilizing a natural product having a non-toxic oral health function.
The method according to claim 9,
The composition is an antimicrobial bristle composition utilizing a natural product having a nontoxic oral health function, characterized in that it further comprises a rosemarinic acid (Rosmarinic Acid) having a concentration of 1-100mg / ㎖.
The method according to claim 9,
The composition is an antimicrobial bristle composition utilizing a natural product having a nontoxic oral health function, characterized in that it further comprises EGCG (Epigallocatechin gallate) having a concentration of 1-10mg / ㎖.
The method according to claim 9,
The composition is an antimicrobial bristle composition utilizing a natural product having a nontoxic oral health function, characterized in that it further comprises EC (Epicatechin) having a concentration of 1-10mg / ㎖.
The method according to claim 9,
The composition includes Rosemarinic Acid having a concentration of 1-100 mg / ml, Epigallocatechin gallate (EGCG) having a concentration of 1-10 mg / ml, and EC (Epicatechin having a concentration of 1-10 mg / ml). Antimicrobial bristle composition utilizing natural products having a nontoxic oral health function, characterized in that it further comprises).
Preparing an antimicrobial bristle composition comprising a yellow filament extract having a concentration of 1-5% dissolved in alcohol alcohol, and a polyester filament on which the yellow strip extract is carried;
Making a master batch after adding a functional additive to the antimicrobial bristle composition, Rosemarinic Acid, Rosegalic Acid, or EGCG (Epigallocatechin gallate) or EC (Epicatechin) alone or in a mixture of two or more;
Step of manufacturing the bristles by melting, spinning, cooling the masterbatch to produce a bristles, characterized in that consisting of antibacterial bristle composition using a natural product having a non-toxic oral health function.
Brush head manufactured using the bristle composition of any one of Claims 1-13.

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