CN112402443A

CN112402443A - Biological polysaccharide for preventing and treating oral diseases and application thereof

Info

Publication number: CN112402443A
Application number: CN202010849985.0A
Authority: CN
Inventors: 郭宏亮; 叶榛; 吴佳迪; 庄秀园; 王轩; 朱斌念
Original assignee: Zhejiang Gllion Bioscience Co ltd
Current assignee: Zhejiang Gllion Bioscience Co ltd
Priority date: 2019-08-22
Filing date: 2020-08-21
Publication date: 2021-02-26

Abstract

The invention provides a biological polysaccharide for preventing and treating oral diseases and application thereof. In particular, the invention provides the use of beta-glucan for the preparation of a formulation or composition for the prevention and/or treatment of an oral disease. The beta-glucan has the functions of obviously relieving and improving oral diseases and controlling the occurrence and development of the oral diseases, and can be used for preparing oral cleaning products.

Description

Biological polysaccharide for preventing and treating oral diseases and application thereof

Technical Field

The invention relates to the technical field of biology, in particular to a biological polysaccharide with the effects of preventing and treating oral diseases and application thereof.

Background

The mouth is the initial part of the digestive tract, one of the vital organs of the human body, which, in addition to its own function, is a "window" of general health. Some systemic diseases may manifest themselves in the oral cavity, even when characterized first in the oral cavity. Periodontal tissue and oral mucosal system can cause various diseases, such as acute and chronic non-specific infectious inflammation, some specific infection (such as syphilis, tuberculosis, AIDS, etc.), wound, deformity, tumor, and some systemic diseases (including some congenital diseases) also can occur in oral cavity.

Oral mucosal disease refers to a variety of infectious and non-infectious diseases that occur in the oral mucosa and soft tissues of the oral cavity. The disease can be divided into two main categories, one is the disease which is originally generated in the oral mucosa, and the other is the manifestation of systemic disease in the oral cavity, mainly manifested as oral mucosa damage. The common diseases include ulcer, lichen planus, leukoplakia, discoid lupus erythematosus, stomatitis, glossitis, etc. The oral mucosa is usually found on the mucosa of cheek, tongue, lip, soft palate, etc., and can also attack the disease with the skin. In addition, diseases such as oral ulcer are difficult to cure radically and easy to attack repeatedly. At present, the treatment means mainly comprise anti-inflammation, pain relief and ulcer healing promotion, and antibiotics, hormone drugs and vitamins, such as tetracycline, dexamethasone, betamethasone, vitamin C, vitamin B complex, watermelon crystal and the like, are commonly used.

Periodontal disease includes two major diseases, gum disease and periodontitis. The most common of gum diseases is chronic inflammation caused by plaque, i.e., gingivitis, and some gum diseases that are 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 with incompletely identical pathogenic factors and body reactivity, different disease courses, and different responses to treatment. China is a big country with high incidence of periodontal diseases, 80-90% of adults suffer from periodontal diseases with different degrees, but most people lack oral health care consciousness, have poor oral hygiene, and lack understanding of periodontal diseases, so that the people have to pull out teeth or wait for loose teeth to fall off automatically when taking care of the opportunity. The medicine for treating periodontal disease mainly comprises antibiotic medicine and anti-inflammatory medicine, such as nitroimidazole, tetracycline, penicillin, macrolides medicine, non-steroidal anti-inflammatory medicine, etc.

The oral disease treatment drug has an unobvious treatment effect or has certain side effects, so that the development of a novel drug for preventing and treating oral diseases has very important significance.

Disclosure of Invention

The invention aims to provide a biological polysaccharide with the efficacy of preventing and treating oral diseases and application thereof.

In a first aspect of the invention, there is provided the use of β -glucan for the preparation of a formulation or composition for the prevention and/or treatment of an oral disease.

In another preferred embodiment, the oral diseases include periodontal diseases, tooth body diseases and oral mucosa diseases.

In another preferred embodiment, the periodontal disease comprises gingivitis, periodontitis.

In another preferred embodiment, the tooth body disease includes caries, dentin sensitivity, tooth stains.

In another preferred embodiment, the oral mucosal disease comprises leukoplakia, oral lichen planus, oral ulcer, discoid lupus erythematosus, stomatitis, glossitis.

In another preferred embodiment, the formulation or composition is used for preventing alveolar bone loss, preventing dental caries, inhibiting the formation of dental plaque, removing dental calculus, resisting dentinal hypersensitivity, improving gingival bleeding, removing tooth stain, whitening teeth, regulating oral micro-ecology, improving oral hygiene, preventing and treating canker sores, restoring and enhancing oral mucosal immunity, or a combination thereof.

In another preferred embodiment, the formulation or composition is also used for post-oral care (promoting wound healing), prevention and/or treatment of oral diseases in diabetic patients.

In another preferred example, the alveolar bone loss includes alveolar bone loss caused by periodontal disease and alveolar bone loss of a dental implant.

In another preferred embodiment, the beta-glucan is beta-D-glucan.

In another preferred embodiment, the beta-glucan is a beta-1, 3-glucan, preferably a beta-1, 3-glucan having beta-1, 6-branches.

In another preferred example, the structure of the beta-glucan is shown as the formula I,

wherein l is an integer of 0 or more, preferably 0 to 50, preferably 0 to 10, more preferably 0 to 3, more preferably 1 to 2, more preferably 1; m is an integer of 0 or more, preferably 0 to 19, more preferably 0 to 4, still more preferably 0 to 1, still more preferably 0; n is an integer of 3 or more, preferably 30 to 60000, more preferably 100-10000.

In another preferred embodiment, the beta-glucan has a Degree of Branching (DB) of 0.02 to 0.8, preferably 0.1 to 0.5, preferably 0.2 to 0.4.

In another preferred embodiment, the beta-glucan comprises a beta-glucan having a triple helix stereo structure.

In another preferred embodiment, the beta-glucan of the triple helix stereo structure is contained in an amount of 80%, 90%, 95% based on the total molar amount of the beta-glucan.

In another preferred embodiment, the β -1, 3-backbone of the β -glucan is the main body of the triple helix steric structure.

In another preferred embodiment, the beta-1, 6-branch of the beta-glucan is located outside the triple helix steric structure.

In another preferred embodiment, the molecular weight of said beta-glucan is 2kD or more, preferably 2kD to 40000kD, more preferably 20kD to 20000 kD.

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

In another preferred embodiment, the β -glucan is selected from the group consisting of: schizophyllum commune beta-glucan, shiitake mushroom beta-glucan, sclerotium rolfsii beta-glucan, grifola frondosa beta-glucan, pleurotus ostreatus polysaccharide, mushroom beta-glucan, yeast beta-glucan, oat beta-glucan, or a combination thereof.

In another preferred embodiment, the beta-glucan is Schizophyllum commune beta-glucan.

In another preferred example, the lentinus edodes beta-glucan is a beta-glucan with 2 beta-1, 6-branches per 5 beta-1, 3-main chains and 1 glucose residue per branch.

In another preferred embodiment, the purity of the beta-glucan is 70% or more, preferably 90% or more, more preferably 95% or more, and still more preferably 99% or more.

In another preferred embodiment, the beta-glucan has good stability.

In another preferred embodiment, the beta-glucan is in a solid form or a liquid form, such as solid particles or powder of beta-glucan, or an aqueous solution of beta-glucan.

In another preferred embodiment, the beta-glucan particles or powder have a particle size of 20mm or less, preferably 0.001 to 10mm, more preferably 0.01 to 5mm, still more preferably 0.1 to 2 mm.

In another preferred embodiment, the beta-glucan is a fully water-soluble beta-glucan.

In another preferred embodiment, the beta-glucan (granule or powder) has good water solubility and/or natural solubility.

In another preferred embodiment, the solubility of the beta-glucan (granules or powder) in water (100g) at 25 ℃ is 0.0001g or more, preferably 0.01 to 50g, more preferably 0.1 to 10 g.

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

In another preferred example, the beta-glucan solution is a solution of beta-glucan in water, i.e., an aqueous solution of beta-glucan.

In another preferred example, the beta-glucan (water) solution has a high viscosity; preferably, the viscosity of the aqueous solution of beta-glucan at a mass concentration of 0.5% (at 25 ℃) is not less than 40 mPas, more preferably 100-10000 mPas, still more preferably 500-2000 mPas.

In another preferred example, the viscosity of the beta-glucan aqueous solution (25 ℃) with the mass concentration of 0.5% can be 50-10000 mPa.s; 100-9000 mPas; 200-; 300-7000 mPas; 400-6000 mPas; 450-; 500-5000 mPas; 550-4000mPa s; 600-3000mPa · s; 650-2000 mPas; 700-1500 mPas.

In another preferred embodiment, the aqueous solution of beta-glucan with the mass concentration of 1% has high clarity or high light transmittance, and the light transmittance of the aqueous solution of beta-glucan with the mass concentration of 1% is greater than or equal to 50%, preferably greater than or equal to 80%, preferably greater than or equal to 85%, and more preferably greater than or equal to 95%;

in another preferred embodiment, the beta-glucan solution has good stability.

In a second aspect of the invention, there is provided a formulation comprising β -glucan.

In another preferred embodiment, the preparation is Schizophyllum commune beta-glucan, and the beta-glucan has good water solubility, natural solubility and/or resolubility.

In another preferred embodiment, the β -glucan in the formulation is in solid form or liquid form.

In another preferred embodiment, the formulation comprises (a) β -glucan; and optionally (b) a pharmaceutically, cosmetically, or device acceptable carrier or excipient.

In another preferred embodiment, the formulation comprises (a) Schizophyllum commune beta-glucan; and optionally (b) a pharmaceutically, cosmetically, or device acceptable carrier or excipient.

In another preferred embodiment, the formulation contains 0.0001-99 wt%, preferably 0.001-90 wt%, more preferably 0.01-50 wt%, more preferably 0.05-10 wt% of beta-glucan, based on the total weight of the formulation.

In another preferred embodiment, the mass concentration of the beta-glucan in the preparation is more than or equal to 1 mu g/mL, and specifically can be 1 mu g/mL-200mg/mL, or 1 mu g/mL-5mg/mL, or 1 mu g/mL-1 mg/mL.

In another preferred embodiment, the composition or formulation is in the form of a solid dosage form, a semi-solid dosage form, or a liquid dosage form.

In another preferred embodiment, the formulation may be formulated as an oral cleaning product, preferably a toothpaste, a mouthwash, an oral gel, a chewing gum.

In another preferred embodiment, the formulation (gargle) may be a liquid formulation, aerosol, powder, tablet, concentrate.

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

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

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

In another preferred embodiment, the β -glucan in the formulation has one or more characteristics selected from the group consisting of:

(1) the purity of the beta-glucan is more than or equal to 70 percent, preferably more than or equal to 90 percent, more preferably more than or equal to 95 percent, and more preferably more than or equal to 99 percent;

(2) the beta-glucan has good water solubility, redissolution and/or natural solubility;

(3) the solubility of the beta-glucan (solid particles or powder) in water at 25 ℃ is more than or equal to 0.0001g/100g of water, preferably 0.01-50g/100g of water, more preferably 0.1-10 g/100g of water;

(4) the aqueous solution of beta-glucan has high clarity or high light transmittance; preferably, the light transmittance of the beta-glucan aqueous solution with the mass concentration of 1% is more than or equal to 50%, preferably more than or equal to 80%, preferably more than or equal to 85%, and more preferably more than or equal to 95%;

(5) the beta-glucan solution has high viscosity; preferably, the viscosity of the beta-glucan aqueous solution (at 25 ℃) with the mass concentration of 0.5 percent is more than or equal to 40 mPas, more preferably 100-10000, more preferably 600-2000 mPas;

(6) the aqueous solution of the beta-glucan has good stability; and/or

(7) The molecular weight of the beta-glucan is more than or equal to 2kD, preferably 2kD-40000kD, and more preferably 20kD-20000 kD.

In another preferred embodiment, the formulation comprises (a) β -glucan; and (b) a pharmaceutically, cosmetically, or device acceptable carrier or excipient.

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

In another preferred embodiment, the second active ingredient is selected from the group consisting of: anticaries agents (including sodium fluoride, stannous fluoride, SMFP, erythritol, and anticariogenic enzymes), antiplaque and gingivitis compounds (including triclosan, zinc citrate + triclosan, sanguinarine sodium lauryl sulfate, chlorhexidine, domiphen bromide, cetylpyridinium chloride), anticalculus agents (including pyrophosphate + NaF, pyrophosphate + PVM/MA/NaF, zinc citrate + sodium monofluorophosphate, zinc salts + sodium fluoride), anti-inflammatory agents (including dihydrocholesterol, allantoin, glycyrrhizin, glycine, azulene), desensitizing agents (including potassium nitrate, strontium chloride, disodium hydrogen citrate), and deodorizing agents (including sodium copper chlorophyllin).

In another preferred embodiment, the formulation is for use in the prevention and/or treatment of oral diseases.

In a third aspect of the invention, an oral cleaning product is provided, wherein the oral cleaning product comprises beta-glucan and an acceptable auxiliary material in the cleaning product.

In another preferred embodiment, the adjuvants include a base (abrasive, humectant, surfactant, thickener, preservative), flavor enhancer (flavoring agent, sweetener), functional additive (colorant, corrosion inhibitor, anticaries agent, antiplaque and gingivitis compound, anticalculus agent, anti-inflammatory agent, desensitizing agent, deodorant), solvent (deionized water), or combinations thereof.

In another preferred embodiment, the oral cleaning product is a toothpaste, and the toothpaste comprises a component selected from the group consisting of: beta-glucan (functional additive), xanthan gum (thickener), sodium carboxymethylcellulose (thickener), hydrated silica (abrasive), sorbitol (humectant), sodium monofluorophosphate (anticalculus agent), sodium benzoate (preservative), sodium saccharin (sweetener), SDS (surfactant), glycerin (humectant), flavor (fragrance), or a combination thereof.

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

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

In a fourth aspect of the present invention, there is provided a method for preventing and/or treating oral diseases, comprising the steps of: (a) administering beta-glucan to a subject in need thereof.

In another preferred embodiment, the subject comprises a patient with oral disease or a normal population.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.

Drawings

Fig. 1 shows the determination of the plaque index from the extent of staining by the plaque disclosing agent. Scoring criteria: 0, no plaque exists on the tooth surface; 1, scattered punctate bacterial plaque is arranged at the gingival margin of the neck part; 2, the width of the continuous narrow-band dental plaque at the neck of the tooth is not more than 1 mm; 3 ═ plaque coverage area of the neck of the tooth exceeds 1mm, but is less than the tooth surface 1/3; 4, the plaque coverage area at least occupies the tooth surface 1/3 but does not exceed 2/3; the 5-dental plaque coverage area is more than the tooth surface 2/3 or 2/3.

Fig. 2 shows the condition of dental plaque before and after the volunteer applied the toothpaste containing β -glucan, and it is apparent from the photographs that the dental plaque was significantly improved and the teeth were also whitened after the volunteer applied the toothpaste containing β -glucan for one month.

Fig. 3 shows the results of scoring and statistics of plaque on the volunteer's teeth. Specifically, the plaque on the teeth of the volunteers in fig. 2 was scored according to the method for determining plaque index shown in fig. 1, and statistical results showed that the plaque shown on the photograph gave a significant inhibitory effect after using the toothpaste containing β -glucan (P value ═ 0.0018, ﹡ ﹡)

Fig. 4 shows the basis for the determination of the Bleeding Index (BI). Wherein the scoring criteria are: 0 is healthy gum, no inflammation and bleeding; 1, the color of the gum is changed in an inflammatory way, and bleeding is not caused in probing; 2, a little bleeding after probing; probing bleeding spreads along the gingival margin; 4, the bleeding flows to the full and overflows the gingival sulcus; 5 automatic bleeding.

Fig. 5 shows the statistical results of gingival bleeding of volunteers. Specifically, the dentist judges the basis of the bleeding index according to fig. 4, and records the gingival bleeding of the volunteers, and through statistical analysis, the gingival bleeding of the volunteers is obviously improved after the volunteers use the toothpaste containing the beta-glucan, and although the gingival bleeding is not significant (the number of samples is insufficient) in the diagnosis of the exploratory bleeding, the gingival bleeding problem is obviously improved in the questionnaire of the volunteers, wherein 8 volunteers are accompanied by the gingival bleeding before using the toothpaste, but the gingival bleeding problem is obviously improved after using the toothpaste containing the beta-glucan for two weeks on average, and the gingival bleeding problem is completely solved after using the toothpaste for two months by 4 volunteers.

Figure 6 shows the scoring criteria for the amount of tartar in the cervical region of the tooth. Specifically, when examining the dental calculus, the probe was inserted into the far-middle surface gingival sulcus and then moved medially along the gingival sulcus, scored according to the amount of dental calculus in the neck portion. 0 is no tartar on and under the gum; 1, the covered area of the supragingival dental calculus occupies the tooth surface 1/3 for one time; 2, the area covered by the supragingival dental calculus is between the tooth surfaces 1/3 and 2/3, or the neck of the tooth is provided with the scattered subgingival dental calculus; the 3-supragingival calculus covers more than the tooth surface 2/3, or the neck of the tooth has continuous and thick subgingival calculus.

Figure 7 shows the improvement of tartar in volunteers after use of a toothpaste containing beta-glucan. The experimental results show that the tartar is remarkably improved after the volunteer uses the toothpaste for one month, and the condition of the tartar is further improved after the volunteer uses the toothpaste for two months. In fact, gingivitis and the subsequent development of periodontitis is primarily caused by plaque and calculus on the teeth, which directly improves the level of gingival inflammation if oral hygiene is improved.

Figure 8 shows various periodontal probes. Among them, FIG. 8A shows the UNC-15 probe: scale marks are arranged every 1mm, and thickened color marks are arranged every 5 mm; fig. 8B shows a non-metallic probe: scale marks are arranged every 1mm, and thickened color marks are arranged every 5 mm; FIG. 8C shows a Williams probe: the scale marks are respectively 1, 2, 3, 5, 7, 8, 9 and 10 mm; FIG. 8D shows WHO (CPITN) probes: the tip is 0.5mm sphere, and the scale marks are 3.5, 5.5, 8.5 and 11.5mm respectively.

Figure 9 shows the periodontal pocket probing sites and methods. Wherein, FIG. 9A shows probing by lip and tongue at six points; FIG. 9B shows three points of lifting-inserting type detection for near, middle and far

Figure 10 shows a schematic diagram of an adjacent visit. Wherein fig. 10A shows that the vertical insertion probe may not probe the gingival valley; fig. 10B shows the probe approaching the contact area, slightly angled into the trough, and reaching the deepest part of the gingival valley.

Figure 11 shows the results of periodontal probing. Specifically, according to the change of the probing depth, the conditions of gingival inflammation before and after the volunteer uses the toothpaste containing the beta-glucan are known, and the probing measurement of periodontal probing and data analysis show that the depth of periodontal probing is obviously improved after the volunteer uses the toothpaste containing the beta-glucan. Since the depth of the normal healthy periodontal probing should be 3mm or less, but the gingival attachment ability is decreased due to the occurrence of gingival inflammation, which in turn leads to the depth of the periodontal probing to be increased. After the volunteers used the toothpaste containing beta-glucan, the periodontal probing depth was reduced, indicating that the gingival attachment ability was enhanced and possibly that the gingival inflammation was improved.

FIG. 12 shows a Fourier transform infrared spectrum of Schizophyllum commune beta-glucan prepared in example 1.

Figure 13 shows the effect of a toothpaste containing Schizophyllum commune beta-glucan on the tooth sensitivity index.

Figure 14 shows the effect of the healing cycle of a schizophyllum beta-glucan containing mouthwash dental ulcer.

Figure 15 shows the effect of a toothpaste containing Schizophyllum commune beta-glucan on the healing cycle of canker sores.

Detailed Description

The present inventors have made extensive and intensive studies and, for the first time, have unexpectedly found that beta-glucan, particularly beta-glucan having a naturally soluble property, a high molecular weight and a high viscosity (e.g., Schizophyllum commune beta-glucan), is very effective in preventing and/or treating oral diseases. Experiments show that the Schizophyllum commune beta-glucan can prevent and treat oral diseases; preventing alveolar bone loss caused by periodontal diseases; regulating oral microecology, improving oral hygiene, inhibiting dental plaque formation and removing dental calculus; repairing damaged oral mucosa, and preventing and treating oral ulcer; improving the immunity of oral mucosa and resisting the infection of external pathogenic microorganisms. On the basis of this, the present invention has been completed.

Description of the terms

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.

As used herein, the term "about" when used in reference to a specifically recited value 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 99 and 101 and all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

As used herein, the term "comprising" or "includes" can be open, semi-closed, and closed. In other words, the term also includes "consisting essentially of …," or "consisting of ….

As used herein, the term "completely water-soluble" means that the beta-glucan in the solid form can be completely dissolved in water to form an aqueous beta-glucan solution, i.e., the solubility of beta-glucan in 100g of water at 25 ℃ is 0.0001g or more, preferably 0.01 to 50g, more preferably 0.1g to 10 g.

As used herein, the term "naturally soluble" refers to the property of β -glucan in its native state that is itself completely soluble in water to form an aqueous solution. By "native state β -glucan" is meant β -glucan produced (e.g., biofermented) using natural processes without any chemical modification and without any physical and/or chemical and/or biological process to break down long chain molecules and reduce their molecular mass. In another preferred embodiment, the beta-glucan of the present invention is a natural beta-glucan.

Diseases of oral mucosa

Oral mucosal disease refers to a variety of infectious and non-infectious diseases that occur in the oral mucosa and soft tissues of the oral cavity. The disease can be divided into two main categories, one is the disease which is originally generated in the oral mucosa, and the other is the manifestation of systemic disease in the oral cavity, mainly manifested as oral mucosa damage. The common diseases include ulcer, lichen planus, leukoplakia, discoid lupus erythematosus, stomatitis, glossitis, etc. The oral mucosa is usually found on the mucosa of cheek, tongue, lip, soft palate, etc., and can also attack the disease with the skin.

The white spot world health organization defined the definition of white spots in 1979, which refers to white lesions that occur on the oral mucosa, cannot be wiped off, and cannot be clinically and histologically diagnosed as other diseases. A large number of epidemiological surveys show that the parts where white spots occur are mostly seen in the buccal mucosa and the upper and lower lips. The white spot is a precancerous lesion, smoking is a main risk factor for causing the white spot, the white spot has the possibility of causing oral cancer, the canceration rate of the white spot is 3% -6% by investigation, and the white spot can be eliminated after smoking is stopped.

(II) oral lichen planus is a keratinizing lesion with chronic superficial inflammation that occurs on the skin and mucous membranes. The oral cavity is mainly represented by white linear, reticular or annular stripes on the mucous membrane. In epidemiological investigations, the evaluation index of lichen planus is mainly the prevalence.

Dentin sensitivity

Dentin sensitivity refers to short and sharp pain of exposed dentin caused by external stimulus, and cannot be attributed to tooth body defects or lesions caused by other specific reasons. Common external stimuli include temperature, blow-up, mechanical or chemical stimuli. The reasons for the development of dentinal hypersensitivity are explained in several ways, such as the theory of neurology, the theory of dentinal fibre transmission and the theory of hydrodynamics. The theory of hydrodynamics is now the most accepted etiology of dentinal sensitivity. Anatomically, dentin sensitivity mainly appears in enamel loss, after dentin exposure, dentin tubules positioned in dentin are exposed at both ends of a pulp cavity and a mouth, and liquid in the tubules flows under external stimulation to press nerve fibers in the tubules to generate pain. The loss of enamel is caused by many reasons, such as tooth erosion, tooth abrasion, and tooth neck damage. Dentinal hypersensitivity may also be caused by gingival recession and dentinal exposure due to several causes, such as periodontal disease, improper brushing, etc

Method for evaluating dentin sensitivity

Methods for checking dentinal sensitivity generally employ temperature testing, cold air blowing, probe probing, pressure testing, and the like. A more common method is the Schiff cold air sensitivity index.

Cold air blowing sensitivity evaluation an air gun of a dental comprehensive treatment table is used for blowing for 1 second at a distance of 1cm away from sensitive teeth, the blowing temperature is about 19-21 ℃, and fingers are placed on adjacent teeth during blowing so as to avoid the symptoms of the adjacent teeth from influencing the accuracy of results. The results were evaluated using the Schiff cold air sensitivity index and scored as follows:

0 ═ teeth and subjects did not respond to air stimuli;

1 ═ tooth and subject responded to air stimulation, but did not request cessation of stimulation;

2 ═ tooth and subject respond to air stimuli, requesting cessation or removal of stimuli;

3 ═ tooth and subject responded to air irritation, which caused pain, requesting cessation;

a low score for this parameter indicates low tooth sensitivity and vice versa.

Dental caries

Caries is a disease in which the hard tissues of the tooth body undergo chronic progressive destruction under the influence of various factors, mainly bacteria. The caries risk factor refers to a potential factor which may cause caries, also called a susceptible factor or a harmful factor, and is included in bacteria, hosts and food factors which promote caries.

Laboratory prediction of caries: the test for detecting caries risk factors using cariogenic bacteria and acidic products as indicators is called the Caries Activity Test (CAT). The currently more mature methods are as follows:

dentocult SM test

The purpose is as follows: caries activity was assessed by observing the number of Streptococcus mutans per milliliter colony forming unit (CFU/ml) in saliva.

The kit comprises: 5ml of nutted culture tubes containing light saliva selection medium, standard plastic attachment plates, bacitracin paper and paraffin.

The detection method comprises the following steps: the test subject chews a paraffin wax pill for 1 minute, turns and coats the attachment plate on the back of the tongue for 10 times, immediately places the plate in a culture tube, screws a nut, cultures for 48 hours at 37 ℃, and counts the density of streptococcus mutans (blue) on the attachment plate.

And (5) judging a result: four stages are divided: "0 and 1" < 10⁵；“2”＜10⁵—10⁶；“3”＞10⁶. "3" is high cariogenic activity.

Tooth stains

Dental stains are accumulated, not intrinsic to the teeth, and are caused by discoloration of the pellicle or plaque. These extrinsic tooth stains may include at least three mechanisms of formation: colour-producing bacteria produce coloured material in dental plaque. ② some produce color (such as fruit pulp, tea and coffee) by oral ingestion of some colored material, or by smoking. And forming colored matter through chemical change of the obtained film. In addition, tea, wine, cigarettes, other substances that impart color to the acquired film, antimicrobial agents, etc., all affect tooth staining.

Beta-glucan

Beta-glucan is a natural polysaccharide found in a wide variety of natural environments, usually in the cell walls of specific species of bacteria, yeasts, fungi (Ganoderma lucidum) and also in the coating of higher plant seeds. The production method of beta-glucan mainly comprises two methods, one is that the beta-glucan is directly extracted from grain such as oat or fruiting body fungi such as mushroom; and the other is to obtain the beta-glucan by extracting and processing fermentation liquor through liquid fermentation of fungi or bacteria.

As used herein, "beta-glucan of the invention", "biopolysaccharide of the invention" are used interchangeably and refer primarily to beta-glucan according to the first aspect of the invention, which is selected from the group consisting of: schizophyllum commune beta-glucan, shiitake mushroom beta-glucan, sclerotium rolfsii beta-glucan, grifola frondosa beta-glucan, pleurotus ostreatus polysaccharide, mushroom beta-glucan, yeast beta-glucan, oat beta-glucan, or a combination thereof; preferably Schizophyllum commune beta-glucan.

As used herein, "schizophyllum beta-glucan" refers to beta-glucan derived from schizophyllum.

In another preferred example, the structure of the beta-glucan is shown as the formula I.

In another preferred embodiment, the beta-glucan has good stability.

in another preferred embodiment, the beta-glucan solution has good stability.

In another preferred example, the beta-glucan is derived from higher plants or various bacteria and fungi.

The embodiment of the present invention specifically takes the fermentation product of Schizophyllum commune as an example, but is not limited thereto.

Formulations or compositions

The present invention provides an agent or composition for preventing and/or treating oral diseases, which comprises (a) β -glucan; and optionally (b) a pharmaceutically, cosmetically, or device acceptable carrier or excipient.

In another preferred embodiment, the formulation or composition comprises (a) Schizophyllum commune beta-glucan; and optionally (b) a pharmaceutically, cosmetically, or device acceptable carrier or excipient.

In another preferred embodiment, the formulation or composition comprises 0.0001-99 wt%, preferably 0.001-90 wt%, more preferably 0.01-50 wt%, more preferably 0.05-10 wt% of beta-glucan (preferably schizophyllum commune beta-glucan), based on the total weight of the formulation or composition.

In another preferred embodiment, the mass concentration of the beta-glucan in the preparation or the composition is more than or equal to 1 mu g/mL, and specifically can be 1 mu g/mL-200mg/mL, or 1 mu g/mL-5mg/mL, or 1 mu g/mL-1 mg/mL.

In another preferred embodiment, the preparation comprises an aqueous solution of Schizophyllum commune beta-glucan, wherein the content of the aqueous solution of Schizophyllum commune beta-glucan in the preparation is more than or equal to 80 wt%, preferably more than or equal to 90 wt%, more preferably more than or equal to 95 wt%, more preferably more than or equal to 99 wt%, more preferably more than or equal to 99.5 wt%, based on the total weight of the preparation.

In another preferred embodiment, the concentration by mass of the Schizophyllum commune beta-glucan in the aqueous Schizophyllum commune beta-glucan solution is 0.0001-50 wt%, preferably 0.02-10 wt%, more preferably 0.05-5 wt%, based on the total weight of the aqueous Schizophyllum commune beta-glucan solution.

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:

TABLE 1 toothpaste having effects of preventing and/or treating oral diseases

The "active ingredient (first active ingredient)" in the preparation or composition of the present invention refers to the β -glucan according to the present invention (preferably schizophyllum commune β -glucan).

The "active ingredient (first active ingredient)", the preparation and/or the composition of the present invention can be used for the prevention and/or treatment of oral diseases.

By "second active ingredient" is meant an active ingredient for the treatment of oral diseases other than the beta-glucan in the present invention.

In a preferred embodiment, the second active ingredient is selected from the group consisting of: anticaries agents (including sodium fluoride, stannous fluoride, SMFP, erythritol, and anticariogenic enzymes), antiplaque and gingivitis compounds (including triclosan, zinc citrate + triclosan, sanguinarine sodium lauryl sulfate, chlorhexidine, domiphen bromide, cetylpyridinium chloride), anticalculus agents (including pyrophosphate + NaF, pyrophosphate + PVM/MA/NaF, zinc citrate + sodium monofluorophosphate, zinc salts + sodium fluoride), anti-inflammatory agents (including dihydrocholesterol, allantoin, glycyrrhizin, glycine, azulene), desensitizing agents (including potassium nitrate, strontium chloride, disodium hydrogen citrate), and deodorizing agents (including sodium copper chlorophyllin).

"safe and effective amount" means: the amount of active ingredient is sufficient to significantly ameliorate the condition or symptom without causing serious side effects.

Typically, the pharmaceutical composition contains 1-2000mg of active ingredient per dose, more preferably, 10-200mg of active ingredient per dose. Preferably, the dose is a tablet or an injection.

"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 of sufficient purity and sufficiently low toxicity.

By "compatible" is meant herein that the components of the composition are capable of being combined with the active ingredients of the present invention and with each other without significantly diminishing the efficacy of the active ingredient.

Examples of pharmaceutically acceptable carrier moieties are cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), and the like

) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

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

In another preferred embodiment, the β -glucan of the present invention can be linked to a macromolecular compound or polymer by a chemical bond. The macromolecular compounds may be biological macromolecules such as homoglycans, proteins, nucleic acids, polypeptides, and the like.

The mode of administration of the active ingredient or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to: topical, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and the like.

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

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:

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

(b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia;

(c) humectants, for example, glycerol;

(d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate;

(e) slow solvents, such as paraffin;

(f) absorption accelerators, e.g., quaternary ammonium compounds;

(g) wetting agents, such as cetyl alcohol and glycerol monostearate;

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

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

In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

The solid dosage forms may also be prepared using coatings and shells, such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active ingredient in such compositions may be delayed in a certain portion of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances.

Liquid dosage forms include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly employed in the art such as water or other solvents, solubilizing agents and emulsifiers, e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide, and oils, especially cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like. In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active ingredients, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these materials, and the like.

The compositions may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

When the pharmaceutical composition is used, a safe and effective amount of the composition of the present invention is administered to a mammal (such as a human) in need of treatment, wherein the administration dose is a pharmaceutically acceptable effective administration dose, and for a human with a weight of 60kg, the daily administration dose is usually 1 to 10000mg, preferably 10 to 2000mg, more preferably 20 to 1000 mg. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.

The present invention also provides a composition product comprising:

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

(2) a second pharmaceutical composition which is a different oral disease treatment drug from the first pharmaceutical composition.

The compositions of the invention may be administered alone or in combination with (e.g. formulated in the same pharmaceutical composition as) other therapeutic agents.

The pharmaceutical compositions of the present invention may also be combined with other agents known to treat or ameliorate similar conditions. When administered in combination, the mode of administration and dosage of the original drug is maintained, while the pharmaceutical composition of the present invention is used simultaneously or subsequently. The combination also includes the use of the pharmaceutical composition of the invention in combination with one or more other known drugs for an overlapping period of time. When the pharmaceutical composition of the present invention is administered in combination with one or more other drugs, the dosage of the pharmaceutical composition of the present invention or known drugs may be lower than the dosage when they are administered alone.

The main advantages of the invention include:

(a) the beta-glucan of the invention (such as Schizophyllum commune beta-glucan) can be very effective in preventing and/or treating oral diseases, such as gingivitis and periodontitis.

(b) The beta-glucan of the invention can be used for preventing alveolar bone loss, preventing dental caries, inhibiting the generation of dental plaque, removing dental calculus, resisting dentin sensitivity, removing dental stain, whitening teeth, regulating oral micro-ecology, improving oral hygiene state, preventing and treating oral ulcer, and improving oral mucosa immunity, or the combination thereof.

(c) The beta-glucan of the invention can also be used for oral postoperative care (promotion of wound healing), prevention and/or treatment of oral diseases in diabetic patients.

(d) The beta-glucan of the invention can be combined with other medicines for treating oral diseases, can further enhance the prevention and/or treatment effect, and has synergistic effect.

(e) The beta-glucans (preferably schizophyllum commune beta-glucans) of the invention are biopolysaccharides of purely natural origin, are completely or naturally soluble and have not been subjected to any chemical and/or physical modification or modification.

(f) The beta-glucan completely reserves the three-dimensional conformation of the triple helix, and has better activity for preventing and/or treating oral diseases.

(g) The beta-glucan has excellent stability, can coexist with most substances to keep the activity of the beta-glucan, has wide application field, can be combined with other oral disease treatment medicines for use, improves the oral hygiene condition and whitens teeth while preventing and treating the oral diseases.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Experimental procedures without specific conditions noted in the following examples, generally followed by conventional conditions, such as Sambrook et al, molecular cloning: the conditions described in the Laboratory Manual (New York: Cold Spring Harbor Laboratory Press,1989), or according to the manufacturer's recommendations. Unless otherwise indicated, percentages and parts are percentages and parts by weight.

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Materials, reagents, equipment and the like used in the following examples are commercially available unless otherwise specified.

Example 1

Beta-glucan acquisition and assay

This embodiment is only an example of beta-glucan obtained by Schizophyllum commune fermentation, but is not limited thereto.

First, obtaining the fermentation liquor of Schizophyllum commune strain

The fermentation broth in the following examples was prepared by fermentation of Schizophyllum commune strain (Schizophyllum commune Fr-1003, catalog:

38548^TMpurchased from American type culture Collection [ ATCC ]]) The method comprises the following steps:

1. and (3) activation of the Schizophyllum commune strain: preparing a plate culture medium from 200g/L of potato, 30g/L of glucose, 10g/L of sodium chloride and 20g/L of agar, inoculating Schizophyllum commune strains on the plate culture medium, and culturing in a constant-temperature incubator at 25 ℃ for 7 days to obtain plate mycelia;

2. seed activation: putting a liquid culture medium prepared from 100g/L of potato starch, 40g/L of glucose, 2g/L of yeast extract powder and water into a shake flask, wherein the liquid filling amount is 1/3, inoculating the flat plate mycelium obtained in the step 1 into the shake flask, and performing shake culture at 160rpm in a constant temperature shaking table at 25 ℃ for 7 days to obtain a seed solution;

3. fermentation culture: adding fermentation culture medium prepared from 50g/L glucose, 50g/L sucrose, 5g/L soybean flour (Wechkoku corporation, Shandong Shanyuan), 2g/L yeast extract powder, 0.5g/L potassium dihydrogen phosphate, 0.5g/L magnesium sulfate heptahydrate, 0.5g/L ammonium sulfate, 6g/L potassium nitrate and water into a fermentation tank, sterilizing at 121 deg.C for 15 min, inoculating the seed solution obtained in step 2 into the fermentation tank, stirring at 25 deg.C and 300rpm, and performing aeration fermentation culture for 8 days to obtain Schizophyllum commune fermentation broth.

Separation and purification of beta-glucan and preparation of beta-glucan solution

(1) Mixing the first part of the obtained Schizophyllum commune fermentation liquor with distilled water of 4 times of the volume, and soaking and boiling at 60 deg.C for 8 hr to obtain a soaking and boiling solution;

(2) centrifuging the digestion solution obtained in the step (1) at 4,000rpm for 5min, and taking supernatant; filtering the supernatant with 300 mesh filter cloth under negative pressure, and collecting the filtrate to obtain the decoction clear solution;

(3) and (3) heating the clear liquid obtained after filtration in the step (2) to 50 ℃, and simultaneously adding 200-mesh wood activated carbon and 8-16-mesh coconut shell activated carbon into the clear liquid, wherein the adding volume of each activated carbon is 1% of the volume of the clear liquid. Continuously stirring for 4h at 50 ℃ and 350rpm, and cooling for later use to obtain a soaking and boiling clear liquid mixed with activated carbon; then, sequentially performing negative pressure suction filtration on the boiled clear liquid mixed with the activated carbon by using 300-mesh filter cloth and an SCP-321# filter plate (the aperture is about 1.5 mu m), and taking the filtered clear liquid for later use;

(4) dissolving Solarbio lipase (L8070, the enzyme activity is 100-400U/mg) in a physiological phosphate buffer solution, adding the lipase solution into the filtrate prepared in the step (3) according to the enzyme dosage of 10U/mL, uniformly stirring, and carrying out enzymolysis for 2 hours at 40 ℃; dissolving Solarbio neutral protease (Z8030, the enzyme activity is more than 60U/mg) in physiological phosphate buffer solution, adding the neutral protease solution into the enzymatic hydrolysate according to the enzyme dosage of 60U/mL, uniformly stirring, and carrying out enzymolysis for 2 hours at 40 ℃. Heating in 90 deg.C water bath for 30min after enzymolysis to inactivate enzyme, vacuum filtering with SCP-321# filter plate (pore size of about 1.5 μm), and collecting the filtered clear liquid;

(5) quickly mixing the clear liquid filtered in the step (4) with edible ethanol with the concentration of 95% (volume ratio is 1: 3), and stirring until precipitates are obtained; dissolving the precipitate to original volume, rapidly mixing with 95% edible ethanol (volume ratio of 1: 3), and stirring to obtain precipitate;

(6) placing the precipitate obtained in the step (5) in a tray with holes, and drying the precipitate by using an electric heating oven at 40 ℃ until the weight is constant to obtain a dried product;

(7) crushing the dried beta-glucan obtained in the step (6), weighing 5g of crushed beta-glucan, dissolving the crushed beta-glucan in 1,000mL of ultrapure water, and stirring the crushed beta-glucan at 600rpm for 2 hours until the beta-glucan is fully dissolved to obtain a beta-glucan solution; and (3) carrying out negative pressure suction filtration on the beta-glucan solution by using a 5-micron filter membrane, and adding a proper amount of preservative for cosmetics to obtain the beta-glucan solution with high viscosity and high light transmittance and concentration of 0.5%.

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

Identification and detection of tri, beta-glucan

1. Infrared spectroscopic identification of beta-glucan

The 0.5% beta-glucan solution was identified by performing full-wavelength scanning using a fourier transform infrared spectrometer on a sample dried at 105 ℃ by using infrared spectroscopy according to method C in appendix IV of pharmacopoeia of the people's republic of china (2010 version). As a result, the 0.5% solution obtained in the second part (7) of example 1 was a β -glucan solution.

Compared with the infrared spectrogram of the yeast beta-glucan in the yeast beta-glucan industry standard QBT 4572-2013, the functional group position is basically consistent. As shown in fig. 12, the main functional group positions are:

1)3301cm^-1nearby stronger and wider absorption peak (sugar O-H bond stretching vibration absorption peak)

2)2921cm^-1Nearby weak absorption peak (sugar C-H bond stretching vibration absorption peak)

3)886cm^-1Nearby weaker absorption peak (sugar beta configuration characteristic vibration absorption peak)

4)1076cm^-1Nearby stronger absorption peak (sugar C-OH, C-O-C stretching vibration absorption peak)

Fourier transform infrared spectroscopy shows that the product obtained in this example is beta-glucan.

2. Content detection of beta-glucan

The dried product obtained in the second part (6) of the example 1 is subjected to beta-glucan quantitative detection, specifically, according to the method for determining the content of the yeast beta-glucan in the yeast beta-glucan industry standard QBT 4572-2013, the obtained beta-glucan dried sample is ground to about 1.0mm in diameter. The product was found to have a beta-glucan content of 99.23%.

3. Hormone detection of beta-glucan solutions

The above-mentioned 0.5% β -glucan solution was subjected to 48 hormone measurements, specifically, according to the first method of measuring 7 components, such as "cosmetic safety specifications" (2015 edition), chapter 2.4 estriol, chapter iv, and the like, and the liquid chromatography/tandem mass spectrometry and thin layer chromatography methods for measuring forty-one glucocorticoid in GB/T24800.2-2009 cosmetics. As a result, the hormone was not detected in the 0.5% ss-glucan solution obtained in the second part (7) of example 1 (see Table 2).

TABLE 248 hormone assay

4. Viscosity measurement of beta-glucan solution

According to the method of the second part (7) of example 1, aqueous solutions of beta-glucan of 0.3%, 0.5%, 0.8%, and 1.0% (mass/volume) were prepared, and the kinetic viscosity was measured at 25 ℃. The results are detailed in Table 3, and the viscosity of each sample increases with increasing β -glucan content, 472, 740, 2150 and 3100 mPas, respectively.

The commercially available dispersible (water insoluble) yeast beta-glucan particles (purchased from Wellmune) were weighed accurately to 2g, mixed with deionized water and made up to 200mL to give a yeast beta-glucan suspension with a mass to volume ratio of 1.0%, and the dynamic viscosity was measured at 25 ℃. As a result, the dynamic viscosity of the 1.0% yeast β -glucan suspension was 0 mPas (see Table 3).

Weighing 2g of commercially available soluble yeast beta-glucan powder (purchased from Wellmune company), adding deionized water to dissolve the powder, fixing the volume to 200mL to obtain a yeast beta-glucan aqueous solution with the mass-volume ratio of 1.0%, and detecting the dynamic viscosity at 25 ℃. As a result, the dynamic viscosity of the 1.0% yeast β -glucan aqueous solution was 0 mPas (see Table 3).

The dynamic viscosity is measured as follows:

(1) taking 200mL of the solution sample, and placing the solution sample in a 250mL beaker;

(2) placing the beaker containing the solution/mixed solution sample in a water bath kettle, and keeping the temperature at 25 ℃ for 1 h;

(3) the kinematic viscosity of each sample was measured at 25 ℃ using a rotational viscometer.

5. Measurement of light transmittance of beta-glucan solution

According to the method of the second part (7) of example 1, 0.3%, 0.5%, 0.8%, 1.0% (mass/volume) of aqueous solution of β -glucan was prepared, and the light transmittance of the solution was measured at a wavelength of 600nm, and as shown in table 3, the light transmittance of each sample was 96.5%, 93.1%, 87.5%, and 81.1%, respectively.

The transmittance of a commercially available 1.0% oat β -glucan solution (available from Symrise) was measured at a wavelength of 600nm and found to be 59.7% (see table 3).

The method comprises the steps of accurately weighing 2g of commercially available dispersible yeast beta-glucan particles, adding deionized water for mixing, and fixing the volume to 200mL to obtain yeast beta-glucan suspension with the mass-volume ratio of 1.0%, and measuring the light transmittance of the suspension at the wavelength of 600nm to obtain the light transmittance of only 1.3% (see table 3).

The method comprises the steps of accurately weighing 2g of commercially available soluble yeast beta-glucan powder, adding deionized water to dissolve the powder, fixing the volume to 200mL to obtain a yeast beta-glucan aqueous solution with the mass-volume ratio of 1.0%, and measuring the light transmittance of the solution at a wavelength of 600nm to obtain the light transmittance of 68.4% (see table 3).

The method for measuring the light transmittance is as follows:

(1) 10mL of the solution sample is placed in a centrifuge tube;

(2) centrifuging at 1000rpm for 1min to remove air bubbles (two yeast beta-glucan solutions/suspensions are bubble-free and not centrifuged);

(3) carefully taking 3mL to 1cm of glass cuvette to avoid bubbles;

(4) the transmittance of the sample was measured at a wavelength of 600nm using a spectrophotometer with deionized water as a blank reference (the transmittance of deionized water is 100%).

6. Stability assay for beta-glucan solutions

According to the method of the second part (7) of the example 1, 0.5%, 0.8%, 1.0% (mass/volume ratio) of the aqueous solution of β -glucan is prepared, after the preservative is added, the solution is left at room temperature (without shielding from light) for 24 months, the stability of the solution is observed, and the dynamic viscosity and light transmittance of the solution are measured. As a result, the three solution states were stable, and the viscosity and transmittance were not greatly changed, and the transmittance was even improved (Table 3).

A commercially available 1.0% oat beta-glucan solution was taken, left at room temperature (not protected from light) for 24 months, the solution stability was observed, and the solution dynamic viscosity and light transmittance were measured. As a result, the 1.0% oat β -glucan solution was very unstable and solid substances were precipitated after 3 months of storage at room temperature, resulting in undetectable viscosity and transmittance (table 3).

TABLE 3 various beta-glucan solution dynamic viscosity data

Note: a is commercially available dispersible yeast beta-glucan particles (insoluble in water) and b is commercially available soluble yeast beta-glucan powder.

EXAMPLE 2 toothpaste composition and method of making

(1) Toothpaste formulation (%)

TABLE 4 toothpaste formulations for periodontal disease

(2) Preparation method

Dispersing xanthan gum, sodium carboxymethylcellulose and essence in glycerol according to a formula table;

② adding 20ml of 1 percent schizophyllum beta-dextran solution and 25g of sorbitol, fully stirring and dissolving;

③ adding 20g of hydrated silica, and homogenizing by a homogenizer to ensure that the hydrated silica is fully dispersed in the system;

dissolving the components of the phase B in water, adding the water into the system, and fully stirring;

and (4) defoaming by using a vacuum pump, and then subpackaging.

Example 3 Schizophyllum commune beta-glucan containing toothpaste for improving periodontal disease

One, 20 volunteers were recruited to test the amelioration of periodontal disease by Schizophyllum commune beta-glucan. The toothpaste containing Schizophyllum commune beta-glucan prepared in example 2 was used by 20 volunteers to compare the improvement of periodontal inflammation and oral hygiene before and after use.

(II) experimental method:

1. medical history collection

The patient is asked for the history of periodontal disease, and the data obtained by performing a careful clinical examination and searching for risk factors are analyzed comprehensively, which is the basis of periodontal disease diagnosis. It should be noted that periodontal disease is closely related to the whole body, and during the examination and diagnosis, the patient's whole body condition, changes in other parts of the oral cavity, etc. should be included.

(1) Systematic medical history

Periodontal-related systemic diseases are more commonly acute and chronic blood diseases (leukemia, thrombocytopenic purpura, etc.), cardiovascular diseases, diabetes or other endocrine diseases, nervous system diseases, immune function deficiency, certain genetic diseases or genetic susceptibility factors, etc. There are also some infectious diseases such as hepatitis, AIDS, etc.

(2) History of oral diseases

The oral diseases except periodontal tissue are asked, especially some diseases can occur in oral mucosa and periodontal tissue simultaneously, such as oral leukoplakia, lichen planus, pemphigus, pemphigoid, etc.

(3) History of periodontal disease and oral hygiene habits

The record of the chief complaints is concerned with the current chief complaints, the occurrence time and the changes; the current medical history needs to inquire and record the possible causes, the development process of the disease, the treatment course and the curative effect in detail.

In the examination and communication of periodontal disease patients, the oral health consciousness, oral health habits, daily oral health measures and the like of the patients need to be known in detail as much as possible, which has important reference values for the diagnosis, treatment plan, prognosis judgment and the like of periodontal disease.

(4) Family history

Parents, siblings or other immediate relatives are asked about periodontal health, especially some periodontal diseases that may be genetically related, such as aggressive periodontitis, gingival fibroma, etc. In addition, a family history should be asked for when abnormal manifestations of dentition, tooth and maxillofacial development are found and genetic predisposition is suspected.

2. Oral hygiene conditions

The health of the oral cavity and the health of the periodontal tissue are closely related, and the examination is performed for patients in both initial and follow-up visits, including examination of plaque, soft calculus, tartar and stain deposition, food impaction, abnormal tooth structure, halitosis, etc.

Examination of plaque, using visual inspection in combination with visual observation with a plaque disclosing agent, the latter typically using a 2% basic fuchsin solution. The patient rinses mouth with clear water, then dips the liquid medicine with a cotton swab or a small cotton ball, coats the liquid medicine on the tooth surface near the gingival margin, and rinses mouth again, wherein the stained area of the tooth surface is the attached bacterial plaque.

Plaque index (PLI, platue index)

The scoring criteria for the plaque index are shown in figure 1. It was proposed by Quigley and Hein (1962) and improved by Turesky et al, which required application of a plaque disclosing agent to the tooth surface, followed by rinsing and examination of the location and extent of the stained plaque on the tooth surface, and this scoring method was relatively objective. The index mainly reflects the oral hygiene condition, checks whether the measures for controlling the self bacterial plaque of the patient are effective or not and clinically observes the effect of certain antibacterial plaque agents, and the patient can also check the mirror by himself, so the application is more convenient.

3. Gingival condition

(1) Bleeding index (BI, blue index)

Scoring criteria for Bleeding Index (BI) are shown in fig. 4. Mazza used an improved bleeding index in a clinical study in 1981. The gingival sulcus or the bottom of the bag is slightly probed with a blunt periodontal probe, and the presence and degree of bleeding are observed after the probe is taken out for 30 s. Scoring in a scale of 0-5: 0 is healthy gum, no inflammation and bleeding; 1, the color of the gum is changed in an inflammatory way, and bleeding is not caused in probing; 2, a little bleeding after probing; probing bleeding spreads along the gingival margin; 4, the bleeding flows to the full and overflows the gingival sulcus; 5 automatic bleeding.

4. Periodontal probing

Periodontal probing (periodontal probing) is the most important examination method in the diagnosis of periodontal disease, particularly periodontitis.

(1) Purpose of periodontal probing

Knowing the loss condition of periodontal support tissue, detecting whether the perialveolar sacs are formed on each face of all teeth of the whole dentition, the depth of the periodontal sacks and the attachment level of the periodontal ligament; in addition, the bifurcation lesion and the presence or absence of gingival sulcus/pocket bleeding after probing should be probed and responded with a numerical record.

(2) Periodontal probing tool

Periodontal probing is done clinically with a common periodontal scaling probe or electronic probe. The periodontal probe has a plurality of types, the types are common to the periodontal probe and are provided with scales, but the design characteristics of the periodontal probe are different according to different inspection purposes, the bending angle and the scale interval of the periodontal probe are different, each scale is 1mm or 2-3 mm, the working end is basically cylindrical and is tapered, and the insertion is facilitated; the tip is blunt and has a diameter of 0.5 mm. FIG. 8 shows various periodontal probes

(3) Method of periodontal probing

Periodontal probes should be probed on all sides along the long axis of the tooth, usually measured in the buccal (labial), lingual mesial, central, mesial sides of the tooth, respectively, and 6 sites of probing depth are recorded for each tooth. During probing, the patient should walk along the bottom of the periodontal pocket to locate periodontal pockets of different depths on the same tooth surface. The periodontal pocket probe site and method are shown in fig. 9.

When measuring the periodontal pocket, the periodontal probe tip is always close to the tooth surface, the probe is parallel to the long axis of the tooth, but due to the blocking of the adjacent surface contact area, the probe cannot enter the gingival valley area if being parallel to the long axis of the tooth, and the deepest point of the adjacent surface pocket is usually at the gingival valley. Thus, when probing the adjacent surface, the probe may be allowed to abut the contact point and be slightly inclined toward the center of the adjacent surface, so that the deepest part of the adjacent pocket can be probed (fig. 10).

(4) Record and evaluation index of periodontal probing

Periodontal probing is the most important method for clinically evaluating the loss degree of periodontal supporting tissues, and two indexes are mainly adopted in clinical examination:

5. degree of tooth looseness

Normally, teeth have slight physiological mobility, mainly in the horizontal direction. The physiological mobility of a single tooth is slightly greater than that of multiple teeth. In the morning, the mobility of the teeth is the largest, and the teeth are slightly pushed out probably because the upper teeth and the lower teeth are not contacted at night; in daytime, teeth can be pressed into the alveolus due to chewing and swallowing actions. This subtle change is not evident in normal periodontal tissues. In periodontitis, the mobility of teeth exceeds the range of physiological mobility due to alveolar bone resorption, occlusal trauma, acute inflammation and other destruction of periodontal support structures, resulting in pathological tooth loosening.

(1) Examination of tooth loosening degree

The anterior teeth clamp the incisal margin by a dental forceps and shake in the direction of the lip and tongue; closing the forceps at the posterior teeth, and holding the posterior teeth with the tips of the forceps

The facial fossa is shaken in the buccolingual or mesial-distal directions. Often classified as three degree recording.

The I degree loosening exceeds the physiological mobility, but the amplitude is within 1 mm.

The loosening amplitude of II degrees is 1-2 mm.

The looseness amplitude of the III degree is more than 2 mm.

The looseness can also be determined according to the looseness direction, wherein the looseness in the cheek (lip) tongue direction is I degree, the looseness in the cheek (lip) tongue and the far, middle and near directions is II degree, and the looseness in the cheek (lip), the far, middle and near directions and the vertical direction is III degree.

6. Radiographic examination

(1) X-ray image of normal periodontal tissue

Alveolar bone

The natural alveolar bone around the root of the tooth appears as a white-line dense image of continuous obstruction, called the dura plate. The marrow cavity of the cancellous bone is transparent, and the trabecular bone is in the form of obstruction and is interwoven into a net shape. Normally, the distance from the alveolar bone top to the enamel cementum boundary is about 1-1.5 mm and is not more than 2mm, which is an important reference mark for determining whether bone absorption exists or not.

Periodontal ligament

The periodontal ligament occupies a certain space on the X-ray sheet, called periodontal ligament space, and is a continuous and uniform linear black transmission band with a width of about 0.18-0.25 mm, and the increase and change of the width of the transmission band is significant for the diagnosis of periodontal diseases.

In periodontitis, the dura mater plate is often incomplete or lost due to destruction of the alveolar bone, and the periodontal ligament space correspondingly shows broadening or significant broadening. In addition, the performance of the hard bone plate can reflect the change of diseases to a certain extent, and the hard bone plate is usually disappeared before the treatment of the diseases or during the active period; after periodontal treatment, the scleral plate is reformed and presents a dense continuous image; the patients and the teeth in the maintenance period should be regularly and repeatedly checked and photographed with X-ray films, and the recovery condition and stability of the disease condition can be indirectly judged by observing the change of the hard bone plate.

The bone condition in the mesial and distal teeth is mainly displayed on the X-ray film, and the alveolar bone on the buccolingual side is not clearly displayed due to the overlapping with the teeth. On the standard apical root, alveolar bone resorption is considered to be present when the distance from the crest of the alveolar ridge to the cementum enamel boundary exceeds 2 mm.

On the X-ray sheet, the types of alveolar bone resorption are expressed as horizontal-type resorption and vertical-type resorption.

Horizontal absorption: the height of the alveolar bone is reduced in a horizontal shape, and the bone absorption surface is in a horizontal or cup-shaped depression. Anterior teeth are mostly absorbed horizontally due to narrow alveolar ridges.

Vertical absorption: x-ray films show that the bone resorption surface forms an acute angle, also called angular resorption, with the root of the tooth, which is frequently found in posterior teeth with widely spaced alveoli.

The degree of bone resorption is generally described in terms of the proportion of the resorptive area to the length of the tooth root, and is usually divided into three degrees.

Degree I: alveolar bone is resorbed within the apex 1/3 of the root.

II degree: alveolar bone resorption exceeds root length 1/3, but is within root length 2/3, or absorbs 1/2 to the root length.

And (3) III degree: alveolar bone resorption is greater than root length 2/3.

7. The requirement of brushing teeth

(a) The tooth brushing sequence is as follows: in order to ensure that certain parts are not missed during tooth brushing, the face brushing is recommended to be achieved by brushing the teeth according to a certain sequence. The set position for placing the toothbrush each time generally occupies the distance of 1-3 tooth surfaces, each part is brushed for at least 5-10 times, then the toothbrush is moved to the next adjacent tooth brushing position, the two brushing positions are overlapped, the lower jaw teeth generally have about 9 brushing positions on the buccal side of the lips, and the tongue is measured as 11 teeth;

(b) time and dosage of tooth brushing: the general population recommends that the brushing time be at least 2 minutes per time; about 1cm of toothpaste per use.

(c) The times of tooth brushing are as follows: after the dental plaque is removed by brushing teeth for several hours, the dental plaque can be attached to the clean tooth surface again and is continuously formed, and particularly after falling asleep at night, the saliva secretion is reduced, the self-cleaning function of the oral cavity is poor, and bacteria are easy to grow. Therefore, brushing teeth at least twice a day is more important before sleeping at night;

(d) hard-to-brush part: when brushing teeth, some parts are often ignored, such as the far middle surface of the last tooth of the upper and lower jaws and the tooth surface adjacent to the edentulous area, the palate surface of the upper and lower teeth, the lingual surface of the lower and upper teeth, the misaligned teeth and the teeth erupting ectopically. These areas are easily overlooked or difficult to reach during brushing, and special attention should be given to brushing during brushing, and some brushing actions or pressure lines or interdental brushes are needed to supplement. Oral cleaning should include the tongue surface, which reduces the amount of food residues and microorganisms in the mouth, delays plaque formation and total plaque deposition, and aids in overall oral cleaning.

(III) experimental results:

experimental results on dental plaque as shown in fig. 2 and 3, the volunteers showed a significant improvement of dental plaque and marked whitening of teeth after one month using the toothpaste containing β -glucan. The plaque on the teeth of the volunteers in fig. 2 was scored according to the method for determining plaque index shown in fig. 1, and statistical results showed that the plaque shown on the photographs was significantly inhibited after using the toothpaste containing β -glucan (P value ═ 0.0018 ×).

As shown in fig. 5, the results of experiments on gingival bleeding were significantly improved after the application of the toothpaste containing β -glucan to volunteers, and although there was no significant diagnosis (insufficient number of samples) from the bleeding from probing, from the questionnaire of volunteers, 8 volunteers among them had the bleeding from the teeth while brushing the teeth, but the bleeding from the teeth was significantly improved after two weeks on average using the toothpaste containing β -glucan, and 4 volunteers among them had the bleeding from the teeth after two months using the toothpaste.

The experimental results on the amount of tartar are shown in fig. 7, and the condition of tartar is further improved after volunteers use the toothpaste for one month, and the condition of tartar is further improved after the volunteers continuously use the toothpaste for two months. In fact, gingivitis and the subsequent development of periodontitis is primarily caused by plaque and calculus on the teeth, which directly improves the level of gingival inflammation if oral hygiene is improved.

The results of the periodontal probing are shown in fig. 11, and the depth of periodontal probing is significantly improved after the volunteers use the toothpaste containing β -glucan. Since the depth of the normal healthy periodontal probing should be 3mm or less, but the gingival attachment ability is decreased due to the occurrence of gingival inflammation, which in turn leads to the depth of the periodontal probing to be increased. After the volunteers used the toothpaste containing beta-glucan, the periodontal probing depth was reduced, indicating that the gingival attachment ability was enhanced and possibly that the gingival inflammation was improved.

Example 4 Schizophyllum commune beta-glucan containing toothpaste improves dentinal sensitivity

One, 20 dentin sensitivity volunteers were collected and evaluated for dentin sensitivity.

Preparation of toothpaste

(1) Formulation of

TABLE 5 toothpaste formulations for dentinal sensitivity

(2) Preparation method

(a) Dispersing xanthan gum, sodium carboxymethylcellulose and essence in glycerol according to a formula table;

(b) then adding 10ml of 1% Schizophyllum commune beta-glucan solution and 35g of sorbitol, and fully stirring and dissolving;

(c) adding 18g of hydrated silica, and homogenizing by using a homogenizer to ensure that the hydrated silica is fully dispersed in the system;

(d) dissolving the components of phase B in water, adding into the system, and stirring;

(e) and (4) defoaming by using a vacuum pump, and then subpackaging.

(III) experimental results:

the toothpaste containing Schizophyllum commune beta-glucan can effectively improve dentin sensitivity. The sensitivity of the volunteers is reduced after the volunteers use the feed for 3 days, the sensitivity of cold air blowing is reduced by 1-2 minutes and 30 days before the volunteers use the cold air blowing for 7 days, and the sensitivity of cold air blowing is reduced by 2-3 minutes before the volunteers use the cold air blowing for 30 days, so that dentin sensitivity is basically avoided. As shown in fig. 13.

In addition, 5 volunteers had sensitive and acid-tingling part of their teeth during brushing. After 3-5 days using a toothpaste containing 0.1% Schizophyllum commune beta-glucan, the conscious tooth sensitivity phenomenon disappeared.

Example 5 Schizophyllum commune beta-glucan amelioration of oral mucosal disease

One, 30 canker sore volunteers were collected and divided into three groups on average. The degree of ulcer healing was observed by daily visual observation and self-evaluation.

(II) experimental grouping:

group 1 Using 5mg/mL sterile Schizophyllum commune beta-glucan prepared in example 1, 1-20mL was retained in the mouth for at least 10 seconds; gargle 3 times daily.

Group 2 used 1mg/mL sterile Schizophyllum commune beta-glucan prepared in example 1, with 1-20mL in the mouth, retained for at least 10 seconds; gargle 3 times daily.

Group 3 was buccally rinsed with 1-20mL of sterile water for at least 10 seconds 3 times daily.

(III) experimental results:

after 5mg/mL sterile Schizophyllum commune beta-glucan is used for 24 hours by the volunteers in the group 1, the area of the oral ulcer is obviously reduced, and the pain is relieved; after 48 hours, 7 volunteers healed; after 72 hours, 10 volunteers all recovered.

After the group 2 uses 1mg/mL sterile schizophyllum commune beta-glucan for 24 hours, the area of the oral ulcer is obviously reduced, and the pain is relieved; after 48 hours, 4 volunteers healed; after 72 hours, 8 volunteers healed; after 94 hours, 10 volunteers all recovered.

In group 3, there was still no significant improvement after 72 hours of use of sterile water, with some volunteers having an increased ulcer.

Specific results are shown in Table 6

TABLE 6 Schizophyllum commune beta-glucan amelioration of oral mucosal disease

Example 6 mouth rinse containing Schizophyllum commune beta-glucan for ameliorating oral mucosal disorders

(I) 60 persons were collected and divided into three groups, group 1 was a control group, and the mouth wash without Schizophyllum commune beta-glucan was used, and

groups

2 and 3 were experimental groups, and mouth wash with Schizophyllum commune beta-glucan at different concentrations was used. Volunteers gargle the dispensed mouthwash at least 2 times a day and evaluated the improvement of oral mucosal disease.

(II) preparing the mouth wash containing the schizophyllum commune beta-glucan and the mouth wash not containing the schizophyllum commune beta-glucan.

TABLE 7 mouthwash formulations

Ingredients in the su saliva	Formulation	1	Formulation 2	Formulation 3
					Beta-glucan	0	0.05	0.1
Glycerol	9	9	9
				Propylene glycol	9	9	9
Sorbitol	5	5	5
				Essence (tea extract)	0.25	0.25	0.25
Saccharin sodium salt	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 (W)	To 100 of	To 100 of	To 100 of

(III) experimental results:

the mouth wash without the schizophyllum commune beta-glucan can improve tooth cleanliness indexes such as dental plaque and the like, but has no obvious influence on oral mucosa diseases, and the healing period is generally 6.88 days; the mouth wash containing the schizophyllum commune beta-glucan has an excellent treatment effect on oral mucosa diseases while cleaning the oral cavity, and has an effect of preventing the recurrence of the oral mucosa diseases after long-term use. After the oral liquid containing 0.05% of Schizophyllum commune beta-glucan is used, the healing cycle of the oral ulcer is averagely 6.18 days, and the healing cycle is shortened by 10.2%; after the oral liquid containing 0.1% of Schizophyllum commune beta-glucan is used, the healing period of the oral ulcer is averagely 4.5 days, and the healing period is shortened by 34.6%. As shown in fig. 14.

Example 7 Schizophyllum commune beta-glucan containing toothpaste for ameliorating oral mucosal disorders

And (I) 30 volunteers with oral mucosa diseases are collected.

Preparation of toothpaste

(1) Toothpaste formula

TABLE 8 toothpaste formula for oral mucosa diseases

(2) Preparation method

(a) Dispersing xanthan gum, sodium carboxymethylcellulose and essence in glycerol and polyethylene glycol 400 according to a formula table;

(b) then adding 25ml of 1% Schizophyllum commune beta-glucan solution and 35g of sorbitol, and fully stirring and dissolving;

(c) adding 15g of hydrated silica, and homogenizing by using a homogenizer to ensure that the hydrated silica is fully dispersed in the system;

(e) and (4) defoaming by using a vacuum pump, and then subpackaging.

(III) experimental results:

the toothpaste containing the Schizophyllum commune beta-glucan has excellent treatment effect on oral mucosa diseases, and has the effect of preventing the recurrence of the oral mucosa diseases after long-term use. The effect of the schizophyllum commune β -glucan containing toothpaste on the recovery time of canker sores is shown in figure 15.

Example 8 toothpaste and chewing gum containing Schizophyllum commune beta-glucan to ameliorate periodontal inflammation problems caused by dental caries

(one) 30 carious volunteers were recruited. The 30 patients had varying degrees of caries, and caries had developed red, swollen gums, pain, difficulty chewing, varying degrees of sensitivity to cold, hot, sour and sweet. Pain of five volunteers can be relieved by taking anti-inflammatory and analgesic medicines, but the pain can recur after stopping taking the medicines. It is recommended to brush teeth in the morning and evening, and use chewing gum after three meals.

Preparation of toothpaste and chewing gum

(1) Toothpaste formula

TABLE 9 toothpaste formula for dental caries-induced periodontal disease

(2) Preparation method

(c) adding 20g of hydrated silica, and homogenizing by using a homogenizer to ensure that the hydrated silica is fully dispersed in the system;

(e) and (4) defoaming by using a vacuum pump, and then subpackaging.

(III) preparation of chewing Gum

(1) Chewing gum formulation

TABLE 10 toothpaste formulations for dentinal sensitivity

Chewing gum composition	Composition (I)
		Gum bases	30％
Mixed sugar alcohol (xylitol: maltitol 7:1)	60％
		1% Schizophyllum commune beta-glucan solution	10％

(2) Preparation method

(a) Softening of the gum base: the gum base was placed in a constant temperature oven for 2h softening at 60 ℃. The gum base is kept moist when being softened, and is taken in time after being softened, otherwise, the gum base is hardened;

(b) mixing and blending the ingredients: firstly, crushing xylitol and sieving the crushed xylitol with a 200-mesh sieve for later use. During operation, firstly, the softened gum base is stirred in a water bath kettle at about 60 ℃ for several minutes and gradually melted into mucus, namely, liquid maltitol and the gum base are added and mixed for 6-8 min, and then xylitol powder and the gum base are added and mixed for 5-6 min. And after the sugar alcohol powder and the gum base are mixed basically and uniformly, adding 10g of 1% schizophyllum commune beta-glucan solution, stirring and mixing for 3-5 min.

(c) Extruding and molding: heating the blended gum base ingredients to 85-90 ℃ for condensation, cooling to below 40 ℃ after 1-2 min, putting the gum base ingredients into a noodle press for repeated extrusion to form a banded candy blank with compact tissue and smooth surface, and repeatedly extruding for several times; cutting and molding the sugar sheet extruded into a certain thickness.

(d) Packaging: the finished product of the chewing gum is aged for 10-20 hours at the temperature of about 20 ℃ and under the relative humidity of less than 55%, so that the moisture of the chewing gum is balanced and the hardness of the chewing gum is moderate.

(III) experimental results:

toothpaste and chewing gum containing Schizophyllum commune beta-glucan are effective in relieving gingival swelling, pain, chewing difficulty caused by dental caries and sensitivity to cold, hot, sour and sweet, and preventing dental caries from further development.

TABLE 11 improvement of periodontal disease due to dental caries with a toothpaste containing Schizophyllum commune beta-glucan

	Tooth sensitivity of 10 persons	26 persons with chewing pain	Tooth loosening 5 persons	3 patients with pus discharge
					Positive feedback	8	26	1	3
Negative feedback	0	0	0	0
					Invalid feedback	2	0	4	0

Wherein the feedback rate of improvement of pain and purulence of teeth due to inflammation is 100%, and the feedback rate of improvement of tooth sensitivity is 80%, but there is no excellent improvement effect on the loosened teeth, so that the loosened teeth cannot be restored, and it is recommended to go to hospital for treatment.

Example 9 Schizophyllum commune beta-glucan containing toothpaste for ameliorating tooth stains

(one) 30 carious volunteers were recruited. Brushing the teeth in the morning and evening.

(II) preparation of toothpaste

(1) Toothpaste formula

TABLE 12 toothpaste formulations for improving tooth stain

(2) Preparation method

(b) then adding 15ml of 1% Schizophyllum commune beta-glucan solution and 35g of sorbitol, and fully stirring and dissolving;

(c) adding 5g of hydrated silica and 35g of calcium carbonate, and homogenizing by using a homogenizer to ensure that the grinding agent is fully dispersed in the system;

(e) and (4) defoaming by using a vacuum pump, and then subpackaging.

(III) experimental results:

the toothpaste containing the schizophyllum commune beta-glucan can improve tooth stains, but has no obvious effect on patients with fluorosis, has a certain effect on removing smoke stains, but cannot completely remove the smoke stains. Therefore, the toothpaste containing the schizophyllum commune beta-glucan has better effect mainly on milder tooth stains, and if the tooth stains are deposited seriously, the toothpaste containing the schizophyllum commune beta-glucan is recommended to be used for preventing the tooth pigments from being deposited again after the teeth are washed.

TABLE 13 improvement of dental stains by a toothpaste containing Schizophyllum commune beta-glucan

	Smoking 8 people	Dental fluorosis 2 people	Dark tooth color and luster for 20 people
				Positive feedback	5	0	18
Negative feedback	0	0	0
				Invalid feedback	3	2	2

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims

1. Use of beta-glucan for the preparation of a formulation or composition for the prevention and/or treatment of an oral disease.

2. The use according to claim 1, wherein the beta-glucan has the structure shown in formula I,

3. Use according to claim 1, wherein the beta-glucan has a degree of branching of 0.02 to 0.8, preferably 0.1 to 0.5, preferably 0.2 to 0.4.

4. Use according to claim 1, wherein the molecular weight of the β -glucan is 2kD or more, preferably 2kD to 40000kD, more preferably 20kD to 20000 kD.

5. The use of claim 1, wherein the β -glucan is selected from the group consisting of: schizophyllum commune beta-glucan, shiitake mushroom beta-glucan, sclerotium rolfsii beta-glucan, grifola frondosa beta-glucan, pleurotus ostreatus polysaccharide, mushroom beta-glucan, yeast beta-glucan, oat beta-glucan, or a combination thereof.

6. The use of claim 1, wherein the oral disease comprises periodontal disease, tooth body disease and oral mucosal disease.

7. The use of claim 1, wherein the oral disease comprises a canker sore.

8. The use according to claim 1, wherein the formulation or composition is used for preventing alveolar bone loss, preventing and treating periodontal diseases related to dental caries, inhibiting the formation of dental plaque, removing dental calculus, resisting dentinal hypersensitivity, removing tooth stain, whitening teeth, regulating oral micro-ecology, improving oral hygiene, preventing and/or treating oral ulcer, restoring and enhancing oral mucosal immunity, or a combination thereof.

9. The use of claim 1, wherein the formulation or composition is for the prevention and/or treatment of canker sores, restoring and improving oral mucosal membrane resistance, or a combination thereof.

10. A formulation comprising (a) β -glucan; and (b) a pharmaceutically, cosmetically, or device acceptable carrier or excipient.

11. The formulation of claim 10, further comprising (c) a second active ingredient, wherein the second active ingredient is an active ingredient for treating an oral disease different from the beta-glucan,

preferably, the second active ingredient is selected from the group consisting of: anticaries agents, antiplaque and gingivitis compounds, anticalculus agents, anti-inflammatory agents, desensitizing agents, and deodorizing agents.

12. An oral cleaning product, which is characterized in that the oral cleaning product comprises beta-glucan and acceptable auxiliary materials in the cleaning product,

preferably, the auxiliary materials comprise a base material, a flavoring agent, a functional additive, a solvent, or a combination thereof.