A composite probiotic for regulating oral microecological environment and its application

Abstract

The present invention relates to a composite probiotic for regulating oral microecological environment and application thereof. The composite probiotic for regulating oral microecological environment comprises inactivated Lactobacillus paracasei with a preservation number of CGMCC No.30464. Lacticaseibacillus paracasei FB97 strain and inactivated Lactobacillus rhamnosus with a deposit number of CGMCC No.31054 Lacticaseibacillus rhamnosus When the RH22 strain is used in combination, the two can cooperate and promote each other, synergizing in promoting the health of the oral microecological environment. Specifically, they have significant effects in inhibiting the growth of harmful oral bacteria, promoting the proliferation of beneficial bacteria, and inhibiting the formation of dental plaque.

Description

Technical Field

[0001] The invention belongs to the technical field of oral care products and relates to a composite probiotic for regulating oral microecological environment and application thereof. Background Art

[0002] As the starting point of the respiratory tract and gastrointestinal tract, the oral cavity is the first barrier to human health. Harmful oral bacteria can cause lesions in the nasal cavity and throat, and in severe cases can lead to problems with organs such as the kidneys and heart. As the saying goes, "diseases come from the mouth." Most oral problems stem from the imbalance of the oral microecological environment. The bacteria in the mouth are simply divided into two categories: beneficial bacteria and harmful bacteria. The disruption of the microecological environment is mainly due to the colonization and growth of a large number of harmful bacteria, mainly represented by Fusobacterium nucleatum, Actinomycetemcomitans, and Streptococcus mutans. These harmful bacteria exist in our mouths and decompose the remaining food residues in the mouth into harmful compounds, corroding our teeth and gums while releasing hydrogen sulfide gas, which causes bad breath. Beneficial bacteria are a type of bacteria that are beneficial to the oral cavity, such as Streptococcus salivarius. They hinder the growth of harmful bacteria through competitive inhibition to achieve the purpose of balancing the oral flora.

[0003] Dental plaque is a bacterial ecological environment that exists on the tooth surface or in the periodontal pocket, where bacteria grow, develop and die, and carry out complex material metabolism activities. Under certain conditions, bacteria and their products will damage teeth and periodontal tissues. Plaque is a soft and unmineralized bacterial plaque that adheres to the surface of the teeth. It is not easy to detect when the number is small. However, as time goes by, if the plaque is not removed in time, it will cause stubborn spots and stains that cannot be removed by daily oral care, making the tooth surface lose its luster and turn yellow and dark. The current methods for treating dental plaque mainly include mechanical removal and chemical treatment, but both have some limitations. Traditional mechanical removal methods, such as brushing and flossing, are the most common ways to remove dental plaque. However, these methods rely on personal cleaning habits and skills, and cannot guarantee the complete removal of all dental plaque, especially in the gaps between teeth and other hard-to-reach areas. In addition, if the brushing force is too strong or used improperly, it may also cause gum damage. Chemical treatment methods such as mouthwashes inhibit bacterial growth by using antibacterial ingredients, but due to the fluidity of saliva, most drugs are removed from the mouth very quickly and cannot be effective for a long time, and cannot fundamentally solve the problem of dental plaque formation. In addition, long-term use of mouthwashes containing strong antibacterial ingredients may cause an imbalance in the oral flora and cause other oral health problems. Therefore, it is of great significance to provide an oral care product containing probiotics that can effectively regulate the oral microecological environment, inhibit the excessive growth of harmful bacteria, promote the proliferation of beneficial bacteria, and effectively inhibit the formation of dental plaque. Summary of the invention

[0004] In view of the deficiencies in the prior art, the object of the present invention is to provide a composite probiotic for regulating the oral microecological environment and its application, and in particular to provide a composite probiotic for regulating the oral microecological environment and its application in the preparation of oral care products.

[0005] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a composite probiotic for regulating the oral microecological environment, wherein the composite probiotic for regulating the oral microecological environment comprises an inactivated Lactobacillus paracasei FB97 strain with a preservation number of CGMCC No. 30464 and an inactivated Lactobacillus rhamnosus RH22 strain with a preservation number of CGMCC No. 31054.

[0007] The present invention develops a new probiotic compounding method and a new strategy for regulating the oral microecological environment, that is, compounding the inactivated Lacticaseibacillus paracasei FB97 strain and the inactivated Lacticaseibacillus rhamnosus RH22 strain, and finds that there is a potential interaction between the two postbiotics, which can cooperate with each other and synergize. When the amount of bacteria used is the same, it has a more significant and excellent effect than a single postbiotic or other postbiotic compounding methods. Specifically, it is manifested in: (1) inhibiting the excessive growth of harmful oral bacteria; (2) promoting the proliferation of beneficial oral bacteria and maintaining a healthy oral microecological environment; (3) inhibiting the formation of dental plaque. Therefore, the composite probiotic has a good application prospect for the preparation of oral care products.

[0008] Preferably, the ratio of the bacterial counts of the Lactobacillus paracasei FB97 strain and the Lactobacillus rhamnosus RH22 strain is 1:10-10:1, for example, 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, etc. Other specific point values ​​within this numerical range can be selected, and they will not be described one by one here.

[0009] Based on the potential interaction between the two strains, the present invention also found that when the two strains are used in combination at the above-mentioned specific bacterial count ratio, the effects of inhibiting the growth of harmful oral bacteria, promoting the proliferation of beneficial bacteria and inhibiting the formation of dental plaque are more significant.

[0010] In a second aspect, the present invention provides a probiotic for regulating the oral microecological environment, wherein the strains in the probiotic include the composite probiotic described in the first aspect.

[0011] Preferably, the concentration of the composite probiotics in the probiotic agent is not less than 1×10 10 / mL or 1×10 10 Pieces / g, for example 1×10 10 pieces / mL (pieces / g), 2×10 10 pieces / mL (pieces / g), 5×10 10 pieces / mL (pieces / g), 8×10 10 pieces / mL (pieces / g), 1×10 11 pieces / mL (pieces / g), 5×10 11 pieces / mL (pieces / g), 1×10 12 Pieces / mL (pieces / g), etc. Other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0012] Preferably, the dosage form of the probiotic includes liquid, lyophilized powder, capsule, tablet or granule.

[0013] In a third aspect, the present invention provides a use of the composite probiotic according to the first aspect or the probiotic according to the second aspect in the preparation of an oral care product.

[0014] The oral care products include toothpaste, mouthwash, oral lozenges, and oral gel.

[0015] In a fourth aspect, the present invention provides a mouthwash for preventing or improving the formation of dental plaque, wherein the raw materials for preparing the mouthwash include the probiotic agent and solvent described in the second aspect.

[0016] Preferably, the raw materials for preparing the mouthwash further include any one of flavoring agents, sweeteners, moisturizers, surfactants, colorants, preservatives or functional additives, or a combination of at least two thereof.

[0017] Preferably, the mass percentage of the probiotic in the mouthwash is 0.05%-2%, for example, it can be 0.05%, 0.1%, 0.2%, 0.4%, 0.5%, 0.4%, 0.8%, 1%, 1.2%, 1.4%, 1.5%, 1.6%, 1.8%, 2%, etc. Other specific point values ​​within this numerical range can be selected, which will not be described here one by one.

[0018] In a fifth aspect, the present invention provides a toothpaste for preventing or improving the formation of dental plaque, wherein the raw materials for preparing the toothpaste include: the probiotic agent, moisturizer, abrasive, foaming agent, thickener, and taste regulator described in the second aspect.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The present invention develops a new probiotic compounding method and a new strategy for regulating the oral microecological environment, that is, compounding the inactivated Lacticaseibacillus paracasei FB97 strain and the inactivated Lacticaseibacillus rhamnosus RH22 strain, and finds that there is a potential interaction between the two postbiotics, which can cooperate with each other and synergize. When the amount of bacteria used is the same, it has a more significant and excellent effect than a single postbiotic or other postbiotic compounding methods. Specifically, it is manifested in: (1) inhibiting the excessive growth of harmful oral bacteria; (2) promoting the proliferation of beneficial oral bacteria and maintaining a healthy oral microecological environment; (3) inhibiting the formation of dental plaque. Therefore, the composite probiotic has a good application prospect for the preparation of oral care products.

[0021] The FB97 strain involved in the present invention is classified and named as Lactobacillus paracasei, and the preservation unit is the General Microbiology Center of China Microorganism Culture Collection Administration, the preservation time is April 28, 2024, the preservation number is CGMCC No.30464, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0022] The RH22 strain involved in the present invention is classified and named as Lactobacillus rhamnosus, and the preservation unit is the General Microbiology Center of China Microorganism Culture Collection Administration, the preservation time is June 24, 2024, the preservation number is CGMCC No. 31054, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. DETAILED DESCRIPTION

[0023] In order to further explain the technical means and effects adopted by the present invention, the technical solution of the present invention is further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0024] The FB97 strain involved below is named as Lactobacillus paracasei, and its deposit number is CGMCC No.30464;

[0025] The RH22 strain involved below is classified and named as Lacticaseibacillus rhamnosus, and its deposit number is CGMCC No. 31054;

[0026] The ATCC 25302 strain referred to below is Lactobacillus paracasei ATCC 25302.

[0027] The preparation method of the inactivated bacterial suspension or inactivated bacterial powder involved below is: inoculate the required strain into MRS medium at a concentration of 2%, shake culture at 37°C, obtain bacterial solution of different concentrations after culture for different time periods as required, adjust the pH of the bacterial solution to 7.0 using NaOH, centrifuge at 5000 rpm for 5 min, discard the supernatant, and collect the centrifugal precipitate (bacterial mud). Add heat shock protective agent to the bacterial mud in proportion (bacterial mud: 30% trehalose solution = 1:1, W / W). Preheat the obtained mixture to 80°C, then stir continuously at 80°C and 200 rpm for 30 min to complete inactivation, and obtain an inactivated bacterial suspension. Alternatively, the inactivated bacterial suspension is further spray-dried and crushed to obtain inactivated bacterial powder, which is stored at room temperature and protected from light for later use.

[0028] The components of the BHI medium involved below are: 4.0 g / L of bovine brain extract powder, 4.0 g / L of bovine heart extract powder, 5.0 g / L of peptone, 16.0 g / L of casein peptone, 5.0 g / L of sodium chloride, 2.0 g / L of glucose, 2.5 g / L of disodium hydrogen phosphate dodecahydrate, and pH value is 7.2~7.4.

[0029] The components of the MRS medium involved below are: yeast powder 5.0 g / L, beef extract 10.0 g / L, peptone 10.0 g / L, glucose 20.0 g / L, anhydrous sodium acetate 2.0 g / L, diammonium hydrogen citrate 2.0 g / L, dipotassium hydrogen phosphate trihydrate 2.6 g / L, manganese sulfate monohydrate 0.25 g / L, magnesium sulfate heptahydrate 0.5 g / L and Tween-80 1 mL / L, pH 6.2~6.4.

[0030] Preparation Example 1

[0031] This preparation example provides a probiotic agent, which is composed of inactivated FB97 bacterial powder and inactivated SY23 bacterial powder, wherein the total content of FB97 strain and RH22 strain in the probiotic agent is 1×10 10 / g, and the ratio of the bacterial counts of FB97 strain and RH22 strain was 8:1.

[0032] Preparation Example 2

[0033] This preparation example provides a probiotic agent, which is composed of inactivated FB97 bacterial powder and inactivated RH22 bacterial powder, wherein the total content of FB97 strain and RH22 strain in the probiotic agent is 1×10 10 / g, and the ratio of the bacterial counts of FB97 strain and RH22 strain was 1:1.

[0034] Preparation Example 3

[0035] This preparation example provides a probiotic agent, which is composed of inactivated FB97 bacterial powder and inactivated RH22 bacterial powder, wherein the total content of FB97 strain and RH22 strain in the probiotic agent is 1×10 10 / g, and the ratio of the bacterial counts of FB97 strain and RH22 strain was 1:5.

[0036] Comparative Preparation Example 1

[0037] This comparative preparation example provides a probiotic agent, which is composed of inactivated ATCC 25302 bacterial powder and inactivated RH22 bacterial powder, wherein the total content of ATCC 25302 strain and RH22 strain in the probiotic agent is 1×10 10 / g, and the ratio of the bacterial counts of ATCC 25302 strain and RH22 strain was 1:1.

[0038] Comparative Preparation Example 2

[0039] This comparative preparation example provides a probiotic agent, which is an inactivated FB97 bacterial powder, wherein the number of bacteria contained in the FB97 bacterial powder is 1×10 10 Pieces / g.

[0040] Comparative Preparation Example 3

[0041] This comparative preparation example provides a probiotic agent, which is an inactivated RH22 bacterial powder, wherein the number of bacteria contained in the RH22 bacterial powder is 1×10 10 Pieces / g.

[0042] Test Example 1

[0043] Test on the ability to inhibit harmful oral bacteria

[0044] (1) Preparation of test bacteria: Oral pathogenic bacteria mutans Streptococcus was inoculated into BHI liquid culture medium at 2% (v / v), cultured at 37°C for 24 h, and then the bacterial solution concentration was adjusted to 10 6 CFU / mL.

[0045] (2) Experimental sample preparation: The probiotic samples prepared in Preparation Examples 1-3 and Comparative Preparation Examples 1-3 were diluted with PBS buffer to a concentration of 0.1%.

[0046] (3) Take 5 mL of sample and place them in sterile test tubes. After 5 min in a 20°C water bath, add 0.1 mL of oral pathogenic Streptococcus mutans suspension to each tube. Mix and react for 1 min. Take 1 mL of the reaction solution and inoculate it into a plate and pour the culture medium. At the same time, use PBS buffer instead of sample to perform parallel experiments as a positive control. All groups of samples were cultured at 36°C for 48 h and the final results were observed: the experiment was repeated 3 times and the inhibition rate was calculated according to the following formula.

[0047]

[0048] Among them, X is the inhibition rate, %; A0 is the amount of bacteria recovered in the positive control group, CFU / mL; A1 is the amount of bacteria recovered in the experimental sample group, CFU / mL.

[0049] The results are shown in Table 1.

[0050] Table 1

[0051]

[0052] From the data in the table, it can be seen that each experimental sample group has different degrees of inhibitory effect on the oral pathogenic bacteria Streptococcus mutans, and the FB97 strain and the RH22 strain have a synergistic effect in inhibiting the efficacy of oral pathogens.

[0053] Test Example 2

[0054] Test on the ability to promote the growth of beneficial oral bacteria

[0055] (1) Preparation of test bacteria: 2% (v / v) of Streptococcus salivarius, a beneficial oral bacteria, was inoculated into MRS liquid culture medium and cultured at 37°C for 24 h. The concentration of the culture medium was adjusted to 10 8 CFU / mL.

[0056] (2) Experimental sample preparation: Weigh 0.1 g of the probiotic samples prepared in Preparation Examples 1-3 and Comparative Preparation Examples 1-3, respectively, add them into 100 mL of sterilized MRS liquid culture medium, mix well, and then test.

[0057] (3) The Streptococcus salivarius suspension prepared in step (1) was inoculated at an inoculum rate of 2% (v / v) into the MRS liquid culture medium containing the probiotic prepared in step (2), and cultured at 37°C with a shaker at 100 rpm for 20 h. At the same time, PBS buffer was used instead of the probiotic to perform a parallel experiment as a blank control. The experiment was repeated 3 times, and the growth promotion rate was calculated according to the following formula.

[0058]

[0059] Among them, Y is the growth promotion rate, %; N0 is the number of colonies in the experimental sample group; N1 is the number of colonies in the blank group.

[0060] The results are shown in Table 2.

[0061] Table 2

[0062]

[0063] From the data in the table, it can be seen that each experimental sample group has a different degree of promoting effect on the beneficial oral bacteria Streptococcus salivarius, and the FB97 strain and the RH22 strain have a synergistic effect in promoting the proliferation of beneficial oral bacteria.

[0064] Test Example 3

[0065] Biofilm inhibition test

[0066] (1) Preparation of test bacteria: Oral pathogenic bacteria mutans Streptococcus was inoculated into BHI liquid culture medium at 2% (v / v), cultured at 37°C for 24 h, and then diluted with BHI liquid culture medium to an OD of 600 =0.5.

[0067] (2) Experimental sample preparation: Weigh 0.2 g of the probiotic samples prepared in Preparation Example 1-3 and Comparative Preparation Example 1-3, respectively, add them into 100 mL of sterilized BHI liquid culture medium, mix and dilute, and then test.

[0068] (3) The test bacterial solution prepared in step (1) and the test sample dilution prepared in step (2) were mixed evenly at a volume ratio of 1:1, and 200 μL of the mixture was added to a 96-well plate. The test bacterial solution in step (1) was diluted by one half using BHI liquid culture medium, and 200 μL was added to a 96-well plate as a positive control group. Three parallels were set for each group. The sample group mixture and the positive control group test bacterial solution were placed at 37°C for 24 h for anaerobically incubated to form a biofilm.

[0069] (4) Use crystal violet staining to test the amount of biofilm formation. After the culture is completed, carefully aspirate the culture medium of each well to remove free bacteria and maintain the integrity of the biofilm. Wash three times with sterile PBS solution and add methanol to fix for 15 minutes. Discard the methanol and let it stand to dry. Then add 100 μL of 0.1% (m / v) crystal violet solution to each well and stain for 15 minutes at 25°C. After staining, remove excess stain and wash three times with sterile PBS solution. After drying, add 200 μL of 95% ethanol to dissolve the stained biofilm, shake at low speed at 25°C for 20 minutes to decolorize. After decolorization, use an enzyme reader at OD 550 Read the absorbance value at nm and calculate the biofilm inhibition rate according to the following formula.

[0070]

[0071] Among them, P0 is the absorbance value of the positive control group after staining, and P1 is the absorbance value of the sample group after staining.

[0072] The results are shown in Table 3.

[0073] Table 3

[0074]

[0075] From the data in the table, it can be seen that each experimental sample group has different degrees of inhibitory effect on the formation of biofilm of Streptococcus mutans, and the FB97 strain and the RH22 strain have a synergistic effect in inhibiting the formation of biofilm of oral harmful bacteria.

[0076] Application Example 1

[0077] This application example provides a toothpaste product, wherein the raw materials for preparation are composed of the following by mass percentage: 0.1% of the probiotic prepared in Preparation Example 1, 50% of calcium carbonate, 10% of glycerol, 5% of sorbitol, 2% of sodium lauryl sulfate, 1% of flavor, 1% of sodium carboxymethyl cellulose, 0.5% of calcium hydrogen phosphate, 0.25% of saccharin, 0.1% of water glass, and the balance is water. The preparation method is: after mixing the raw materials, homogenizing them with a homogenizer to obtain the toothpaste product.

[0078] Application Example 2-3

[0079] This application example provides two toothpaste products, and the difference in the raw materials for their preparation from those in application example 1 is only that equal amounts of the probiotics prepared in preparation example 1 are replaced by the probiotics prepared in preparation examples 2-3, respectively, and other conditions remain unchanged.

[0080] Comparative Application Examples 1-3

[0081] This comparative application example provides three toothpaste products, the preparation raw materials of which differ from those of application example 1 only in that equal amounts of the probiotics prepared in preparation example 1 are replaced by probiotics prepared in comparative preparation examples 1-3, respectively, and other conditions remain unchanged.

[0082] Test Example 4

[0083] Plaque inhibition experiment - toothpaste slurry sample

[0084] Materials preparation:

[0085] (1) Glass rod: Use silicon carbide sandpaper No. 240, 320, 400 and 600 on a lathe to grind the glass rod 25 mm from the thick end. Before each experiment, use dilute hydrochloric acid to ultrasonically clean the glass rod, rinse it, dry it, grind it with No. 600 silicon carbide sandpaper, rinse it with deionized water and dry it before use.

[0086] (2) Collection and preparation of saliva: Collect saliva from 10 people each time, mix thoroughly, and refrigerate at 4°C for later use.

[0087] (3) Preparation of plaque growth medium: Dissolve 15 g of BBL tryptic soy peptone and 50 g of sucrose in 402 g of deionized water that has been sterilized by high-pressure steam for 15 min and cooled, mix thoroughly, and add 33 g of freshly collected saliva before use.

[0088] (4) Preparation of glycolysis medium: Dissolve 15 g of BBL tryptic soy peptone and 25 g of sucrose in deionized water that has been sterilized by high-pressure steam for 15 min and cooled, and make up to 500 mL.

[0089] Experimental process:

[0090] (1) Fix the glass rod and immerse it in 7 mL of fresh saliva containing 0.1% sucrose. Place it in a 37°C incubator and immerse it for 10 h. The immersion depth is 1.5 cm and the rotation speed is 1 r / min.

[0091] (2) Transfer the glass rod to 7 mL of plaque growth medium (with freshly collected saliva added), soak it in a 37°C incubator for 8 h to a depth of 1.5 cm, and then place it in freshly collected saliva for 10 h.

[0092] (3) Transfer the glass rod to 7 mL of the toothpaste prepared in Application Examples 1-3, Comparative Application Examples 1-3 and the negative control group, and soak it once every 2 s for 0.5 s each time, for a total of 60 s. Then, wash the glass rod with deionized water for 10 s each time. After washing twice, transfer the glass rod to 7 mL of glycolysis medium and soak it at 37°C for 6 h. Measure and record the pH of the glycolysis buffer of the control group and the experimental sample group, and calculate the relative efficacy of the toothpaste slurry samples according to the following formula.

[0093] Among them, step (3) of the positive control group sample is to directly transfer the glass rod to the glycolysis medium for immersion without being treated with toothpaste slurry.

[0094]

[0095] In the above formula, X is the relative efficacy of the experimental sample group and the positive control group, %; A0 is the mean pH value of the positive control group, A1 is the mean pH value of the experimental sample group, and A2 is the mean pH value of the negative control group.

[0096] The results are shown in Table 4.

[0097] Table 4

[0098]

[0099] From the data in the table, it can be seen that each experimental sample group has different degrees of inhibitory effect on dental plaque formation, and the FB97 strain and the RH22 strain have a synergistic effect in inhibiting the formation of dental plaque.

[0100] Test Example 5

[0101] Inhibition of dental plaque formation test - mouthwash

[0102] Material preparation: The raw materials for the preparation of mouthwash products are calculated by mass percentage by mixing 0.1% of the probiotic samples prepared in Preparation Examples 1-3 and Comparative Preparation Examples 1-3, 1% sorbitol, 2% propylene glycol and 96.9% water to prepare 6 groups of mouthwash products.

[0103] The experiment adopted a randomized, double-blind, placebo-controlled before-after population trial. Volunteers were screened one week before the trial, and then subjects were selected according to the inclusion criteria. The subjects' mouths were cleaned to establish the same oral environment, and the same commercially available toothbrush and toothpaste were used. After a 7-day recovery period, 10 people were randomly assigned to each group for trial testing.

[0104] Experimental process: Each group of subjects brushed their teeth with the same commercially available toothbrush and toothpaste every night, then spit out 15mL of mouthwash after 30s, and were not allowed to take in any food or water afterwards. After 21 days of the experiment, the improvement of the gingival index (GI) and plaque index (PLI) of each group of subjects was tested.

[0105] Among them, the gingival index (GI) examination method: the examination uses a blunt-tipped periodontal probe, combined with visual examination and probing. Check the entire mouth or a few selected teeth. The gums around each tooth must be checked, and the surrounding gums are divided into the mesial lip (cheek) papilla, the median lip (cheek) margin, the distal lip (cheek) papilla and the lingual gingival margin. The score for each tooth is the average of the scores of the four tooth surfaces, and the score for each person is the average of the scores of all the teeth examined. The evaluation method is: healthy gums, 0 points; mild gingival inflammation (the color of the gums has changed slightly and there is mild edema, and there is no bleeding on probing), 1 point; moderate gingival inflammation (the gums are red, the edema is bright, and there is bleeding on probing), 2 points; severe gingival inflammation (the gums are obviously red and swollen or there are ulcers, and there is a tendency to bleed automatically), 3 points.

[0106] The plaque index (PLI) is evaluated by staining the plaque with a plaque stain and then evaluating the area of ​​the plaque. The evaluation method is: no plaque on the tooth surface, 0 points; scattered dot-shaped plaque at the gingival margin of the neck of the tooth, 1 point; the width of the plaque at the neck of the tooth does not exceed 1mm, 2 points; the plaque coverage area of ​​the neck of the tooth exceeds 1mm 2 , but less than 1 / 3 of the tooth surface, 3 points; plaque coverage area accounts for 1 / 3 to 2 / 3 of the tooth surface, 4 points; plaque coverage area accounts for more than 2 / 3 of the tooth surface, 5 points. The results are shown in Table 5.

[0107] Table 5

[0108]

[0109] From the data in the table, it can be seen that each experimental sample group has different degrees of inhibitory effect on dental plaque formation, and the FB97 strain and the RH22 strain have a synergistic effect in inhibiting the formation of dental plaque.

[0110] In summary, the present invention has developed a new probiotic compounding method and a new strategy for regulating the oral microecological environment, that is, compounding the inactivated Lacticaseibacillus paracasei FB97 strain and the inactivated Lacticaseibacillus rhamnosus RH22 strain, and found that there is a potential interaction between the two postbiotics, which can cooperate with each other and synergize. When the amount of bacteria used is the same, it has a more significant and excellent effect than a single postbiotic or other postbiotic compounding methods. Specifically, it is manifested in: (1) inhibiting the excessive growth of harmful oral bacteria; (2) promoting the proliferation of beneficial oral bacteria and maintaining a healthy oral microecological environment; (3) inhibiting the formation of dental plaque. Therefore, the composite probiotic has a good application prospect for the preparation of oral care products.

[0111] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0112] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0113] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A probiotic for regulating the oral microecological environment, characterized in that: The strain in the probiotic agent is Lactobacillus paracasei with a preservation number of CGMCC No.30464. Lacticaseibacillus paracasei FB97 strain postbiotics and inactivated Lactobacillus rhamnosus with a deposit number of CGMCC No. 31054 Lacticaseibacillus rhamnosus Postbiotic composition of RH22 strain; The ratio of the bacterial counts of the Lactobacillus paracasei FB97 strain to the Lactobacillus rhamnosus RH22 strain is 1:10-10:1; The concentration of the strain in the probiotic is not less than 1×10 10 / mL or 1×10 10 Pieces / g.

2. The probiotic agent for regulating the oral microecological environment according to claim 1, characterized in that: The dosage form of the probiotics includes liquid, lyophilized powder, capsule, tablet or granule.

3. Use of the probiotic according to claim 1 or 2 in the preparation of oral care products.

4. The use according to claim 3, characterized in that: The oral care products include toothpaste, mouthwash, oral lozenges, and oral gel.

5. A mouthwash for preventing or improving dental plaque formation, characterized in that: The raw materials for preparing the mouthwash include the probiotic agent according to claim 1 or 2 and a solvent.

6. The mouthwash according to claim 5, characterized in that The raw materials for preparing the mouthwash also include any one of flavoring agents, sweeteners, moisturizers, surfactants, colorants, preservatives or functional additives, or a combination of at least two of them.

7. The mouthwash according to claim 5, characterized in that The mass percentage of the probiotic in the mouthwash is 0.05%-2%.

8. A toothpaste for preventing or improving dental plaque formation, characterized in that: The raw materials for preparing the toothpaste include: the probiotic agent according to claim 1 or 2, a moisturizer, an abrasive, a foaming agent, a thickener and a taste regulator.

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