Antioxidant oral care composition and gingival fibroblast protective agent

Abstract

To provide an antioxidant oral care composition and a gingival fibroblast-protecting agent in which an antioxidant action and a cell-protecting action are recognized by bringing the composition into contact with oral cells in a relatively short time in the oral cavity, and a specific mineral composition contributes to care and health maintenance in the oral cavity.SOLUTION: An anti-oxidative oral care composition or a gingival fibroblast-protecting agent comprises an amorphous mineral composition as an active ingredient, wherein the amorphous mineral composition is a concentrate of amorphous mineral components of brackish water and has a magnesium ion (Mg2 +) concentration of 0.8-12mg / mL, a chloride ion (Cl -) concentration of 2.5-30mg / mL, a sulfate ion (SO42-) concentration of 0.18-2.4mg / mL, a sodium ion (Na +) concentration of 0.002-0.4mg / mL and a potassium ion (K +) concentration of 0.017-0.24mg / mL.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an antioxidant oral care composition and a gingival fibroblast protectant. [Background technology]

[0002] Generally, oral care compositions for maintaining gum health include toothpastes, mouthwashes, lozenges, chewable tablets, chewing gums, floss coating agents, denture care products, and the like, which contain some active ingredient.

[0003] Known ingredients that are effective for the health of the gums and other gingiva to be contained in such oral care compositions include amino acids and mucoadhesive ingredients for inhibiting gingival recession due to aging, metal ions such as tin and zinc (Patent Document 1), and vitamins such as ascorbic acid phosphate esters (Patent Document 2).

[0004] Furthermore, oral hygiene compositions containing rock salt together with amino acids and used in toothpastes and the like are known, with the expectation that they will also have therapeutic effects such as preventing gingivitis and alveolar pyorrhea (Patent Document 3). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Publication No. 2024-28801 [Patent Document 2] Patent No. 5644591 [Patent Document 3] Japanese Patent Application Laid-Open No. 2007-254315 Summary of the Invention [Problem to be solved by the invention]

[0006] However, the antioxidant effect of the ascorbic acid phosphate ester described in Patent Document 2 requires that the decomposition of the ascorbic acid phosphate ester be sustained for a long period of time by phosphatase in the body, and it takes a long time, about 24 to 48 hours, for the effect to be fully achieved. Therefore, sufficient antioxidant effect cannot be expected in a short period of time from short-term oral care in daily treatments or from the short cell contact time.

[0007] Furthermore, the oral hygiene composition containing rock salt described in Patent Document 3 contains 0.1 to 35% by weight of rock salt, which contains sodium chloride, a crystalline mineral component, at a high concentration of approximately 80% (paragraph 0016 of the same document).

[0008] Such oral hygiene compositions have been evaluated for their ability to remove germs from the oral cavity, prevent tooth decay, stimulate the gums in monitor tests, provide a pleasant taste, have a refreshing feeling, provide a clean feeling, eliminate bad breath, and have a pleasant texture in the mouth, but their antioxidant properties have not been confirmed.

[0009] Furthermore, among the Sicilian rock salt, German rock salt, Mongolian rock salt, red rock salt, and sea salt described in Patent Document 3, Mongolian rock salt, which is considered to be particularly desirable because it has no bitterness, contains virtually no magnesium or contains extremely low amounts of magnesium (paragraph 0009 of the same document).

[0010] Incidentally, rock salt, which has a low magnesium content, has the advantage of being not bitter in flavor but has a slight sweetness (paragraph 0011 of the same document), but it has been necessary to supplement it with essential amino acids and medicinal ingredients as active ingredients for oral care.

[0011] Therefore, the object of this invention is to solve the problems of conventional oral care compositions described above, and to provide an antioxidant oral care composition and gingival fibroblast protective agent that have antioxidant and cell protective effects when brought into contact with oral cells in the oral cavity for a relatively short period of time, and whose specific mineral composition contributes to oral care and health maintenance. [Means for solving the problem]

[0012] In order to solve the above problems, the present invention uses an amorphous mineral composition, which is a concentrate of amorphous mineral components from salt lake water, as an active ingredient, and the magnesium ions (Mg 2+ ) The resulting composition was an antioxidant oral care composition or gingival fibroblast protective agent having a concentration of 0.8 to 12 mg / mL.

[0013] The antioxidant oral care composition or gingival fibroblast protective agent of the present invention, configured as described above, contains as an active ingredient an amorphous mineral composition in which amorphous mineral components are concentrated, rather than crystalline mineral components such as sodium chloride, and in particular the magnesium concentration is adjusted to fall within a predetermined concentration range.

[0014] That is, the antioxidant oral care composition or gingival fibroblast protective agent of the present invention contains magnesium (Mg) as an antioxidant amorphous mineral, and also contains magnesium ions (Mg 2+ ) concentration of 0.8 to 12 mg / mL is thought to sufficiently promote the radical scavenging reaction of the antioxidant substance by the redox active ions, resulting in excellent antioxidant properties.

[0015] Therefore, as is clear from the test results described below, the antioxidant oral care composition or gingival fibroblast protective agent of this invention is shown to have a strong antioxidant effect, with a specific ability to provide radical scavenging activity due to antioxidant capacity and cell protection for human gingival fibroblasts (hGFs).

[0016] In order to fully exert such antioxidant properties, the amorphous mineral composition as a concentrate of amorphous mineral components of specific salt lake water contains magnesium ions (Mg 2+ ) concentration is 0.8 to 12 mg / mL, and chloride ions (Cl - ) concentration 2.5-30mg / mL, sulfate ion (SO4 2- ) concentration 0.18 to 2.4 mg / mL, sodium ion (Na + ) concentration of 0.02 to 0.4 mg / mL and potassium ion (K +) The amorphous mineral composition preferably has a concentration of 0.017 to 0.24 mg / mL.

[0017] The above-mentioned amorphous mineral composition is more preferably an amorphous mineral composition having an osmotic pressure of 50 to 1500 mOSM / kg, since it does not impose stress on the cells in the mouth due to osmotic pressure and exerts the above-mentioned antioxidant effect more sufficiently and stably. The saltwater lake water used in the present invention is particularly preferably the saltwater lake water of the Great Salt Lake in the United States.

[0018] The saltwater lake water is evaporated under sunlight, preferably over a period of about a year, and the crystallized salt (crystalline mineral components) is removed to obtain an amorphous mineral composition, which is a concentrate of amorphous mineral components rich in magnesium.

[0019] The magnesium ions (Mg 2+ The concentration is adjusted by diluting with water or the like so that the concentration is 0.8 to 12 mg / mL to form an antioxidant oral care composition, which is used as an active ingredient in a gingival fibroblast protective agent or the like. [Effects of the Invention]

[0020] This invention uses an amorphous mineral composition, which is a concentrate of amorphous mineral components from salt lake water, as an active ingredient, and contains magnesium (Mg) as an antioxidant amorphous mineral, and also contains magnesium ions (Mg 2+ ) concentration of 0.8 to 12 mg / mL, antioxidant and cell protective effects can be observed by contacting oral cells in the oral cavity for a relatively short period of time, and this has the advantage of making it an antioxidant oral care composition or gingival fibroblast protective agent whose specific mineral composition contributes to oral care and health maintenance. [Brief explanation of the drawings]

[0021] [Figure 1] ESR spectrum used to measure hydroxyl radical scavenging activity in the example [Figure 2] Diagram showing the relationship between the dilution rate of the original solution and the elimination rate of DPPH [Figure 3] Graph showing the relationship between the concentration of the active ingredient in the examples and the survival rate of gingival fibroblasts [Figure 4] Graph showing the relationship between the concentration of the active ingredient in the examples and the survival rate of gingival fibroblasts after cell-damaging treatment DETAILED DESCRIPTION OF THE INVENTION

[0022] The antioxidant oral care composition or gingival fibroblast protective agent according to an embodiment of the present invention contains, as an active ingredient, an amorphous mineral composition that is a concentrate of amorphous mineral components from salt lake water, and the magnesium ions (Mg 2+ ) concentration is 0.8 to 12 mg / mL.

[0023] The saltwater lake water used in this invention contains magnesium ions (Mg 2+ Natural salt lake water with a relatively high salt concentration can be used without any particular restrictions on its origin.

[0024] The saltwater lake water is exposed to sunlight in the atmosphere, or artificially heated and dried at a heat level that does not thermally denature the mineral components, thereby evaporating the water and increasing the concentration, and separating as much of the crystallized salt (crystalline mineral components) as possible, thereby concentrating the amorphous mineral components, including magnesium. The separation rate (removal rate) of the crystalline mineral components in this concentrate is preferably 99% or more, more preferably 99.5% or more.

[0025] Such amorphous mineral compositions contain magnesium ions (Mg 2+ A concentrate of amorphous mineral components from saltwater lake water containing various amorphous minerals is diluted 10 to 100 times to a concentration of 0.8 to 12 mg / mL and used as the active ingredient in an antioxidant oral care composition.

[0026] A preferred example of the ion composition of the amorphous mineral composition that is a concentrate before dilution is magnesium ions (Mg 2+ ) concentration is 0.8 to 12 mg / mL, and chloride ions (Cl - ) concentration 2.5-30mg / mL, sulfate ion (SO4 2- ) concentration 0.18 to 2.4 mg / mL, sodium ion (Na + ) concentration of 0.02 to 0.4 mg / mL and potassium ion (K + ) concentration is 0.017 to 0.24 mg / mL.

[0027] Preferred examples of such amorphous mineral compositions are those using, as raw materials, lake water from the Great Salt Lake in Utah, USA, or saltwater from the Murray River Basin or Deborah Salt Lake in Australia.

[0028] The resulting amorphous mineral composition preferably has an osmotic pressure of 50 to 1500 mOSM / kg so as not to excessively stimulate or stress cells in the oral cavity.

[0029] Such an antioxidant oral care composition can protect cells in the oral cavity through its antioxidant properties, and can also be used as a gingival fibroblast protective agent, for example. The antioxidant oral care composition of the present invention contains magnesium (Mg) as an antioxidant amorphous mineral, and also contains magnesium ions (Mg 2+ The concentration of the antioxidant is 0.8 to 12 mg / mL, so that antioxidant and cytoprotective effects are observed even after a relatively short contact time with oral cells, such as 1 to 3 minutes. Such an antioxidant oral care composition can be used not only without rinsing, but also as a main ingredient in various oral care products such as mouthwashes, toothpastes, and mouth sprays.

[0030] In addition to containing the amorphous mineral composition as an active ingredient, the antioxidant oral care composition of the present invention may contain additives depending on the purpose of the product, to the extent that the intended purpose of the present invention is not impaired. Additives include aqueous components such as water, as well as well-known additives suitable for various formulations, such as flavorings, thickeners, surfactants, emulsifiers, abrasives, bactericidal or antibacterial components, anti-inflammatory components, oily components, and coating agents. [Example]

[0031] [Examples 1-4] An amorphous mineral composition was produced by naturally drying and concentrating lake water from the Great Salt Lake in Utah, USA, for one year under sunlight, and removing 99.5% of crystalline mineral components such as sodium chloride. The mineral composition was measured and confirmed to be the composition shown in Table 1 below.

[0032] [Table 1]

[0033] The osmotic pressure of the above amorphous mineral composition diluted with pure water 10 times (Example 1), 40 times (Example 2), 100 times (Example 3), and 1000 times (Reference Example) (mouthwash or liquid dentifrice in the formulation examples) was measured, and Examples 1, 3, and the Reference Example are shown in Table 2.

[0034] [Table 2]

[0035] The above amorphous mineral composition was diluted 10 times with water to prepare the antioxidant oral care composition of Example 1, and similarly diluted 40 times to prepare Example 2, and diluted 100 times to prepare Example 3. Other dilutions were used as reference examples. The radical scavenging activity as an antioxidant capacity and the cytoprotective effect on human gingival fibroblasts (hGFs) of Examples 1-3 were investigated by the following tests.

[0036] <Test 1: Measurement of hydroxyl radical scavenging activity (Examples 1 and 2)> Hydroxyl radicals (·OH) generated by the Fenton reaction were measured by electron spin resonance (ESR)-spin trapping. The Fenton reaction was carried out by adding ferrous sulfate to hydrogen peroxide solution, as shown in the following formula: H2O2+Fe 2+ → OH+OH - +Fe 3+

[0037] Using amorphous mineral compositions (reference examples) of various concentrations, and for comparison, dimethyl sulfoxide (DMSO), a typical ·OH scavenger, 80 μL of each was mixed for 10 seconds with 50 μL of 2 mM H2O2, 20 μL of 8.9 M DMPO, and 50 μL of 0.2 mM FeSO4. Immediately after mixing, the mixture was transferred to an ESR spectrometry cell, and the DMPO-OH spin adducts (derived from ·OH) were quantified after 60 seconds using an X-band ESR spectrometer with a 20 μM TEMPOL standard solution.

[0038] FIG. 1 shows representative ESR spectra of the pure water control and the stock solution at different dilution ratios (Examples 1 and 2), and the signal intensity of DMPO-OH was clearly reduced in a concentration-dependent manner by the stock solution. The signal intensity of DMPO-OH was observed as an ESR signal with hyperfine coupling constants aN = 1.49 and aH = 1.49 mT, which is attributed to DMPO-OH (a spin adduct of DMPO and ·OH) when DMPO was added.

[0039] The observed decrease in the signal intensity of DMPO-OH may be induced not only by the elimination of OH but also by the interference of the Fenton reaction, whereas Mg 2+ It was clear that the radical scavenging reaction by redox-active metal ions such as these could not be ruled out. Therefore, it was found that Examples 1 and 2 had the ability to eliminate ·OH when the original solution was diluted 10 to 40 times.

[0040] <Test 2: Confirmation test of scavenging effect against stable radical DPPH (Examples 1, 4 and Reference Example)> The DPPH scavenging rate of the above amorphous mineral composition (undiluted solution) against the stable radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) was investigated for a 2.5-fold diluted solution (Reference Example), a 5.0-fold diluted solution (Reference Example), a 10-fold diluted solution (Example 1), and a 20-fold diluted solution (Example 4), and the results are shown in Figure 2.

[0041] That is, 80 μL of the amorphous mineral composition at each of the above diluted concentrations was mixed with 16 μL of 100 mM Tris-HCl buffer (pH 7.5), 64 μL of 100% ethanol, and 40 μL of 1 mM DPPH in wells of a 96-well microplate.

[0042] The plate was then left in a light-shielded location for 20 minutes, after which the absorbance at 520 nm was measured using a microplate reader. The DPPH scavenging rate was calculated according to the following formula and shown in Figure 2. In the formula, A520 is the absorbance at 520 nm. ((A520 of solvent control - A520 of sample) / A520 of solvent control) x 100 As is clear from the results shown in FIG. 2, the amorphous mineral composition eliminated DPPH in a concentration-dependent manner.

[0043] <Test 3: Test to confirm proliferation of subconfluent cells after exposure to pure water or hydrogen peroxide (Examples 1 and 3)> Human gingival fibroblasts (hGFs) were cultured in a culture vessel to a subconfluent state of approximately 30%, and the cells (hGFs) that had room to grow were washed with PBS and exposed to the above amorphous mineral composition diluted 100 times (Example 3) and 10 times (Example 1) for 3 minutes.

[0044] The amorphous mineral composition was then removed, washed with PBS, and exposed to pure water or 100 μM hydrogen peroxide (HO) for 3 minutes. After washing with PBS, fresh medium was added and the cells were cultured at 37°C under humidified 5% CO for 24 hours. Cell proliferation was evaluated by MTT assay using DMEM as a control. The results are shown in Figure 3.

[0045] As is clear from the results shown in Figure 3, cell (hGFs) proliferation 24 hours after exposure was significantly reduced compared to the untreated control group, but was improved by pretreatment with the amorphous mineral composition diluted 100-fold (Example 3) and 10-fold (Example 1).

[0046] <Test 4: Test to confirm viability of confluent cells after exposure to hydrogen peroxide (Examples 1 and 3)> Human gingival fibroblasts (hGFs) were grown to confluence in a culture vessel, washed with PBS, and exposed to the above amorphous mineral composition diluted 100-fold (Example 3) and 10-fold (Example 1) for 3 minutes.

[0047] The amorphous mineral composition was then removed, washed with PBS, and exposed to 12.5 mM, 25 mM, 50 mM, or 100 mM hydrogen peroxide (HO) for 3 minutes. After washing with PBS, fresh medium was added and the cells were cultured at 37°C under humidified 5% CO for 24 hours. Cell viability was assessed by MTT assay using PBS as a control. The results are shown in Figure 4.

[0048] As can be seen from the results shown in Figure 4, the cell (hGFs) viability 24 hours after exposure was reduced to 67%, 88%, 96%, and 96% in the 12.5 mM, 25 mM, 50 mM, and 100 mM hydrogen peroxide (H2O2)-treated groups, respectively, compared to the untreated control group.

[0049] When these hydrogen peroxide (H2O2)-treated groups were pretreated with 100-fold diluted (Example 3) and 10-fold diluted (Example 1) amorphous mineral compositions, the reduction rates of the 12.5 mM and 25 mM H2O2-treated groups were significantly improved.

[0050] Each value shown in Figure 3 or Figure 4 is the mean + standard deviation (n = 6), and * and # in the figures indicate statistically significant differences (p < 0.05) from the untreated control and the corresponding DMEM, respectively.

[0051] From the above test results, it is clear that the water contains magnesium derived from the chloride lake water, as well as magnesium ions (Mg 2+ Examples 1 to 3, which consist of amorphous mineral compositions with a hydroxyl radical (·OH) concentration of 0.8 to 12 mg / mL, have the ability to scavenge hydroxyl radicals (·OH). When exposed to human gingival fibroblasts (hGFs) for 3 minutes, they were found to improve the cell viability of hGFs that had been exposed to pure water or hydrogen peroxide (H2O2) and suffered proliferation disorders and oxidative cell damage, and to exhibit a cytoprotective effect as a gingival fibroblast protective agent. [Industrial Applicability]

[0052] The present invention provides a general-purpose oral care composition for maintaining the health of cells in the oral cavity, particularly the gums, and can be used, for example, as a toothpaste, mouthwash, lozenge, chewable tablet, chewing gum, floss coating agent, denture care product, or as a gingival fibroblast protective agent.

Claims

1. The present invention uses an amorphous mineral composition, which is a concentrate of amorphous mineral components from salt lake water, as an active ingredient, and the magnesium ions (Mg 2＋ ) an antioxidant oral care composition having a concentration of 0.8 to 12 mg / mL.

2. The amorphous mineral composition contains chloride ions (Cl － ) concentration 2.5 to 30 mg / mL, sulfate ions (SO ４ ２－ ) concentration 0.18-2.4 mg / mL, sodium ion (Na ＋ ) concentration 0.02 to 0.4 mg / mL and potassium ion (K ＋ 2. The antioxidant oral care composition according to claim 1, which is an amorphous mineral composition having a concentration of 0.017 to 0.24 mg / mL.

3. 3. The antioxidant oral care composition according to claim 1, wherein the amorphous mineral composition has an osmotic pressure of 50 to 1500 mOSM / kg.

4. 3. The antioxidant oral care composition according to claim 1, wherein the saltwater lake water is lake water from the Great Salt Lake in the United States.

5. 4. The antioxidant oral care composition according to claim 3, wherein the saltwater lake water is lake water from the Great Salt Lake in the United States.

6. A gingival fibroblast protective agent comprising the antioxidant oral care composition according to claim 1 or 2.

7. A gingival fibroblast protective agent comprising the antioxidant oral care composition according to claim 5.

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