Biopolysaccharides for preventing and treating oral diseases and their applications

Abstract

The present invention provides a biopolysaccharide having the function of preventing and treating oral diseases and its application. Specifically, the present invention provides the use of β-glucan for preparing a preparation or composition, wherein the preparation or composition is used for preventing and / or treating oral diseases. The β-glucan has the function of significantly alleviating and improving oral diseases and controlling the occurrence and development of oral diseases, and can be used for preparing oral cleaning products.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to a biopolysaccharide having the efficacy of preventing and treating oral diseases and an application thereof. Background Art

[0002] The oral cavity is the starting point of the digestive tract and one of the important organs of the human body. In addition to its own functions, it is also a "window" to the overall health status. Some systemic diseases can manifest in the oral cavity, or even first manifest in the oral cavity. Various diseases can occur in the periodontal tissue and oral mucosal system, such as acute and chronic non-specific infectious inflammation, certain special infections (such as syphilis, tuberculosis, AIDS, etc.), trauma, deformity, tumors, and some systemic diseases (including certain congenital diseases) can also occur in the oral cavity.

[0003] Oral mucosal diseases refer to a variety of infectious and non-infectious diseases that occur in the oral mucosa and oral soft tissues. They can be divided into two categories, one is the disease that originates in the oral mucosa, and the other is the manifestation of systemic diseases in the oral cavity, mainly manifested as oral mucosal damage. Common diseases include ulcers, lichen planus, leukoplakia, discoid lupus erythematosus, stomatitis, glossitis, etc. Oral mucosa is more prone to mucosa such as cheeks, tongue, lips, and soft palate, and can also develop at the same time as the skin. In addition, diseases such as oral ulcers are difficult to cure and are prone to recurrence. The current treatment methods are mainly anti-inflammatory, analgesic, and promote ulcer healing. Commonly used antibiotics, hormone drugs and vitamins, such as tetracycline, dexamethasone, betamethasone, vitamin C, vitamin B complex, watermelon frost, etc.

[0004] Periodontal disease includes two major diseases: gum disease and periodontitis. The most common type of gum disease is chronic inflammation caused by dental plaque, namely gingivitis, as well as some gum diseases affected by systemic factors (endocrine, blood diseases, drugs, etc.) and local irritation. Periodontitis is a group of multifactorial diseases with similar clinical manifestations and histological changes, but different pathogenic factors and body reactivity, different course of disease, and different responses to treatment. China is a country with a high incidence of periodontal disease. 80% to 90% of adults suffer from periodontal disease to varying degrees, but most people lack oral health awareness, poor oral hygiene, and lack of knowledge about periodontal disease, which leads to missed treatment opportunities and forced tooth extraction or waiting for loose teeth to fall off on their own. The treatment drugs for periodontal disease mainly include antibiotics and anti-inflammatory drugs, such as nitroimidazoles, tetracyclines, penicillins, macrolides, non-steroidal anti-inflammatory drugs, etc.

[0005] The above-mentioned oral disease treatment drugs either have insignificant treatment effects or have certain side effects. Therefore, it is of great significance to develop new drugs for preventing and treating oral diseases. Summary of the invention

[0006] The purpose of the present invention is to provide a biopolysaccharide having the efficacy of preventing and treating oral diseases and application thereof.

[0007] The first aspect of the present invention provides a use of β-glucan for preparing a preparation or composition for preventing and / or treating oral diseases.

[0008] In another preferred embodiment, the oral diseases include periodontal diseases, tooth diseases and oral mucosal diseases.

[0009] In another preferred embodiment, the periodontal disease includes gingivitis and periodontitis.

[0010] In another preferred embodiment, the tooth diseases include caries, dentin hypersensitivity, and tooth stains.

[0011] In another preferred embodiment, the oral mucosal diseases include leukoplakia, oral lichen planus, oral ulcers, discoid lupus erythematosus, stomatitis, and glossitis.

[0012] In another preferred embodiment, the preparation or composition is used to prevent alveolar bone loss, prevent tooth decay, inhibit the formation of dental plaque, remove dental calculus, resist dentin sensitivity, improve gingival bleeding, remove tooth stains, whiten teeth, regulate oral microecology, improve oral hygiene, prevent and treat oral ulcers, repair and enhance oral mucosal immunity, or a combination thereof.

[0013] In another preferred embodiment, the preparation or composition is also used for oral postoperative care (promoting wound healing), prevention and / or treatment of oral diseases in diabetic patients.

[0014] In another preferred embodiment, the alveolar bone loss includes alveolar bone loss caused by periodontal disease and alveolar bone loss caused by dental implants.

[0015] In another preferred embodiment, the β-glucan is β-D-glucan.

[0016] In another preferred embodiment, the β-glucan is β-1,3-glucan, preferably β-1,3-glucan having β-1,6-branching.

[0017] In another preferred embodiment, the structure of the β-glucan is as shown in Formula I,

[0018]

[0019] Wherein, l is an integer ≥0, preferably 0-50, preferably 0-10, more preferably 0-3, more preferably 1-2, more preferably 1; m is an integer ≥0, preferably 0-19, preferably 0-4, more preferably 0-1, more preferably 0; n is an integer ≥3, preferably 30-60000, more preferably 100-10000.

[0020] In another preferred embodiment, the branching degree (DB) of the β-glucan is 0.02-0.8, preferably 0.1-0.5, and more preferably 0.2-0.4.

[0021] In another preferred embodiment, the β-glucan includes β-glucan having a triple helical structure.

[0022] In another preferred embodiment, the content of the β-glucan with triple helical structure is 80%, 90%, or 95%, calculated based on the total molar amount of β-glucan.

[0023] In another preferred embodiment, the β-1,3-main chain of the β-glucan is the main body of a triple helical structure.

[0024] In another preferred embodiment, the β-1,6-branch of the β-glucan is located outside the triple helical structure.

[0025] In another preferred embodiment, the molecular weight of the β-glucan is ≥2kD, preferably 2kD-40000kD, and more preferably 20kD-20000kD.

[0026] In another preferred embodiment, the molecular weight of the β-glucan can be 5kD-35000kD; 10kD-30000kD; 50kD-25000kD; 100kD-20000kD; 200kD-18000kD; 400kD-16000kD; 500kD-14000kD; 1000kD-12000kD; 2000kD-4000kD; 3000kD-5000kD; 4000kD-6000kD; 5000kD-7000kD; 6000kD-8000kD; 7000kD-9000kD; or 8000kD-10000kD.

[0027] In another preferred embodiment, the β-glucan is selected from the group consisting of Schizophyllum β-glucan, Lentinula edodes β-glucan, Sclerotium β-glucan, Grifola frondosa β-glucan, Pleurotus ostreatus polysaccharide, mushroom β-glucan, yeast β-glucan, oat β-glucan, or a combination thereof.

[0028] In another preferred embodiment, the β-glucan is Schizophyllum β-glucan.

[0029] In another preferred embodiment, the Lentinus edodes β-glucan is a β-glucan with 5 β-1,3-main chains, 2 β-1,6-branches, and 1 glucose residue in each branch.

[0030] In another preferred embodiment, the purity of the β-glucan is ≥70%, preferably ≥90%, more preferably ≥95%, and even more preferably ≥99%.

[0031] In another preferred embodiment, the β-glucan has good stability.

[0032] In another preferred embodiment, the β-glucan is in solid or liquid form, such as β-glucan solid particles or powder, or β-glucan aqueous solution.

[0033] In another preferred embodiment, the particle size of the β-glucan particles or powder is ≤20 mm, preferably 0.001-10 mm, more preferably 0.01-5 mm, and even more preferably 0.1-2 mm.

[0034] In another preferred embodiment, the β-glucan is completely water-soluble β-glucan.

[0035] In another preferred embodiment, the β-glucan (granules or powder) has good water solubility and / or natural solubility.

[0036] In another preferred embodiment, the solubility of the β-glucan (granules or powder) in 25°C water (100g) is ≥0.0001g, preferably 0.01-50g, more preferably 0.1-10g.

[0037] In another preferred embodiment, the solubility of the β-glucan (granules or powder) in 25°C water (100g) may be 0.1-100g; 0.2-90g; 0.5-80g; 1-50g; or, the solubility may be 0.1-0.3g; 0.2-0.4g; 0.3-0.5g; 0.4-0.6g; 0.5-0.7g; 0.6-0.8g; 0.7-0.9g; 0.8-1g; or 1-3g; 2-4g; 3-5g; 4-6g; 5-7g; 6-8g; 7-9g; 8-10g.

[0038] In another preferred embodiment, the β-glucan solution is a solution of β-glucan in water, that is, a β-glucan aqueous solution.

[0039] In another preferred embodiment, the β-glucan (water) solution has a high viscosity; preferably, the viscosity of the β-glucan aqueous solution (at 25° C.) with a mass concentration of 0.5% is ≥40 mPa·s, more preferably 100-10000 mPa·s, and more preferably 500-2000 mPa·s.

[0040] In another preferred embodiment, the viscosity of the β-glucan aqueous solution (25°C) with a mass concentration of 0.5% can be 50-10000mPa·s; 100-9000mPa·s; 200-8000mPa·s; 300-7000mPa·s; 400-6000mPa·s; 450-5000mPa·s; 500-5000mPa·s; 550-4000mPa·s; 600-3000mPa·s; 650-2000mPa·s; 700-1500mPa·s.

[0041] In another preferred embodiment, the aqueous solution of β-glucan with a mass concentration of 1% has high clarity or high transmittance, and the transmittance of the aqueous solution of β-glucan with a mass concentration of 1% is ≥50%, preferably ≥80%, preferably ≥85%, and more preferably ≥95%;

[0042] In another preferred embodiment, the β-glucan solution has good stability.

[0043] According to a second aspect of the present invention, a preparation is provided, wherein the preparation comprises β-glucan.

[0044] In another preferred embodiment, the β-glucan is selected from the group consisting of Schizophyllum β-glucan, Lentinula edodes β-glucan, Sclerotium β-glucan, Grifola frondosa β-glucan, Pleurotus ostreatus polysaccharide, mushroom β-glucan, yeast β-glucan, oat β-glucan, or a combination thereof.

[0045] In another preferred embodiment, the β-glucan is Schizophyllum β-glucan.

[0046] In another preferred embodiment, the β-glucan is completely water-soluble β-glucan.

[0047] In another preferred embodiment, the preparation is Schizophyllum β-glucan, and the β-glucan has good water solubility, natural solubility and / or resolubility.

[0048] In another preferred embodiment, the β-glucan in the preparation is in solid form or liquid form.

[0049] In another preferred embodiment, the preparation contains (a) β-glucan; and optionally (b) a pharmaceutically, cosmetically, or device-acceptable carrier or excipient.

[0050] In another preferred embodiment, the preparation contains (a) Schizophyllum β-glucan; and optionally (b) a pharmaceutically, cosmetically, or device-acceptable carrier or excipient.

[0051] In another preferred embodiment, the preparation contains 0.0001-99wt%, preferably 0.001-90wt%, more preferably 0.01-50wt%, more preferably 0.05-10wt% of β-glucan, based on the total weight of the preparation.

[0052] In another preferred embodiment, the mass concentration of the β-glucan in the preparation is ≥1 μg / mL, specifically 1 μg / mL-200 mg / mL, or 1 μg / mL-5 mg / mL, or 1 μg / mL-1 mg / mL.

[0053] In another preferred embodiment, the dosage form of the composition or preparation is a solid dosage form, a semisolid dosage form, or a liquid dosage form.

[0054] In another preferred embodiment, the preparation can be made into oral cleaning products, preferably toothpaste, mouthwash, oral gel, and chewing gum.

[0055] In another preferred embodiment, the preparation (gargle) can be a liquid preparation, an aerosol, a powder, a tablet, or a concentrate.

[0056] In another preferred embodiment, the composition is a pharmaceutical composition or a cosmetic composition.

[0057] In another preferred embodiment, the preparation comprises a β-glucan aqueous solution, and the mass concentration of the β-glucan is 0.0001-50wt%, preferably 0.02-10wt%, more preferably 0.05-5wt%, based on the total weight of the β-glucan aqueous solution.

[0058] In another preferred embodiment, the content of the β-glucan aqueous solution in the preparation is ≥80wt%, preferably ≥90wt%, more preferably ≥95wt%, more preferably ≥99wt%, more preferably ≥99.5wt%, based on the total weight of the preparation.

[0059] In another preferred embodiment, the β-glucan in the preparation has one or more characteristics selected from the following group:

[0060] (1) The purity of the β-glucan is ≥70%, preferably ≥90%, more preferably ≥95%, and even more preferably ≥99%;

[0061] (2) The β-glucan has good water solubility, redissolubility and / or natural solubility;

[0062] (3) The solubility of the β-glucan (solid particles or powder) in water at 25°C is ≥ 0.0001 g / 100 g water, preferably 0.01-50 g / 100 g water, and more preferably 0.1 g-10 g / 100 g water;

[0063] (4) The β-glucan aqueous solution has high clarity or high light transmittance; preferably, the light transmittance of the β-glucan aqueous solution with a mass concentration of 1% is ≥50%, preferably ≥80%, preferably ≥85%, and more preferably ≥95%;

[0064] (5) The β-glucan solution has a high viscosity; preferably, the viscosity of a 0.5% β-glucan aqueous solution (at 25° C.) is ≥40 mPa·s, more preferably 100-10000, and more preferably 600-2000 mPa·s;

[0065] (6) The aqueous solution of β-glucan has good stability; and / or

[0066] (7) The molecular weight of the β-glucan is ≥ 2 kD, preferably 2 kD-40000 kD, and more preferably 20 kD-20000 kD.

[0067] In another preferred embodiment, the preparation contains (a) β-glucan; and (b) a pharmaceutically, cosmetically, or device-acceptable carrier or excipient.

[0068] In another preferred embodiment, the preparation contains 0.0001-99wt%, preferably 0.001-90wt%, more preferably 0.01-50wt%, more preferably 0.05-10wt% of β-glucan, based on the total weight of the preparation.

[0069] In another preferred embodiment, the preparation further contains (c) a second active ingredient, which is an active ingredient for preventing and / or treating oral diseases other than the β-glucan.

[0070] In another preferred embodiment, the second active ingredient is selected from the following group: anticaries agents (including sodium fluoride, stannous fluoride, SMFP, erythritol and anticaries enzymes), anti-plaque and gingivitis compounds (including triclosan, zinc citrate + triclosan, sanguinarine lauryl sulfate sodium salt, chlorhexidine, domiphene, cetylpyridinium chloride), anticalculus agents (including pyrophosphate + NaF, pyrophosphate + PVM / MA / NaF, zinc citrate + sodium monofluorophosphate, zinc salt + sodium fluoride), anti-inflammatory agents (including dihydrocholesterol, allantoin, glycyrrhizin, aminoacetic acid, azulene), desensitizers (including potassium nitrate, strontium chloride, disodium hydrogen citrate), deodorants (including sodium copper chlorophyllin).

[0071] In another preferred embodiment, the preparation is used for preventing and / or treating oral diseases.

[0072] The third aspect of the present invention provides an oral cleaning product, which comprises β-glucan and an auxiliary material acceptable in cleaning products.

[0073] In another preferred embodiment, the auxiliary materials include base materials (abrasives, humectants, surfactants, thickeners, preservatives), flavor enhancers (flavoring agents, sweeteners), functional additives (colorants, corrosion inhibitors, caries prevention agents, anti-plaque and gingivitis compounds, anti-tartar agents, anti-inflammatory agents, desensitizers, deodorants), solvents (deionized water), or combinations thereof.

[0074] In another preferred embodiment, the oral cleaning product is a toothpaste, and the toothpaste comprises components selected from the following group: β-glucan (functional additive), xanthan gum (thickener), sodium carboxymethyl cellulose (thickener), hydrated silica (abrasive), sorbitol (humectant), sodium monofluorophosphate (antitatarculus agent), sodium benzoate (preservative), sodium saccharin (sweetener), SDS (surfactant), glycerin (humectant), flavor (flavoring agent), or a combination thereof.

[0075] In another preferred embodiment, the oral cleaning product is a mouthwash.

[0076] In another preferred embodiment, the oral cleaning product is chewing gum.

[0077] In a fourth aspect, the present invention provides a method for preventing and / or treating oral diseases, comprising the steps of: (a) administering β-glucan to a subject in need thereof.

[0078] In another preferred embodiment, the β-glucan is selected from the group consisting of Schizophyllum β-glucan, Lentinula edodes β-glucan, Sclerotium β-glucan, Grifola frondosa β-glucan, Pleurotus ostreatus polysaccharide, mushroom β-glucan, yeast β-glucan, oat β-glucan, or a combination thereof.

[0079] In another preferred embodiment, the β-glucan is Schizophyllum β-glucan.

[0080] In another preferred embodiment, the subjects include patients with oral diseases or normal people.

[0081] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0082] Figure 1The plaque index is determined according to the range of plaque display agent staining. Scoring criteria: 0 = no plaque on the tooth surface; 1 = scattered dot-shaped plaque at the gingival margin of the tooth neck; 2 = continuous narrow band plaque at the tooth neck with a width of no more than 1mm; 3 = plaque coverage area at the tooth neck exceeds 1mm, but less than 1 / 3 of the tooth surface; 4 = plaque coverage area at least 1 / 3 of the tooth surface, but no more than 2 / 3; 5 = plaque coverage area 2 / 3 or more of the tooth surface.

[0083] Figure 2 The photo shows the dental plaque of volunteers before and after using toothpaste containing β-glucan. It can be clearly seen from the photo that after the volunteers used toothpaste containing β-glucan for one month, their dental plaque was significantly improved and their teeth were whitened.

[0084] Figure 3 The results of scoring and statistics of the plaque on the teeth of volunteers are shown. Figure 1 The method for determining the plaque index shown in Figure 2 The dental plaque on the volunteers' teeth was scored. The statistical results showed that the dental plaque shown in the photo was significantly inhibited after using the toothpaste containing β-glucan (P value = 0.0018, ﹡﹡)

[0085] Figure 4 The basis for judging the bleeding index (BI) is shown. The scoring criteria are as follows: 0 = healthy gums, no inflammation or bleeding; 1 = inflammatory changes in gum color, no bleeding on probing; 2 = punctate bleeding after probing; 3 = bleeding on probing spreads along the gum margin; 4 = bleeding overflows the gingival sulcus; 5 = spontaneous bleeding.

[0086] Figure 5 The results show the statistical results of the volunteers' gum bleeding. Figure 4 The basis for judging the bleeding index was the recording of the volunteers' gingival bleeding. Through statistical analysis, it was found that the volunteers' gingival bleeding was significantly improved after using the toothpaste containing β-glucan. Although there was no significance in the diagnosis of bleeding on probing (insufficient sample size), from the volunteer questionnaire, 8 of the volunteers had gingival bleeding when brushing their teeth before use. However, after using the toothpaste containing β-glucan for an average of two weeks, the gingival bleeding problem was significantly improved, and 4 of the volunteers had their gingival bleeding problem completely solved after using the toothpaste for two months.

[0087] Figure 6The scoring criteria for the amount of calculus in the cervical region of the tooth are shown. Specifically, when checking calculus, the probe is inserted into the gingival sulcus on the distal surface of the tooth, and then moved along the gingival sulcus toward the mesial region, and the amount of calculus in the cervical region of the tooth is scored. 0 = no calculus above or below the gingival surface; 1 = the area covered by calculus above the gingival surface is less than 1 / 3 of the tooth surface; 2 = the area covered by calculus above the gingival surface is between 1 / 3 and 2 / 3 of the tooth surface, or there is scattered subgingival calculus in the cervical region of the tooth; 3 = the area covered by calculus above the gingival surface is more than 2 / 3 of the tooth surface, or there is continuous and thick subgingival calculus in the cervical region of the tooth.

[0088] Figure 7 The results show that the volunteers' dental calculus improved after using toothpaste containing β-glucan. The experimental results show that the volunteers' dental calculus was significantly improved after using the toothpaste for one month, and the dental calculus was further improved after two months of continuous use. In fact, the main cause of gingivitis and subsequent periodontitis is dental plaque and calculus on the teeth. If the oral hygiene is improved, the level of gingival inflammation can be directly improved.

[0089] Figure 8 Various periodontal probes are shown. Among them, Figure 8 A shows the UNC-15 probe: there are scale marks every 1 mm and bold color marks every 5 mm; Figure 8 B shows a non-metallic probe: there are scale marks every 1 mm and bold color marks every 5 mm; Figure 8 C shows the Williams probe: the scale markings are 1, 2, 3, 5, 7, 8, 9, and 10 mm; Figure 8 D shows the WHO (CPITN) probe: the tip is 0.5 mm spherical and the scale marks are 3.5, 5.5, 8.5, and 11.5 mm.

[0090] Fig. 9 The location and method of periodontal pocket detection are shown. Fig. 9 A shows six-point detection by lips and tongue; Fig. 9 B shows the lifting and plugging detection of near, middle and far points

[0091] Fig.10 A schematic diagram of interproximal probing is shown. Fig.10 A shows that the gingival valley may not be detected when the probe is inserted vertically; Fig.10 B shows that the probe is close to the contact area and inserted at a slight angle to reach the deepest part of the gingival valley.

[0092] Fig.11The results of periodontal probing are shown. Specifically, based on the changes in probing depth, the conditions of gingival inflammation before and after the volunteers used toothpaste containing β-glucan were understood. Through probing measurement of periodontal probing and data analysis, it was found that the depth of periodontal probing was significantly improved after the volunteers used toothpaste containing β-glucan. Because the normal and healthy periodontal probing depth should be 3mm or less, but the occurrence of gingival inflammation leads to a decrease in gingival attachment ability, which in turn leads to a deepening of periodontal probing depth. After the volunteers used toothpaste containing β-glucan, the periodontal probing depth decreased, indicating that the gingival attachment ability was enhanced, and it also reflects that gingival inflammation has improved.

[0093] Fig.12 The Fourier transform infrared spectrum of the Schizophyllum β-glucan prepared in Example 1 is shown.

[0094] Fig.13 The effect of toothpaste containing Schizophyllum β-glucan on tooth sensitivity index is shown.

[0095] Fig.14 The effect of a mouthwash containing Schizophyllum β-glucan on the healing period of oral ulcers was shown.

[0096] Fig.15 The effect of toothpaste containing Schizophyllum β-glucan on the healing period of oral ulcers was shown. DETAILED DESCRIPTION

[0097] After extensive and in-depth research, the inventor unexpectedly discovered for the first time that β-glucan, especially β-glucan with natural solubility, high molecular weight and high viscosity (such as Schizophyllum β-glucan), can very effectively prevent and / or treat oral diseases. Experiments show that Schizophyllum β-glucan can prevent and treat oral diseases; prevent alveolar bone loss caused by periodontal disease; regulate oral microecology, improve oral hygiene, inhibit the formation of dental plaque and remove tartar; repair damaged oral mucosa, prevent and treat oral ulcers; enhance oral mucosal immunity and resist infection by external pathogenic microorganisms. On this basis, the present invention was completed.

[0098] Terminology

[0099] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0100] As used herein, when used in reference to a specific recited numerical value, the term "about" means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0101] As used herein, the term "comprising" or "including (comprising)" may be open, semi-closed and closed. In other words, the term also includes "consisting essentially of" or "consisting of".

[0102] As used herein, the term "completely water-soluble" means that the solid form of β-glucan can be completely dissolved in water to form a β-glucan aqueous solution, that is, the solubility of β-glucan in 100g of water at 25°C is ≥0.0001g, preferably 0.01-50g, and more preferably 0.1g-10g.

[0103] As used herein, the term "naturally soluble" refers to the property of β-glucan in its natural state that it completely dissolves in water to form an aqueous solution. The "natural β-glucan" refers to β-glucan produced by natural methods (such as biological fermentation), and the β-glucan has not undergone any chemical modification, and has not undergone any physical and / or chemical and / or biological methods to break its long chain molecules and reduce its molecular weight. In another preferred embodiment, the β-glucan of the present invention is a natural β-glucan.

[0104] Oral mucosal diseases

[0105] Oral mucosal diseases refer to a variety of infectious and non-infectious diseases that occur in the oral mucosa and oral soft tissues. They can be divided into two categories, one is the disease that originates in the oral mucosa, and the other is the manifestation of systemic diseases in the oral cavity, which mainly manifests as oral mucosal damage. Common diseases include ulcers, lichen planus, leukoplakia, discoid lupus erythematosus, stomatitis, glossitis, etc. Oral mucosal diseases are more likely to occur in the cheeks, tongue, lips, soft palate and other mucosa, and can also occur at the same time as the skin.

[0106] (I) Leukoplakia The World Health Organization defined leukoplakia in 1979. Leukoplakia refers to white lesions on the oral mucosa that cannot be wiped off and cannot be diagnosed as other diseases clinically and histologically. A large number of epidemiological surveys have shown that leukoplakia often occurs on the buccal mucosa and the upper and lower lips. Leukoplakia is a precancerous lesion. Smoking is the main risk factor for leukoplakia. Leukoplakia may lead to oral cancer. According to surveys, the canceration rate is 3%-6%. Leukoplakia can be eliminated after stopping smoking.

[0107] (II) Oral lichen planus Lichen planus is a keratotic lesion that occurs on the skin and mucous membranes and is accompanied by chronic superficial inflammation. The main manifestation in the oral cavity is white linear, reticular or annular stripes on the mucous membranes. In epidemiological surveys, the evaluation index of lichen planus is mainly the prevalence rate.

[0108] Dentin sensitivity

[0109] Dentin sensitivity refers to the short and sharp pain caused by external stimuli in the exposed dentin, and it cannot be attributed to tooth defects or lesions caused by other specific reasons. Common external stimuli include temperature stimulation, blowing stimulation, mechanical stimulation or chemical stimulation. There are many explanations for the causes of dentin sensitivity, such as the neural theory, the dentin fiber conduction theory and the fluid dynamics theory. The fluid dynamics theory is currently the most accepted etiology of dentin sensitivity. Anatomically, dentin sensitivity mainly occurs when enamel is lost. After the dentin is exposed, the dentinal tubules in the dentin are exposed at both ends of the pulp cavity and the oral cavity. The fluid in the tubules flows under external stimuli, compressing the nerve fibers in the tubules and causing pain. There are many reasons for the loss of enamel, such as tooth erosion, tooth wear, and tooth neck injury. There are also some reasons that cause gingival recession and dentin exposure, which can also cause dentin sensitivity, such as periodontal disease and improper brushing.

[0110] Evaluation methods for dentin hypersensitivity

[0111] The methods for checking dentin sensitivity usually include temperature testing, cold air blowing, probe detection and pressure testing, etc. The more commonly used method is the Schiff cold air sensitivity index.

[0112] Cold air sensitivity evaluation uses an air gun on a dental unit to blow air for 1 second at a distance of 1 cm from the sensitive tooth. The blowing temperature is about 19-21°C. When blowing, place your fingers on the adjacent teeth to avoid the symptoms of the adjacent teeth affecting the accuracy of the results. The Schiff cold air sensitivity index is used for evaluation, and the scores are as follows:

[0113] 0 = teeth and subjects did not respond to air stimulation;

[0114] 1 = The subject responded to the air stimulation but did not request to stop the stimulation;

[0115] 2 = The tooth and the subject responded to the air stimulation and requested to stop the stimulation or remove the stimulation;

[0116] 3 = The tooth and the subject responded to the air stimulation, the stimulation caused pain, and requested to stop;

[0117] A low score on this parameter indicates low tooth sensitivity and vice versa.

[0118] Dental caries

[0119] Dental caries is a disease in which the hard tissue of the tooth is destroyed chronically and progressively under the influence of multiple factors, mainly bacteria. Dental caries risk factors refer to potential factors that may cause dental caries, also known as susceptibility factors or harmful factors, which are included in the bacteria, host and food factors that promote the occurrence of dental caries.

[0120] Laboratory prediction of dental caries: The test that uses cariogenic bacteria and acid products as indicators to detect the risk factors for dental caries is called the caries activity test (CAT). The more mature methods are as follows:

[0121] Dentocult SM test

[0122] Objective: To observe the number of mutans Streptococcus per milliliter (CFU / ml) in saliva to determine the activity of caries.

[0123] Kit: 5 ml culture tube with screw cap containing light saliva selection medium, standard plastic attachment plate, bacitracin paper and paraffin.

[0124] Detection method: First, let the subject chew a paraffin pellet for 1 minute, then hold the attachment plate and smear it on the back of the tongue 10 times, then immediately place the plate in a culture tube, screw on the nut, and culture at 37℃ for 48 hours, then count the density of Streptococcus mutans (blue) on the attachment plate.

[0125] Result judgment: divided into four levels: "0 and 1" <10 5 ; "2"<10 5 —10 6 ; "3">10 6 . “3” indicates high caries activity.

[0126] Tooth stains

[0127] Tooth stains are accumulated and not inherent to the teeth. They are caused by discoloration of the dental pellicle or dental plaque. These external tooth stains may include at least three formation mechanisms: ① Color-producing bacteria produce colored substances in dental plaque. ② Some colored substances are ingested through the mouth to produce color (such as pulp, tea and coffee), or caused by smoking. ③ Chemical changes in the acquired film form colored substances. In addition, tea, alcohol, cigarettes, other substances that produce colored substances in the acquired film, antibacterial agents, etc. all affect tooth stains.

[0128] β-Glucan

[0129] β-glucan is a natural polysaccharide. Many types of β-glucan can be found in the natural environment. It is usually found in the cell walls of special types of bacteria, yeasts, fungi (Ganoderma lucidum), and can also be found in the coating of higher plant seeds. There are two main methods for producing β-glucan. One is to directly extract it from cereals such as oats or fruiting fungi such as mushrooms; the other is to extract and process the fermentation liquid through liquid fermentation of fungi or bacteria to obtain β-glucan.

[0130] As used herein, "β-glucan of the present invention" and "biopolysaccharide of the present invention" are used interchangeably, and mainly refer to the β-glucan described in the first aspect of the present invention, wherein the β-glucan is selected from the following group: Schizophyllum β-glucan, Lentinula edodes β-glucan, Sclerotium β-glucan, Grifola frondosa β-glucan, Pleurotus ostreatus β-glucan, mushroom β-glucan, yeast β-glucan, oat β-glucan, or a combination thereof; preferably Schizophyllum β-glucan.

[0131] As used herein, "Schizophyllum β-glucan" refers to β-glucan derived from Schizophyllum.

[0132] In another preferred embodiment, the structure of the β-glucan is as shown in Formula I.

[0133] In another preferred embodiment, the molecular weight of the β-glucan is ≥2kD, preferably 2kD-40000kD, and more preferably 20kD-20000kD.

[0134] In another preferred embodiment, the molecular weight of the β-glucan can be 5kD-35000kD; 10kD-30000kD; 50kD-25000kD; 100kD-20000kD; 200kD-18000kD; 400kD-16000kD; 500kD-14000kD; 1000kD-12000kD; 2000kD-4000kD; 3000kD-5000kD; 4000kD-6000kD; 5000kD-7000kD; 6000kD-8000kD; 7000kD-9000kD; or 8000kD-10000kD.

[0135] In another preferred embodiment, the purity of the β-glucan is ≥70%, preferably ≥90%, more preferably ≥95%, and even more preferably ≥99%.

[0136] In another preferred embodiment, the β-glucan has good stability.

[0137] In another preferred embodiment, the β-glucan is in solid or liquid form, such as β-glucan solid particles or powder, or β-glucan aqueous solution.

[0138] In another preferred embodiment, the particle size of the β-glucan particles or powder is ≤20 mm, preferably 0.001-10 mm, more preferably 0.01-5 mm, and even more preferably 0.1-2 mm.

[0139] In another preferred embodiment, the β-glucan (granules or powder) has good water solubility and / or natural solubility.

[0140] In another preferred embodiment, the solubility of the β-glucan (granules or powder) in 25°C water (100g) is ≥0.0001g, preferably 0.01-50g, more preferably 0.1-10g.

[0141] In another preferred embodiment, the solubility of the β-glucan (granules or powder) in 25°C water (100g) may be 0.1-100g; 0.2-90g; 0.5-80g; 1-50g; or, the solubility may be 0.1-0.3g; 0.2-0.4g; 0.3-0.5g; 0.4-0.6g; 0.5-0.7g; 0.6-0.8g; 0.7-0.9g; 0.8-1g; or 1-3g; 2-4g; 3-5g; 4-6g; 5-7g; 6-8g; 7-9g; 8-10g.

[0142] In another preferred embodiment, the β-glucan solution is a solution of β-glucan in water, that is, a β-glucan aqueous solution.

[0143] In another preferred embodiment, the β-glucan (water) solution has a high viscosity; preferably, the viscosity of the β-glucan aqueous solution (at 25° C.) with a mass concentration of 0.5% is ≥40 mPa·s, more preferably 100-10000 mPa·s, and more preferably 500-2000 mPa·s.

[0144] In another preferred embodiment, the viscosity of the β-glucan aqueous solution (25°C) with a mass concentration of 0.5% can be 50-10000mPa·s; 100-9000mPa·s; 200-8000mPa·s; 300-7000mPa·s; 400-6000mPa·s; 450-5000mPa·s; 500-5000mPa·s; 550-4000mPa·s; 600-3000mPa·s; 650-2000mPa·s; 700-1500mPa·s.

[0145] In another preferred embodiment, the aqueous solution of β-glucan with a mass concentration of 1% has high clarity or high transmittance, and the transmittance of the aqueous solution of β-glucan with a mass concentration of 1% is ≥50%, preferably ≥80%, preferably ≥85%, and more preferably ≥95%;

[0146] In another preferred embodiment, the β-glucan solution has good stability.

[0147] In another preferred embodiment, the β-glucan is derived from higher plants or various bacteria and fungi.

[0148] The embodiments of the present invention specifically take the fermentation product of Schizophyllum as an example, but are not limited thereto.

[0149] Preparation or composition

[0150] The present invention provides a preparation or composition for preventing and / or treating oral diseases, wherein the preparation or composition contains (a) β-glucan; and optionally (b) a pharmaceutically, cosmetically, or device-acceptable carrier or excipient.

[0151] In another preferred embodiment, the preparation or composition contains (a) Schizophyllum β-glucan; and optionally (b) a pharmaceutically, cosmetically, or device-acceptable carrier or excipient.

[0152] In another preferred embodiment, the preparation or composition contains 0.0001-99wt%, preferably 0.001-90wt%, more preferably 0.01-50wt%, more preferably 0.05-10wt% of β-glucan (preferably Schizophyllum β-glucan), based on the total weight of the preparation or composition.

[0153] In another preferred embodiment, the mass concentration of the β-glucan in the preparation or composition is ≥1 μg / mL, specifically 1 μg / mL-200 mg / mL, or 1 μg / mL-5 mg / mL, or 1 μg / mL-1 mg / mL.

[0154] In another preferred embodiment, the preparation contains a Schizophyllum β-glucan aqueous solution, and the content of the Schizophyllum β-glucan aqueous solution in the preparation is ≥80wt%, preferably ≥90wt%, more preferably ≥95wt%, more preferably ≥99wt%, and more preferably ≥99.5wt%, based on the total weight of the preparation.

[0155] In another preferred embodiment, in the preparation, the mass concentration of Schizophyllum β-glucan in the Schizophyllum β-glucan aqueous solution is 0.0001-50wt%, preferably 0.02-10wt%, more preferably 0.05-5wt%, based on the total weight of the Schizophyllum β-glucan aqueous solution.

[0156] In a preferred embodiment, the present invention provides a toothpaste for preventing and / or treating oral diseases, the components of which are shown in Table 1:

[0157] Table 1 Toothpastes with the efficacy of preventing and / or treating oral diseases

[0158]

[0159]

[0160] The "active ingredient (first active ingredient)" in the preparation or composition of the present invention refers to the β-glucan of the present invention (preferably Schizophyllum β-glucan).

[0161] The "active ingredient (first active ingredient)", preparation and / or composition described in the present invention can be used to prevent and / or treat oral diseases.

[0162] The "second active ingredient" refers to an active ingredient for treating oral diseases that is different from the β-glucan in the present invention.

[0163] In a preferred embodiment, the second active ingredient is selected from the following group: anticaries agents (including sodium fluoride, stannous fluoride, SMFP, erythritol and anticaries enzymes), anti-plaque and gingivitis compounds (including triclosan, zinc citrate + triclosan, sanguinarine lauryl sulfate sodium salt, chlorhexidine, domiphene, cetylpyridinium chloride), anticalculus agents (including pyrophosphate + NaF, pyrophosphate + PVM / MA / NaF, zinc citrate + sodium monofluorophosphate, zinc salt + sodium fluoride), anti-inflammatory agents (including dihydrocholesterol, allantoin, glycyrrhizin, aminoacetic acid, azulene), desensitizers (including potassium nitrate, strontium chloride, disodium hydrogen citrate), deodorants (including sodium copper chlorophyllin).

[0164] "Safe and effective amount" means: the amount of active ingredient is sufficient to significantly improve the condition or symptoms without causing serious side effects.

[0165] Typically, the pharmaceutical composition contains 1-2000 mg of active ingredient per dose, more preferably, 10-200 mg of active ingredient per dose. Preferably, the "one dose" is one tablet or one injection.

[0166] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must be sufficiently pure and sufficiently low in toxicity.

[0167] "Compatibility" herein means that the components in the composition can be blended with the active ingredient of the present invention and with each other without significantly reducing the efficacy of the active ingredient.

[0168] Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0169] In another preferred embodiment, the β-glucan of the present invention can form a complex with a macromolecular compound or a polymer through a non-bonding reaction.

[0170] In another preferred embodiment, the β-glucan of the present invention can be connected to a macromolecular compound or a polymer through a chemical bond. The macromolecular compound can be a biological macromolecule such as a polymer, protein, nucleic acid, polypeptide, etc.

[0171] There is no particular limitation on the administration of the active ingredient or pharmaceutical composition of the present invention. Representative administration methods include, but are not limited to, topical application, oral administration, intratumoral administration, rectal administration, parenteral administration (intravenous, intramuscular or subcutaneous administration), and the like.

[0172] Solid dosage forms include capsules, tablets, pills, powders and granules.

[0173] In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient or carrier, such as sodium citrate or dicalcium phosphate, or with one or more of the following ingredients:

[0174] (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid;

[0175] (b) binders, for example, hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic;

[0176] (c) humectants, for example, glycerin;

[0177] (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate;

[0178] (e) a buffering solvent, such as paraffin;

[0179] (f) absorption accelerators, for example, quaternary ammonium compounds;

[0180] (g) wetting agents, such as cetyl alcohol and glyceryl monostearate;

[0181] (h) adsorbents, for example, kaolin; and / or

[0182] (i) Lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof.

[0183] In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0184] The solid dosage forms can also be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They can contain opacifying agents, and the release of the active ingredient in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes.

[0185] Liquid dosage forms include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active ingredient, the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, etc. In addition to these inert diluents, the composition may also contain adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0186] In addition to the active ingredients, suspensions may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances.

[0187] The composition may comprise a physiologically acceptable sterile aqueous or anhydrous solution, dispersion, suspension or emulsion, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0188] When using the pharmaceutical composition, a safe and effective amount of the composition of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage. For a person weighing 60 kg, the daily dosage is usually 1 to 10,000 mg, preferably 10 to 2,000 mg, and more preferably 20 to 1,000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill range of skilled physicians.

[0189] The present invention also provides a composite product, comprising:

[0190] (1) a first pharmaceutical composition, comprising (a) a first active ingredient, wherein the first active ingredient is β-glucan; and (b) a pharmaceutically, cosmetically, or device-acceptable carrier or excipient; and

[0191] (2) A second pharmaceutical composition, wherein the second pharmaceutical composition is a drug for treating oral diseases that is different from the first pharmaceutical composition.

[0192] The composition of the present invention can be administered alone or in combination with other therapeutic drugs (eg, formulated in the same pharmaceutical composition).

[0193] The pharmaceutical composition of the present invention may also be used in combination with other drugs known to treat or improve similar conditions. When the drug is administered in combination, the administration method and dosage of the original drug remain unchanged, and the pharmaceutical composition of the present invention is used simultaneously or subsequently. Drug combination also includes the use of the pharmaceutical composition of the present invention and one or more other known drugs in overlapping time periods. When the pharmaceutical composition of the present invention is used in combination with one or more other drugs, the dosage of the pharmaceutical composition of the present invention or the known drug may be lower than the dosage when they are used alone.

[0194] The main advantages of the present invention include:

[0195] (a) The β-glucan of the present invention (such as Schizophyllum β-glucan) can effectively prevent and / or treat oral diseases, such as gingivitis and periodontitis.

[0196] (b) The β-glucan of the present invention can be used to prevent alveolar bone loss, prevent tooth decay, inhibit the formation of dental plaque, remove dental calculus, resist dentin sensitivity, remove tooth stains, whiten teeth, regulate oral microecology, improve oral hygiene, prevent and treat oral ulcers, enhance oral mucosal immunity, or a combination thereof.

[0197] (c) The β-glucan of the present invention can also be used for oral postoperative care (promoting wound healing), and preventing and / or treating oral diseases in diabetic patients.

[0198] (d) The β-glucan of the present invention can be combined with other drugs for treating oral diseases to further enhance the preventive and / or therapeutic effects and have a synergistic effect.

[0199] (e) The β-glucan of the present invention (preferably Schizophyllum β-glucan) is a biological polysaccharide of pure natural origin, which is completely soluble or naturally soluble and has not been subjected to any chemical and / or physical modification or modification.

[0200] (f) The β-glucan of the present invention completely retains the three-dimensional conformation of the triple helix and has better activity in preventing and / or treating oral diseases.

[0201] (g) The β-glucan of the present invention has excellent stability and can coexist with most substances while maintaining their activity. Therefore, it has a wide range of applications and can be used in combination with other oral disease treatment drugs to improve oral hygiene and whiten teeth while preventing and treating oral diseases.

[0202] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. Unless otherwise indicated, percentages and parts are weight percentages and weight parts.

[0203] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0204] Unless otherwise specified, the materials, reagents, equipment, etc. used in the following examples can be obtained from commercial sources.

[0205] Example 1

[0206] Acquisition and determination of β-glucan

[0207] This embodiment only takes the β-glucan obtained by fermentation of Schizophyllum as an example, but is not limited thereto.

[0208] 1. Obtaining the fermentation liquid of Schizophyllum

[0209] The fermentation broth in the following examples was fermented by Schizophyllum commune Fr-1003, product catalog: 38548 TM , purchased from the American Type Culture Collection [ATCC]), as follows:

[0210] 1. Activation of Schizophyllum strains: Prepare a plate culture medium with 200 g / L potato, 30 g / L glucose, 10 g / L sodium chloride and 20 g / L agar, inoculate the Schizophyllum strains onto the plate culture medium, and culture in a constant temperature incubator at 25°C for 7 days to obtain plate mycelium;

[0211] 2. Seed activation: put a liquid culture medium made of 100 g / L potato starch, 40 g / L glucose, 2 g / L yeast extract, 2 g / L yeast extract powder and water into a shake flask, with the liquid volume being 1 / 3, inoculate the shake flask with the plate mycelium obtained in step 1, and culture it in a constant temperature shaker at 25°C and 160 rpm for 7 days as the seed liquid;

[0212] 3. Fermentation culture: add a fermentation medium made of 50 g / L glucose, 50 g / L sucrose, 5 g / L soybean powder (Shandong Zhaoyuan Wenji Food Co., Ltd.), 2 g / L yeast extract powder, 0.5 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 0.5 g / L ammonium sulfate, 6 g / L potassium nitrate and water into a fermenter and sterilize at 121°C for 15 minutes. Then inoculate the seed solution obtained in step 2 into the fermenter, and culture at a constant temperature of 25°C, 300 rpm, 4 Lpm, and aeration for 8 days to obtain a Schizophyllum fermentation liquid.

[0213] 2. Isolation and purification of β-glucan and preparation of β-glucan solution

[0214] (1) mixing the Schizophyllum fermentation liquid obtained in the first part with 4 times the volume of distilled water, and boiling at 60° C. for 8 h to obtain a boiling liquid;

[0215] (2) centrifuging the cooking liquid obtained in step (1) at 4,000 rpm for 5 min, taking the supernatant; filtering the supernatant with a 300-mesh filter cloth under negative pressure, and taking the filtered clear liquid for standby use, which is the cooking clear liquid;

[0216] (3) The filtered cooking liquid in step (2) is heated to 50°C, and 200-mesh wood activated carbon and 8-16-mesh coconut shell activated carbon are added thereto at the same time, with the added volume of each activated carbon being 1% of the volume of the cooking liquid. The mixture is stirred at 50°C and 350 rpm for 4 hours, cooled and set aside to obtain a cooking liquid mixed with activated carbon; the cooking liquid mixed with activated carbon is then filtered under negative pressure using a 300-mesh filter cloth and a SCP-321# filter plate (pore size of about 1.5 μm) in turn, and the filtered liquid is taken for use;

[0217] (4) Dissolve Solarbio lipase (L8070, enzyme activity 100-400U / mg) in physiological phosphate buffer, add the lipase solution to the filtrate prepared in step (3) at an enzyme dosage of 10U / mL, stir evenly and perform enzymolysis at 40°C for 2h; then dissolve Solarbio neutral protease (Z8030, enzyme activity>60U / mg) in physiological phosphate buffer, add the neutral protease solution to the above enzymolysis solution at an enzyme dosage of 60U / mL, stir evenly and perform enzymolysis at 40°C for 2h. After the enzymolysis is completed, heat in a 90°C water bath for 30min to inactivate the enzyme, and then perform negative pressure filtration with an SCP-321# filter plate (pore size of about 1.5μm), and take the filtered clear liquid for standby use;

[0218] (5) the clear liquid filtered in step (4) is quickly mixed with 95% edible ethanol (volume ratio is 1:3), and stirred until a precipitate is obtained; the precipitate is redissolved to the original volume, and then quickly mixed with 95% edible ethanol (volume ratio is 1:3), and stirred until a precipitate is obtained;

[0219] (6) placing the precipitate obtained in step (5) in a perforated tray and drying it in an electric oven at 40° C. until constant weight is obtained to obtain a dry product;

[0220] (7) The dried β-glucan obtained in step (6) is pulverized, 5 g of the pulverized product is weighed, dissolved in 1,000 mL of ultrapure water, and stirred at 600 rpm for 2 h until the β-glucan is fully dissolved to obtain a β-glucan solution; the β-glucan solution is filtered under negative pressure using a 5 μm filter membrane, and an appropriate amount of a cosmetic preservative is added to obtain a β-glucan solution with a high viscosity and high transmittance at a concentration of 0.5%.

[0221] The light transmittance of the beta-glucan solution with a mass concentration of 0.5% at a wavelength of 600nm (detected by a spectrophotometer) can reach 90%, and the viscosity can reach more than 600mPa·s at 40°C.

[0222] 3. Identification and Detection of β-Glucan

[0223] 1. Infrared spectroscopy identification of β-glucan

[0224] The above 0.5% β-glucan solution was identified by infrared spectroscopy according to Method C in Appendix IV of Part II of the Pharmacopoeia of the People's Republic of China (2010 Edition), and the sample dried at 105°C was scanned at full wavelength using a Fourier transform infrared spectrometer. Results The 0.5% solution obtained in step (7) of the second part of Example 1 above was a β-glucan solution.

[0225] Compared with the infrared spectrum of yeast β-glucan in the yeast β-glucan industry standard QBT 4572-2013, the functional group positions are basically consistent. Fig.12 As shown, the main functional group positions are:

[0226] 1)3301cm -1 There is a strong and broad absorption peak nearby (sugar OH bond stretching vibration absorption peak)

[0227] 2)2921cm -1 There is a weaker absorption peak nearby (sugar CH bond stretching vibration absorption peak)

[0228] 3) 886cm -1 There is a weaker absorption peak nearby (the characteristic vibration absorption peak of the β configuration of sugars)

[0229] 4)1076cm -1 There are strong absorption peaks nearby (sugar C-OH, COC stretching vibration absorption peaks)

[0230] The Fourier transform infrared spectroscopy test results show that the product obtained in this example is β-glucan.

[0231] 2. Detection of β-glucan content

[0232] The dried product obtained in step (6) of the second part of the above Example 1 was subjected to quantitative detection of β-glucan, specifically according to the method for determining the content of yeast β-glucan in the yeast β-glucan industry standard QBT 4572-2013, after grinding the obtained β-glucan dried product sample to a diameter of about 1.0 mm. The result showed that the content of β-glucan in the product was 99.23%.

[0233] 3. Hormone detection of β-glucan solution

[0234] The above 0.5% β-glucan solution was subjected to 48 hormone tests, specifically in accordance with the first method of high pressure liquid chromatography-diode array detector method for 7 components including estriol in Chapter 4, 2.4 of the "Technical Specifications for Safety of Cosmetics" (2015 edition) and the determination of 41 glucocorticoids in cosmetics by liquid chromatography / tandem mass spectrometry and thin layer chromatography in GB / T24800.2-2009. Results In the 0.5% β-glucan solution obtained in step (7) of the second part of the above Example 1, the above hormones were not detected (the results are shown in Table 2).

[0235] Table 2 48 hormone tests

[0236]

[0237]

[0238]

[0239] 4. Determination of viscosity of β-glucan solution

[0240] According to the method of step (7) of the second part of Example 1, 0.3%, 0.5%, 0.8%, and 1.0% (mass volume ratio) β-glucan aqueous solutions were prepared, and the dynamic viscosity was measured at 25°C. The results are shown in Table 3. As the β-glucan content increases, the viscosity of each sample also increases, which are 472, 740, 2150, and 3100 mPa·s, respectively.

[0241] Take commercially available dispersible (water-insoluble) yeast β-glucan particles (purchased from Wellmune), accurately weigh 2 g, add deionized water to mix, and dilute to 200 mL to obtain a yeast β-glucan suspension with a mass volume ratio of 1.0%, and test the dynamic viscosity at 25°C. Results The dynamic viscosity of the 1.0% yeast β-glucan suspension was 0 mPa·s (see Table 3).

[0242] Take commercially available soluble yeast β-glucan powder (purchased from Wellmune), accurately weigh 2 g, add deionized water to dissolve, and dilute to 200 mL to obtain a yeast β-glucan aqueous solution with a mass volume ratio of 1.0%, and test the dynamic viscosity at 25°C. The results show that the dynamic viscosity of the 1.0% yeast β-glucan aqueous solution is 0 mPa·s (see Table 3).

[0243] The above dynamic viscosity is determined as follows:

[0244] (1) Take 200 mL of the above solution sample and place it in a 250 mL beaker;

[0245] (2) Place the beaker containing the above solution / mixture sample in a water bath and keep it at 25°C for 1 h;

[0246] (3) Using a rotational viscometer, measure the dynamic viscosity of each sample at 25°C.

[0247] 5. Determination of light transmittance of β-glucan solution

[0248] According to the method of step (7) of the second part of Example 1, 0.3%, 0.5%, 0.8%, and 1.0% (mass volume ratio) of β-glucan aqueous solutions were prepared, and the transmittance of the solutions was measured at a wavelength of 600nm. The results are shown in Table 3. The transmittance of each sample was 96.5%, 93.1%, 87.5% and 81.1%, respectively.

[0249] A commercially available 1.0% oat β-glucan solution (purchased from Symrise) was taken and the light transmittance of the solution was measured at a wavelength of 600 nm. The result showed that the light transmittance was 59.7% (see Table 3).

[0250] Take commercially available dispersible yeast β-glucan particles, accurately weigh 2 g, add deionized water to mix, and make up to 200 mL to obtain a yeast β-glucan suspension with a mass volume ratio of 1.0%. The transmittance of the suspension was measured at a wavelength of 600 nm, and the result showed that the transmittance was only 1.3% (see Table 3).

[0251] Take commercially available soluble yeast β-glucan powder, accurately weigh 2 g, add deionized water to dissolve, and make up to 200 mL to obtain a yeast β-glucan aqueous solution with a mass volume ratio of 1.0%. The transmittance of the solution was measured at a wavelength of 600 nm, and the result was 68.4% (see Table 3).

[0252] The above light transmittance is measured as follows:

[0253] (1) Take 10 mL of the above solution sample and place it in a centrifuge tube;

[0254] (2) low-speed centrifugation at 1000 rpm for 1 min to remove bubbles (the two yeast β-glucan solutions / suspensions had no bubbles and were not centrifuged);

[0255] (3) Carefully transfer 3 mL into a 1 cm glass cuvette, avoiding air bubbles;

[0256] (4) Using a spectrophotometer, with deionized water as a blank reference (the transmittance of deionized water is 100%), measure the transmittance of the sample at a wavelength of 600 nm.

[0257] 6. Determination of stability of β-glucan solution

[0258] According to the method of step (7) of the second part of Example 1, 0.5%, 0.8%, and 1.0% (mass volume ratio) of β-glucan aqueous solutions were prepared, and after adding preservatives, they were placed at room temperature (not protected from light) for 24 months, and the stability of the solutions was observed, and the dynamic viscosity and transmittance of the solutions were tested. As a result, the above three solutions were all very stable, and their viscosity and transmittance did not change much, and the transmittance even increased (Table 3).

[0259] A commercially available 1.0% oat β-glucan solution was placed at room temperature (not protected from light) for 24 months to observe the stability of the solution and test the solution dynamic viscosity and transmittance. As a result, the 1.0% oat β-glucan solution was very unstable, and solids precipitated after being placed at room temperature for 3 months, resulting in its viscosity and transmittance being undetectable (Table 3).

[0260] Table 3 Dynamic viscosity data of various β-glucan solutions

[0261]

[0262] Note: a is commercially available dispersible yeast β-glucan granules (insoluble in water), b is commercially available soluble yeast β-glucan powder.

[0263] Example 2 Composition of toothpaste and preparation method thereof

[0264] (1) Toothpaste formula (%)

[0265] Table 4 Toothpaste formula for periodontal disease

[0266]

[0267]

[0268] (2) Preparation method

[0269] ① Disperse xanthan gum, sodium carboxymethyl cellulose and flavor in glycerin according to the formula table;

[0270] ②Add 20ml of 1% Schizophyllum β-glucan solution and 25g of sorbitol, stir thoroughly to dissolve;

[0271] ③ Add 20g of hydrated silica and homogenize with a homogenizer to fully disperse the hydrated silica in the system;

[0272] ④ Dissolve the components of phase B in water first and then add them into the above system and stir thoroughly;

[0273] Use a vacuum pump to remove bubbles and then repack.

[0274] Example 3 Toothpaste containing Schizophyllum β-glucan improves periodontal disease

[0275] (I) 20 volunteers were recruited to test the improvement of Schizophyllum β-glucan on periodontal disease. 20 volunteers used the toothpaste containing Schizophyllum β-glucan prepared in Example 2, and the improvement of periodontal inflammation and oral hygiene before and after use were compared.

[0276] (II) Experimental methods:

[0277] 1. Medical history collection

[0278] The basis for the diagnosis of periodontal disease is to fully inquire about the patient's periodontal disease history, conduct a careful clinical examination and find risk factors, and conduct a comprehensive analysis of the obtained data. It should be noted that periodontal disease is closely related to the whole body. During the examination and diagnosis process, the patient's general condition and changes in other parts of the oral cavity should be included.

[0279] (1) Systemic medical history

[0280] Systemic diseases related to periodontal disease include acute and chronic blood diseases (leukemia, thrombocytopenic purpura, etc.), cardiovascular diseases, diabetes or other endocrine diseases, nervous system diseases, immune deficiency, and certain genetic diseases or genetic susceptibility factors. There are also some infectious diseases such as hepatitis and AIDS.

[0281] (2) Oral history

[0282] Ask about oral diseases other than periodontal tissues, especially some diseases that can occur in both the oral mucosa and periodontal tissues, such as oral leukoplakia, lichen planus, pemphigus, pemphigoid, etc.

[0283] (3) Periodontal disease history and oral hygiene habits

[0284] The record of the chief complaint should focus on the current main symptoms, time of occurrence and changes; the current medical history should inquire in detail and record the possible causes and the development process of the disease, treatment course and efficacy.

[0285] During the examination and communication with periodontal disease patients, it is necessary to understand their oral health awareness, oral hygiene habits, and daily oral hygiene measures as detailed as possible. This has important reference value for the diagnosis, treatment planning and prognosis of periodontal disease.

[0286] (4) Family history

[0287] Ask about the periodontal health of parents, siblings or other immediate family members, especially some periodontal diseases that may be related to heredity, such as aggressive periodontitis, gingival fibroma, etc. In addition, if abnormalities in dentition, teeth and maxillofacial development are found and a genetic tendency is suspected, family history should be inquired.

[0288] 2. Oral hygiene

[0289] Oral hygiene is closely related to the health of periodontal tissues. Whether it is a first visit or a follow-up visit, the patient must first undergo this examination, which includes checking the deposition of plaque, soft plaque, tartar and stains, and whether there is food impaction, abnormal tooth structure, bad breath, etc.

[0290] Plaque inspection is a combination of visual inspection and plaque display agent, the latter of which is generally 2% alkaline fuchsin solution. The patient first rinses the mouth with water, then uses a cotton swab or a small cotton ball to dip the solution and apply it to the tooth surface near the gingival margin. After rinsing the mouth again, the stained area on the tooth surface is the attached plaque.

[0291] Plague index (PLI)

[0292] The scoring criteria for plaque index are as follows: Figure 1 It was proposed by Quigley and Hein (1962) and improved by Turesky et al. It requires applying a plaque display agent to the tooth surface, and then checking the distribution and range of the colored plaque on the tooth surface after rinsing the mouth. This scoring method is relatively objective. This index mainly reflects the oral hygiene status, checks whether the patient's self-plaque control measures are effective, and clinically observes the effects of certain anti-plaque agents. Patients can also check themselves in the mirror, so it is relatively convenient to use.

[0293] 3. Gum condition

[0294] (1) Bleeding index (BI)

[0295] The scoring criteria for the bleeding index (BI) are as follows: Figure 4As shown in Figure 1. Mazza used a modified bleeding index in a clinical study in 1981. Use a blunt-tipped periodontal probe to gently probe into the gingival sulcus or pocket bottom. After removing the probe for 30 seconds, observe whether there is bleeding and the degree of bleeding. Scoring is based on a scale of 0 to 5: 0 = healthy gums, no inflammation and bleeding; 1 = inflammatory changes in gum color, no bleeding on probing; 2 = punctate bleeding after probing; 3 = bleeding on probing spreads along the gingival margin; 4 = bleeding fills and overflows the gingival sulcus; 5 = automatic bleeding.

[0296] 4. Periodontal probing

[0297] Periodontal probing is the most important examination method in the diagnosis of periodontal disease (especially periodontitis).

[0298] (1) Purpose of periodontal probing

[0299] Understand the loss of periodontal supporting tissues, detect the formation of periodontal pockets, the depth of periodontal pockets, and the level of periodontal attachment on each surface of all teeth in the entire dentition; in addition, the root bifurcation lesions and the presence of gingival sulcus / periodontal pocket bleeding after probing should also be explored and reflected in numerical records.

[0300] (2) Tools for periodontal probing

[0301] Periodontal probing is performed clinically using a common periodontal scale probe or an electronic probe. There are many types of periodontal probes, all of which have scales, but the design features vary depending on the purpose of the examination, as reflected in the different bending angles and scale spacing of the probes. Each scale is 1mm or 2-3mm, and the working end is basically cylindrical, gradually tapering to facilitate insertion; the tip is blunt and has a diameter of 0.5mm. Figure 8 Various periodontal probes are shown

[0302] (3) Methods of periodontal probing

[0303] The periodontal probe should be used to probe along the long axis of the tooth on all surfaces, usually measuring the distal, central, and mesial of the buccal (labial) and lingual surfaces of the tooth. The probing depth of each tooth should be recorded at 6 locations. During the probing process, the probe should be moved along the bottom of the periodontal pocket in order to detect periodontal pockets of different depths on the same tooth surface. Fig. 9 shown.

[0304] When measuring periodontal pockets, the tip of the periodontal probe is always close to the tooth surface, and the probe should be parallel to the long axis of the tooth. However, due to the obstruction of the interproximal contact area, if the probe is parallel to the long axis of the tooth, it cannot enter the gingival valley area, and the deepest point of the interproximal pocket is often in the gingival valley. Therefore, when probing the interproximal surface, the probe can be allowed to be close to the contact point and slightly tilted toward the center of the interproximal surface, so that the deepest point of the interproximal pocket can be detected ( Fig.10 ).

[0305] (4) Records and evaluation indicators of periodontal probing

[0306] Periodontal probing is the main method for clinically evaluating the extent of loss of periodontal supporting tissues. Clinical examination mainly uses two indicators:

[0307] 5. Tooth mobility

[0308] Under normal circumstances, teeth have slight physiological mobility, mainly in the horizontal direction. The physiological mobility of single-root teeth is slightly greater than that of multi-root teeth. When you wake up in the morning, the mobility of teeth is the greatest, which may be due to the lack of contact between the upper and lower teeth at night, causing the teeth to protrude slightly; during the day, the teeth may be pressed into the alveolar socket due to chewing and swallowing. This subtle change is not obvious in normal periodontal tissues. In periodontitis, due to alveolar bone absorption, occlusal trauma, acute inflammation and destruction of other periodontal supporting structures, the mobility of teeth exceeds the range of physiological mobility, resulting in pathological tooth loosening.

[0309] (1) Tooth mobility check

[0310] Hold the incisal edge of the front teeth with dental forceps and shake it in the direction of the lips and tongue; for the back teeth, close the forceps and press the tip of the forceps against the The facial fossa is rocked in the buccolingual or mesiodistal direction. It is usually recorded in three degrees.

[0311] Grade I loosening exceeds the physiological range of motion, but the amplitude is within 1 mm.

[0312] Grade II loosening: The loosening amplitude is between 1 and 2 mm.

[0313] Grade III loosening: the loosening amplitude is more than 2mm.

[0314] The degree of loosening can also be determined based on the direction of loosening. Loosening in the buccal (lip) and lingual direction is degree I, loosening in both the buccal (lip) and lingual and distal-mesial directions is degree II, and loosening in the buccal (lip), distal-mesial and vertical directions is degree III.

[0315] 6. Radiographic examination

[0316] (1) X-ray image of normal periodontal tissue

[0317] Alveolar bone

[0318] The alveolar bone around the tooth root appears as a continuous radiopaque white line-like dense shadow, called a hard bone plate. The cancellous bone marrow cavity is radiopaque, and the trabeculae are radiopaque and intertwined into a mesh. Under normal circumstances, the distance from the alveolar bone top to the enamel-cementum junction is about 1 to 1.5 mm, not more than 2 mm, which is an important reference mark for determining whether there is bone resorption.

[0319] Periodontal ligament

[0320] The periodontal ligament occupies a certain gap on the X-ray film, which is called the periodontal ligament space. It is a continuous and uniform linear black transmission band with a width of about 0.18 to 0.25 mm. The increase in its width is of great significance for the diagnosis of periodontal disease.

[0321] In periodontitis, due to the destruction of the alveolar bone, the bone plate is often incomplete or disappears, and the periodontal ligament space also appears to be widened or significantly widened accordingly. In addition, the performance of the bone plate can reflect the changes of the disease to a certain extent. Before the treatment or in the active stage of the disease, the bone plate often disappears; after periodontal treatment, the bone plate is re-formed and appears as a dense continuous shadow; for patients and teeth in the maintenance period, X-rays should be taken regularly. By observing the changes in the bone plate, the recovery and stability of the disease can be indirectly judged.

[0322] X-rays mainly show the bone condition of the teeth in the mesiodistal and mesiodistal directions, while the buccal and lingual alveolar bones are not clearly displayed because they overlap with the teeth. On the standard root apex, when the distance from the alveolar ridge to the cementoenamel junction exceeds 2 mm, it can be considered that there is alveolar bone absorption.

[0323] On X-rays, the types of alveolar bone resorption are horizontal resorption and vertical resorption.

[0324] Horizontal absorption: The height of the alveolar bone decreases horizontally, and the bone absorption surface is horizontal or cup-shaped. The front teeth are mostly horizontally absorbed because of the narrow alveolar ridge.

[0325] Vertical absorption: X-rays show that there is an acute angle between the absorption surface of the bone and the tooth root, also known as angular absorption, which often occurs in the posterior teeth with wider alveolar intervals.

[0326] The degree of bone resorption is generally described by the ratio of the resorbed area to the length of the tooth root and is usually divided into three degrees.

[0327] Grade I: Alveolar bone absorption is within the top 1 / 3 of the tooth root.

[0328] Grade II: Alveolar bone resorption exceeds 1 / 3 of the root length but is within 2 / 3 of the root length, or resorption reaches 1 / 2 of the root length.

[0329] Grade III: Alveolar bone absorption is greater than 2 / 3 of the root length.

[0330] 7. Toothbrushing requirements

[0331] (a) Order of brushing: To ensure that no part is missed when brushing, it is recommended to brush in a certain order to ensure that all parts are brushed. The set position of the toothbrush is generally 1 to 3 tooth surfaces. Each part should be brushed at least 5 to 10 times, and then move to the next adjacent tooth brushing position. There should be overlap between the two brushing positions. There are generally about 9 brushing positions on the labial and buccal sides of the mandibular teeth, and 11 on the tongue.

[0332] (b) Time and amount of toothbrushing: It is recommended that the general population brush their teeth for at least 2 minutes each time and use about 1 cm of toothpaste each time.

[0333] (c) How often you brush your teeth: After brushing your teeth to remove dental plaque for several hours, plaque can reattach to the cleaned tooth surface and continue to form, especially at night when saliva secretion decreases and the oral cavity's self-cleaning function is poor, making it easier for bacteria to grow. Therefore, you should brush your teeth at least twice a day, and it is even more important to brush your teeth before going to bed at night;

[0334] (d) Areas that are difficult to brush: When brushing teeth, some areas are often neglected, such as the distal surface of the last tooth of the upper and lower jaws and the tooth surface adjacent to the edentulous area, the palatal surface of the maxillary teeth and the lingual surface of the mandibular teeth, uneven teeth and ectopic teeth. These areas are easily neglected or difficult to reach when brushing teeth. Special attention should be given when brushing teeth. Some brushing actions need to be supplemented or they need to be supplemented with pressure lines or interdental brushes. Oral cleaning should include the tongue surface. Cleaning the tongue surface can reduce the amount of food residues and microorganisms in the mouth, delay plaque formation and total plaque deposition, and help clean the entire mouth.

[0335] (III) Experimental results:

[0336] The experimental results on dental plaque are as follows Figure 2 and Figure 3 As shown in the study, after volunteers used toothpaste containing β-glucan for one month, dental plaque was significantly improved and teeth became noticeably whiter. Figure 1 The method for determining the plaque index shown in Figure 2 The dental plaque on the volunteers' teeth was scored, and the statistical results showed that the dental plaque shown in the photo was significantly inhibited after using the toothpaste containing β-glucan (P value = 0.0018, **).

[0337] The experimental results on gingival bleeding are as follows Figure 5 As shown, the volunteers' gingival bleeding was significantly improved after using toothpaste containing β-glucan. Although there was no significance in the diagnosis of bleeding on probing (insufficient sample size), from the volunteer questionnaire, 8 of the volunteers had gingival bleeding when brushing their teeth before use. However, after using toothpaste containing β-glucan for an average of two weeks, the gingival bleeding problem was significantly improved, and 4 of the volunteers had their gingival bleeding problem completely solved after using the toothpaste for two months.

[0338] The experimental results on the amount of dental plaque are as follows Figure 7As shown in the study, after using the toothpaste for one month, the tartar of the volunteers was significantly improved, and after using it for two months, the tartar was further improved. In fact, the main cause of gingivitis and subsequent periodontitis is dental plaque and tartar on the teeth. If the oral hygiene is improved, the level of gingival inflammation can be directly improved.

[0339] The experimental results of periodontal probing are as follows Fig.11 As shown in the figure, after the volunteers used toothpaste containing β-glucan, the periodontal probing depth was significantly improved. Because the normal healthy periodontal probing depth should be 3mm or less, but due to the occurrence of gingival inflammation, the gingival attachment ability decreased, which in turn led to a deeper periodontal probing depth. After the volunteers used toothpaste containing β-glucan, the periodontal probing depth decreased, indicating that the gingival attachment ability was enhanced, and it also reflected that the gingival inflammation was improved.

[0340] Example 4: Toothpaste containing Schizophyllum β-glucan improves dentin sensitivity

[0341] (I) Recruit 20 volunteers with dentin sensitivity and conduct an evaluation of dentin sensitivity.

[0342] (II) Preparation of toothpaste

[0343] (1) Formulation

[0344] Table 5 Toothpaste formulations for dentin sensitivity

[0345]

[0346]

[0347] (2) Preparation method

[0348] (a) Disperse xanthan gum, sodium carboxymethyl cellulose and flavor in glycerin according to the formula table;

[0349] (b) Add 10 ml of 1% Schizophyllum β-glucan solution and 35 g of sorbitol and stir thoroughly to dissolve;

[0350] (c) adding 18 g of hydrated silica and homogenizing with a homogenizer to fully disperse the hydrated silica in the system;

[0351] (d) Dissolve the components of phase B in water and then add them to the above system and stir thoroughly;

[0352] (e) Use a vacuum pump to remove bubbles and then repack.

[0353] (III) Experimental results:

[0354] Toothpaste containing Schizophyllum β-glucan can effectively improve dentin sensitivity. After using it for 3 days, volunteers reported that the sensitivity level decreased. After using it for 7 days, the sensitivity of cold air spray decreased by 1-2 points compared with before use. After using it for 30 days, the sensitivity of cold air spray decreased by 2-3 points compared with before use. Dentin sensitivity basically disappeared. Fig.13 shown.

[0355] Another 5 volunteers experienced tooth sensitivity and numbness due to mechanical effects when brushing their teeth. After using toothpaste containing 0.1% Schizophyllum β-glucan for 3-5 days, they felt that the tooth sensitivity disappeared.

[0356] Example 5: Schizophyllum β-glucan improves oral mucosal diseases

[0357] (I) Thirty volunteers with oral ulcers were recruited and divided into three groups. The degree of ulcer healing was observed daily by visual inspection and self-evaluation.

[0358] (II) Experimental Grouping:

[0359] Group 1 used 5 mg / mL sterile Schizophyllum β-glucan prepared in Example 1, and contained 1-20 mL in the oral cavity for at least 10 seconds; gargled 3 times a day.

[0360] Group 2 used 1 mg / mL sterile Schizophyllum β-glucan prepared in Example 1, and contained 1-20 mL in the oral cavity for at least 10 seconds; gargling 3 times a day.

[0361] Group 3 used sterile water, gargled 1-20 mL, retained it for at least 10 seconds, and took it 3 times a day.

[0362] (III) Experimental results:

[0363] After using 5 mg / mL sterile Schizophyllum β-glucan for 24 hours, the oral ulcer area of ​​volunteers in Group 1 was significantly reduced and the pain was relieved; after 48 hours, 7 volunteers were cured; after 72 hours, all 10 volunteers were cured.

[0364] In group 2, 24 hours after using 1 mg / mL sterile Schizophyllum β-glucan, the area of ​​oral ulcers was significantly reduced and the pain was relieved; after 48 hours, 4 volunteers were cured; after 72 hours, 8 volunteers were cured; after 94 hours, all 10 volunteers were cured.

[0365] Group 3 showed no significant improvement after using sterile water for 72 hours, and the ulcers of some volunteers worsened.

[0366] The specific results are shown in Table 6

[0367] Table 6 Schizophyllum β-glucan improves oral mucosal diseases

[0368]

[0369] Example 6 Mouthwash containing Schizophyllum β-glucan improves oral mucosal diseases

[0370] (I) 60 volunteers with oral mucosal diseases were recruited and divided into three groups. Group 1 was the control group, which used mouthwash without Schizophyllum β-glucan. Groups 2 and 3 were the experimental groups, which used mouthwash containing different concentrations of Schizophyllum β-glucan. The volunteers rinsed the assigned mouthwash at least twice a day, and the improvement of oral mucosal diseases was evaluated.

[0371] (ii) preparing a mouthwash containing Schizophyllum β-glucan and a mouthwash not containing Schizophyllum β-glucan.

[0372] Table 7 Mouthwash formula

[0373] Saliva ingredients Recipe 1 Recipe 2 Recipe 3 β-Glucan 0 0.05 0.1 glycerin 9 9 9 Propylene glycol 9 9 9 Sorbitol 5 5 5 Flavor (including tea extract) 0.25 0.25 0.25 Saccharin Sodium 0.1 0.1 0.1 Sodium monofluorophosphate 0.05 0.05 0.05 Sodium Benzoate 0.2 0.2 0.2 Menthol 0.03 0.03 0.03 water Add to 100 Add to 100 Add to 100

[0374] (III) Experimental results:

[0375] Mouthwash that does not contain Schizophyllum β-glucan can improve dental plaque and other tooth cleanliness indicators, but has no obvious effect on oral mucosal diseases, and the general healing period is 6.88 days; while mouthwash containing Schizophyllum β-glucan has an excellent therapeutic effect on oral mucosal diseases while cleaning the mouth, and long-term use can prevent the recurrence of oral mucosal diseases. After using mouthwash containing 0.05% Schizophyllum β-glucan, the healing period of oral ulcers is 6.18 days on average, and the healing period is shortened by 10.2%; after using mouthwash containing 0.1% Schizophyllum β-glucan, the healing period of oral ulcers is 4.5 days on average, and the healing period is shortened by 34.6%. Fig.14 shown.

[0376] Example 7 Toothpaste containing Schizophyllum β-glucan improves oral mucosal diseases

[0377] (1) Recruit 30 volunteers with oral mucosal diseases.

[0378] (II) Preparation of toothpaste

[0379] (1) Toothpaste formula

[0380] Table 8 Toothpaste formula for oral mucosal diseases

[0381]

[0382] (2) Preparation method

[0383] (a) Dispersing xanthan gum, sodium carboxymethyl cellulose and flavor in glycerin and polyethylene glycol 400 according to the formula table;

[0384] (b) Add 25 ml of 1% Schizophyllum β-glucan solution and 35 g of sorbitol and stir thoroughly to dissolve;

[0385] (c) adding 15 g of hydrated silica and homogenizing with a homogenizer to fully disperse the hydrated silica in the system;

[0386] (d) Dissolve the components of phase B in water and then add them to the above system and stir thoroughly;

[0387] (e) Use a vacuum pump to remove bubbles and then repack.

[0388] (III) Experimental results:

[0389] Toothpaste containing Schizophyllum β-glucan has an excellent therapeutic effect on oral mucosal diseases, and long-term use can prevent the recurrence of oral mucosal diseases. Fig.15 shown.

[0390] Example 8 Toothpaste and chewing gum containing Schizophyllum β-glucan improve periodontal inflammation caused by dental caries

[0391] (I) Recruit 30 volunteers with dental caries. These 30 people all have dental caries to varying degrees, and dental caries have caused gum redness and swelling, pain, difficulty chewing, and varying degrees of sensitivity to hot, cold, sour, and sweet foods. Five of the volunteers had to take anti-inflammatory and analgesic drugs to relieve the pain, but the pain recurred after stopping the drugs. It is recommended to brush your teeth every morning and evening, and use chewing gum after meals.

[0392] (II) Preparation of toothpaste and chewing gum

[0393] (1) Toothpaste formula

[0394] Table 9 Toothpaste formula for periodontal disease caused by dental caries

[0395]

[0396] (2) Preparation method

[0397] (a) Dispersing xanthan gum, sodium carboxymethyl cellulose and flavor in glycerin and polyethylene glycol 400 according to the formula table;

[0398] (b) Add 10 ml of 1% Schizophyllum β-glucan solution and 35 g of sorbitol and stir thoroughly to dissolve;

[0399] (c) adding 20 g of hydrated silica and homogenizing with a homogenizer to fully disperse the hydrated silica in the system;

[0400] (d) Dissolve the components of phase B in water and then add them to the above system and stir thoroughly;

[0401] (e) Use a vacuum pump to remove bubbles and then repack.

[0402] (III) Preparation of chewing gum

[0403] (1) Chewing gum formula

[0404] Table 10 Toothpaste formulations for dentin sensitivity

[0405] Chewing gum ingredients Element Gum Base 30％ Mixed sugar alcohols (xylitol: maltitol 7:1) 60％ 1% Schizophyllum β-glucan solution 10％

[0406] (2) Preparation method

[0407] (a) Softening of the gum base: Place the gum base in a constant temperature oven at 60°C to soften for 2 hours. Keep the gum base moist while softening and use it promptly after softening, otherwise the gum base will harden;

[0408] (b) Mixing and blending of ingredients: First, crush the xylitol and pass it through a 200-mesh sieve for later use. During operation, first, stir the softened gum base in a water bath at about 60°C for several minutes until it gradually melts into a viscous liquid, then add liquid maltitol and mix with the gum base for 6 to 8 minutes, then add xylitol powder and mix with the gum base for 5 to 6 minutes. After the sugar alcohol powder and the gum base are basically mixed, add 10g of 1% Schizophyllum β-glucan solution and stir for 3 to 5 minutes.

[0409] (c) Extrusion and molding: The prepared gum base ingredients are heated to 85-90°C for coagulation, cooled to below 40°C after 1-2 minutes, and then put into a dough press for repeated extrusion to form a ribbon-shaped candy blank with a tight texture and smooth surface. The extrusion is repeated several times; the candy sheets extruded into a certain thickness are cut and molded.

[0410] (d) Packaging: The finished chewing gum is aged for 10 to 20 hours at a temperature of about 20°C and a relative humidity of less than 55% to ensure that the chewing gum has a balanced moisture content and a moderate hardness and softness.

[0411] (III) Experimental results:

[0412] Toothpaste and chewing gum containing Schizophyllum β-glucan can effectively improve gum redness, pain, chewing difficulties and sensitivity to cold, hot, sour and sweet caused by caries, and can prevent further development of caries.

[0413] Table 11 Improvement of periodontal diseases caused by caries by toothpaste containing Schizophyllum β-glucan

[0414] Teeth sensitivity 10 people Chewing pain 26 people Loose teeth 5 people 3 people with pus discharge Positive Feedback 8 26 1 3 Negative Feedback 0 0 0 0 Invalid feedback 2 0 4 0

[0415] Among them, the improvement feedback rate of tooth pain and pus discharge caused by inflammation was 100%, and the improvement feedback rate of tooth sensitivity was 80%. However, there was no excellent improvement effect on loose teeth. Therefore, there is no way to restore loose teeth. It is recommended to go to the hospital for treatment.

[0416] Example 9: Toothpaste containing Schizophyllum β-glucan improves tooth stains

[0417] (I) Recruit 30 caries volunteers. Brush your teeth every morning and evening.

[0418] (II) Toothpaste preparation

[0419] (1) Toothpaste formula

[0420] Table 12 Toothpaste formula for improving tooth stains

[0421]

[0422]

[0423] (2) Preparation method

[0424] (a) Dispersing xanthan gum, sodium carboxymethyl cellulose and flavor in glycerin and polyethylene glycol 400 according to the formula table;

[0425] (b) Add 15 ml of 1% Schizophyllum β-glucan solution and 35 g of sorbitol and stir thoroughly to dissolve;

[0426] (c) adding 5 g of hydrated silica and 35 g of calcium carbonate, and homogenizing with a homogenizer to fully disperse the abrasive in the system;

[0427] (d) Dissolve the components of phase B in water and then add them to the above system and stir thoroughly;

[0428] (e) Use a vacuum pump to remove bubbles and then repack.

[0429] (III) Experimental results:

[0430] Toothpaste containing Schizophyllum β-glucan can improve tooth stains, but it has no obvious effect on patients with fluorosis. It has a certain effect on removing smoke stains, but it cannot completely remove them. Therefore, toothpaste containing Schizophyllum β-glucan is mainly effective for milder tooth stains. If the tooth stains are more serious, it is recommended to wash your teeth first, and then continue to use toothpaste containing Schizophyllum β-glucan to prevent tooth pigments from re-depositing.

[0431] Table 13 Improvement of tooth stains by toothpaste containing Schizophyllum β-glucan

[0432] Smoke stains 8 people 2 people with fluorosis 20 people with dark teeth Positive Feedback 5 0 18 Negative Feedback 0 0 0 Invalid feedback 3 2 2

[0433] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A use of Schizophyllum β-glucan, characterized in that: As an active ingredient, it is used to prepare a preparation or composition, and the preparation or composition is used to remove tooth stains and whiten teeth. The structure of the Schizophyllum β-glucan is shown in Formula I. Wherein, l is 1; m is 0; n is an integer from 100 to 10000; The branching degree of the β-glucan is 0.1-0.5; The molecular weight of the β-glucan is 3000kD-6000kD; The solubility of the β-glucan in 100g of water at 25°C is 0.0001g-10g; The viscosity of the aqueous solution of the β-glucan at a mass concentration of 0.5% is 500-2000 mPa·s at 25°C; and The light transmittance of the aqueous solution of the beta-glucan with a mass concentration of 1% is ≥80%.

2. The use according to claim 1, characterized in that The branching degree of the β-glucan is 0.2-0.

4.

3. The use according to claim 1, characterized in that The molecular weight of the β-glucan is 3000kD-5000kD.

4. The use according to claim 1, characterized in that The solubility of the β-glucan particles or powder in 100g of water at 25°C is 0.1-10g.

5. The use according to claim 1, characterized in that The β-glucan solution has good stability.

6. Use of a preparation for preparing a composition for removing tooth stains and whitening teeth, characterized in that: The preparation is composed of: (a) β-glucan as an active ingredient; and (b) a pharmaceutically, cosmetically, or device-acceptable carrier or excipient. The β-glucan is Schizophyllum β-glucan, and its structure is shown in Formula I. Wherein, l is 1; m is 0; n is an integer from 100 to 10000; The branching degree of the β-glucan is 0.1-0.5; The molecular weight of the β-glucan is 3000kD-6000kD; The solubility of the β-glucan in 100g of water at 25°C is 0.0001g-10g; The viscosity of the aqueous solution of the β-glucan at a mass concentration of 0.5% is 500-2000 mPa·s at 25°C; and The light transmittance of the aqueous solution of the beta-glucan with a mass concentration of 1% is ≥80%.

7. The use according to claim 6, characterized in that The preparation comprises a β-glucan aqueous solution, wherein the mass concentration of the β-glucan is 0.02-10 wt %, based on the total weight of the β-glucan aqueous solution.

8. The use according to claim 6, characterized in that The preparation comprises a β-glucan aqueous solution, wherein the mass concentration of the β-glucan is 0.05-5wt%, based on the total weight of the β-glucan aqueous solution.

9. An oral cleaning product, characterized in that: The oral cleaning product comprises β-glucan as an active ingredient and an auxiliary material acceptable in cleaning products. The β-glucan is Schizophyllum β-glucan, and its structure is shown in Formula I. Wherein, l is 1; m is 0; n is an integer from 100 to 10000; The branching degree of the β-glucan is 0.1-0.5; The molecular weight of the β-glucan is 3000kD-6000kD; The solubility of the β-glucan in 100g of water at 25°C is 0.0001g-10g; The viscosity of the aqueous solution of the β-glucan at a mass concentration of 0.5% is 500-2000 mPa·s at 25°C; and The light transmittance of the aqueous solution of the beta-glucan with a mass concentration of 1% is ≥80%, and the auxiliary material is selected from the following group: base material, flavor enhancer, functional additive, solvent, or a combination thereof.

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