KR101742482B1 - Composition for preventing or alleviating periodontal disease - Google Patents

Abstract

The present invention relates to a composition for the prevention and improvement of periodontal disease. The present invention relates to a composition for prevention and improvement of periodontal disease, which comprises at least one composition selected from the group consisting of Thistle extract, Glycyrrhiza uralensis extract, Sepharose extract, Seokgukjuk extract, An extract of perennial plant belonging to any one of Asteraceae belonging to the family of Asteraceae and a mixture thereof, and has an effect of preventing and improving periodontal disease including gingivitis and periodontitis.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing or ameliorating periodontal disease,

The present invention relates to a composition for preventing or improving periodontal disease. More particularly, the present invention relates to a method for preventing or eradicating periodontal disease by removing the inflammation caused by periodontal disease by using an extract derived from natural materials, promoting osteoblast proliferation and reducing osteoclast production and activity, will be.

It has been reported that periodontal disease is directly or indirectly correlated with diseases such as gastrointestinal diseases, lipid metabolic diseases, and stroke, and the cause of periodontal disease can not be explained by a single cause. It is usually a chronic disease caused by multiple causes can see.

Streptococcus mutans, one of the many microorganisms in the oral cavity, secretes an enzyme called Glucosyl transferase (GTase) on the surface of cells or cells. This enzyme forms plaque, which is a bacterial coat on the surface of the teeth, and calcium ions (Ca2 +) in teeth are lost by the accumulation of high concentrations of lactic acid produced from lactic acid bacteria that grow in the plaque. This causes tooth decay along with the dissolution of tooth enamel enamel and toxic substances induced by prague penetrate into the crevices between teeth and gums to cause inflammation in the cells, resulting in periodontal diseases such as gingivitis and periodontal disease.

Bacterial antigens, represented by endotoxin or lipopolysaccharide (LPS), from the plaque enter the gingival tissue, and the neutrophils and macrophages induced by this are activated by the digestion of bacterial antigens from the plaque, It releases enzymes such as collagenase, gelatinase, elastase, and Cathepsin G to the outside of the cell. These enzymes work together to decompose the organic components of gingival tissue, such as collagen and proteoglycans. Neutrophils or macrophages release inflammatory cytokines and stimulate the production of matrix metalloproteinase by stimulating the surrounding gingival fibroblasts. However, the production of tissue inhibitor of metalloprotease (TIMF), an inhibitor of these enzymes, The substrate (collagen) is degraded because it does not change or is suppressed. Therefore, softening and breakdown of the gingival tissue and continuous bleeding occur.

As an important supplier of enzymes involved in collagen degradation, periodontal disease-causing bacteria such as porphylomonas gingivalis, which is distributed in prags, can be mentioned. These bacteria secrete proteolytic enzymes similar to collagen hydrolase and trypsin, and these enzymes break down the connective tissues such as collagen and fibronectin.

Bacterial enzymes also exhibit MMP activity by activating inactive precursors or degrading TIMF. Proteoglycans in periodontal tissues can be degraded by various bacterial enzymes and macromolecules such as collagen, proteoglycan and hyarulonic acid are destroyed by normal cellular metabolites such as free radicals.

Thus, periodontal disease is caused by the direct or indirect action of bacteria in the oral cavity, by the free radicals, which are normal metabolites, and by the signaling substances delivered from other body tissues. Therefore, it is possible to prevent and improve periodontal disease by suitably using components or formulations having functions such as antibacterial action, anti-inflammatory action, anti-plaque formation action, and antioxidative action.

Currently, antimicrobial agents are used to supplement these oral diseases. Generally, in the case of dental caries, chlorhexidine 0.2% toothpaste is used for the management of dental plaque, but it is used only for short-term therapy because of side effects and unpleasant taste of tooth stain. In periodontal disease, firstly dental decompression is performed, but in the case of antimicrobial treatment with adjuvant therapy, more diverse microorganisms are involved than in caries. Therefore, in addition to chlorhexidine,

Tetracycline, metronidazole, and the like. However, these antimicrobial agents may cause side effects such as generation of resistant bacteria and non-selective sterilization, and it is necessary to find an effective substance that can replace the existing antimicrobial agent.

Natural remedies have been attracting attention as a substitute product that has less side effects and improved therapeutic effect. Therapeutic agents using these natural products can overcome tolerance and safety problems caused by conventional therapeutic agents. Therefore, in order to prevent or ameliorate periodontal disease, it is required that a composition derived from a wide variety of natural materials be discovered and developed.

KR 10-2002-0082308 A KR 10-2000-0047165 A

The object of the present invention is to provide a composition for preventing and alleviating periodontal disease, which inhibits inflammation caused by postural disease, promotes osteoblast proliferation, inhibits osteoclast formation and activity, and prevents alveolar bone from being destroyed .

It is another object of the present invention to provide a composition for preventing or improving periodontal disease, which can increase the activity of periodontal ligament cells to enhance the activity of the periodontal ligament, thereby enabling effective recovery of periodontal ligament or prevention and improvement of related diseases.

In order to achieve the above object, according to one embodiment of the present invention, there is provided a composition for preventing or improving periodontal disease, which comprises at least one selected from the group consisting of thistle extract, ginkgo biloba extract, Prevention and improvement of periodontal disease, including gingivitis and periodontitis, including perennial plant extracts belonging to any one selected from the group consisting of extracts, dandelion extracts, dandelion extracts, extracts and mixtures thereof Lt; / RTI >

In addition, the composition for preventing and improving periodontal disease according to the present invention is useful for preventing and improving periodontal disease, which comprises extracts of Ganoderma lucidum, , A mixture of plant extracts of perennial plants and thistle extracts belonging to one kind of asteraceae selected from the group consisting of gingivitis and periodontitis, and prevention and improvement of periodontal disease.

The composition for preventing and improving periodontal disease according to the present invention may further comprise at least one selected from the group consisting of Thistle extract, Ginkgo biloba extract, Quercus acutissima extract, Ginkgo biloba extract, Ginkgo biloba extract, Tochomari extract, Extracts, and mixtures thereof. The plant extracts of the perennial plant belonging to the family Asteraceae may be effective for preventing and improving periodontal ligament diseases by periodontal ligament cell activation and proliferation.

Wherein the perennial plant extract belonging to the Asteraceae family is selected from the group consisting of water, an organic solvent having 1 to 10 carbon atoms, a subcritical fluid, a supercritical fluid, and a mixture thereof, It may be an extract.

The food composition according to another embodiment of the present invention may include the composition for preventing and improving periodontal disease.

The pharmaceutical composition according to another embodiment of the present invention may include the composition for preventing or improving periodontal disease.

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

According to one embodiment of the present invention, there is provided a composition for preventing and improving periodontal disease, which comprises at least one selected from the group consisting of Thistle extract, Ginkgo biloba extract, Quercus acutissima extract, Ginkgo biloba extract, Ginkgo biloba extract, Extract of perennial herb belonging to any one of the group consisting of extracts, extracts of leaves, and mixtures thereof, and may be one having the effect of preventing and improving periodontal disease including gingivitis and periodontitis.

Throughout this specification, when an element is referred to as " including " an element, it is understood that the element may include other elements as well, without departing from the other elements unless specifically stated otherwise.

The composition for prevention and improvement of periodontal disease according to one embodiment of the present invention may contain thistle extract and may have the effect of preventing and improving periodontal disease including gingivitis and periodontitis.

The periodontal diseases referred to in the present invention are defined as inflammation including gingivitis limited to the soft tissues and inflammation including periodontitis that progresses to the vicinity of the gums and gums, including periodontal ligament diseases.

The periodontal ligament disease referred to in the present invention is defined as an inflammatory disease accompanied by bone loss and periodontal ligament inflammation, which is formed deeply into the periodontal pouch when the periodontal disease deepens. In addition, restoration of periodontal ligament is defined as meaning that the periodontal ligament disease is improved or the inflammatory disease developed in the periodontal ligament is improved, as well as activation of the recipient cell proliferation.

In general, periodontal disease is an infectious disease caused by periodontal pathogens in the plaque, and it is common that it involves tissue destruction by chronic inflammation. Therefore, periodontal disease can be improved through elimination of germs which are the cause of periodontal disease, regeneration of periodontal tissue through regeneration of periodontal ligament cells and bone tissue, and recovery of its function.

Therefore, there is a need to promote anti-inflammation, osteoblast activity and proliferation of periodontal ligament associated with periodontal disease, and inhibit osteoclast activity.

In addition, in the periodontal ligament disease, the periodontal ligament itself can be restored to prevent the generation of periodontal ligament inflammation and bone loss. Therefore, through the activity and proliferation effect of periodontal ligament cells, the recovery of periodontal ligament can enjoy the effective restoration of periodontal ligament disease.

The thistle (Cirsium japonicum) is a perennial plant of Cercopithecus chinensis and is also called a spiny herb. It stands on a mountain or field, its stem stands straight, 50 to 100 cm high, and has spiderlike hairs with white hairs on the whole. The root leaves of the thistles remain to the petals and larger than the stem leaves. The stem leaves are bulbous, elliptical, and split like feathers. The underside surrounds the base of the stem. The cracked edges are split again, and there are thorns with sawtooths that are deeply deflected. Thistle is distributed in Korea, Japan, northeastern part of China, and Wisdom. It is narrow and green with a little bit of thorn. It is a narrow leaf thistle. It has a lot of thorns. Thorny thorns are white thorns. White thorns are white flowers. It is also classified.

When the thistle extract is used, it inhibits the proliferation of inflammatory cells of periodontal silver and promotes the activity and proliferation of osteoblasts as well as anti-inflammatory effect against periodontal disease, thereby improving the alveolar bone, thereby preventing and improving periodontal disease . In addition, the thistle extract promotes the activity and proliferation of periodontal ligament cells, thereby improving the periodontal ligament diseases.

In addition to the above thistle extract, Asteraceae (Inula britannica var. Chinensis), Chrysanthemum zawadskii var. Alpinum, Coreopsis drummondii L., Bidens bipinnata L., (Xanthium strumarium), Helianthus tuberosus L, Petasites japonicus, Ligularia stenocephala, Rhaponticum uniflorum, Dandelion and Atractylodes japonica.

Preferably, the perennial plant extracts belonging to the composition (Compositae / Asteraceae, Chrysanthemum) may be the extracts of Mandragon. The use of the above-mentioned pudding extract may be superior to the thistle extract as well as other dairy plants in preventing and improving periodontal disease.

Therefore, more preferably, the perennial plant extract belonging to the above-mentioned Asteraceae may be used in combination with Thistle extract and Mandarin extract. In this case, as compared with the case of using the extract of Thistle and the extract of Gandallii alone, the synergistic effect over simple binding can bring about remarkably excellent effect for prevention and improvement of periodontal disease.

Ligularia stenocephala is a perennial herbaceous plant of the dicotyledonous spiny lily of the valley. It grows in the wetlands of deep mountains and is 60-100 cm high. There are no hairs on the whole grass of the grass, the roots are thick, and the leaves on the roots remain until the flowers are bloomed, about 24cm long and 20cm wide. It grows along the back vein of the leaf and has pointed serrations on the edge. The petiole is about 40cm long with no wings and broad bottom. The flower is yellow, hanging from the stem at the end of the stem in Aug.-Sep. The fruit is achene, ripe in October, and is inverted. Its color is brownish white and 6 to 7 mm long. Gandalf is distributed in Korea, Japan, Taiwan, China, etc. It uses young leaves for food and roots for gynecological diseases.

The extracts of Ganoderma lucidum extract have much better effect on periodontal disease than those of Thistle extract, Ganoderma lucidum extract, Quercus acutissima extract, Ginkgo biloba extract, Ganoderma lucidum extract, Tocophera extract,

In addition, when the extract of Ganoderma lucidum is used in combination with thistle extract, it has an advantage of enhancing the anti-inflammatory effect of dental caries, increasing the activity and proliferation of osteoblasts, and increasing the effect of decreasing the activity of osteoclasts. In particular, when the thistle extract and the Ganoderma lucidum extract are mixed, the effect of improving periodontal disease is greatly increased. Therefore, compared with the case of using the Ganoderma extract or the Ganoderma extract alone, There is no problem of cytotoxicity and it can be used for periodontal disease. Therefore, the use of mixed extracts can have a high effect on periodontal disease without side effects.

In addition, when the thistle extract and the Ganoderma extract are used in combination, the activity and proliferation of the periodontal ligament cells are promoted, and the prevention and improvement of periodontal ligament diseases is more effective than when the thistle extract or the Ganoderma extract is used alone Can be increased.

The extract can be any one selected from the group consisting of water, an organic solvent having 1 to 10 carbon atoms, a subcritical fluid, a supercritical fluid, and a mixture thereof.

Preferably, the Phellinus lint extract can be extracted using a solvent in which water and ethanol are mixed. When extracting the extract of Ganoderma lucidum using the above-mentioned solvent, it is possible to effectively obtain an effective ingredient for improving the periodontal disease contained in the ganoderma, and it can be mixed with the thistle extract to give a synergistic effect to the improvement of the scum disease.

Preferably, the composition for preventing and / or improving periodontal disease comprises an extract of Mandarin, and may have osteoblast activity and proliferative effect.

More preferably, the composition for preventing and / or improving periodontal disease comprises the extracts of Gandalf and Thistle, and may have osteoblast activity and proliferative effect.

Osteoblast cells generally have a very important effect on oral health. In periodontal disease, it is difficult to achieve a fundamental improvement effect by simple anti-inflammatory action. It promotes osteoblast activity and proliferation, inhibits osteoclast activity, To inhibit the formation of additional inflammation and osteoporosis to effectively remove periodontal inflammation with anti-inflammatory effects.

In case of using the above-mentioned extract, the activity and proliferation effect of osteoblast can prevent and improve periodontal disease.

The use of the above-mentioned Ganoderma lucidum extract may be superior to the perennial plant extracts belonging to other Asteraceae extracts including thistle extract. In particular, when the extract of Ganoderma lucidum is mixed with the thistle extract, it is possible to greatly enhance the activity and proliferation effect of osteoblast compared to the case of using the Ganoderma extract alone or of the extract of Ganoderma lucidum alone, Can be inhibited and the effect of preventing and improving periodontal disease can be enhanced.

Preferably, the composition for preventing and / or improving periodontal disease includes an extract of Ganoderma lucidum, and may be effective for the prevention and improvement of periodontal ligament diseases by periodontal ligament cell activation and proliferation.

More preferably, the composition for prevention and improvement of periodontal disease includes the extracts of Ganoderma lucidum and Thistle, and may be effective for preventing and improving periodontal ligament diseases by periodontal ligament cell activation and proliferation.

The use of the above-mentioned Ganoderma extract has an advantage of preventing and improving the periodontal ligament diseases as compared with the perennial plant extracts belonging to other Asteraceae including thistle extract.

Particularly, in the case where the above-mentioned Ganoderma extract is mixed with Thistle's extract, the use of the Ganoderma extract alone or the combination of the extracts, The effect can be greatly enhanced.

More preferably, the extracts of Gandalf and Thistle may be mixed in a weight ratio of 1:10 to 10:10, more preferably in a weight ratio of 4: 6 to 7: 3.

By the above range, the anti-inflammatory activity and the osteoblast activity of the periodontal ligament are greatly enhanced, and the periodontal ligament cell activity is increased and the osteoclast activity is suppressed remarkably. Furthermore, according to the above-mentioned range, while the fragrance of tangerine is kept silent, a strong flavor unique to thistle can be removed and thus a food composition having excellent texture and flavor can be provided.

If the amount is outside the above range, the synergistic effect of prevention and improvement of periodontal disease due to the use of the mixture of Ganoderma extract and Thistle oil extract is lowered. In addition, the thickened amount of thistle is left intensely or the flavor of ganoderma leaves is lowered. There is a problem to lose.

The thistle extract may be an extract of a solvent selected from the group consisting of water, an organic solvent having 1 to 10 carbon atoms, a subcritical fluid, a supercritical fluid, and a mixture thereof.

Preferably, the thistle extract may be extracted using a solvent in which water and ethanol are mixed. When the thistle extract is extracted using the above-mentioned solvent, an effective ingredient for effecting the improvement of periodontal disease contained in thistle can be effectively obtained and a synergistic effect can be obtained by mixing with the gutral extract.

Preferably, the thistle extract or the ganganous extract may be a mixture of water and ethanol in a weight ratio of 1: 5 to 5: 1. By the above-mentioned range, the effective ingredient can be obtained more effectively, and the flavor of the thistle and the garland can be maintained, and the taste of the food composition can be improved.

Preferably, the solvent may be extracted using 100 to 2500 parts by weight of the thistle, and the solvent may be extracted using 100 to 2500 parts by weight of the seedling 100 parts by weight.

More preferably, the solvent may be extracted from 700 to 1000 parts by weight of the thistle to 100 parts by weight of the thistle, and may be extracted using 700 to 1000 parts by weight of the seedling 100 parts by weight. The effective amount of the effective ingredient effective for improving periodontal disease can be effectively obtained while maintaining a constant concentration.

The thistle extract may be extracted at 15 to 120 ° C, preferably at about 50 to 100 ° C for about 1 to 10 times, preferably about 2 to 8 times, by hot water extraction or reflux cooling extraction. In addition, the extract can be one extracted at 15 to 120 ° C, preferably at about 50 to 100 ° C for about 1 to 10 times, preferably 2 to 8 times for extraction by hot water extraction or reflux cooling extraction . According to the extraction conditions, there is an advantage that the effective ingredient effective for the periodontal disease improvement can be effectively obtained while maintaining a constant concentration without being destroyed.

The thistle extract may be spray dried or lyophilized after extraction, and may be provided as a uniform formulation that can be used by those skilled in the art including a method of mixing dextrin. The extract may be spray-dried or lyophilized after extraction, and may be provided as a uniform formulation that can be used by those skilled in the art including a method of mixing dextrin.

The food composition according to another embodiment of the present invention may include the composition for preventing and improving periodontal disease.

The pharmaceutical composition according to another embodiment of the present invention may include the composition for preventing or improving periodontal disease.

The composition for the prevention and improvement of periodontal disease according to the present invention can inhibit inflammation caused by a holding disease. In addition, the composition for preventing and improving periodontal disease can promote the proliferation of osteoblasts and inhibit the production and activity of osteoclasts, thereby preventing the alveolar bone from being destroyed.

In addition, the composition for preventing and improving periodontal disease according to the present invention can increase the activity of proliferation of periodontal ligament cells, thereby enabling effective recovery of periodontal ligament or prevention and improvement of related diseases.

The present invention provides a method for preparing the composition for preventing and improving periodontal disease.

FIG. 1 shows the anti-inflammatory effect of an extract according to an embodiment of the present invention.
Figure 2 relates to the anti-inflammatory effect of the extract according to one embodiment of the present invention.
FIG. 3 is a graph showing gingival cell proliferation experiment results of an extract according to an embodiment of the present invention.
FIG. 4 is a graph showing the results of an experiment for promoting osteoblast proliferation of an extract according to an embodiment of the present invention.
FIG. 5 is a graph showing the effect of controlling the expression of MMPs in the periodontal bone of an extract according to an embodiment of the present invention.
FIG. 6 is a graph showing an alveolar bone health index test result of an extract according to an embodiment of the present invention. FIG.
FIG. 7 is a graph showing the results of periodontal ligament cell proliferation experiment of an extract according to an embodiment of the present invention.
FIG. 8 is a graph showing the results of gingival fibroblast proliferation experiment of an extract according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

[ Manufacturing example : Preparation of extract]

The pulverized thistle and the solvent are mixed at a weight ratio of 1: 7 to 1:10 and extracted repeatedly for 2 to 8 hours at an extraction temperature of 50 to 100 캜 for 2 to 8 hours, The resulting extract was filtered with a filter paper, concentrated in vacuo, and then mixed with an extract and dextrin and spray dried to prepare a thistle extract.

On the other hand, in the case of the Ganoderma extract, spray drying was carried out in the same manner as above. Other test conditions and solvents were water and ethanol mixtures.

The mixture of thistle extract and sandalwood extract used in the test was a mixture of the above thistle extract and the above-mentioned sandalwood extract in a ratio of 6: 4 to 7: 3.

[ Test Example  1: Anti-inflammatory effect test]

Changes in interleukin-1 (IL-1) and tumor necrosis factor (TNF-α) levels associated with loss of bone through reduction of bone health, which is the basic skeleton of periodontal disease, and destruction of inflammatory and connective tissue of periodontium, the basic tissue of teeth , Human THP-1 was cultured in RPMI 1640 medium supplemented with 10% FBS, and then the cell line was treated with Lipopolysaccharide (LPS), an inflammation inducing substance, , And the amount of TNF-α and IL-1 secreted in the culture solution was measured by enzyme immunoassay (ELISA).

The results are shown in Figs. 1 and 2. Fig. 1 and 2, it can be confirmed that the extract of Ganoderma lucidum is the most effective as a sole extract, and when treated with 50 μg / ml of the extract of Ganoderma lucidum and Thistle of Ganoderma lucidum, TNF-α (FIG. 1) (Fig. 2). Therefore, it was confirmed that the secretion amount of TNF-α and IL-1 is dramatically reduced in the combination extract of Thistle extract and Ganoderma extract. Therefore, it was confirmed that when the complex extract was used, remarkably high anti-inflammatory effect was obtained by significantly reducing inflammation-related factors.

[ Experimental Example  2: Gingivitis cell proliferation experiment]

In order to investigate the effect of the thistle extract, the Ganoderma lucidum extract and the complex extract thereof on the growth of gingival germ cells and the increase in cell activity according to the present invention, the following experiment was conducted.

For the evaluation of cell proliferation, MTT assay was performed to investigate the inhibitory effect of Ganoderma lucidum and Ganoderma complex extracts on gingival cell proliferation. The extracts prepared in Example 1 were prepared at various concentrations and mixing ratios.

YD-38 cell, a gingival cell, was purchased from Korean Cell Line Bank as a human oral squamous cell carcinoma cell line. For cell culture, 10% FBS was added to RPMI and antibiotics were added to the cells. These cell lines were cultured in 5% CO 2 at 37 ° C, and subcultured for 2-3 days.

In order to examine the inhibition of cell growth, thistle extracts, extracts of Ganoderma lucidum, and extracts of Ganoderma lucidum and their extracts were incubated for 24 hours in each experimental group and control group. After incubation for 4 hours at 0 mg / ml with 5 mg / ml MTT reagent, , And DMSO was added to the cell for fusion, and the formazan produced by MTT reduction in the intracellular mitochondria was measured for absorbance at 570 nm using an ELISA microplate reader. The results are shown in FIG.

Referring to FIG. 3, the average absorbance value of each sample group is determined, and it can be confirmed that the gingival saline cells are inhibited from proliferation. In particular, the results showed that the growth of gingival gland cells was inhibited according to the concentration, and that the extract of Ganoderma lucidum was the most effective inhibitor of gingival cell proliferation in the case of the single extract. In particular, it was shown that the treatment of the combination of Ganoderma lucidum and Ganoderma lucidum extract has the effect of inhibiting the proliferation of effective gingival gland cells.

[ Experimental Example  3: Experimental study on proliferation of osteoblast cells]

The following experiment was conducted to investigate the effect of thistle extract, ganoderma extract, and a combination of these extracts on the proliferation of osteoblast cells.

The osteoblast cells of this experiment were osteoblast cells obtained from ATCC and MC3T3-E1 cells, which are mouse cell lines, were used as culture medium. These cell lines were cultured at 37 ° C in 5% CO ₂ and cultured subculture every 2-3 days.

In order to confirm the osteoblast growth promoting effect of thistle extracts, bark extracts and their combination extracts, each experimental group and control group were cultured for 24 hours, MTT reagent of 5 mg / ml was added, and the cells were cultured for 4 hours. After removing the supernatant, After the cells were melted by adding DMSO, formazan produced by MTT reduction in intracellular mitochondria was measured for absorbance at 570 nm using an ELISA microplate reader. The average absorbance value of each sample group was determined and the promoting effect of the gingival germ cells was shown in FIG.

Referring to FIG. 4, it can be confirmed that when using the extract of Ganoderma lucidum as the sole extract, it is most effective for proliferation of osteoblasts. In addition, it was confirmed that the combined extract of Thistle extract and Gandallii extract was more effective for the proliferation of osteoblast than that of the extract of Glycyrrhizae and the extract of Gundalbia, which has the best effect among the single extracts.

It was confirmed that the extract of Ganoderma lucidum could exert an excellent effect on osteoblast formation as a single extract. In addition, in the case of using a combination extract of a mixture of Thistle extract and Ganoderma lucidum extract, the effect of osteoblast cell proliferation on osteoblast activity can be more enhanced by synergistic effect than the use of thistle extract and gutral exudate alone Respectively.

[ Experimental Example  4: From periodontal MMPs  Effect of regulating expression]

Periodontal disease is generally caused by the microbial secretion of toxins through the oral tissue cell hydrolysing enzyme secretion increases collagen-like tissue fibers break down and cause periodontal tissue collapse. Through these actions, osteoclast precursor cells differentiate into osteoclasts, and differentiated osteoclasts exert bone resorption on the surface of alveolar bone to deepen the breakdown of periodontal tissue.

Therefore, the effect of regulating the expression of MMPs (metrix metalloproteinases) in periodontal bone was examined in order to confirm the inhibitory effect of the expression of the proteolytic enzyme through the composition according to the present invention.

For this experiment, osteoblast cells were purchased from ATCC and MC3T3-E1 cells, which are mouse ovarian cell line, were used as culture medium. These cell lines were cultured at 37 ° C in 5% CO ₂ and cultured subculture every 2-3 days. MMP-3 (stromelysins) and MMP-8 (collagenase) expression by the treatment with LPS at a concentration of 100 μg / ml were measured by RT-PCR , Respectively. Each RT (Reverse Transcriptional) -PCR was analyzed using the primers shown in the following table, and the results are shown in FIG.

target Forward primer Reverse primer MMP-2 5'-GATAACCTGGATGCCGTCGTG-3 ' 5'-GGTGTGCAGCGATGAAGATGATA-3 ' MMP-3 5'-CTCATGCCTATGCACCTGGAC-3 ' 5'-TCATGAGCAGCAACCAGGAA-3 ' MMP-8 5'-CCAAGGAGTGTCCAAGCCCAT-3 ' 5'-CCTGCAGGAAAACTGCATCG-3 '

5, when treated with LPS and treated with a concentration of 100 μg / ml of Ganoderma lucidum extract, the expression level of MMP-2 was reduced by about 50% as compared with the case where only LPS alone was treated . In addition, the mRNA expression of MMP-2 was reduced by about 70% when compared with that of the positive control in the case of the combination extract of Ganoderma extract and Thistle extract.

As a result of comparing the expression level of MMP-3, it was confirmed that the increase of mRNA expression by LPS was reduced by about 55% in the two cells treated with the extract of Mandarbicide. In the cells treated with the extract of Mandarin and Thistle, LPS And that the amount of MMP-2 expression was significantly reduced by about 88% or more.

As a result of confirming the expression level of MMP-8, a kind of collagenase, it was confirmed that the increase of mRNA expression by LPS was reduced by about 34% in the two cell lines treated with the extract of Ganoderma lucidum. The inhibition of MMP-8 enzyme expression by LPS was reduced by about 72% in cells treated with the combination extract of Ganoderma extract and Thistle extract.

As a result, it was confirmed that the effect of the extract of Ganoderma lucidum extract was most effective in inhibiting the expression of MMP-8 enzyme. In particular, in the case of the combination extract of Ganoderma extract and Ganoderma extract, And it is confirmed that the inhibition of cell matrix enzymes can help to improve periodontal health and disease.

[ Experimental Example  5: Checking the alveolar bone health index]

In periodontal disease, gingivitis is mainly limited to soft tissue damage, so treatment and recovery is a form of disease. However, periodontal disease is not only the soft tissue, but also the gingival bone of the main surface is damaged. Treatment and recovery are very difficult, And treat them aggressively. Therefore, one of the most important factors in the health of these gum bones is to prevent bone loss and strengthen the bones.

In the present invention, OPG (osteoprotegrin), which is a protein that binds to RANKL and binds to RANKL in osteoblasts, and OPN (osteopontin), which is specifically expressed in the tissue mineralization process as a noncollagen protein in the alveolar bone, ), The combination of thistle extract, ganoderma extract, or thistle extract and ganoderma extract was found to improve gingival bone health.

For the experiment, MG-63 cells (ATCC) derived from human osteosarcoma were used. After culturing in DMEM containing 10% FBS, the cells were cultured in a 96-well plate at 2 × 10 ⁴ cells in an amount of 80% Respectively. 24 hours after the addition of the extracts to each well, OPG-ELISA kit and OPN-ELISA kit (Oscotec Inc.) were used to measure the amount of OPG and OPN secreted from the cells. Respectively.

6, it was confirmed that the OPG secretion amount was improved by about 2.1 times according to the treatment with Ganoderma lucidum extract under the above-mentioned treatment conditions. In the case of the combination extract of Ganoderma extract and Ganoderma extract, about 3.6 times of OPG And the amount of secretion was increased. The combination of Thistle extract and Ganoderma extract, combined with Thistle extract and Ganoderma lucidum extract, has an effect on gum bone health through increased secretion of OPG. In addition, The synergistic effects of the mixed use are shown.

 In addition, as shown in FIG. 6, when the extract of the present invention was treated with osteoblast, the amount of OPN secretion increased by about 1.87 times as compared with that of the control group, , And the combined extract of mixture of Thistle extract and Ganoderma extract showed a synergistic effect that the amount of OPN secretion increased by about 2.94 times.

When MG-63 cells, a cell line derived from human osteosarcoma, are treated with the extract of the present invention, they have an effect on gum bone health through an increase in the secretion amount of OPN, which is a non-collagen protein, This is a result of showing the possibility of being able to do.

[ Experimental Example  7: Gingival fibroblasts Periodontal ligament cells  Propagation experiment]

In order to investigate the effect of the combined extract of Thistle extract, Ganoderma extract and Thistle extract and Ganoderma extract on the proliferation and cellular activity of periodontal ligament cells and gingival fibroblasts according to the present invention, the following experiment was conducted.

The periodontal ligament of the molar teeth extracted for therapeutic purposes was placed on a culture plate using a curette and the periodontal ligament cells were cultured in DMEM supplemented with 10% FBS. Gingival fibroblasts were also detached from the molar teeth and cultured in the same manner. At this time, the cells were continuously observed under a microscope while being cultured, and it was confirmed that the cells were migrated from the tissue. Then, tissues were removed so that the cells could become a monolayer, and subculture was carried out to evaluate the efficacy of the cells according to 7 passages. Human periodontal ligament cells and gingival fibroblasts were cultured in 96-well plates at a concentration of 2 x 10 ⁴ cells per well and cultured for 24 hours. Each of the extracts of the present invention was further treated for 48 hours, and then the number of cells was measured using Alama blue blue). The results are shown in FIGS. 7 and 8, respectively.

Referring to FIG. 7, when the inventive composition was applied to the periodontal ligament cells alone, the cell proliferation efficiency of about 38% was confirmed, and about 85% of the cells treated with the combined extract of the thistle extract and the pituitary extract Cell proliferation effect. It is possible to confirm the effect of enhancing the proliferation rate of gingival ligament cells of the extract of Ganoderma lucidum. Especially. It was confirmed that the combined extract of Ganoderma lucidum extract and Ganoderma lucidum extract significantly enhanced the growth rate of the gingival ligament cells by the synergistic effect compared with the case of the single extract. Therefore, it was confirmed that the combination extract of the present invention can help periodontal health as compared with a single extract.

8, the efficacy of the inventive composition in the gingival fibroblast was evaluated. As a result, it was confirmed that about 43% of gingival fibroblasts proliferation was improved by treating the extract of Ganoderma lucidum, and that the combination of Thistle extract and Ganoderma extract When the extract was used, the cell proliferation efficiency of about 103% was confirmed.

Therefore, it can be confirmed that the use of the above extract improves the growth rate of gingival fibroblast. When using the extract of Ganoderma lucidum extract or Ganoderma extract and Thistle extract, the synergistic effect is higher than that of the single extract, Can be obtained.

[ Experimental Example  8: Flavor test of composite composition]

The extracts of Ganoderma lucidum and Thistle extracts were prepared according to the method for preparing the extracts of Ganoderma lucidum and Thistle, respectively, according to the methods of producing the extracts of Thistle, Thunbergii, .

In addition, 1: 1 weight ratio of the extracts of Gimpungcho extract, Seokjungguk extract, Gimjeongguk extract, Gumijeoki extract, Tadomari extract, Thunb extract, Wumiga extract, Ganoderma extract, Duckweed extract, Dandelion extract and Ganoderma lucidum extract were mixed in Thistle extract , And the flavor of each extract was evaluated by preparing it as a tasting drink. The flavor and texture of each of the mixed extracts was evaluated by an index of 1 to 10. The results are shown in Table 2 below. The higher the number, the better the effect)

extract Indices Thistle extract alone 5 Thistle extract + Quercus acutissima extract 6 Thistle Extract + Rock Quercus Extract 5 Thistle extract + ginseng extract 4 Thistle Extract + Gemini Extract 5 Thistle extract + Tocophera extract 6 Thistle extract + Thyme extract 3 Thistle extract + butterfly extract 3 Thistle extract + bengal extract 9 Thistle Extract + Extra Extract 3 Thistle extract + dandelion extract 8 Thistle extract + gum extract 4

Referring to Table 2, when the Gentian extract was mixed with the thistle extract, the peculiar aftertaste and scent of the thistle were removed, and the flavor specific to the Gentian scent was enhanced to give a superior flavor. In addition, when using the dandelion extract, the strong smell of dandelion warms the incense of the thistle, confirming that the whole dandelion fragrance remains.

Therefore, it can be confirmed that the use of Kandalbi among the Asteraceae plants does not cover the specific fragrance and the flavor of the thistle and the flavor of the Kandarbi can be harmonized to provide a high flavor.

In addition, the following flavors were evaluated while varying the ratio of thistle to ganoderma. The results are shown in Table 3 below.

Thistle extract Bengal extract Indices 10 0 5 8 2 6 6 4 8.5 5 5 9 4 6 8 3 7 8 One 9 4 0 10 4

(Unit: weight ratio)

Referring to Table 3, it can be seen that when the mixture of the thistle extract and the Ganoderma extract is mixed with the Ganoderma extract and the Thistle extract at 4: 6 to 7: 3, the flavor is excellent.

These results are consistent with the range in which the Ganoderma extract and Thistle extract exhibit the best synergistic effect for the improvement of periodontal disease. By the above-mentioned range, it is possible to provide a tea, beverage and other food composition having excellent flavor and high taste, as well as having the best periodontal disease improving effect by the interaction of the extracts.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, Of the right.

Claims (6)

Contains the Gandalf extract
Which has the effect of preventing and improving periodontal disease including gingivitis and periodontitis.
A composition for preventing and improving periodontal disease. Thistle extract and bark extract
Which has the effect of preventing and improving periodontal disease including gingivitis and periodontitis.
A composition for preventing and improving periodontal disease. Thistle extract and bark extract
The effect of periodontal ligament cell activation and proliferation on the prevention and improvement of periodontal ligament disease
A composition for preventing and improving periodontal disease. 4. The method according to any one of claims 1 to 3,
The composition of claim 1, wherein the extract is a solvent extracted from water, an organic solvent having 1 to 10 carbon atoms, a subcritical fluid, a supercritical fluid, or a mixture thereof. A food composition comprising the composition for the prevention and improvement of periodontal disease according to any one of claims 1 to 3. A pharmaceutical composition comprising a composition for the prevention and improvement of periodontal disease according to any one of claims 1 to 3.

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