CN111840126A - Oral care agent with antibacterial effect and preparation method thereof - Google Patents

Abstract

The invention relates to the field of prevention and treatment of oral diseases, and particularly relates to an oral care agent with a bacteriostatic action, which comprises lysozyme and montmorillonite. Specifically discloses lysozyme biological toothpaste, which comprises 0.4% of lysozyme, 1% of montmorillonite, an abrasive, a wetting agent, an adhesive, a surfactant, a preservative, a sweetening agent, a thickening agent, a foaming agent and the balance of deionized water. The lysozyme biological toothpaste has antibacterial effect on oral pathogenic bacteria, has a wide antibacterial spectrum and a high antibacterial rate, can efficiently and briefly inhibit the growth of the oral pathogenic bacteria, has a low minimum antibacterial concentration, and does not generate drug-resistant strains of the oral pathogenic bacteria after long-term use; the prepared toothpaste can be free from the denaturation influence of foaming agent, binder, sweetener, preservative and other substances on lysozyme, and can prevent the activity of biological lysozyme from being damaged or reduced, thereby playing a role in inhibiting oral pathogenic bacteria, effectively playing a role in preventing caries and periodontitis, and having unique advantages and wide application prospect.

Description

Oral care agent with antibacterial effect and preparation method thereof

Technical Field

The invention relates to the field of prevention and treatment of oral diseases, in particular to an oral care agent with a bacteriostatic action and a preparation method thereof.

Background

The common oral diseases such as dental caries, periodontitis, pericoronitis, oral ulcer and the like are important public health problems worldwide and are proved to be related to oral bacterial infection. Oral bacteria, usually in the form of plaque biofilm, adhere to and surround the tooth surface and continue to produce acid, resulting in demineralization of the enamel and eventual caries, periodontal disease and gingivitis. Inhibiting the growth of/killing oral bacteria is a final goal in the prevention and treatment of oral diseases. Fluoride, antibiotics and chemical disinfectants are common medicines for preventing and treating oral diseases at present and achieve good effect. However, in recent years, researchers have found that the wide use of antibiotics has led to the emergence of drug-resistant strains, and there is no antibiotic that prevents drug-resistant strains. Fluoride and chemical disinfectants also achieve certain efficacy, but still have deficiencies. At present, the main method for preventing oral diseases is tooth brushing, and the tooth brushing effect is directly influenced by the components of toothpaste. At present, toothpaste in the market mainly comprises Chinese herbal medicine components, and a plurality of natural Chinese herbal medicines have certain antibacterial effect, such as gallnut, coptis chinensis, rheum officinale, scutellaria baicalensis, honeysuckle, rhizoma drynariae, traditional Chinese medicine Shuanghuangbu, tea leaves and the like. However, because the traditional Chinese medicine components are complex and the prevention and treatment effects of part of the traditional Chinese medicines cannot be expected, people begin to divert attention, add part of biological and chemical components with obvious bacteriostatic effects into toothpaste, and study the effect of inhibiting oral bacteria.

Lysozyme (also called as cell wall lytic enzyme) can selectively decompose microbial cell walls to achieve the bactericidal effect, and the principle is that beta-1, 4 glycosidic bonds between C-1 of N-acetylmuramic acid and C-4 of N-acetylglucosamine are catalyzed to destroy the cell walls of bacteria, and the bacteria can be broken under the action of intracellular pressure. And the lysozyme does not damage other biological tissues. Meanwhile, the bio-enzyme preparation is a natural protein, can be completely digested and absorbed in gastrointestinal tracts, has no toxicity and irritation to human bodies, and is a bio-enzyme preparation with high safety. Meanwhile, research finds that bacterial infection occurs when Recurrent Aphthous Ulcers (RAU) occur, and the infection range is not limited to the ulcer surface. The lysozyme helps to eliminate infection of a lesion part and promote ulcer healing by killing and inhibiting microorganisms. At present, some patents have applied lysozyme to the control of oral pathogenic bacteria, for example, a patent of an oral care product (CN201710922542.8) containing biological lysozyme discloses an oral care product containing biological lysozyme, the oral care product contains 0.1-15% (wt) of a biological lysozyme additive composition, and the biological lysozyme additive composition consists of the following components in percentage by weight: 10-40% of biological lysozyme, 0.01-2.0% of modified vegetable gum, 0.01-0.3% of EDTA disodium and the balance of deionized water. The oral care product can effectively inhibit and kill harmful bacteria causing oral problems such as oral ulcer, gingivitis, halitosis and the like. The lysozyme is used alone, the bacteriostatic spectrum is narrow, the bacteriostatic effect is general, and technicians in the field perform numerous compatibility experiments on the lysozyme so as to obtain a bacteriostatic composition with wide bacteriostatic spectrum and good bacteriostatic effect, but at present, no composition capable of achieving the purpose is obtained. Meanwhile, because the lysozyme is a protein component, the lysozyme is denatured by foaming agents, binding agents, sweetening agents, chemical preservatives and the like which are essential in the conventional oral products in different degrees, so that the activity of the lysozyme is reduced or lost. Therefore, effective protection of lysozyme activity is one of the major and difficult points for its application in oral products.

Natural montmorillonite is a dioctahedral phyllosilicate clay mineral composed of silicon-oxygen tetrahedron and aluminum-oxygen octahedron, and the random substitution of aluminum and magnesium for silicon and aluminum in the structure causes charge imbalance, so that the natural montmorillonite has the characteristics of swelling, adsorption, electrification and ion exchange when meeting water. Is clinically used as a medicament for treating diarrhea and digestive tract ulcer. Montmorillonite has no direct antibacterial effect of antibacterial drugs.

The invention has the advantages that the composition of lysozyme and montmorillonite has better antibacterial effect, wider antibacterial spectrum and lower minimum antibacterial concentration, can be used as a main component to be applied to preventing oral infectious diseases, and can be prepared into toothpaste, and the antibacterial effect of the toothpaste is not influenced by foaming agents, binding agents, sweetening agents, chemical preservatives and the like in the toothpaste.

Disclosure of Invention

Aiming at the technical problems, the invention provides an oral care agent with antibacterial effect, which comprises 0.4% of lysozyme and 1% of montmorillonite.

Preferably, the toothpaste comprises 0.4% of lysozyme, 1% of montmorillonite, an abrasive, a wetting agent, a binding agent, a surfactant, a preservative, a sweetening agent, a thickening agent and a foaming agent, and the balance of deionized water.

Preferably, the abrasive is one or more of calcium carbonate, calcium hydrophosphate, hydrated silica and silicon dioxide; the humectant is one or more of sorbitol, glycerol, propylene glycol and polyethylene glycol; the adhesive is one or more of sodium carboxymethylcellulose, xanthan gum, hydroxyethyl cellulose, guar gum and carrageenan; the foaming agent is sodium dodecyl sulfate or sodium lauryl sulfate; the surfactant is one or more of sodium lauryl sulfate, sodium 2-acyloxy bond sulfonate, trisodium phosphate and sodium lauroyl sarcosinate; the thickener is one or more of carboxymethyl cellulose, antler pectin, hydroxyethyl cellulose and xanthan gum; the sweetener is one or more of glycerol, sodium cyclamate and saccharin sodium; the preservative is one or more of potassium sorbate and sodium benzoate; the balance of deionized water.

Preferably, the toothpaste further comprises pigment and essence.

Preferably, the toothpaste comprises polyethylene glycol-85%, sodium carboxymethylcellulose 1%, hydroxyethyl cellulose 0.3%, sorbitol 45%, sodium saccharin 0.21%, trisodium phosphate 0.15%, titanium dioxide 0.35%, calcium hydrogen phosphate 0.5%, deionized water 18.5%, hydrated silica 24.5%, sodium lauryl sulfate 2.4%, essence 1%, sodium benzoate 0.2%, montmorillonite 1%, lysozyme 0.4%.

The preparation method of the toothpaste comprises the following steps: mixing lysozyme, montmorillonite, abrasive, wetting agent, adhesive and foaming agent according to the weight percentage, and stirring, grinding and vacuum degassing to obtain the toothpaste body.

Preferably, the method comprises the following steps of:

(1) adding lysozyme, montmorillonite, wetting agent and saccharin into deionized water, stirring to obtain deionized water solution, mixing with abrasive, binder, foaming agent, surfactant, antiseptic, sweetener and thickener, stirring, grinding, and vacuum degassing;

(2) adding essence and pigment, stirring, grinding, and vacuum degassing to obtain toothpaste.

The invention has the beneficial effects that: the lysozyme biological toothpaste has antibacterial effects on oral pathogenic bacteria such as Escherichia coli, Streptococcus mutans, Staphylococcus aureus, Porphyromonas gingivalis, Practiella intermedia and other bacteria, has a wide antibacterial spectrum and a high antibacterial rate, can efficiently and briefly inhibit the growth of the oral pathogenic bacteria, has a low minimum antibacterial concentration, and does not generate oral pathogenic bacteria drug-resistant strains after long-term use; the antibacterial effect of the lysozyme biological toothpaste prepared by the invention is equivalent to that of a composition of lysozyme and montmorillonite, which shows that the toothpaste prepared by the invention can be prevented from being influenced by the denaturation of the lysozyme by substances such as foaming agents, binders, sweeteners, preservatives and the like, and the activity of the biological lysozyme is prevented from being damaged or reduced, so that the function of inhibiting oral pathogenic bacteria is achieved.

Drawings

FIG. 1 shows the bacteriostatic effect of Streptococcus mutans measured by Oxford cup method.

A is experimental group 1: lysozyme treated group, B is experimental group 3: lysozyme and montmorillonite composition treatment group, C is experimental group 4: lysozyme biological toothpaste, D is experimental group 2: montmorillonite treatment group, E experimental group 5: blank toothpaste treatment group.

FIG. 2 is a graph showing the bacteriostatic effect of Escherichia coli measured by Oxford cup method.

FIG. 3 is a graph showing the inhibition effect of Porphyromonas gingivalis by Oxford cup method.

FIG. 4 shows the bacteriostatic effect of Staphylococcus aureus measured by Oxford cup method.

FIG. 5 shows the bacteriostatic effect of Prevotella intermedia measured by the Oxford cup method.

FIG. 6 WST-8 kit for testing the bacteriostatic effect of Streptococcus mutans at different times, p is less than 0.05.

FIG. 7 WST-8 kit for testing the bacteriostatic effect of E.coli at different times, p < 0.05.

FIG. 8 WST-8 kit for testing the inhibitory effect of Porphyromonas gingivalis at different times, p is less than 0.05.

FIG. 9 WST-8 kit for testing the inhibitory effect of Staphylococcus aureus at different times, p < 0.05.

FIG. 10 WST-8 kit for testing the inhibitory effect of Prevotella intermedia at different times, p < 0.05.

Detailed Description

The technical solutions of the present invention are described in detail below by using specific examples, but the scope of the present invention is not limited by the following examples, and any technical solutions obtained by modifying the technical solutions of the present invention by those skilled in the art belong to the scope of the present invention.

The sources of the strains used in the experiment of the invention are as follows:

streptococcus mutans (Streptococcus mutans), Escherichia coli (Escherichia coli), Porphyromonas gingivalis (Porphyromonas gingivalis), Staphylococcus aureus (Staphyloccocusareus) and Prevotella intermedia (Prevotella intermedia) used in the present invention were purchased from China general microbiological culture Collection center.

The sources of the drugs used in the experiment of the invention are as follows:

lysozyme (Shanghai Zhonghua pharmaceutical Co., Ltd., China pharmacopoeia, batch number: 190202);

montmorillonite (Beijing Solibao, cat # YZ-100860);

the dragon tiger lysozyme biological toothpaste (Shanghai Zhonghua pharmaceutical Co., Ltd., batch No. 181220).

An experimental instrument:

anaerobic incubators (GeneScience, E500, usa), bacteria thermostatted incubators (shanghai xin instrument, XY-DR-360), biosafety cabinets (shanghai liking, HFsafe-1200a2), full-wavelength plate readers (TECAN, Infinite 200Pro, switzerland), fluorescence microscopes (olympus, BX53), adjustable pipetting guns (mettler, XLS +), electronic balances (BT 25S, germany seudoli); bovine heart-brain broth medium (Beijing Huayuyo, GU9-2814), Brucella broth medium (Beijing Soilebao, LA3560), LB medium (Beijing Soilebao, L1010), Columbia blood agar medium (Jiangmen Kailin trade Co., Ltd.), hemin (Beijing Soilebao, H8130), WST-8 kit (Japanese colleague, M439)

Example one inhibition of oral bacteria by an oral Care agent with bacteriostatic action

In this experiment, the inhibitory effect of the lysozyme oral care agent on the common oral bacteria Streptococcus mutans (Streptococcus mutans), Escherichia coli (Escherichia coli), Porphyromonas gingivalis (Porphyromonas gingivalis), Staphylococcus aureus (Staphylococcus aureus) and Prevotella intermedia (Prevotella intermedia) was examined by an in vitro assay method. Because the common colony counting method has strong subjectivity, and a sample to be detected cannot be completely dispersed into single bacteria, the result accuracy of the plate colony counting is low, and a grown single colony can come from 2-3 or more cells in the sample, so that accurate counting is not easy to obtain. Therefore, the oxford cup method is adopted in the experiment, and the bacteriostatic effect of each experimental group is directly observed. The Oxford cup method is a common method for detecting the antibacterial effect, and the grouped antibacterial effect is judged by measuring the diameter of the antibacterial zone.

1. Preparation of culture Medium

Preparing a BHI culture medium: 3.7 percent of bovine heart brain infusion medium powder is placed in an ultra-clean bench to be cooled for standby after being sterilized at high temperature and high pressure for 25 minutes. Adding 1.5% agar, preparing agar plate, cooling to 50 deg.C, and pouring. Used for culturing the streptococcus mutans.

Preparing an LB culture medium: and (3) sterilizing the LB culture medium with the concentration of 2.5% at high temperature and high pressure for 25 minutes, and then placing the LB culture medium on a clean bench for cooling for later use. Adding 1.5% agar, preparing agar plate, cooling to 50 deg.C, and pouring. Used for culturing Escherichia coli.

Preparing a BHI blood agar culture medium: 3.7% bovine heart brain infusion medium powder, autoclaving at high temperature for 25 minutes, and adding hemin to a final concentration of 5 μ g/mL. Adding 1.5% agar, preparing agar plate, cooling to 50 deg.C, and pouring. Is used for culturing the porphyromonas gingivalis.

Preparing MH culture medium: weighing 21.0g of culture medium powder, heating and dissolving in 1000ml of distilled water, sterilizing at high temperature and high pressure for 25 minutes, and then placing in an ultra-clean bench for cooling for later use. Adding 1.5% agar, preparing agar plate, cooling to 50 deg.C, and pouring. The culture medium is used for culturing staphylococcus aureus.

Preparing a Columbia culture medium: weighing 39 g of culture medium powder, heating and dissolving in 1000ml of distilled water, subpackaging, sterilizing at high temperature and high pressure for 25 minutes, cooling to about 45-50 ℃, adding sterile gentamicin sulfate containing 20mg of gentamicin, and mixing uniformly for later use. Adding 1.5% agar, preparing agar plate, cooling to 50 deg.C, and pouring. The culture medium is used for culturing the prevotella intermedia.

2. Recovery and culture of strain

And (3) recovering the streptococcus mutans: streptococcus mutans lyophilized strains were inoculated on BHI agar plates and incubated in a 37 ℃ bacterial incubator (containing 5% CO)₂) The culture was carried out for 24 hours.

Culturing streptococcus mutans: selecting recovered Streptococcus mutans strain, inoculating into BHI liquid culture medium, culturing for 24 hr, and preparing into bacterial suspension (10) with PBS by McLeod's turbidimetry⁸CFU/mL), ready for use.

And (3) escherichia coli recovery: the lyophilized strain of Escherichia coli was inoculated on an LB agar plate and incubated at 37 ℃ in a bacteria incubator (containing 5% CO)₂) The culture was carried out for 24 hours.

And E, culturing escherichia coli: selecting recovered colibacillus strain, inoculating it in LB liquid culture medium, culturing for 24 hr,bacterial suspension (10) was prepared from malt turbidimetry with PBS⁸CFU/mL), ready for use.

And (3) recovering the porphyromonas gingivalis: the lyophilized strain of Porphyromonas gingivalis was inoculated on BHI blood agar plates and anaerobically (volume ratio: 80% N) at 37 deg.C₂，10％H₂，10％CO₂) The culture was carried out for 48 hours.

Culturing porphyromonas gingivalis: selecting recovered Porphyromonas gingivalis strain, inoculating into blood-containing BHI liquid culture medium, anaerobically culturing for 48 hr, and preparing into bacterial suspension (10) with PBS by McLeod turbidimetry⁸CFU/mL), ready for use.

And (3) recovering staphylococcus aureus: inoculating lyophilized Staphylococcus aureus strain on MH agar plate, and culturing in 37 deg.C bacteria incubator (containing 5% CO)₂) The culture was carried out for 24 hours.

Culturing staphylococcus aureus: selecting recovered Staphylococcus aureus strain, inoculating to MH liquid culture medium, culturing for 24 hr, and preparing into bacterial suspension (10) with PBS by McLeod turbidimetry⁸CFU/mL), ready for use.

And (3) recovering the intermediate prevotella: the lyophilized strain of Prevotella intermedia was inoculated on a Columbia blood agar plate and anaerobically incubated at 37 ℃ (volume: 85% N)₂，5％H₂，10％CO₂) The culture was carried out for 48 hours.

Culturing intermediate prevotella: selecting recovered strain, inoculating into Columbia blood agar culture medium, placing in 37 deg.C anaerobic incubator, vacuumizing the incubator with vacuum pump, and charging oxygen-free mixed gas (5% H)₂，10％CO₂，85％N₂). After 48 hours of incubation, bacterial suspension (10) was prepared using PBS by McLeod's turbidimetry⁸CFU/mL), ready for use.

3. Preparation and grouping of drugs to be tested

Experimental group 1: accurately weighing 0.4g of lysozyme, ultrasonically dispersing the lysozyme in 100mL of 5 bacterial culture media to be detected, taking 2mL of the solution, adding 2 mu L of bacterial suspension to be detected, wherein the final concentration of the medicine is as follows: 0.004g/ml lysozyme;

experimental group 2: 1.0g of montmorillonite is accurately weighed, the montmorillonite is ultrasonically dispersed in 100mL of 5 bacteria culture media to be detected, 2mL of the solution is taken, 2 mu L of bacteria suspension to be detected is added, and the final concentration of the medicine is as follows: 0.01g/ml montmorillonite;

Experimental group 3: accurately weighing 0.4g of lysozyme and 1.0g of montmorillonite, ultrasonically dispersing in 100mL of 5 bacterial culture media to be detected, taking 2mL of the solution, adding 2 mu L of bacterial suspension to be detected, wherein the final concentration of the medicine is as follows: 0.004g/ml lysozyme and 0.01g/ml montmorillonite;

experimental group 4: accurately weighing biological toothpaste of the dragon and tiger lysozyme, which is equivalent to 0.4g of lysozyme and 1.0g of montmorillonite, ultrasonically dispersing the biological toothpaste in 100mL of 5 bacteria culture media to be detected, taking 2mL of the solution, adding 2 mu L of bacterial suspension to be detected, wherein the final concentration of the medicine is as follows: 0.004g/ml lysozyme and 0.01g/ml montmorillonite;

experimental group 5: blank toothpaste in the same weight as the toothpaste in the experimental group 4 was precisely weighed, ultrasonically dispersed in 100mL of 5 bacterial culture media to be tested, and 2mL of this solution was taken and 2. mu.L of bacterial suspension to be tested was added.

4. Procedure of experiment

And (3) culturing each experimental group for 12h under corresponding culture conditions, and testing the antibacterial effect of each group of samples by adopting an Oxford cup method: and uniformly coating corresponding bacteria on the surface of a corresponding agar culture plate by using an aseptic coating rod, after the corresponding bacteria are basically dried in an ultra-clean bench, coating the surface of the culture plate in a partition mode, placing a plurality of aseptic oxford cups, and injecting corresponding samples in each group into the aseptic cups. After culturing for 12h under corresponding bacteria culture conditions, observing and evaluating the diameter of the bacteriostatic zone, and judging the reference standard of the antibacterial effect: the National Committee for Clinical Laboratory Standardization (NCCLS).

SPSS 21.0 software is used for data processing, single-factor variance analysis is adopted, and statistical significance is achieved when P is less than 0.05.

5. Results of the experiment

Each group 10⁸Diluting the CFU/mL bacterial solution by 1000 times by using a corresponding drug-containing culture medium, uniformly coating a culture dish, adding drugs, and culturing for 12 h. The bacteriostatic effect of different oral pathogenic bacteria of each experimental group is shown in figures 1-5, and the measurement data is shown in table 1: :

TABLE 1 inhibition effect of each experimental group on oral pathogenic bacteria

As can be seen from FIGS. 1 to 5 and Table 1, montmorillonite itself has no bacteriostatic ability; the lysozyme has an inhibiting effect on streptococcus mutans, escherichia coli, porphyromonas gingivalis, staphylococcus aureus and prevotella intermedia; the montmorillonite and lysozyme composition can also inhibit streptococcus mutans, escherichia coli, porphyromonas gingivalis, staphylococcus aureus and prevotella intermedia, compared with lysozyme and montmorillonite, the antibacterial effect of the montmorillonite and lysozyme composition is remarkably increased, auxiliary materials are added into the montmorillonite and lysozyme composition to prepare the lysozyme biological toothpaste, the antibacterial effect of the obtained lysozyme biological toothpaste is equivalent to that of the montmorillonite and lysozyme composition, and the blank toothpaste has no antibacterial effect. Compared with lysozyme, montmorillonite and blank group toothpastes, the lysozyme biological toothpaste has more obvious antibacterial effect which is equivalent to the antibacterial effect of montmorillonite and lysozyme composition groups, and shows that the toothpaste prepared by the invention can be prevented from being influenced by the denaturation of the lysozyme by substances such as foaming agents, binders, sweetening agents, preservatives and the like, and the activity of the biological lysozyme is prevented from being damaged or reduced, so that the function of inhibiting oral pathogenic bacteria is achieved.

EXAMPLE two WST-8 test of bacteriostatic Effect of each group

Compared with the common colony counting method, the WST-8 method has the advantages of high sensitivity, reliable data, good repeatability, simple and convenient operation and the like, and can quantify the antibacterial effect more accurately.

1. Experiment grouping

The experimental groups were the same as in example one.

2. Procedure of experiment

Each group of samples cultured for 6h and 12h was tested using the WST-8 kit. After each experimental group was cultured for a predetermined period of time under the corresponding bacterial culture conditions and left to stand for 30 minutes, 3 samples, each 0.15mL, were added to a 96-well plate from the supernatant of each group. To 96-well plateAdding 15 mu L of WST-8 reagent into each well, mixing uniformly, incubating at 37 ℃ for 1h, and measuring OD by using a full-wavelength microplate reader_450nm. The calculation formula of the antibacterial rate is as follows:

(OD) antibacterial rate_{Positive control}-OD_{Experimental group})/(OD_{Positive control}-OD_{Negative control})×100％

The OD positive control is the absorbance value of the liquid to be detected of the positive control group, the OD experimental group is the absorbance value corresponding to the liquid to be detected of each experimental group, and the OD negative control is the absorbance value corresponding to the liquid to be detected of the negative control group. The experiment takes the antibacterial rate of more than 95 percent according to the national standard to have the antibacterial effect, and the antibacterial rate of more than 80 percent has the antibacterial effect.

3. Bacteriostatic effect

The antibacterial effect of the lysozyme toothpaste is shown in figures 6-10, and can be obtained from figures 6-10, the lysozyme can inhibit the growth of oral pathogenic bacteria streptococcus mutans, escherichia coli, porphyromonas gingivalis, staphylococcus aureus and prevotella intermedia, the antibacterial rate is close to or reaches 80%, the antibacterial effect is achieved, and the antibacterial rate is obviously increased along with the increase of the culture time; montmorillonite has no inhibiting effect on oral pathogenic bacteria such as Streptococcus mutans, Escherichia coli, Porphyromonas gingivalis, Staphylococcus aureus and Primella intermedia; the lysozyme and montmorillonite composition group has the antibacterial rate of more than 85 percent on oral pathogenic bacteria streptococcus mutans, escherichia coli, porphyromonas gingivalis, staphylococcus aureus and prevotella intermedia, and the antibacterial rate is obviously increased along with the increase of the culture time; the antibacterial effect of the lysozyme and montmorillonite composition group is obviously better than that of the lysozyme group and the montmorillonite group; the antibacterial rate of the lysozyme biological toothpaste group to oral pathogenic bacteria streptococcus mutans, escherichia coli, porphyromonas gingivalis, staphylococcus aureus and prevotella intermedia reaches more than 85%, and the antibacterial rate is obviously increased along with the increase of the culture time, and is equivalent to the antibacterial effect of the lysozyme and montmorillonite composition group without obvious difference. The blank toothpaste group had no bacteriostatic effect. Compared with lysozyme, montmorillonite and blank toothpaste, the lysozyme biological toothpaste can obviously inhibit oral pathogenic bacteria.

In conclusion, the lysozyme biological toothpaste disclosed by the invention has an antibacterial effect on oral pathogenic bacteria such as escherichia coli, streptococcus mutans, staphylococcus aureus, porphyromonas gingivalis, prevotella intermedia and other bacteria, is wide in antibacterial spectrum and high in antibacterial rate, can efficiently and briefly inhibit the growth of the oral pathogenic bacteria, is low in minimum antibacterial concentration, cannot generate oral pathogenic bacteria drug-resistant strains after long-term use, most toothpaste preparations on the market still use chemical antibacterial substances (chlorhexidine, antibiotics and the like) as effective components of the toothpaste, and the lysozyme biological toothpaste shows unique advantages when the widely existing oral pathogenic bacteria drug-resistant strains exist; the antibacterial effect of the lysozyme biological toothpaste prepared by the invention is equivalent to that of a composition of lysozyme and montmorillonite, so that the toothpaste prepared by the invention can be prevented from being influenced by the denaturation of lysozyme by substances such as a foaming agent, a binder, a sweetening agent, a preservative and the like, the activity of the biological lysozyme is prevented from being damaged or reduced, the function of inhibiting oral pathogenic bacteria is achieved, the lysozyme biological toothpaste disclosed by the invention can effectively achieve the functions of preventing caries and preventing periodontitis, and has unique advantages and wide application prospects.

Claims (7)

1. An oral care agent with antibacterial effect is characterized in that the oral care agent comprises 0.4% of lysozyme and 1% of montmorillonite.

2. The toothpaste with the antibacterial effect is characterized by comprising 0.4% of lysozyme, 1% of montmorillonite, an abrasive, a wetting agent, an adhesive, a surfactant, a preservative, a sweetening agent, a thickening agent, a foaming agent and the balance of deionized water.

3. The toothpaste of claim 2, wherein said abrasive is one or more of calcium carbonate, calcium hydrogen phosphate, hydrated silica and silicon dioxide; the humectant is one or more of sorbitol, glycerol, propylene glycol and polyethylene glycol; the adhesive is one or more of sodium carboxymethylcellulose, xanthan gum, hydroxyethyl cellulose, guar gum and carrageenan; the foaming agent is sodium dodecyl sulfate or sodium lauryl sulfate; the surfactant is one or more of sodium lauryl sulfate, sodium 2-acyloxy bond sulfonate, trisodium phosphate and sodium lauroyl sarcosinate; the thickener is one or more of carboxymethyl cellulose, antler pectin, hydroxyethyl cellulose and xanthan gum; the sweetener is one or more of glycerol, sodium cyclamate and saccharin sodium; the preservative is one or more of potassium sorbate and sodium benzoate; the balance of deionized water.

4. The toothpaste of claim 2 or 3, further comprising a pigment and a flavor.

5. The toothpaste of claim 4, wherein said toothpaste comprises polyethylene glycol-85%, sodium carboxymethylcellulose 1%, hydroxyethylcellulose 0.3%, sorbitol 45%, sodium saccharin 0.21%, trisodium phosphate 0.15%, titanium dioxide 0.35%, calcium hydrogen phosphate 0.5%, deionized water 18.5%, hydrated silica 24.5%, sodium lauryl sulfate 2.4%, flavor 1%, sodium benzoate 0.2%, montmorillonite 1%, lysozyme 0.4%.

6. A method of preparing a toothpaste as claimed in claim 3, wherein said toothpaste is prepared by a method comprising the steps of: mixing lysozyme, montmorillonite, abrasive, wetting agent, adhesive and foaming agent according to the weight percentage, and stirring, grinding and vacuum degassing to obtain the toothpaste body.

7. The preparation method according to claim 6, wherein the method comprises the following steps of:

(1) adding lysozyme, montmorillonite, wetting agent and saccharin into deionized water, stirring to obtain deionized water solution, mixing with abrasive, binder, foaming agent, surfactant, antiseptic, sweetener and thickener, stirring, grinding, and vacuum degassing;

(2) Adding essence and pigment, stirring, grinding, and vacuum degassing to obtain toothpaste.

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