Use of a plankton extract in the preparation of an oral care composition

Abstract

The present invention relates to the use of a plankton extract in the preparation of an oral care composition, belonging to the technical field of oral care. The plankton extract of the present invention can accelerate the healing rate of acute or chronic wounds by upregulating fibroblast growth factor and keratinocyte growth factor, thereby applying recombinant FGF-2 to the skin wound, and can further promote wound healing by stimulating epithelial proliferation, altering the process of migration and differentiation through KGF. In addition, using the plankton extract of the present invention to prepare an oral care cleaning solution can not only accelerate the reduction of the wound surface diameter of oral ulcers and accelerate wound healing, but also effectively relieve the pain of oral ulcers.

Description

Technical Field

[0001] The present invention relates to the technical field of oral care, and in particular to the application of a plankton extract in an oral care composition. Background Art

[0002] Oral care compositions are essential cleaning products in people's daily lives. There are various types of products in this category, including toothpaste, mouthwash, etc., which are mainly used to keep the oral cavity clean, make the breath fresh, and at the same time care for oral health. Due to people's frequent staying up late affecting the immune system, the bad living habit of eating takeout affecting the microbiome, and the lack of attention to oral cleaning and hygiene during orthodontics, it is easy for oral soft tissues to become inflamed, which will not only cause gum diseases in the gums, but also cause symptoms such as mucosal damage, epidermal exfoliation, and ulcers in other oral tissues.

[0003] Oral ulcer is the most common disease affecting the oral cavity, which is characterized by repeated destruction of the oral mucosa in the form of painful ulcers. Oral ulcers can occur anywhere in the oral cavity, including the floor of the mouth, inner cheeks, gums, lips, and tongue, and 20% of people are affected. The causes of oral ulcers are considered to vary from person to person and may be related to local trauma, systemic diseases, genetic factors, viral and bacterial infections, and nutritional deficiencies. Aphthous ulcers are often associated with them. Among existing oral care products, there is less research on accelerating the healing of oral ulcers and reducing pain.

[0004] At the same time, scientific research has found that plasmin is an abundant plasma protease that can cleave and inactivate clot-related protein fibrin. The lack of plasmin and its inactive zymogen form plasminogen (PLG) in humans will lead to severe inflammation in mucosal tissues such as the oral cavity and eyes. When fibrinolysis is blocked, fibrin accumulation leads to continuous activation of neutrophils and promotes local immunopathology. In the oral mucosa, the commensal microbiota triggers homeostatic inflammation, fibrin deposition, and related neutrophil activation, which physiologically subsides in the case of effective plasminogen-mediated fibrinolysis.

[0005] Therefore, there is an urgent need to develop an oral care composition that can effectively accelerate the healing of oral ulcers and reduce pain. Summary of the Invention

[0006] The purpose of the present invention is to overcome the deficiencies of the prior art and provide an application of a plankton extract in the preparation of an oral care composition, which can effectively promote the rapid healing of oral ulcers and reduce pain. At present, plankton extracts are mostly applied in cosmetics. The present invention discloses its application in the preparation of oral care compositions, which is a new application of plankton extracts.

[0007] To achieve the above purpose, the technical solution adopted by the present invention is:

[0008] The present invention provides an application of a plankton extract in the preparation of an oral care composition. The preparation method of the plankton extract comprises the following steps:

[0009] S1. Fermenting a mixed bacterial mass of Flavobacterium aquatile, Rhizomonas suberifaciens, and Flectobacillus sp. in an aqueous solution of seaweed polysaccharide to obtain a fermentation product;

[0010] S2. Sterilizing, aging, and filtering the fermentation product obtained in step S1 to obtain the plankton extract;

[0011] In step S1, the stirring speed of fermentation is 250-300 r / min, and the fermentation time is 72-120 h;

[0012] In step S2, the aging temperature is 45-60 °C, and the aging time is 72-120 h.

[0013] The inventors publicly studied a plankton metabolite and a plankton extract in 2018. The plankton metabolite and the plankton extract can, by down-regulating the expression of three major inflammatory mediator genes (-MAP-Kp38, -NF-κB, ERK), achieve anti-inflammatory performance, and also have good effects of increasing muscle mass and reducing fat, reshaping the facial contour, and lifting and tightening the skin, having good application value. Before that, the application of plankton extract was for the anti-inflammatory, anti-aging, and lipid-lowering effects of cosmetic formulations on the skin. After adjusting the fermentation process parameters, the inventors found that the fermentation depth can be increased, so that the target product contains more Zn 2+ , K + , F- and Mg 2+ , which is beneficial to inhibiting the generation of reactive oxygen species, thereby reducing COX-2 expression and PGE-2 release, and can also reduce the solubility of the enamel on the tooth surface and promote enamel remineralization. In addition, by adjusting the aging process, the inventors can completely remove the macromolecular proteins in the target product to prevent flocculation during storage, which is more conducive to the application requirements of oral care products.

[0014] In a second aspect, the present invention also provides a plankton extract. The preparation method of the plankton extract comprises the following steps:

[0015] S1. Ferment a mixed bacterial mass of Flavobacterium aquatile, Rhizomonas suberifaciens, and Flectobacillus sp. in an aqueous solution of algal polysaccharide to obtain a fermentation product;

[0016] S2. Sterilize, ripen, and filter the fermentation product obtained in step S1 to obtain the plankton extract;

[0017] In step S1, the stirring speed for fermentation is 250 - 300 r / min, and the fermentation time is 72 - 120 h;

[0018] In step S2, the ripening temperature is 45 - 60 °C, and the ripening time is 72 - 120 h.

[0019] The inventors of the present invention have found through research that the plankton extract prepared by the above method of the present invention can obtain a higher content of the target product, and the product is more clarified and transparent, which is beneficial to its application in oral care products; at the same time, the plankton extract obtained by the preparation method of the present invention can also up-regulate the expression of the plasminogen gene, promote epithelial-mesenchymal transition in wound healing by enhancing the expression of growth factors, which is beneficial to accelerating the wound healing of oral ulcers and effectively reducing the pain of oral ulcers, and can be applied to the preparation of oral care compositions.

[0020] As a preferred embodiment of the plankton extract of the present invention, in step S1, the stirring speed for fermentation is 300 r / min, and the fermentation time is 96 h.

[0021] The inventors of the present invention have found through research that using the above stirring speed and fermentation time is beneficial to increasing the pH value drop and viable cell count of the target product, and at the same time, the target product contains more Zn 2+ , K + , F - and Mg 2+ , which is beneficial to improving the performance of the plankton extract in promoting wound healing and repair.

[0022] As a preferred embodiment of the plankton extract of the present invention, in step S2, the ripening temperature is 45 °C, and the ripening time is 96 h.

[0023] The inventors of the present invention have found through research that using the above ripening temperature and time is beneficial to removing macromolecular proteins in the target product to prevent flocculation during storage, which is more conducive to the application requirements of oral care products.

[0024] In a third aspect, the present invention also provides an oral care composition that promotes the healing of oral ulcers. The oral care composition includes the above-mentioned plankton extract; the oral care composition is toothpaste or an oral care cleaning solution.

[0025] The inventors of the present invention have found through research that the oral care composition prepared with the plankton extract of the present invention can not only up-regulate the expression of the plasminogen gene and up-regulate fibroblast growth factor (FGF-2) and keratinocyte growth factor (KGF). Among them, FGF-2 can accelerate the healing of acute or chronic wounds, and the increase in KGF can stimulate epithelial proliferation and change the processes of migration and differentiation, further promoting wound healing, thereby achieving epithelial-mesenchymal transition during wound healing. In addition, through the application of the plankton extract of the present invention in the preparation of an oral care cleaning solution, it has been found that the oral care cleaning solution containing the plankton extract of the present invention can reduce the wound diameter of oral ulcers and effectively relieve the pain of oral ulcers, which is beneficial to improving oral health.

[0026] As a preferred embodiment of the oral care composition of the present invention, the pH value of the oral care composition is 3.0 to 9.0.

[0027] The inventors of the present invention have found through research that controlling the pH value of the oral care composition of the present invention within the above range can not only enable the plankton extract to exert better efficacy, but also enhance its stability in the oral care composition, thereby enabling the oral care composition to have better physical and chemical properties and stability.

[0028] As a preferred embodiment of the oral care composition of the present invention, the oral care composition is toothpaste;

[0029] The toothpaste includes the following components in parts by weight: 15 to 60 parts of a humectant, 15 to 50 parts of an abrasive, 1 to 4 parts of a foaming agent, 0.8 to 2 parts of a fragrance, 0.5 to 2 parts of a thickener, 0.1 to 0.3 parts of a sweetening agent, and 0.9 to 9 parts of a plankton extract.

[0030] The inventors of the present invention have found through research that by compounding the components with the above specific ratios with the plankton extract, the paste of the toothpaste can have better physical and chemical properties and the stability of the appearance quality.

[0031] As a more preferred embodiment of the oral care composition of the present invention, the weight part of the plankton extract is 1.8 parts.

[0032] The inventors of the present invention have found through research that when the weight portion of the plankton extract is 1.8 parts, the toothpaste prepared not only has better physical and chemical properties and appearance quality stability, but also is more beneficial to oral health care. As a preferred embodiment of the oral care composition of the present invention, the oral care composition is an oral care cleaning solution; the oral care cleaning solution is a mouthwash or an oral spray;

[0033] The oral care cleaning solution comprises the following components in parts by weight: 5-15 parts of a humectant, 0.1-0.3 parts of a sweetener, 0.2-1.5 parts of a fragrance, 0.2-5 parts of a solubilizer, 0.02-0.5 parts of a preservative, and 0.9-9 parts of a plankton extract.

[0034] As a more preferred embodiment of the oral care composition of the present invention, the weight portion of the plankton extract is 1.8 parts.

[0035] The inventors of the present invention have found through research that when the weight portion of the plankton extract in the oral care cleaning solution of the present invention is within the above range, it has a better effect of reducing the wound surface diameter of oral ulcers and can also make the pain of oral ulcers weaker.

[0036] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0037] (1) The fermentation process parameters of the plankton extract of the present invention can improve the fermentation depth, so that the target product contains more Zn 2+ , K + , F - and Mg 2+ , which is beneficial to inhibiting the generation of reactive oxygen species, thereby reducing COX-2 expression and PGE-2 release, and can also reduce the solubility of the enamel on the tooth surface and promote enamel remineralization; and the aging process of the present invention can completely remove the macromolecular proteins in the target product to prevent flocculation during storage, which is more conducive to the application requirements of oral care products;

[0038] (2) The present invention provides the application of the plankton extract in the preparation of an oral care composition. The plankton extract of the present invention can up-regulate fibroblast growth factor (FGF-2) and keratinocyte growth factor (KGF). Among them, FGF-2 can accelerate the healing of acute or chronic wounds, and the increase of KGF can stimulate epithelial proliferation, change the process of migration and differentiation, and further promote wound healing, so as to achieve epithelial-mesenchymal transformation in wound healing. In addition, the oral care cleaning solution prepared with the plankton extract of the present invention can not only reduce the wound surface diameter of oral ulcers, but also effectively relieve the pain of oral ulcers;

[0039] (3) The oral care composition prepared from the plankton extract of the present invention can adjust its pH value, enabling the plankton extract to exert better efficacy, and can also improve the stability of the physical and chemical properties and appearance quality of the oral care composition, having better application performance. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 It is a comparison chart of the influence of the plankton extracts of Examples 1-2 and Comparative Examples 1-5 of the present invention on the pH value decrease;

[0041] Figure 2 It is a comparison chart of the influence of the plankton extracts of Examples 1-2 and Comparative Examples 1-5 of the present invention on the viable bacteria count;

[0042] Figure 3 It is a comparison chart of the influence of the plankton extracts of Example 1, Examples 3-4 and Comparative Examples 6-7 of the present invention on the metabolite Zn 2+ ;

[0043] Figure 4 It is a comparison chart of the influence of the plankton extracts of Example 1, Examples 3-4 and Comparative Examples 6-7 of the present invention on the metabolite K + ;

[0044] Figure 5 It is a comparison chart of the influence of the plankton extracts of Example 1, Examples 3-4 and Comparative Examples 6-7 of the present invention on the metabolite F - ;

[0045] Figure 6 It is a comparison chart of the influence of the plankton extracts of Example 1, Examples 3-4 and Comparative Examples 6-7 of the present invention on the metabolite Mg 2+ ;

[0046] Figure 7 It is a chart of the in vitro expression content of the fibrinolytic enzyme gene by the plankton extract of the present invention;

[0047] Figure 8 It is a toxicity test chart of the plankton extract of the present invention on NHDF cells;

[0048] Figure 9 It is a chart of the in vitro expression content of FGF-2 by the plankton extract of the present invention;

[0049] Figure 10 It is a chart of the in vitro expression content of KGF by the plankton extract of the present invention;

[0050] Figure 11 It is a chart of the ulcer size test of the oral care composition of Example 13 of the present invention;

[0051] Figure 12It is a pain score test chart of the oral care composition of Example 13 of the present invention. Detailed implementation manners

[0052] The technical solutions of the present invention will be further described below in conjunction with the embodiments and the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts fall within the scope of protection of the present invention. The methods or operations used in the embodiments are conventional methods or conventional operations in the art unless otherwise specified.

[0053] Examples 1-7 and Comparative Examples 1-12

[0054] Examples 1-7 and Comparative Examples 1-12 of the present invention are the plankton extracts described in the present invention.

[0055] The preparation methods of the plankton extracts of Examples 1-7 and Comparative Examples 1-12 include the following steps:

[0056] S1. Ferment a mixed bacterial mass of Flavobacterium aquatile, Rhizomonas suberifaciens, and Flectobacillus sp. in an aqueous solution of algal polysaccharide to obtain a fermentation product;

[0057] S2. Sterilize and ripen the fermentation product obtained in step S1, cool it to 25°C, and then filter it through a double-layer 300-mesh filter cloth to remove flocculent precipitates to obtain the plankton extract.

[0058] The process parameters of the plankton extracts of Examples 1-7 and Comparative Examples 1-12 are shown in Table 1 below.

[0059] Table 1

[0060]

[0061]

[0062] Effect Example 1

[0063] To verify the influence of the fermentation process parameters of the present invention on the performance of the plankton extract, the plankton extracts of Examples 1-4 and Comparative Examples 1-7 were tested for pH value, viable bacteria count, and metabolite content.

[0064] The specific test methods are as follows.

[0065] (1) pH value test

[0066] Test method: Use a pH meter to detect the pH value of the sample before and after fermentation;

[0067] Calculation method: Degree of pH decrease = pH value before fermentation - pH value after fermentation.

[0068] (2) Viable cell count test

[0069] Test principle: The fermentation process of the plankton extract described in the present invention belongs to coupled fermentation, conforms to first-order fermentation kinetics, and there is a positive correlation between the growth rate of cells (i.e., the number of viable cells) and the increase in metabolic rate. The more the number of viable cells, the higher the degree of fermentation.

[0070] Test method: The fermentation broth after fermentation of the plankton extract during preparation enters a flow-through thin-layer colorimetric ring, and the optical density (OD value) of the fermentation broth is measured with a light beam of 550 nm. The measured OD value of the fermentation broth has a linear relationship with the cell concentration, and the number of viable cells in the fermentation broth is obtained according to the standard curve.

[0071] (3) Ion content test

[0072] Test method: Use an inductively coupled plasma optical emission spectrometer (ICP-OES) to measure the contents of Zn 2+ , K + and Mg 2+ in the fermentation product respectively; Refer to the test method for free fluorine in GB / T 8372-2017 "Toothpaste", and use an ion meter equipped with a fluoride ion selective electrode and a reference electrode and a pH meter to test the content of F - in the fermentation product.

[0073] The test results are as Figures 1 - 6 shown.

[0074] Figure 1 This is a comparison chart of the influence of the plankton extracts of Examples 1-2 and Comparative Examples 1-5 of the present invention on the degree of pH decrease. It can be seen from the figure that by fermenting at the stirring speed of the plankton extracts of Examples 1-2 of the present invention, the degree of pH decrease of the target product can be effectively improved, and the degree of pH decrease of the plankton extract of Example 1 is the highest.

[0075] Figure 2 This is a comparison chart of the influence of the plankton extracts of Examples 1-2 and Comparative Examples 1-5 of the present invention on the number of viable cells. It can be seen from the figure that by fermenting at the stirring speed of the plankton extracts of Examples 1-2 of the present invention, the number of viable cells of the target product can be effectively improved, and the number of viable cells of the plankton extract of Example 1 is the highest.

[0076] Figures 3 - 6Plankton extracts of Example 1, Examples 3-4 and Comparative Examples 6-7 on metabolites Zn 2 + , K + , F - , Mg 2+ The comparative diagram of the influence. It can be seen from the figure that by controlling the fermentation time, the content of metabolites Zn 2 + , K + , F - , Mg 2+ can be effectively increased, and the metabolite content of the plankton extract in Example 1 is the highest among them.

[0077] Effect Example 2

[0078] In order to verify the influence of the ripening process of the present invention on the performance of the plankton extract, the plankton extracts of Example 1, Examples 5-7 and Comparative Examples 8-12 were subjected to protein precipitation tests.

[0079] The specific test method is as follows:

[0080] The test results are shown in Table 2 below.

[0081] Table 2

[0082]

[0083]

[0084] It can be seen from Table 2 that by using the ripening process of the plankton extract of Example 1, the protein precipitation rate can reach 100% at a shorter ripening time and a lower ripening temperature, which is beneficial to improving the production process efficiency.

[0085] Effect Example 3

[0086] In order to verify the applicable pH value range of the plankton extract of the present invention in the oral care composition, the plankton extract prepared in Example 1 of the present invention was subjected to stability tests under different pH values and different test conditions.

[0087] The test method is as follows: Prepare aqueous solutions of plankton extracts with different pH values using citric acid and sodium hydroxide. The mass percentage of the plankton extract in the aqueous solution of the plankton extract is 1.8%. The preservative is sodium benzoate, and the mass ratio of sodium benzoate to the aqueous solution of the plankton extract is sodium benzoate: aqueous solution of the plankton extract = 0.004:1. Place the aqueous solutions of the plankton extracts with different pH values under the conditions of 48°C, 5°C, 25°C, UV, and -10°C respectively, and continuously observe for 7 days to observe the changes in the appearance, color, and odor of the aqueous solutions of the plankton extracts on the 7th day.

[0088] The test results are shown in Tables 3 to 7 below.

[0089] Table 3 Test conditions: 25°C

[0090]

[0091] Table 4 Test conditions: 48°C

[0092]

[0093]

[0094] Table 5 Test conditions: 5°C

[0095]

[0096] Table 6 Test conditions: UV

[0097]

[0098] Table 7 Test conditions: -10°C

[0099]

[0100]

[0101] As can be seen from Tables 3 to 7, when the pH value of the aqueous solution of the plankton extract of the present invention is 3.0 to 9.0, the appearance, color, and odor of the aqueous solution of the plankton extract are normal after being placed for 7 days under different test conditions. However, when the pH value is too low or too high, the stability of the aqueous solution of the plankton extract decreases, and the appearance, color, and odor of the product change to varying degrees under different test conditions, which is not conducive to the application of the product.

[0102] Effect Example 4

[0103] In order to verify the effect of the plankton extract of the present invention on oral mucosal inflammation, using human epidermal keratinocytes as the culture substrate, the effect of the plankton extract prepared in Example 1 on the activity of plasminogen was tested.

[0104] Test method: After washing human epidermal keratinocytes with phosphate buffer solution, the cells were incubated with 100 μL of 0.5 mM plasminogen (Roche Molecular Biochemicals, Mannheim, Germany) at 37 °C for 30 min. Then the cells were incubated with the chromogenic fibrin-specific substrate L1450 (Bachem, Heidelberg, Germany) in 50 μL of 1.5 mM solution for 20 min. The production of cleaved substrate was photometrically evaluated in an enzyme-linked immunosorbent assay (ELISA) reader by normalizing the optical density (OD) at the maximum extinction of L1450 (405 nm) to the extinction at an irrelevant wavelength (614 nm).

[0105] The concentrations of the plankton extract in the aqueous solution of the plankton extract used for the test were respectively: 10 -9 mol / L, 10 -6 mol / L, 10 -3 mol / L.

[0106] The test results are as Figure 7 shown. It can be seen from Figure 7 that compared with the untreated cells, treatment of keratinocytes with plankton extracts at different concentrations led to plasminogen activation, and the effect of significantly increased (P < 0.05) plasminogen activation was dose-dependent.

[0107] Effect Example 5

[0108] To further verify the effect of the plankton extract of the present invention on wound healing performance, the cytotoxic effect of the plankton extract of the present invention on dermal fibroblasts and the expression effect on growth factors (FGF-2 and KGF) were tested.

[0109] Test samples: The test group was the plankton extract of Example 1 of the present invention (recorded as Bucocean), and the control group was a blank group without the plankton extract of the present invention (recorded as Control).

[0110] Cell culture: Normal human dermal fibroblasts (NHDF cells, Asahi Techno Glass, Japan) were used and cultured in Dulbecco's modified Eagle's medium (DMEM; GIBCO-BRL, Bethesda, MD) supplemented with 10% heat-inactivated fetal bovine serum (FBS; GIBCO-BRL) and antibiotics (including penicillin 100 units / mL - streptomycin 100 mg / mL). The NHDF cells were stored in a humidified incubator at 37 °C and 5% CO 2 2.

[0111] The cytotoxicity of the plankton extract of the present invention against NHDF cells was evaluated by cell MTT assay. The specific test method was as follows:

[0112] S1. NHDF cells were seeded into 96-well plates at a density of 13,000 cells / 100 μL, 100 μL per well, to allow cell attachment, and 150 μL of growth medium was added to each well;

[0113] S2. The plankton extract was diluted with water to obtain dilution solutions with concentrations of 0.03 mg / mL, 0.06 mg / mL, 0.13 mg / mL, 0.25 mg / mL, 0.50 mg / mL, and 1.00 mg / mL of the plankton extract. 50 μL of the sample dilution solution was added to each well described in step S1, and incubated for 24 hours;

[0114] S3. After incubation, the medium in each well was completely removed, then replaced with 200 μL of fresh growth medium. 20 μL of MTT solution was added to each well and incubated at 37 °C for 2 hours;

[0115] S4. After incubation, the medium in each well was removed, and 200 μL of dimethyl sulfoxide was added to each well to dissolve the formazan crystals precipitated in the wells. The absorbance of each well was measured at 570 nm using a microplate reader (Infinite M200Pro, Tecan, Switzerland). The wells incubated with growth medium were used as negative controls. The test results are as shown. It can be seen that the cell viability of the plankton extract of the present invention at the 6 test concentrations was > 100%, indicating that the plankton extract of the present invention is non-toxic to NHDF cells.

[0116] The specific test method for the expression of growth factors (FGF-2 and KGF) was as follows: Figure 8 as shown Figure 8 It can be seen that the cell viability of the plankton extract of the present invention at the 6 test concentrations was > 100%, indicating that the plankton extract of the present invention is non-toxic to NHDF cells.

[0117] The specific test method for the expression of growth factors (FGF-2 and KGF) was as follows:

[0118] S1. 200,000 NHDF cells were seeded onto 35 mm culture dishes. After culturing for 3 days, the conditioned medium was taken out, centrifuged at 1000 rpm for 2 min, and then washed with 0.5 mL of PBS to obtain a cell suspension;

[0119] S2. The cell suspension was sonicated on ice 3 times, 10 s each time, and both the conditioned medium and the cell suspension were stored at -20 °C;

[0120] S3. Measure the levels of FGF-2 and KGF in cell-related samples and supernatant samples using a commercially available ELISA kit. Quantify the protein levels by comparing the optical density of each measured protein with its standard curve and normalizing the cell count.

[0121] The test results of growth factor expression are as Figure 9 and Figure 10 shown. Figure 9 Figure Figure 9 shows the in vitro expression of FGF-2 in the test group containing the plankton extract of the present invention and the control group without the plankton extract in a culture dish. It can be seen from Figure 10 Figure Figure 10 that the total level of FGF-2 in the test group is higher than that in the control group, indicating that the plankton extract prepared from the plankton extract of the present invention can promote the expression of FGF-2.

[0122] Examples 8 to 11 and Comparative Example 13

[0123] Examples 8 to 11 and Comparative Example 13 are oral care compositions of the present invention. The oral care compositions of Examples 8 to 11 and Comparative Example 13 are toothpastes.

[0124] The components and their parts by weight of the toothpastes of Examples 8 to 11 and Comparative Example 13 are shown in Table 8 below, and the total parts by weight are all 100 parts. In the toothpastes of Examples 8 to 11 and Comparative Example 13: the plankton extracts are all prepared in Example 1, the thickener is xanthan gum, the abrasive is silica, the sweetener is sodium saccharin, the foaming agent is sodium lauryl sulfate, and the humectants are sorbitol and glycerol. The pH values of the toothpastes of Examples 8 to 11 and Comparative Example 13 are all 6.5 to 7.5.

[0125] The preparation method of the toothpastes of Examples 8 to 11 and Comparative Example 13 includes the following steps:

[0126] S1. Weigh the abrasive and the thickener, and stir until evenly mixed to obtain mixture A;

[0127] S2. Weigh the sweetener, the plankton extract, the humectant and water, and stir until evenly mixed to obtain mixture B;

[0128] S3. Add the mixture B described in step S2 and the mixture A described in step S1 to the paste-making machine in sequence, and stir until evenly mixed;

[0129] S4. Add a foaming agent and a flavor to the paste-making machine, stir under vacuum until evenly mixed, and degas to obtain toothpaste.

[0130] Table 8

[0131]

[0132]

[0133] Effect Example 6

[0134] To further explore the product stability of the plankton extract of the present invention when applied to an oral care composition, the toothpastes of Examples 8 to 11 and Comparative Example 13 were subjected to a stability performance test.

[0135] Test method: (1) Place the prepared toothpastes under the conditions of 45°C and 25°C respectively, continuously observe for 8 weeks (56 days), observe the changes in the appearance, color and smell of the toothpastes, and use a consistency rack to test their consistency changes; (2) Place the prepared toothpastes at a low temperature of -8°C and continuously observe for 4 weeks (28 days), observe the changes in the appearance, color and smell of the toothpastes, and use a consistency rack to test their consistency changes; (3) Perform 4 cycles of hot and cold (-15 to 45°C) on the prepared toothpastes, observe the changes in the appearance, color and smell of the toothpastes, and use a consistency rack to test their consistency changes.

[0136] The test results are shown in Tables 9 to 12 below.

[0137] Table 9 Test condition: 25°C

[0138]

[0139] Table 10 Test condition: 45°C

[0140]

[0141]

[0142] Table 11 Test condition: -8°C

[0143]

[0144] Table 12 Test condition: 4 cycles of hot and cold

[0145]

[0146] As can be seen from Tables 9 to 12, the toothpastes prepared in Examples 8 to 11 of the present invention have good stability under different test conditions, while when the addition amount of the plankton extract is relatively large, the stability of the toothpaste prepared in Comparative Example 13 is poor.

[0147] Examples 12 to 15 and Comparative Example 14

[0148] Examples 12 to 15 and Comparative Example 14 are oral care compositions of the present invention. The oral care compositions of Examples 12 to 15 and Comparative Example 14 are oral care cleaning liquids, and the oral care cleaning liquids are mouthwashes.

[0149] The components and their parts by weight of the oral care compositions of Examples 12 to 15 and Comparative Example 14 are shown in Table 13 below. The total parts by weight are all 100 parts, and the unit of the parts by weight of each component is: part. In the mouthwashes of Examples 12 to 15 and Comparative Example 14 of the present invention: the plankton extract is all prepared in Example 1, the humectant is all sorbitol, the sweetener is all saccharin sodium, the solubilizer is all Tween-20, and the preservative is sodium benzoate. The pH value of the mouthwashes of Examples 12 to 15 and Comparative Example 14 of the present invention is 6 to 7.

[0150] The preparation method of the mouthwashes of Examples 12 to 15 and Comparative Example 14 includes the following steps:

[0151] S1. Weigh the essence and solubilizer, and stir until evenly mixed to obtain mixture C;

[0152] S2. Weigh the humectant, sweetener and water, and stir until evenly mixed to obtain mixture D;

[0153] S3. Add the plankton extract, preservative and mixture C in step S1 to the mixture D in step S2, and stir until evenly mixed to obtain the mouthwash.

[0154] Table 13

[0155]

[0156] Effect Example 7

[0157] In order to further explore the product stability of the plankton extract of the present invention applied to oral care compositions, the mouthwashes of Examples 12 to 15 and Comparative Example 14 were subjected to a stability performance test.

[0158] Test method: (1) Place the prepared mouthwashes at 45°C, -8°C, and 25°C respectively, and continuously observe for 4 weeks (28 days) to observe the changes in the appearance, color and odor of the mouthwashes; (2) Perform 4 cycles of hot and cold cycling (-15 to 45°C) on the prepared mouthwashes, and observe the changes in the appearance, color and odor of the mouthwashes.

[0159] The test results are shown in Tables 14 to 17 below.

[0160] Table 14 Test conditions: 25°C

[0161]

[0162] Table 15 Test conditions: 45°C

[0163]

[0164] Table 16 Test conditions: -8°C

[0165]

[0166] Table 17 Test conditions: 4 cycles of thermal cycling

[0167]

[0168]

[0169] As can be seen from Tables 14 to 17, the mouthwashes prepared in Examples 12 to 15 of the present invention have good stability under different test conditions. When the addition amount of the plankton extract is relatively large, the stability of the mouthwash prepared in Comparative Example 14 is poor.

[0170] Effect Example 8

[0171] In order to further verify the effect of the plankton extract of the present invention on the healing of oral ulcer wounds in the preparation of oral care compositions, the mouthwash of the present invention was subjected to in vivo testing.

[0172] Test samples: The test group was the mouthwash of Example 13 of the present invention, denoted as BM; the placebo was a mouthwash without plankton extract (the only difference between the placebo and Example 13 was that the plankton extract was replaced with the same amount of water, and the other components and their amounts were the same as those in Example 13), denoted as Placebo.

[0173] Subject population: 40 healthy non-smoking adult subjects, aged 19 to 45 years old, all diagnosed with aphthous oral ulcers, without any other known diseases, randomly divided into 2 groups, 20 subjects in each group, corresponding to the products of the test group and the control group respectively as a group, and not receiving any anti-inflammatory or antibiotic treatment within two weeks from the start of the test.

[0174] Test method: Use it once in the morning and once in the evening every day, 30 ml each time, gargle for 1 minute, and conduct a double-blind test for 2 weeks.

[0175] Pain score: The subjects were pain-scored using the VAS scoring method on the 0th day, 7th day, and 14th day of the test. It consisted of a 10-cm long horizontal line, fixed by 2 oral descriptors, each descriptor representing extreme symptoms ("no pain" scored 0, "very severe pain" scored 10). As the degree of pain increased, the pain score became higher.

[0176] Ulcer size: The ulcer size of the subjects was measured on the 0th, 7th, and 14th days of the test respectively. The maximum diameter of the white area of the aphthous ulcer in the subjects was measured using a dental caliper.

[0177] The test results of the ulcer size are as Figure 11 shown. It can be seen from Figure 11 that in the test group containing the plankton extract of the present invention, the ulcer size decreased significantly on the 7th and 14th days, while in the test group of the placebo, the ulcer size increased significantly on the 7th and 14th days. There was a statistically significant difference between the two groups of volunteers in terms of the complete healing of the ulcer lesions. Because 5 patients in the test group of the plankton extract healed completely on the 14th day, while there was no case of complete healing in the placebo group, indicating that the oral ulcer composition prepared from the plankton extract of the present invention can accelerate the wound healing of oral ulcers.

[0178] The test results of the pain score are as Figure 12 shown. It can be seen from Figure 12 that in the test group containing the plankton extract of the present invention, the pain level of the subjects decreased significantly on the 7th and 14th days, and the pain score of the subjects decreased to below 2.5 on the 14th day, while in the test group of the placebo, the pain level only decreased slightly on the 14th day, indicating that the oral ulcer composition prepared from the plankton extract of the present invention can effectively relieve the pain of oral ulcers.

[0179] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. Use of a plankton extract in the preparation of an oral care composition having the effect of promoting the healing of oral ulcers, characterized in that, the preparation method of the plankton extract comprises the following steps: S1. Ferment a mixed bacterial mass of Flavobacterium aquatile, Rhizomonas suberifaciens and Flectobacillus sp. in an aqueous solution of algal polysaccharide to obtain a fermentation product; S2. Sterilize, ripen and filter the fermentation product obtained in step S1 to obtain the plankton extract; in step S1, the stirring speed of fermentation is 250-300 r / min, and the fermentation time is 72-120 h; in step S2, the ripening temperature is 45-60 °C, and the ripening time is 72-120 h.

2. The use according to claim 1, characterized in that, in step S1, the stirring speed of fermentation is 300 r / min, and the fermentation time is 96 h.

3. The use according to claim 1, characterized in that, in step S2, the ripening temperature is 45 °C, and the ripening time is 96 h.

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