Preparation method of dental plaque display type anti-caries invisible appliance based on synergism of iron-based nano-enzyme catalysis and traditional Chinese medicine caries inhibition

Abstract

The invention discloses a dental plaque display type anti-caries invisible appliance based on synergism of iron-based nano-enzyme catalysis and traditional Chinese medicine caries inhibition, and belongs to the field of oral medical instruments. The preparation method of the appliance diaphragm comprises the following steps: preparing a composite material with the functions of catalytically degrading a bacterial biofilm matrix barrier and killing internal bacteria from polyethylene glycol terephthalate-1, 4-cyclohexanedimethanol (PETG) and a Fe3O4 nano-enzyme filler in a solution blending manner; and after the composite material is subjected to thermoplastic molding, a layer of molecularly imprinted polymer (MIP)-spiropyrane and chitosan-magnolol are coated with the fluorescent display agent which is prepared by mixing the molecularly imprinted polymer (MIP)-spiropyrane and the chitosan-magnolol. The iron-based nano-enzyme can catalytically degrade a bacterial biofilm matrix barrier and kill internal bacteria, and the MIP-spiropyrane fluorescent display agent can specifically display the accumulation condition of cariogenic bacterial plaque in the oral cavity under ultraviolet irradiation. The invisible orthodontic appliance can reduce or prevent caries while having the same orthodontic effect as other orthodontic appliances in the market.

Description

Technical Field

[0001] This patent relates to the field of oral medical devices, and particularly to a cariostatic invisible aligner with plaque display function based on the synergistic effect of iron-based nanozyme catalysis and traditional Chinese medicine for caries inhibition, and its preparation method. Background Art

[0002] Oral health has an important impact on overall health. Dental caries (tooth decay) is one of the most common oral diseases, which is mainly caused by the formation and development of bacterial biofilms (dental plaque). As an important tool for modern orthodontic treatment, invisible aligners are widely used due to their aesthetic appearance, comfort, and removable nature. However, prolonged wearing of invisible aligners may lead to plaque accumulation, thereby increasing the risk of dental caries. In addition, the surface of invisible aligner materials is prone to becoming a breeding ground for bacterial attachment and biofilm formation, further exacerbating the accumulation of dental plaque. Therefore, how to add a cariostatic function to invisible aligners and reduce the risk of dental caries during wearing is an urgent problem to be solved currently.

[0003] Existing cariostatic methods mainly include mechanical cleaning, fluoride toothpaste, antibacterial mouthwash, and antibacterial coatings, etc. However, these methods have certain limitations during the wearing of invisible aligners. For example, mechanical cleaning is difficult to completely remove bacteria on the surface of invisible aligners, the use of fluoride products depends on the patient's awareness, the action time of antibacterial mouthwash is limited, and the long-term stability and biosafety of antibacterial coatings still need to be further optimized. Therefore, there is an urgent need for a functionalized invisible aligner that can effectively inhibit plaque formation and reduce the risk of dental caries.

[0004] PETG (polyethylene terephthalate-1,4-cyclohexanedimethanol ester) is widely used in the medical device field due to its excellent transparency, mechanical properties, and biocompatibility. However, traditional PETG materials lack antibacterial ability and are difficult to effectively inhibit the growth of oral bacteria and the formation of biofilms. Therefore, functional modification of PETG materials to endow them with antibacterial and plaque color display functions is expected to improve the cariostatic performance of invisible aligners.

[0005] Iron-based nanozymes have gradually attracted attention in the biomedical field due to their peroxidase-like activity and antibacterial properties. Since iron-based nanozymes can catalyze hydrogen peroxide to generate highly reactive free radicals in weakly acidic sites (such as bacterial infection sites), and these free radicals attack the phospholipid bilayer of the bacterial cell membrane and oxidize proteins and DNA inside the bacteria, resulting in the destruction of the cell membrane and metabolic functions, iron-based nanozymes can play a very effective antibacterial role. In addition, although the catalytic process of iron-based nanozymes follows enzymatic reaction kinetics, its active center is still iron ions rather than protein cofactors, so it is a nano metal material with enzyme-like catalytic activity rather than a traditional enzyme.

[0006] On the other hand, molecularly imprinted polymers (MIPs) are highly selective recognition materials synthesized based on template molecules. By using the surface antigens or metabolites of cariogenic bacteria as templates, polymer materials with specific capture ability for target bacteria can be prepared to achieve efficient detection and recognition of dental plaque. Spiropyran is an intelligent responsive fluorescent material, and its color development intensity changes in a specific microenvironment. Therefore, by combining molecular imprinting technology with spiropyran, the distribution of dental plaque can be shown under ultraviolet light irradiation in the presence of cariogenic bacteria, enabling users to intuitively judge the oral hygiene status.

[0007] In addition, Magnolia officinalis, as a traditional Chinese medicine, can further achieve the function of preventing caries by interfering with the expression of surface adhesion proteins on the lipid bilayer of the cell membrane of Streptococcus mutans. Moreover, the polar groups of magnolol interact with the bacterial phospholipid bilayer, which can also enhance the penetration of inner-layer bacteria, thereby removing cariogenic bacteria wrapped by the matrix. In addition to the anti-caries effect, Magnolia officinalis can inhibit the generation of inflammatory enzymes / cytokines, the activation of NF-κB and MAPKs, and the activation of white blood cells, showing certain anti-inflammatory properties. Chitosan-magnolol drug-loading layers can be prepared using Magnolia officinalis and chitosan. The drug-loading layer can protect magnolol from the oral environment. In the acidic environment of bacterial infection, the amino groups in chitosan are protonated and swollen, accelerating drug release, thereby achieving the functions of protecting the drug and intelligent controlled release.

[0008] Based on the above research, by combining the catalytic degradation of bacterial biofilms by iron-based nanozymes, the anti-caries effect of traditional Chinese medicine, and the molecular imprinting-fluorescent color development technology, the PETG material is functionally modified to make it have antibacterial, catalytic degradation of biofilms, and dental plaque visualization functions, as well as the combined application of Magnolia officinalis, which becomes an innovative means to improve the anti-caries performance of invisible orthodontic appliances. The application of this technology can not only effectively reduce the formation of bacterial biofilms during orthodontic treatment, but also reduce the risk of caries, thereby enhancing the clinical application value of invisible orthodontic appliances. Summary of the Invention

[0009] In view of the possible indirect caries risk caused by bracketless invisible orthodontic appliances during orthodontics, the present invention proposes a PETG invisible orthodontic appliance with both dental plaque display and anti-caries functions and its preparation method. The present invention uses PETG as the matrix and Fe 3 O 4 nanozymes as fillers to prepare an invisible orthodontic appliance that can catalytically degrade the matrix barrier of bacterial biofilms and kill internal bacteria; a mixture of MIP-spiropyran and magnolol is used as a coating to enable it to show the accumulation of cariogenic bacterial plaque biofilms in the oral cavity under ultraviolet light irradiation and have an anti-caries function.

[0010] To achieve the above object, the present invention adopts the following technical solutions.

[0011] A cariostatic invisible aligner with plaque display based on the synergistic effect of iron-based nanozyme catalysis and traditional Chinese medicine for caries inhibition, comprising raw materials in the following weight parts: 0.2 parts of FeCl 2 、0.4 parts of FeCl 3 ·6H 2 O, 10 parts of ammonia water, 10 - 15 parts of PETG pellets, 50 - 100 parts of organic solvent, 100 parts of amino-modified silica nanoparticles (SiO 2 -NH 2 , with a particle size of 100 nm), 0.4 mmol of spiropyran-methacrylate (SPMA), 100 parts of phosphate buffer solution (PBS, pH 7.4), 1 - 5 parts of purified glucosyltransferase (Gtase, concentration ≥ 1 mg / mL, dissolved in PBS with pH 7.4), 10 parts of chitosan solution (pH 4.0, adjusted with acetic acid), 5 parts of 0.1% SDS + 1M NaCl solution, 10 parts of 0.5% Tween-20 solution, 10 - 15 parts of magnolol.

[0012] Furthermore, the preparation steps of the Fe 3 O 4 nanozyme are as follows: Step 1: Dissolve 0.2 parts of FeCl 2 and 0.4 parts of FeCl 3 ·6H 2 O in deionized water and mix well using a magnetic stirrer; Step 2: After heating to 60 °C, add 10 parts of ammonia water at one time, continue stirring and heating to 80 °C, and increase the rotation speed and continue the reaction for 2 h; Step 3: After the reaction is completed, separate the Fe 3 O 4 nano-particles with a magnet, and wash them alternately with distilled water and absolute ethanol 3 times, and dry them in a vacuum drying oven at 60 °C for 4 - 6 h to obtain the Fe 3 O 4 nanozyme filler powder.

[0013] Furthermore, the organic solvent is one of dichloromethane (DCM), tetrahydrofuran (THF), chloroform, dimethylformamide (DMF) or dimethyl sulfoxide (DMSO).

[0014] Furthermore, the preparation steps of the Fe 3 O 4 / PETG composite are as follows: Step 1: Dissolve 0.2 - 0.5 parts of Fe 3 O 4 nanozyme filler powder in a separate beaker containing an organic solvent, and place it on a heating plate at (80 °C) and stir for 20 minutes to obtain Fe3 O 4 Nanozyme solution; Step 2: Put 10 - 15 parts of PETG pellets into a container filled with an organic solvent, stir in a water bath at 35 - 50 °C until the PETG pellets are completely dissolved to obtain a PETG solution; Step 3: Subsequently, mix the Fe 3 O 4 nanozyme solution and the PETG solution, then place the mixture in an ultrasonic bath (ultrasonic power: 200 - 500 W, time: 30 - 60 minutes), and vibrate vigorously at 80 °C for 2 hours; Step 4: After the solution in Step 3 cools down, inject the uniformly mixed solution in Step 3 into a mold, then transfer the mold to a constant temperature oven at 50 - 60 °C for 12 - 24 hours for solvent volatilization treatment. After the film formation is completed, place the film in a vacuum drying oven and heat - dry it at 60 °C for 4 - 6 hours. Finally, peel the completely cured composite material from the mold substrate to obtain the PETG composite film with Fe 3 O 4 nanozyme.

[0015] Furthermore, the organic solvent used for preparing the mixed developer is one of methacrylic acid (MAA, purity ≥ 99%), ethylene glycol dimethacrylate (EGDMA, purity ≥ 98%), ammonium persulfate (APS), tetramethylethylenediamine (TEMED), acetonitrile, 2 - hydroxy - 2 - methylpropiophenone (HMPP, suitable for 365 nm ultraviolet light initiation).

[0016] Furthermore, the preparation steps of MIP are as follows (this process needs to be operated in the dark): Step 1: Take 100 parts of SiO 2 -NH 2 Disperse it in 10 parts of PBS, add 1 part of glutaraldehyde (5% v / v), stir at room temperature for 2 hours, and then centrifuge (8000 rpm, 10 min); Step 2: Wash it 3 times with PBS to remove the unreacted glutaraldehyde; Step 3: Mix the activated SiO 2 -NH 2 with 1 - 5 parts of Gtase solution with a concentration of 1 mg / mL, incubate with shaking at 4 °C for 12 hours, then centrifuge to remove the unbound enzyme, and wash with PBS until the absorbance of the eluate at 280 nm is close to the baseline.

[0017] Furthermore, the preparation steps of the MIP - spiropyran developer are as follows (this process needs to be operated in the dark): Step 1: Fix the Gtase - modified SiO 2 -NH 2Disperse in 20 parts of PBS / acetonitrile mixed solvent, add 0.4 mmol of MAA and 0.4 mmol of spiropyran-SPMA, and under red light environment, react with shaking at 4 °C for 2 hours to enable the monomers and spiropyran-SPMA to synergistically bind to the template enzyme; Step 3: Add 3.6 mmol of EGDMA (crosslinking degree 90%), 10 parts of APS and 0.02 part of tetramethylethylenediamine (TEMED), deoxygenate by passing nitrogen for 10 minutes, and seal the reaction vessel; Step 4: Place the prepolymer solution under a 365 nm ultraviolet light source (at a distance of 10 cm), irradiate at 25 °C for 30 minutes to complete the polymerization (Note: Use a water cooling device or intermittent irradiation (stop for 1 minute every 5 minutes) to control the temperature ≤ 25 °C); Step 5: Centrifuge to collect the polymer, ultrasonically wash with 0.1% SDS + 1 M NaCl solution (30 min × 3 times), and further wash with 0.5% Tween-20 twice to remove the residual surfactant; Step 6: Verify that the template removal rate > 95% through a bovine serum albumin (BCA) protein detection kit.

[0018] Furthermore, the preparation steps of chitosan-magnolol nanoparticles are as follows: Step 1: Dissolve 10 - 15 parts of magnolol in 1 part of absolute ethanol, and slowly drop it into a 0.5% chitosan solution (pH 4.0, adjusted with acetic acid); Step 2: Perform ultrasonic emulsification (200 W, 2 minutes) to form a primary emulsion, and gradually dropwise add a 0.1% sodium tripolyphosphate solution, and stir magnetically for 30 minutes; Step 3: Centrifuge (10,000 rpm, 15 minutes), collect the nanoparticles, and disperse them in phosphate buffer for standby.

[0019] Furthermore, the preparation steps of the MIP-spiropyran and chitosan-magnolol mixed indicator are as follows: Step 1: Disperse the dried MIP-spiropyran in ethanol; Step 2: Add 1 - 5 parts of chitosan-magnolol nanoparticles, and adsorb with shaking at room temperature for 12 hours under dark conditions; Step 3: Centrifuge to collect the precipitate, wash it twice with PBS to remove the unadsorbed chitosan-magnolol, and obtain the MIP-spiropyran and chitosan-magnolol mixed indicator.

[0020] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. By introducing iron-based nanozymes into PETG materials, these materials can catalyze hydrogen peroxide to generate highly reactive free radicals, directly damaging bacterial cell membranes and their internal metabolic functions, thus achieving efficient antibacterial effects. At the same time, the enzymatic catalytic properties of iron-based nanozymes can effectively degrade bacterial biofilms in the oral cavity, reduce plaque accumulation, thereby reducing the risk of dental caries during the wearing of invisible orthodontic appliances, significantly superior to traditional single anti-caries measures such as mechanical cleaning, fluoride-containing products, or antibacterial coatings.

[0021] 2. By adopting the combination technology of molecularly imprinted polymers (MIP) and spiropyran fluorescent materials, invisible orthodontic appliances are equipped with the ability to highly selectively capture specific cariogenic bacteria and develop color under ultraviolet light irradiation, thus realizing the visual detection of plaque distribution. This technology enables users to intuitively judge oral hygiene conditions. Compared with traditional mouthwashes or antibacterial coatings, its detection and protection functions are more intelligent, providing an innovative solution for the personalized care of invisible orthodontic appliances.

[0022] 3. Through the design of a chitosan-magnolol drug-loading layer, this material can respond intelligently in the acidic environment of bacterial infection. Chitosan protonates and swells, accelerating the release of magnolol, thereby effectively interfering with the expression of adhesion proteins of Streptococcus mutans and further enhancing antibacterial penetration ability. In addition, the anti-inflammatory properties of magnolol can reduce the inflammatory response caused by oral flora imbalance. Compared with traditional antibacterial coatings or mouthwashes, its drug effect is more persistent and has biological safety, providing a better solution for the long-term wearing of invisible orthodontic appliances. Detailed implementation methods

[0023] To make the technical means, creative features, achieved purposes, and effects of the present invention easy to understand, the present invention will be further described below in conjunction with specific implementation methods.

[0024] Example 1 Fe 3 O 4 The preparation steps of the nanozyme are as follows: Dissolve 0.2 parts of FeCl 2 and 0.4 parts of FeCl 3 ·6H 2 O in deionized water and mix well using a magnetic stirrer. After heating to 60 °C, add 10 parts of ammonia water at one time, continue stirring and heating to 80 °C, and increase the rotation speed to continue reacting for 2 h. After the reaction is completed, separate the Fe 3 O 4 nano-particles with a magnet, and wash them alternately with distilled water and absolute ethanol 3 times, and dry them in a vacuum drying oven at 60 °C for 4 - 6 h for standby.

[0025] Fe 3 O 4 The preparation steps of the Fe Dissolve 0.2 parts of Fe 3 O 4 nanozyme filler powder in a separate beaker containing an organic solvent, and place it on a heating plate at (80 °C) and stir for 20 minutes to obtain Fe 3 O 4 nanozyme solution. Put 15 parts of PETG pellets into a container containing an organic solvent, stir in a water bath at 35 °C until the PETG pellets are completely dissolved to obtain a PETG solution; then mix the Fe 3 O 4 nanozyme solution and the PETG solution, and place them in an ultrasonic bath (ultrasonic power is 300 W, time is 40 minutes) and vibrate violently at 80 °C for 2 hours. After the solution in step three is cooled, inject the uniformly mixed solution in step three into a mold, and then transfer the mold to a constant temperature oven at 60 °C for 24 hours for solvent evaporation treatment. After the film formation is completed, place the film in a vacuum drying oven and heat dry it at 60 °C for 6 hours. Finally, peel the completely cured composite material from the mold substrate to obtain a PETG composite film sheet of Fe 3 O 4 nanozyme.

[0026] The preparation steps of MIP-spiropyran are as follows (this process needs to be operated in the dark): Take 100 parts of SiO 2 -NH 2 Disperse it in 10 parts of PBS, add 1 part of glutaraldehyde (5% v / v), stir at room temperature for 2 hours and then centrifuge (8000 rpm, 10 min); wash it 3 times with PBS to remove unreacted glutaraldehyde; the activated SiO 2 -NH 2 Mix with 1 - 5 parts of Gtase solution with a concentration of 1 mg / mL, incubate with shaking at 4 °C for 12 hours, then centrifuge to remove unbound enzyme, and wash with PBS until the absorbance of the eluate at 280 nm is close to the baseline. The SiO 2 -NH 2Disperse in 20 parts of PBS / acetonitrile mixed solvent, add 0.4 mmol of MAA and 0.4 mmol of spiropyran-SPMA, and under a red light environment, react with shaking at 4 °C for 2 hours to enable the monomers and spiropyran-SPMA to synergistically bind to the template enzyme; add 3.6 mmol of EGDMA (crosslinking degree 90%), 10 parts of APS and 0.02 part of tetramethylethylenediamine (TEMED), purge with nitrogen for 10 minutes to remove oxygen, and seal the reaction vessel; place the prepolymer solution under a 365 nm ultraviolet light source (distance 10 cm), irradiate at 25 °C for 30 minutes to complete the polymerization (Note: Use a water cooling device or intermittent irradiation (stop for 1 minute every 5 minutes) to control the temperature ≤ 25 °C); centrifuge to collect the polymer, wash ultrasonically with 0.1% SDS + 1 M NaCl solution (30 min × 3 times), and further wash with 0.5% Tween-20 twice to remove residual surfactants; verify that the template removal rate > 95% through a bovine serum albumin (BCA) protein detection kit.

[0027] The preparation steps of the MIP-spiropyran and magnolol mixed indicator are as follows: Disperse the dried MIP-spiropyran in ethanol; add 1 part of chitosan-magnolol nanoparticles, and adsorb with shaking at room temperature in the dark for 12 hours; centrifuge to collect the precipitate, wash twice with PBS to remove unadsorbed chitosan-magnolol, and obtain the MIP-spiropyran and chitosan-magnolol mixed indicator.

[0028] Example 2 Fe 3 O 4 The preparation steps of the Fe Dissolve 0.5 part of Fe 3 O 4 nanozyme filler powder in a separate beaker containing an organic solvent, and stir on a heating plate at (80 °C) for 20 minutes to obtain an Fe 3 O 4 nanozyme solution. Put 15 parts of PETG pellets into a container containing an organic solvent, stir in a water bath at 50 °C until the PETG pellets are completely dissolved to obtain a PETG solution; then mix the Fe 3 O 4 nanozyme solution and the PETG solution, and place them in an ultrasonic bath (ultrasonic power 500 W, time 60 minutes) and vibrate violently at 80 °C for 2 hours. After the solution in step three cools, inject the uniformly mixed solution in step three into a mold, and then transfer the mold to a constant temperature oven at 60 °C for 24 hours for solvent evaporation treatment. After the film formation is completed, place the film in a vacuum drying oven and heat-dry at 60 °C for 6 hours. Finally, peel the completely cured composite material from the mold substrate to obtain Fe 3 O 4PETG composite film of nanozyme.

[0029] The preparation steps of MIP-spiropyran are as follows (this process needs to be operated under light protection): Take 100 parts of SiO 2 -NH 2 Disperse it in 10 parts of PBS, add 1 part of glutaraldehyde (5% v / v), stir at room temperature for 2 hours and then centrifuge (8000 rpm, 10 min); wash with PBS 3 times to remove unreacted glutaraldehyde; disperse the activated SiO 2 -NH 2 Mix with 3 parts of Gtase solution with a concentration of 1 mg / mL, incubate with shaking at 4°C for 12 hours, then centrifuge to remove unbound enzyme, and wash with PBS until the absorbance of the eluate at 280 nm is close to the baseline. Disperse the SiO 2 -NH 2 fixed with Gtase in 20 parts of PBS / acetonitrile mixed solvent, add 0.4 mmol MAA and 0.4 mmol spiropyran-SPMA, react with shaking at 4°C for 2 hours in a red light environment to make the monomer and spiropyran-SPMA synergistically bind to the template enzyme; add 3.6 mmol EGDMA (crosslinking degree 90%), 10 parts of APS and 0.02 part of tetramethylethylenediamine (TEMED), deoxygenate with nitrogen for 10 minutes, and seal the reaction vessel; place the prepolymer solution under a 365 nm ultraviolet light source (distance 10 cm), irradiate at 25°C for 30 minutes to complete the polymerization (note: use a water cooling device or intermittent irradiation (stop for 1 minute every 5 minutes) to control the temperature ≤ 25°C); centrifuge to collect the polymer, wash ultrasonically with 0.1% SDS + 1 M NaCl solution (30 min × 3 times), and further wash with 0.5% Tween-20 2 times to remove residual surfactants; verify that the template removal rate > 95% through a bovine serum albumin (BCA) protein detection kit.

[0030] The preparation steps of the MIP-spiropyran and magnolol mixed indicator are as follows: Disperse the dried MIP-spiropyran in ethanol; add 5 parts of chitosan-magnolol nanoparticles, and adsorb with shaking at room temperature for 12 hours under light protection; centrifuge to collect the precipitate, wash with PBS 2 times to remove unadsorbed chitosan-magnolol, and obtain the MIP-spiropyran and chitosan-magnolol mixed indicator.

[0031] Comparative Example 1 Dissolve 0.2 part of Fe 3 O 4 nanozyme filler powder in a separate beaker containing an organic solvent, and stir on a hot plate at (80°C) for 20 minutes to obtain Fe 3 O 4Nanozyme solution. Put 10 parts of PETG pellets into a container filled with organic solvent, stir in a water bath at 40 °C until the PETG pellets are completely dissolved to obtain a PETG solution; then mix the Fe 3 O 4 nanozyme solution and the PETG solution, place them in an ultrasonic bath (ultrasonic power is 400 W, time is 50 minutes), and vibrate vigorously at 80 °C for 2 hours. After the solution in step three is cooled, inject the uniformly mixed solution in step three into a mold, and then transfer the mold to a constant temperature oven at 60 °C for 24 hours for solvent evaporation treatment. After film formation is completed, place the film in a vacuum drying oven and heat-dry it at 60 °C for 6 hours. Finally, peel the completely cured composite material from the mold substrate to obtain a PETG composite film with Fe 3 O 4 nanozyme.

[0032] Comparative Example 2 Put 10 parts of PETG pellets into a container filled with organic solvent, stir in a water bath at 35 °C until the PETG pellets are completely dissolved to obtain a PETG solution; then mix the same amount of organic solvent and PETG solution used to dissolve Fe 3 O 4 in the example, place them in an ultrasonic bath (ultrasonic power is 300 W, time is 40 minutes), and vibrate vigorously at 80 °C for 2 hours. After cooling, inject the uniformly mixed solution into a mold, and then transfer the mold to a constant temperature oven at 60 °C for 24 hours for solvent evaporation treatment. After film formation is completed, place the film in a vacuum drying oven and heat-dry it at 60 °C for 6 hours. Finally, peel the completely cured composite material from the mold substrate to obtain a PETG film.

Claims

1. A method for preparing a plaque-displaying anti-caries invisible orthodontic appliance based on iron-based nanozyme catalysis and traditional Chinese medicine anti-caries synergy, characterized in that: The following steps are involved: The PETG composite braces of Fe3O4 nanozyme were combined with the MIP-spiropyran and magnolol mixed indicator coating to produce a plaque-displaying anti-caries invisible braces based on the synergistic effect of iron-based nanozyme catalysis and traditional Chinese medicine anti-caries.

2. The method according to claim 1, characterized in that: The weight proportions of the raw material components are as follows: 0.2 parts of FeCl2, 0.4 parts of FeCl3·6H2O, 10 parts of ammonia water, 10-15 parts of PETG pellets, 50-100 parts of organic solvents, 100 parts of amino-modified silica nanoparticles (SiO2-NH2), 0.4 mmol of spiropyran-methylacrylate, 100 parts of phosphate buffer, 1-5 parts of purified glucosyltransferase, 10 parts of chitosan solution, 5 parts of 0.1% SDS + 1 M NaCl solution, 10 parts of 0.5% Tween-20 solution, and 10-15 parts of magnolol.

3. The method according to claim 1, characterized in that: The preparation steps of Fe3O4 / PETG composite are as follows: after mixing Fe3O4 nanozyme solution and PETG solution, place them in an ultrasonic treatment bath with an ultrasonic power of 200-500 W, treat for 30-60 minutes, and vibrate vigorously at 80°C for 2 hours. After the solution is cooled, inject the evenly mixed solution into the mold, and then transfer the mold to a constant temperature oven at 50-60°C for 12-24 hours of solvent volatilization treatment. After the film formation is completed, place the membrane in a vacuum drying oven at 60°C for thermal drying for 4-6 hours. Finally, peel off the fully cured composite material from the mold base to obtain a PETG composite membrane of Fe3O4 nanozyme.

4. The method according to claim 3, characterized in that: The preparation steps of Fe3O4 solution and PETG solution are as follows: dissolve 0.2-0.5 parts of Fe3O4 nanozyme filler powder in a separate beaker filled with organic solvent, place it on an 80°C hot plate and stir for 20 minutes to obtain Fe3O4 nanozyme solution; put 10-15 parts of PETG pellets into a container filled with organic solvent, stir in a water bath at 35~50°C until the PETG pellets are completely dissolved to obtain PETG solution.

5. The method according to claim 4, characterized in that: The organic solvent is one of dichloromethane, tetrahydrofuran, chloroform, dimethylformamide or dimethyl sulfoxide.

6. The method according to claim 4, characterized in that: The preparation steps of Fe3O4 nanozyme are as follows: dissolve 0.2 parts of FeCl2 and 0.4 parts of FeCl3·6H2O in deionized water, mix them with a magnetic stirrer, heat them to 60°C, add 10 parts of ammonia water at one time, continue stirring and heat them to 80°C, increase the speed and continue the reaction for 2 hours. After the reaction, separate the Fe3O4 nanoparticles with a magnet, wash them alternately with distilled water and anhydrous ethanol for 3 times, and dry them in a vacuum drying oven at 60°C for 4-6 hours to obtain Fe3O4 nanozyme filler powder.

7. The method according to claim 1, characterized in that: The preparation steps of the MIP-spiropyran and chitosan-magnolia phenol mixed display agent are as follows: disperse the dried MIP-spiropyran in ethanol, add 1-5 parts of chitosan-magnolia phenol nanoparticles, adsorb under shaking at room temperature for 12 hours under light-proof conditions, then collect the precipitate by centrifugation, wash twice with PBS to remove the unadsorbed chitosan-magnolia phenol, and prepare the MIP-spiropyran and chitosan-magnolia phenol mixed display agent.

8. The method according to claim 7, characterized in that: The preparation steps of MIP-spiropyran display agent are as follows: SiO2-NH2 with fixed Gtase is dispersed in 20 parts of PBS / acetonitrile mixed solvent, 0.4mmol MAA and 0.4mmol spiropyran-SPMA are added, and the reaction is oscillated at 4°C for 2 hours under red light environment to make the monomer and spiropyran-SPMA synergistically bind to the template enzyme, 3.6mmol EGDMA, 10 parts APS and 0.02 parts tetramethylethylenediamine are added, nitrogen is passed for deoxygenation for 10 minutes, the reaction container is sealed, and the prepolymer solution is placed under a 365 nm ultraviolet light source at a distance of 10 cm and 25°C for 30 minutes to complete the polymerization, the polymer is collected by centrifugation, and ultrasonically washed with 0.1% SDS + 1 M NaCl solution for 30 min 3 times, and further washed twice with 0.5% Tween-20 to remove the residual surfactant to obtain MIP-spiropyran display agent.

9. The method according to claim 7, characterized in that: The preparation steps of chitosan-magnolia phenol nanoparticles are as follows: 10-15 parts of magnolia phenol are dissolved in 1 part of anhydrous ethanol, slowly dripped into 0.5% chitosan solution, ultrasonic emulsified at 200 W for 2 minutes to form colostrum, 0.1% sodium tripolyphosphate solution is added dropwise, magnetic stirring is performed for 30 minutes, and then centrifuged at 10,000 rpm for 15 minutes to collect nanoparticles to obtain chitosan-magnolia phenol nanoparticles.

10. The method according to claim 8, characterized in that: The preparation steps of MIP are as follows: take 100 parts of SiO2-NH2 and disperse them in 10 parts of PBS, add 1 part of glutaraldehyde, stir at room temperature for 2 hours, then centrifuge at 8000 rpm for 10 min, wash with PBS three times to remove unreacted glutaraldehyde, mix the activated SiO2-NH2 with 1-5 parts of Gtase solution with a concentration of 1 mg / mL, incubate with shaking at 4°C for 12 hours, then centrifuge to remove unbound enzyme, wash with PBS until the absorbance of the eluate at 280 nm is close to the baseline, and obtain MIP.