COMPOSITION FOR PROMOTING PERIODONTAL REGENERATION

Abstract

The present invention relates to a composition for promoting periodontal regeneration. The composition for promoting periodontal regeneration according to the present invention includes any one dicotyledon extract belonging to the order Campanula punctata, selected from the group consisting of Ussuri thistle extract, Helianthus annuus extract, Dandelion extract, Chrysanthemum extract, Arctium lappa extract, Common cosmos extract, Ligularia stenocephala extract, Foremost mugwort extract, Cichorium intybus extract, and mixture thereof.

Claims

1. A composition for promoting periodontal regeneration, the composition comprising any one dicotyledon extract belonging to the order Campanula punctata, selected from the group consisting of Ussuri thistle extract, Helianthus annuus extract, Dandelion extract, Chrysanthemum extract, Arctium lappa extract, Common cosmos extract, Ligularia stenocephala extract, Foremost mugwort extract, Cichorium intybus extract, and mixture thereof.

2. A composition for promoting periodontal regeneration, the composition comprising Ligularia stenocephala extract and any one dicotyledon extract belonging to the order Campanula punctata, selected from the group consisting of Ussuri thistle extract, Helianthus annuus extract, Dandelion extract, Chrysanthemum extract, Arctium lappa extract, Common cosmos extract, Foremost mugwort extract, Cichorium intybus extract, and mixture thereof.

3. The composition of claim 1, wherein the dicotyledon extract belonging to the order Campanula punctata is extracted with any one 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.

4. The composition of claim 2, wherein the dicotyledon extract belonging to the order Campanula punctata is extracted with any one 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.

5. A food composition including the composition for promoting periodontal regeneration according to claim 1.

6. A pharmaceutical composition including the composition for promoting periodontal regeneration according to claim 1.

Description

DESCRIPTION OF DRAWINGS

[0053] FIG. 1 illustrates the anti-inflammatory effect of a composition for promoting periodontal regeneration according to an embodiment of the present invention.

[0054] FIG. 2 illustrates a test result of stem cell proliferation effect of a composition for promoting periodontal regeneration according to an embodiment of the present invention.

[0055] FIG. 3 illustrates mRNA expression effect of a composition for promoting periodontal regeneration according to an embodiment of the present invention.

[0056] FIG. 4 illustrates mRNA expression effect of the composition for promoting periodontal regeneration according to an embodiment of the present invention.

[0057] FIG. 5 illustrates mRNA expression effect of a composition for promoting periodontal regeneration according to an embodiment of the present invention.

[0058] FIG. 6 illustrates mRNA expression effect of a composition for promoting periodontal regeneration according to an embodiment of the present invention.

[0059] FIG. 7 illustrates a test for promotive effect on the damage-recovery ability of periodontal stem cells according to an embodiment of the present invention.

BEST MODE

[0060] The extract obtained by repeatedly extracting 2 to 8 times at an extraction temperature of 100 C. for 2 to 8 hours was filtered with a filter cloth and concentrated in vacuum. Then the extract and dextrin were mixed and spray-dried to prepare Ussuri thistle extract.

[0061] Meanwhile, Ussuri thistle extract, Helianthus annuus extract, Dandelion extract, Chrysanthemum extract, Arctium lappa extract, Common cosmos extract, Ligularia stenocephala extract, Foremost mugwort extract, and Cichorium intybus extract were spray-dried under the same conditions and methods as those of Ussuri thistle extract. For other test conditions, the mixture of water and ethanol was used as a solvent.

Experimental Example 1: Test for Anti-Inflammatory Effect

[0062] For purpose of confirming the anti-inflammatory effect of each extract in addition to the activity for periodontium reconstruction, in order to find effect on change of Interleukin-1 (IL-1) and tumor necrosis factor (TNF-) levels associated with the inflammation of gingivitis and the destruction of connective tissue, human-derived THP-1 was cultured in a culture medium supplemented with 10% FBS in RPMI 1640, and then the cell strains were co-treated with lipopolysaccharide (LPS), an inflammation-inducing substance, and each extract according to the present invention. Then, the secreted amount of TNF- in the culture solution was measured using enzyme-linked immunosorbent assay (ELISA).

[0063] Referring to FIG. 1, as a result thereof, it was confirmed that each extract had a specific anti-inflammatory effect. However, as a result of conducting experiments at a concentration range of 50 to 150 mg/ml, it was found that Helianthus annuus extract, Dandelion extract, Chrysanthemum extract, Arctium lappa extract, Common cosmos extract, Foremost mugwort extract, and Cichorium intybus extract did not exhibit an anti-inflammatory effect of more than 90%. On the other hand, in the case of Ussuri thistle extract and Ligularia stenocephala extract, it was confirmed that the anti-inflammatory effect was increased depending on the concentration range of each extract. In particular, in the case of the group in which 50 mg/ml of the Ligularia stenocephala extract was also put exhibited anti-inflammatory effect with more than 90%. Thus, it was confirmed that the Ligularia stenocephala extract had the best anti-inflammatory effect in each concentration range.

[0064] Therefore, it can be found that the use of the Ligularia stenocephala extract reduces the amount of TNF- secreted, and thus the Ligularia stenocephala extract exhibits a remarkably excellent anti-inflammatory effect in the periodontal disease.

Modes of the Invention

Experimental Example 2: Test for Proliferation Effect of Periodontal Ligament Stem Cells by Ligularia stenocephala Extract

[0065] Stem cells derived from periodontium are adult stem cells, which generally act as maintaining the tissue in vivo and regenerating the damage. Therefore, in order to confirm the effect of Ligularia stenocephala extract on the periodontal ligament stem cells in periodontal reconstruction using the Ligularia stenocephala extract having the best anti-inflammatory effect, the test was conducted for the effect of the Ligularia stenocephala extract on the promotion of proliferation of periodontal ligament stem cells.

[0066] In order to confirm the effect on the reconstruction of oral cavity tissue by healthy self-division of periodontal ligament stem cells, human-derived periodontal stem cells were cultured with a culture medium in which Dulbecco's modified eagle medium (DMEM) was supplemented with 10% heat-inactivated fetal bovine serum. Then, the first cultured stem cells were treated with or without the Ligularia stenocephala extract of the present invention, and then the cell proliferation was confirmed by MTT assay on the periodontal cells. The results are illustrated in FIG. 2.

[0067] Referring to FIG. 2 as described below, it can be seen that the proliferation of periodontal ligament stem cells was improved depending on the concentration thereof when the treatment with the Ligularia stenocephala extract according to the present invention. Therefore, it can be seen that the Ligularia stenocephala extract improves proliferation of periodontal ligament stem cells, thereby promoting periodontal regeneration.

Experimental Example 3: Measurement of Expression of MYC, KLF4, NOTCH1, and PCNA Gene in the Ligularia stenocephala Extract

[0068] MYC, KLF4, NOTCH1, and PCNA genes have very close effects on the proliferation of periodontal ligament stem cells. Therefore, in order to confirm a change in the expression of the above-described genes by the Ligularia stenocephala extract according to the present invention, the expression of the gene mRNA was confirmed and then the results thereof are illustrated in FIGS. 3 to 6 as described below.

[0069] Referring to FIGS. 3 to 6 as described below, it can be confirmed that the expression of the above-described genes is promoted in the case of using the Ligularia stenocephala extract. In particular, referring to FIGS. 3 and 4, the expression of KLF4 and NOTCH1, which are biomarkers of periodontal ligament stem cells, is 2.6 times and 2.1 times, respectively, higher than that of the control group when treated with a concentration of 200 g/ml of Ligularia stenocephala extract. This is the result showing the proliferation of the stem cells when the state of the stem cell is maintained.

[0070] Further, referring to FIGS. 5 and 6, the expression of MYC and PCNA, genes involved in the proliferation of periodontal ligament stem cells was increased in 4.2 times and 3.2 times higher than that of the control group, respectively, when treated with a concentration of 200 g/ml of the Ligularia stenocephala extract. As a result, it is shown that Ligularia stenocephala may enhance the proliferation of stem cells of periodontal ligament through MYC and PCNA genes, which are generally increased when stem cells proliferate.

Experimental Example 4: Test for Effect of Promoting Damage Recovery Ability of Periodontal Stem Cells by Ligularia stenocephala Extract

[0071] In order to confirm whether the Ligularia stenocephala extract according to the present invention can provide a regenerating effect of the real tissue, the scratch damage analysis model was used to conduct a test for the transference of stem cells and ability to transfer to damaged tissue for the Ligularia stenocephala extract. The results are illustrated in FIG. 7 as described below.

[0072] Referring FIG. 7 as described below, the ability to transfer of the periodontal ligament stem cells treated with the Ligularia stenocephala extract having a concentration of 200 g/ml and the periodontal ligament stem cells as a control group was analyzed. As a result, it was confirmed that the periodontal ligament stem cells treated with the Ligularia stenocephala extract were more quickly transferred to the damaged area, thereby reducing the scratch area.

[0073] Further, in the case of the periodontal ligament stem cells treated with the Ligularia stenocephala extract, the damage recovery ability was such that the damaged area was recovered by about 27.34% for 12 hours, while the control group had the damage recovery ability of about 15.33% for 12 hours. Further, it was confirmed that the periodontal ligament stem cells treated with the Ligularia stenocephala extract showed a recovery of 75.32% after 24 hours, whereas the cells of the control group were recovered only 52.12% after 24 hours. In other words, this result shows that the Ligularia stenocephala extract can increase the regeneration effect of damaged tissue through the enhancement of the ability to transfer of stem cells.

[0074] While the preferred embodiments of present invention have been described in detail as described above, the present invention is not limited to the embodiments, and various modifications and improvements using the basic concept of the present invention, defined in the following claims by those skilled in the art are also within the scope of the present invention.