阅读说明：本技术 一种含游动纳米机器人用于口腔主动预防与治疗的漱口水 (Mouth wash containing swimming nano robot for active prevention and treatment of oral cavity ) 是由 贺强 吴英杰 于 2021-08-30 设计创作，主要内容包括：一种含游动纳米机器人用于口腔主动预防与治疗的漱口水,本发明涉及一种含游动纳米机器人的漱口水。本发明要解决现有漱口水无主动性,漱口水中局部应用的抗菌药物无法达到深而窄的牙周袋底,而利用现有纳米机器人于漱口水中,或存在难以实现高效驱动,或存在制备工艺繁琐的问题。含游动纳米机器人用于口腔主动预防与治疗的漱口水,它包含组分A和组分B,所述的组分A由过氧化氢、醇、甘油、羧甲基纤维素、十二醇硫酸钠和余量超纯水组成；所述的组分B为含有游动纳米机器人的水溶液。本发明用于含游动纳米机器人用于口腔主动预防与治疗的漱口水。(The invention discloses mouthwash containing a swimming nano robot for active prevention and treatment of an oral cavity, and relates to mouthwash containing the swimming nano robot. The invention aims to solve the problems that the existing mouthwash is not active, the antibacterial drug locally applied in the mouthwash cannot reach the deep and narrow periodontal pocket bottom, and the existing nano robot is used in the mouthwash, or the efficient driving is difficult to realize, or the preparation process is complicated. The mouth wash containing the swimming nano robot for active prevention and treatment of the oral cavity comprises a component A and a component B, wherein the component A consists of hydrogen peroxide, alcohol, glycerol, carboxymethyl cellulose, sodium lauryl sulfate and the balance of ultrapure water; the component B is an aqueous solution containing a swimming nano robot. The invention is used for the mouth wash containing the swimming nano robot and used for the active prevention and treatment of the oral cavity.)

1. A mouth wash containing a swimming nano robot for active prevention and treatment of oral cavity is characterized by comprising a component A and a component B, wherein the volume ratio of the component A to the component B is (5-10) to 1; the component A consists of 0.5 to 3 percent of hydrogen peroxide, 3 to 5 percent of alcohol, 3 to 5 percent of glycerin, 0.1 to 0.3 percent of carboxymethyl cellulose, 0.01 to 0.03 percent of lauryl sodium sulfate and the balance of ultrapure water according to mass percentage; the component B is 0.5 to 1 mass percent of water solution containing a swimming nano robot; the alcohol is sorbitol or sorbitol;

the swimming nano robot consists of an antibacterial anti-inflammatory drug, a hollow capsule-shaped framework and an enzyme catalyst, wherein the antibacterial anti-inflammatory drug is coated inside the hollow capsule-shaped framework, and the enzyme catalyst is positioned on one side of the hollow capsule-shaped framework; the diameter of the hollow saccular skeleton is 300 nm-100 μm.

2. The mouthwash containing the swimming nanomachines for active oral prevention and treatment according to claim 1, wherein the hollow saccular skeleton is formed of a biocompatible and degradable high molecular polymer; the biocompatible and degradable high molecular polymer is polyvinyl alcohol, polyethylene glycol amphiphilic block polymer, glucan and chitosan.

3. The mouthwash containing the swimming nanotechnology for active prevention and treatment of the oral cavity according to claim 1, wherein the enzyme catalyst is catalase, peroxidase, platinum nanoparticles or ferroferric oxide nanoparticles.

4. The mouthwash containing the swimming nano-robot for the active prevention and treatment of the oral cavity according to claim 1, wherein the antibacterial and anti-inflammatory drug is chlorhexidine, fluosilicic acid or sodium fluosilicate.

5. The mouthwash according to claim 1, wherein when the biocompatible and degradable polymer is polyethylene glycol-polylactic acid amphiphilic triblock copolymer and the enzyme catalyst is catalase, the swimming nanotechnology is prepared by the following steps:

dissolving a polyethylene glycol-polylactic acid amphiphilic triblock copolymer in a solvent and uniformly mixing to obtain an organic phase mixed solution, adding the organic phase mixed solution into an aqueous solution containing polyvinyl alcohol and an antibacterial and anti-inflammatory drug, stirring and emulsifying for 30-60 min under the condition that the rotation speed is 500-1000 rpm to obtain an emulsified solution, adding the emulsified solution into water, volatilizing the organic solvent for 4-5 h under the condition that the magnetic stirring speed is 500-1000 rpm, filtering, collecting and washing to obtain polymer microspheres loaded with antibacterial active ingredients;

the mass-to-volume ratio of the polyethylene glycol-polylactic acid amphiphilic triblock copolymer to the solvent is 1g (50-100) mL; the mass percentage of the polyvinyl alcohol in the aqueous solution containing the polyvinyl alcohol and the antibacterial and anti-inflammatory drug is 5-10%; the volume ratio of the organic phase mixed solution to the aqueous solution containing the polyvinyl alcohol and the antibacterial and anti-inflammatory drugs is 1: 1;

dispersing polymer microspheres loaded with antibacterial active ingredients in deionized water to obtain microsphere dispersion liquid, dropwise adding the microsphere dispersion liquid on a hydrophilic substrate, standing for 10-18 h, naturally spreading into single-layer particles, then semi-modifying gold nanoshells on one side of the surfaces of the microspheres by a vacuum sputtering method under the condition that the current is 20-30 mA, sputtering for 2-3 min to obtain gold-modified microspheres, washing the gold-modified microspheres from the hydrophilic substrate by deionized water, and centrifuging for 2-5 min under the condition that the rotating speed is 4000-8000 r/min to obtain the drug-loaded Au-Janus nano robot;

thirdly, placing the drug-loaded Au-Janus nano robot in PBS buffer solution containing N-hydroxyl thiosuccinimide and 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide hydrochloride, processing for 12-24 h, cleaning, placing in catalase solution with the concentration of 1-2 mg/mL, incubating for 12-18 h at the temperature of 37 ℃, and finally cleaning and storing for later use to obtain the swimming nano robot;

the concentration of the N-hydroxy thiosuccinimide in the PBS buffer solution containing the N-hydroxy thiosuccinimide and the 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide hydrochloride is 100 mmol/L-200 mmol/L, and the concentration of the 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide hydrochloride is 300 mmol/L-600 mmol/L.

6. The mouth wash containing the swimming nano-robot for active prevention and treatment of the oral cavity according to claim 5, wherein the solvent in the step (r) is a mixed solution of dichloromethane and acetone, and the volume ratio of the dichloromethane to the acetone is 1 (1-2).

7. The mouth wash containing the swimming nano-robot for active prevention and treatment of the oral cavity according to claim 5, wherein the mass percentage of the antibacterial and anti-inflammatory drug in the aqueous solution containing the polyvinyl alcohol and the antibacterial and anti-inflammatory drug in the step (r) is 5-10%.

8. The mouth wash containing the swimming nano-robot for active prevention and treatment of the oral cavity according to claim 5, characterized in that the volume ratio of the emulsified solution to water in the step (r) is 1 (5-10).

9. The mouthwash containing the swimming nano-robot for active prevention and treatment of the oral cavity according to claim 5, wherein the concentration of the microsphere dispersion in the step (II) is 0.1mg/mL to 0.5 mg/mL.

10. The mouth wash containing the swimming nano-robot for active prevention and treatment of the oral cavity according to claim 5, wherein the pH of the PBS buffer solution containing the N-hydroxy thiosuccinimide and the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride in the step (c) is 7-7.5; washing for three times by using PBS (phosphate buffer solution) with the pH value of 7-7.5 and the concentration of 0.1-0.5 mol/L; the storage for standby use in the step (III) is to store in PBS buffer solution with the temperature of 2-4 ℃.

Technical Field

The invention relates to a mouth wash containing a swimming nano robot.

Background

Periodontal disease is an inflammatory disease caused by the accumulation of plaque, which if left uncontrolled results in loss of gingival attachment, formation of periodontal pockets, alveolar bone resorption, and ultimately, tooth loosening and even loss. Periodontal disease has become the leading cause of tooth loss in adults today, and severely affects people's health and quality of life. Currently, the toothpaste and the mouthwash which are used most frequently when people clean the oral cavity in daily life are used, and in recent years, the consumption of the mouthwash in people is higher and higher, and the use frequency is increased year by year. However, at present, most of pathogenic bacteria are located in the periodontal pocket or deep part of gingiva, the existing mouthwash cannot effectively permeate and remove the pathogenic bacteria, and the antibacterial effective components cannot reach the deep and narrow periodontal pocket bottom, so that the using effect of the mouthwash is affected.

The swimming nano robot has the core advantages of additional self-driving force, autonomous navigation, drug loading and controllable release capacity and the like, is expected to greatly improve the targeted drug delivery efficiency, and shows great potential in the aspect of biomedicine. If the swimming nano robot moves effectively and autonomously in gingival crevicular fluid, saliva and other different biological media, an important research method and an implementation means can be provided for the delivery of the periodontal targeted drug of the anti-inflammatory active ingredient, and the swimming nano robot has important significance for improving the prevention and treatment effects of the oral periodontal disease. However, the existing glucose oxidase and urease driven nano-robots have good biocompatibility, but mainly depend on an autophoretic driving mechanism, are greatly influenced by electrolytes in a solution, are difficult to realize high-efficiency driving in body fluid (saliva, gingival crevicular fluid and the like), and cannot reach the bottom of a deep and narrow periodontal pocket, and the existing catalase driven swimming nano-robots cannot realize large-scale industrial application at present due to the complexity of a preparation process.

Disclosure of Invention

The invention aims to solve the problems that the existing mouthwash is not active, the antibacterial drug locally applied in the mouthwash cannot reach the deep and narrow periodontal pocket bottom, and the existing nano robot is used in the mouthwash, or the efficient driving is difficult to realize, or the preparation process is complicated, so that the mouthwash containing the swimming nano robot for the active prevention and treatment of the oral cavity is further provided.

A mouth wash containing a swimming nano robot for active prevention and treatment of oral cavity comprises a component A and a component B, wherein the volume ratio of the component A to the component B is (5-10): 1; the component A consists of 0.5 to 3 percent of hydrogen peroxide, 3 to 5 percent of alcohol, 3 to 5 percent of glycerin, 0.1 to 0.3 percent of carboxymethyl cellulose, 0.01 to 0.03 percent of lauryl sodium sulfate and the balance of ultrapure water according to mass percentage; the component B is 0.5 to 1 mass percent of water solution containing a swimming nano robot; the alcohol is sorbitol or sorbitol;

the swimming nano robot consists of an antibacterial anti-inflammatory drug, a hollow capsule-shaped framework and an enzyme catalyst, wherein the antibacterial anti-inflammatory drug is coated inside the hollow capsule-shaped framework, and the enzyme catalyst is positioned on one side of the hollow capsule-shaped framework; the diameter of the hollow saccular skeleton is 300 nm-100 μm.

The invention has the beneficial effects that:

the invention relates to a functional mouthwash containing a catalase-driven swimming micro-nano robot. The low-concentration hydrogen peroxide in the mouthwash is used as a fuel, the swimming micro-nano robot carries and enriches antibacterial ingredients to the deep of periodontal pockets and gingiva efficiently while performing self-driven rapid movement, and continuous and efficient inhibition of periodontitis pathogenic bacteria is achieved. Compared with the prior art, the enzyme-driven swimming nano robot in the mouthwash has better biocompatibility and biodegradability, simple preparation process and can realize industrial batch production. The swimming nano robot generates bubble drive by decomposing hydrogen peroxide, has stronger driving force, is less influenced by electrolyte, can realize high-efficiency drive in body fluid, obviously improves the using effect of the obtained mouthwash, and is safe and efficient functional mouthwash.

The invention is used for the mouth wash containing the swimming nanometer robot and used for the active prevention and treatment of the oral cavity.

Drawings

FIG. 1 is a diagram of a mouth wash containing a swimming nano-robot for active prevention and treatment of oral cavity according to the present invention;

FIG. 2 is a schematic diagram of a mouth wash containing a swimming nano-robot for active prevention and treatment of oral cavity according to the present invention;

FIG. 3 is a scanning electron microscope image of the flying nano-robot prepared in the first embodiment;

FIG. 4 is a diagram of the movement of the swimming nano-robot prepared in the first embodiment in a hydrogen peroxide solution with a mass percentage of 1%;

FIG. 5 is a fluorescence microscope image of the antimicrobial test.

Detailed Description

The technical solution of the present invention is not limited to the specific embodiments listed below, and includes any combination of the specific embodiments.

The first embodiment is as follows: specifically, referring to fig. 1 and fig. 2, the mouth wash for active prevention and treatment of oral cavity containing swimming nano-robot in the embodiment comprises a component a and a component B, wherein the volume ratio of the component a to the component B is (5-10): 1; the component A consists of 0.5 to 3 percent of hydrogen peroxide, 3 to 5 percent of alcohol, 3 to 5 percent of glycerin, 0.1 to 0.3 percent of carboxymethyl cellulose, 0.01 to 0.03 percent of lauryl sodium sulfate and the balance of ultrapure water according to mass percentage; the component B is 0.5 to 1 mass percent of water solution containing a swimming nano robot; the alcohol is sorbitol or sorbitol;

the swimming nano robot consists of an antibacterial anti-inflammatory drug, a hollow capsule-shaped framework and an enzyme catalyst, wherein the antibacterial anti-inflammatory drug is coated inside the hollow capsule-shaped framework, and the enzyme catalyst is positioned on one side of the hollow capsule-shaped framework; the diameter of the hollow saccular skeleton is 300 nm-100 μm.

The component A and the component B in the embodiment are respectively filled and packaged to obtain a finished product.

The purpose of the present embodiment is to provide a mouth wash containing a swimming nano robot, which has efficient effects of preventing and treating periodontal diseases based on active drug targeting delivery of the swimming nano robot.

The swimming nano robot of the specific embodiment has biocompatibility and biodegradability. The preparation method is characterized in that a polymer with biocompatibility and biodegradability, gold and enzyme are selected as assembly materials to prepare the nano-composite material.

Firstly, preparing a drug-loaded microsphere coated by a biocompatible polymer shell by adopting an oil-in-water-solvent volatilization method;

secondly, modifying gold nanoshells on one side of the drug-loaded microspheres obtained in the second step by a micro-contact printing or vacuum sputtering method;

and thirdly, modifying the gold surface by using a cross-linking agent, and cross-linking catalase on the gold surface by using a covalent cross-linking method to obtain the drug-loaded swimming nano robot with one modified side with the enzyme catalyst.

The principle is as follows: compared with the swimming nano robot driven by autophoresis, the catalase driving swimming nano robot of the embodiment generates bubbles through decomposing hydrogen peroxide to drive movement, has stronger driving force, is less influenced by electrolyte, and can realize high-efficiency driving in body fluid (saliva, gingival crevicular fluid and the like). The swimming nanometer machine of the specific embodiment has better biocompatibility and biodegradability, and the hollow capsule cavity can realize the efficient loading of the antibacterial components.

The swimming nano robot enzyme catalyst is positioned on one side of the hollow capsule-shaped framework, and the swimming nano robot can realize chemical driving movement through chemical reaction due to the asymmetric distribution of the catalyst. During the use, through mixing AB component to low concentration hydrogen peroxide in the mouthwash is the fuel, and the medicine carrying moves about nanometer robot catalysis hydrogen peroxide and produces oxygen and carry out autonomous movement, carries the high-efficient carrier of antibacterial component, enriches to periodontal pocket and gum depths when carrying out the self-driven quick motion, reaches and lasts high-efficient suppression to periodontitis pathogenic bacteria ground.

The beneficial effects of the embodiment are as follows:

the embodiment relates to functional mouthwash containing a catalase-driven swimming micro-nano robot. The low-concentration hydrogen peroxide in the mouthwash is used as a fuel, the swimming micro-nano robot carries and enriches antibacterial ingredients to the deep of periodontal pockets and gingiva efficiently while performing self-driven rapid movement, and continuous and efficient inhibition of periodontitis pathogenic bacteria is achieved. Compared with the prior art, the enzyme-driven swimming nano robot in the mouthwash of the embodiment has better biocompatibility and biodegradability, the preparation process is simple, and industrial batch production can be realized. The swimming nano robot generates bubble drive by decomposing hydrogen peroxide, has stronger driving force, is less influenced by electrolyte, can realize high-efficiency drive in body fluid, obviously improves the using effect of the obtained mouthwash, and is safe and efficient functional mouthwash.

The second embodiment is as follows: the first difference between the present embodiment and the specific embodiment is: the hollow saccular skeleton is formed by biocompatible and degradable high molecular polymer; the biocompatible and degradable high molecular polymer is polyvinyl alcohol, polyethylene glycol amphiphilic block polymer, glucan and chitosan. The rest is the same as the first embodiment.

The third concrete implementation mode: this embodiment is different from the first or second embodiment in that: the enzyme catalyst is catalase, peroxidase, platinum nanoparticles or ferroferric oxide nanoparticles. The other is the same as in the first or second embodiment.

The fourth concrete implementation mode: the difference between this embodiment mode and one of the first to third embodiment modes is: the antibacterial and anti-inflammatory drug is chlorhexidine, fluosilicic acid or sodium fluosilicate. The others are the same as the first to third embodiments.

The fifth concrete implementation mode: the difference between this embodiment and one of the first to fourth embodiments is: when the biocompatible and degradable high molecular polymer is polyethylene glycol-polylactic acid amphiphilic triblock copolymer and the enzyme catalyst is catalase, the swimming nano robot is prepared by the following steps:

dissolving a polyethylene glycol-polylactic acid amphiphilic triblock copolymer in a solvent and uniformly mixing to obtain an organic phase mixed solution, adding the organic phase mixed solution into an aqueous solution containing polyvinyl alcohol and an antibacterial and anti-inflammatory drug, stirring and emulsifying for 30-60 min under the condition that the rotation speed is 500-1000 rpm to obtain an emulsified solution, adding the emulsified solution into water, volatilizing the organic solvent for 4-5 h under the condition that the magnetic stirring speed is 500-1000 rpm, filtering, collecting and washing to obtain polymer microspheres loaded with antibacterial active ingredients;

the mass-to-volume ratio of the polyethylene glycol-polylactic acid amphiphilic triblock copolymer to the solvent is 1g (50-100) mL; the mass percentage of the polyvinyl alcohol in the aqueous solution containing the polyvinyl alcohol and the antibacterial and anti-inflammatory drug is 5-10%; the volume ratio of the organic phase mixed solution to the aqueous solution containing the polyvinyl alcohol and the antibacterial and anti-inflammatory drugs is 1: 1;

dispersing polymer microspheres loaded with antibacterial active ingredients in deionized water to obtain microsphere dispersion liquid, dropwise adding the microsphere dispersion liquid on a hydrophilic substrate, standing for 10-18 h, naturally spreading into single-layer particles, then semi-modifying gold nanoshells on one side of the surfaces of the microspheres by a vacuum sputtering method under the condition that the current is 20-30 mA, sputtering for 2-3 min to obtain gold-modified microspheres, washing the gold-modified microspheres from the hydrophilic substrate by deionized water, and centrifuging for 2-5 min under the condition that the rotating speed is 4000-8000 r/min to obtain the drug-loaded Au-Janus nano robot;

thirdly, placing the drug-loaded Au-Janus nano robot in PBS buffer solution containing N-hydroxyl thiosuccinimide and 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide hydrochloride, processing for 12-24 h, cleaning, placing in catalase solution with the concentration of 1-2 mg/mL, incubating for 12-18 h at the temperature of 37 ℃, and finally cleaning and storing for later use to obtain the swimming nano robot;

the concentration of the N-hydroxy thiosuccinimide in the PBS buffer solution containing the N-hydroxy thiosuccinimide and the 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide hydrochloride is 100 mmol/L-200 mmol/L, and the concentration of the 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide hydrochloride is 300 mmol/L-600 mmol/L. The rest is the same as the first to fourth embodiments.

The polyethylene glycol-polylactic acid amphiphilic triblock copolymer is PLLA₁₀₀-PEG₅₀-PLLA₁₀₀。

The sixth specific implementation mode: the difference between this embodiment and one of the first to fifth embodiments is: the solvent in the step I is a mixed solution of dichloromethane and acetone, wherein the volume ratio of dichloromethane to acetone is 1 (1-2). The rest is the same as the first to fifth embodiments.

The seventh embodiment: the difference between this embodiment and one of the first to sixth embodiments is: the mass percentage of the antibacterial and anti-inflammatory drug in the aqueous solution containing the polyvinyl alcohol and the antibacterial and anti-inflammatory drug is 5-10%. The others are the same as the first to sixth embodiments.

The specific implementation mode is eight: the present embodiment differs from one of the first to seventh embodiments in that: the volume ratio of the emulsified solution to water in the step I is 1 (5-10). The rest is the same as the first to seventh embodiments.

The specific implementation method nine: the present embodiment differs from the first to eighth embodiments in that: the concentration of the microsphere dispersion liquid in the step II is 0.1 mg/mL-0.5 mg/mL. The other points are the same as those in the first to eighth embodiments.

The detailed implementation mode is ten: the present embodiment differs from one of the first to ninth embodiments in that: the pH value of the PBS buffer solution containing the N-hydroxy thiosuccinimide and the 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide hydrochloride in the step (c) is 7-7.5; washing for three times by using PBS (phosphate buffer solution) with the pH value of 7-7.5 and the concentration of 0.1-0.5 mol/L; the storage for standby use in the step (III) is to store in PBS buffer solution with the temperature of 2-4 ℃. The other points are the same as those in the first to ninth embodiments.

The following examples were used to demonstrate the beneficial effects of the present invention:

the first embodiment is as follows:

a mouth wash containing a swimming nano robot for active prevention and treatment of oral cavity comprises 450mL of component A and 50mL of component B; the component A consists of 0.5 percent of hydrogen peroxide, 5 percent of sorbitol, 5 percent of glycerin, 0.1 percent of carboxymethyl cellulose, 0.01 percent of sodium lauryl sulfate and the balance of ultrapure water according to mass percentage; the component B is 1% of water solution containing a swimming nano robot in percentage by mass;

the swimming nano robot consists of an antibacterial anti-inflammatory drug, a hollow capsule-shaped framework and an enzyme catalyst, wherein the antibacterial anti-inflammatory drug is coated inside the hollow capsule-shaped framework, and the enzyme catalyst is positioned on one side of the hollow capsule-shaped framework; the diameter of the hollow saccular skeleton is 2 μm.

And respectively filling and packaging the component A and the component B to obtain a finished product.

The hollow saccular skeleton is formed by biocompatible and degradable high molecular polymer; the biocompatible and degradable high molecular polymer is polyethylene glycol-polylactic acid amphiphilic triblock copolymer (PLLA)₁₀₀-PEG₅₀-PLLA₁₀₀) The enzyme catalyst is catalase, and the antibacterial and anti-inflammatory drug is chlorhexidine; the swimming nano robot is prepared by the following steps:

dissolving 2g of polyethylene glycol-polylactic acid amphiphilic triblock copolymer in 200mL of solvent and uniformly mixing to obtain organic phase mixed solution, adding the organic phase mixed solution into 200mL of aqueous solution containing polyvinyl alcohol and antibacterial and anti-inflammatory drugs, stirring and emulsifying for 40min at the rotation speed of 1000rpm to obtain emulsified solution, adding the emulsified solution into 2000mL of water, volatilizing the organic solvent for 5h at the magnetic stirring speed of 800rpm, collecting microspheres by a filtration method, washing for three times by distilled water to obtain polymer microspheres loaded with antibacterial active ingredients, and storing in ultrapure water;

the solvent is a mixed solution of dichloromethane and acetone in a volume ratio of 1: 1; the mass percentage of the polyvinyl alcohol in the aqueous solution containing the polyvinyl alcohol and the antibacterial and anti-inflammatory drug is 10 percent; the mass percentage of the antibacterial and anti-inflammatory drug in the aqueous solution containing the polyvinyl alcohol and the antibacterial and anti-inflammatory drug is 5 percent;

dispersing polymer microspheres loaded with antibacterial active ingredients in deionized water to obtain microsphere dispersion liquid, dropwise adding the microsphere dispersion liquid on a hydrophilic substrate, standing for 10 hours, naturally spreading into single-layer particles, then semi-modifying gold nanoshells on one side of the surfaces of the microspheres by a vacuum sputtering method under the condition that the current is 30mA, wherein the sputtering time is 3min to obtain gold-modified microspheres, washing the gold-modified microspheres from the hydrophilic substrate by using the deionized water, and centrifuging for 2min under the condition that the rotating speed is 4500r/min to obtain the medicine-carrying Au-Janus nano robot;

thirdly, placing the drug-loaded Au-Janus nano robot into PBS buffer solution containing N-hydroxy thiosuccinimide and 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide hydrochloride, processing for 12 hours, washing, placing into catalase solution with the concentration of 2mg/mL, incubating for 12 hours at the temperature of 37 ℃, finally washing and storing for later use to obtain the swimming nano robot;

the concentration of the N-hydroxy thiosuccinimide in the PBS buffer solution containing the N-hydroxy thiosuccinimide and the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is 100mmol/L, and the concentration of the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is 400 mmol/L;

the concentration of the microsphere dispersion liquid in the step two is 0.1 mg/mL;

the pH value of the PBS buffer solution containing the N-hydroxy thiosuccinimide and the 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide hydrochloride in the step (c) is 7.2; washing for three times by using PBS (phosphate buffer solution) with the pH value of 7.2 and the concentration of 0.1 mol/L; and the storage for standby in the step (III) is to store in PBS buffer solution with the temperature of 4 ℃ for standby.

Fig. 3 is a scanning electron microscope image of the swimming nano-robot prepared in the first embodiment, and it can be known from the image that the drug-loaded nano-robot prepared in the first embodiment has a uniform size and a complete structure, catalase is modified on one side of the nano-robot, and the asymmetric distribution of the enzyme can provide a high-efficiency self-driving force for the nano-robot.

Fig. 4 is a diagram illustrating the movement of the swimming nano-robot prepared in the first embodiment in a hydrogen peroxide solution with a mass percentage of 1%. As can be seen from the figure, the drug-loaded nano-robot prepared in the first embodiment has the autonomous movement capability, and can decompose the substrate in the low hydrogen peroxide solution through the enzyme catalytic reaction to enhance diffusion.

Control group: the contrast group differs from the first example in that: the same concentration of the antibacterial drug was dissolved directly in water to give component B, otherwise the same as in example one.

1. Evaluation of antibacterial performance of mouthwash:

to qualitatively evaluate the efficacy of the mouthwash prepared in this example, the antibacterial performance of porphyromonas gingivalis was evaluated. The mouthwash prepared in the first embodiment is used as an experimental group, the mouthwash without a drug-loaded swimming nano robot is used as a control group, and is co-cultured with porphyromonas gingivalis respectively, and the antibacterial performance of the mouthwash is tested by using calcein and Pi double-dying live bacteria. Calcein-stained live bacteria are green, while Pi-stained dead bacteria are red. FIG. 5 is a fluorescence microscope image of the antibacterial test, and it can be seen that after 24 hours of co-culture, the positive fluorescence microscope observation shows that a large number of pi-stained bacteria in the experimental group appear red, almost one hundred percent of Porphyromonas gingivalis are killed, while the control group has only forty percent of sterilization rate, indicating that the mouthwash has obvious antibacterial activity. The researches show that the mouthwash added with the medicine-carrying swimming nanometer machine has good antibacterial activity and can maintain the micro-ecological stability of the oral cavity.

2. Evaluation of the efficacy of the mouthwash:

the mouthwash prepared in the first example is used as an experimental group, and the mouthwash without a drug-loaded swimming nano robot is used as a control group for 20 patients with mild periodontitis respectively, and the using method comprises the following steps: after rinsing with clear water, rinsing with mouthwash for 20 seconds; the medicine is used once in the morning and at night every day, and is continuously used for seven days, and each observation index of a patient is counted: and detecting the periodontal probing depth, the plaque index and the gingival sulcus bleeding index of the patient. The results of the analysis using the SPSS22.0 statistical software are shown in table 1 below. After one week of use, the periodontal probing depth, plaque index and gingival sulcus bleeding index of the experimental group were lower than those of the control group, and the difference was statistically significant (P < 0.05). The mouthwash prepared by the embodiment has good safety and certain effect, and can improve the periodontal health index of patients and reduce inflammatory reaction.

TABLE 1