Zinc silicate loaded lysozyme multifunctional toothpaste and preparation method thereof

Abstract

The invention relates to zinc silicate loaded lysozyme multifunctional toothpaste and a preparation method thereof. The zinc silicate loaded lysozyme multifunctional toothpaste is prepared from the following components in percentage by mass: 40 to 50 weight percent of abrasive, 18 to 25 weight percent of humectant, 2.0 to 2.9 weight percent of surfactant, 1.0 to 1.5 weight percent of adhesive, 0.1 to 0.5 weight percent of preservative, 0.1 to 0.4 weight percent of sweetening agent, 1 to 20 weight percent of lysozyme loaded zinc silicate and the balance of water. The zinc silicate loaded lysozyme multi-effect toothpaste disclosed by the invention has various effects of well preventing and treating decayed teeth, oral ulcers and the like, not only has a good mineralization promoting effect, but also has excellent antibacterial, anti-inflammatory and repair promoting effects, and has a very wide application prospect. The preparation method disclosed by the invention has the advantages of low cost, simplicity and convenience in operation, controllable process, mild reaction conditions and the like.

Description

Technical Field

[0001] This invention belongs to the field of oral care technology, and in particular relates to a zinc silicate-loaded lysozyme multifunctional toothpaste and its preparation method. Background Technology

[0002] Oral diseases have become a major global health problem, with dental caries and oral ulcers being the most common. Dental caries originates from the demineralization of teeth caused by the acid production of cariogenic bacteria's biofilm metabolism, while damage to the oral mucosa easily leads to secondary bacterial infections and delays healing. In the field of antibacterial agents, chlorhexidine (CHX) and triclosan (TCS) can inhibit cariogenic bacteria, but long-term use can cause side effects such as tooth discoloration, microbial resistance, dysbiosis, and potential systemic toxicity. In the field of remineralization, fluoride (such as fluoride toothpaste) can improve the acid resistance of enamel by forming fluorapatite, but the fluorosis caused by excessive fluoride intake and the emergence of fluoride-resistant bacteria cannot be ignored. In-depth analysis reveals that the core defect of existing oral care materials lies in their "functional singularity": antibacterial agents lack mineralization capabilities, and mineralizing agents lack antibacterial activity. This disconnect leads to a treatment dilemma of "treating the symptoms rather than the root cause." For example, while fluoride toothpaste can reduce the incidence of tooth decay, it cannot reduce the occurrence of oral ulcers; chlorhexidine mouthwash can control plaque, but it cannot promote the repair of demineralized enamel. The more fundamental problem lies in the fact that existing materials fail to match the dynamic complexity of the oral microenvironment: the oral cavity undergoes multiple physiological activities daily, including eating, saliva secretion, and chewing friction. Single-function materials cannot simultaneously achieve multiple goals such as antibacterial, mineralization, and mucosal repair in this complex and dynamic environment. This "separate antibacterial and repair" treatment model is ill-suited to the need for synergistic repair of multiple oral tissues, highlighting the urgent need to develop multifunctional integrated materials.

[0003] Chinese patent application CN106924090A discloses the preparation and application of a multi-porous bioactive glass-immobilized lysozyme toothpaste. Although the multi-porous bioactive glass-immobilized lysozyme toothpaste in this patent application is a novel toothpaste with multiple functions, its performance in preventing and treating tooth decay and oral ulcers needs to be further improved.

[0004] In summary, it is essential to provide a zinc silicate-loaded lysozyme multifunctional toothpaste and its preparation method. Summary of the Invention

[0005] To address one or more technical problems existing in the prior art, this invention provides a zinc silicate-loaded lysozyme multi-functional toothpaste and its preparation method. The zinc silicate-loaded lysozyme multi-functional toothpaste of this invention has excellent effects in preventing and treating tooth decay and oral ulcers.

[0006] The present invention provides, in a first aspect, a zinc silicate-supported lysozyme multi-functional toothpaste, wherein the zinc silicate-supported lysozyme multi-functional toothpaste comprises the following components by mass fraction: 40-50 wt% abrasive, 18-25 wt% humectant, 2.0-2.9 wt% surfactant, 1.0-1.5 wt% adhesive, 0.1-0.5 wt% preservative, 0.1-0.4 wt% sweetener, 1-20 wt% zinc silicate carrying lysozyme and balance water.

[0007] Preferably, the preparation of the zinc silicate carrying lysozyme includes the following steps: S1. Dissolve lysozyme in an acetate-sodium acetate buffer solution to obtain a lysozyme solution; S2. Add zinc silicate to the lysozyme solution and oscillate to load it, then centrifuge, wash and dry to obtain zinc silicate loaded with lysozyme.

[0008] Preferably, the pH of the acetate-sodium acetate buffer solution is 5.0-6.0; the shaking loading is carried out at 37°C and 100-150 rpm for 12-24 hours; and / or the concentration of lysozyme in the lysozyme solution is 10-50 mg / mL.

[0009] Preferably, the mass ratio of zinc silicate to the lysozyme in the lysozyme solution is (0.5~3):1.

[0010] Preferably, the zinc silicate in the zinc silicate carrying lysozyme is a flower-shaped porous zinc silicate microsphere and / or a cluster-shaped zinc silicate microsphere.

[0011] Preferably, the zinc silicate-loaded lysozyme multifunctional toothpaste contains 5-15 wt%, more preferably 10-15 wt%, of lysozyme-loaded zinc silicate.

[0012] Preferably, the abrasive is at least one of calcium carbonate, calcium phosphate, and silicon dioxide; and / or the humectant is at least one of sorbitol, glycerin, and polyethylene glycol.

[0013] Preferably, the surfactant is at least one of sodium dodecyl sulfate, sodium lauryl sulfate, sodium lauroyl sarcosinate, and sodium dodecylbenzene sulfonate; and / or the adhesive is at least one of sodium carboxymethyl cellulose, hydroxyethyl cellulose, guar gum, xanthan gum, and carbomer.

[0014] Preferably, the preservative is at least one of parabens, sodium benzoate, and phenoxyethanol; and / or the sweetener is sodium saccharin and / or xylitol.

[0015] In a second aspect, the present invention provides a method for preparing the zinc silicate-loaded lysozyme multifunctional toothpaste described in the first aspect, the method comprising: (1) Add the prescribed amount of humectant, the prescribed amount of surfactant and the prescribed amount of adhesive to water and stir evenly to obtain the first mixture; add the prescribed amount of sweetener and the prescribed amount of preservative to water and stir evenly to obtain the second mixture; (2) Add the second mixture to the first mixture and stir well. After standing, the third mixture is obtained. (3) Add the formulated amount of abrasive and the formulated amount of zinc silicate loaded with lysozyme to the third mixture and stir evenly to obtain zinc silicate loaded with lysozyme multifunctional toothpaste.

[0016] Compared with the prior art, the present invention has at least the following beneficial effects: (1) This invention introduces zinc silicate loaded with lysozyme into toothpaste to construct a sustained-release system. Compared with single lysozyme or zinc silicate materials, the antibacterial rate of this composite material is significantly improved at the same concentration. On the one hand, zinc silicate, by adsorbing Ca... 2+ / PO4 3- It actively induces hydroxyapatite deposition, achieving in-situ regeneration of demineralized dental hard tissue; on the other hand, the anti-inflammatory properties of lysozyme are similar to those of Zn. 2+ Synergistically, its antibacterial effects effectively reduce the accumulation of inflammatory factors at the site of oral ulcers, accelerate mucosal barrier repair, avoid secondary infections, and relieve pain symptoms. Compared with single zinc silicate or lysozyme components, its healing-promoting effect is significantly enhanced. Simultaneously, it abandons traditional antibiotics and chemical preservatives, using biocompatible components such as lysozyme, zinc, and silicon to avoid tooth discoloration and systemic toxicity risks. Utilizing this single composite material of lysozyme-loaded zinc silicate, it simultaneously achieves multi-functional integration of "antibacterial-mineralization-repair," solving the formulation complexity problems caused by multi-component compounding in existing technologies. Breaking through the limitations of traditional toothpaste that only targets teeth or mucosa, the lysozyme-loaded zinc silicate composite system simultaneously promotes enamel and dentin remineralization and oral mucosal repair, constructing a comprehensive oral health protection network and achieving synergistic effects of cleaning and repair (hard tissue regeneration and mucosal repair), demonstrating promising application prospects.

[0017] (2) Compared with the prior art, the toothpaste of the present invention not only has a good mineralization effect, but also has excellent antibacterial, anti-inflammatory and repair-promoting effects. The present invention has verified its multiple functions through in vitro antibacterial experiments, tea stain adsorption experiments, in vitro mineralization experiments and in vivo repair-promoting experiments, and has a very broad application prospect. The preparation method of the present invention has the advantages of low cost, simple operation, controllable process and mild reaction conditions. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 This is the cumulative release rate curve of zinc silicate carrying lysozyme at different pH values ​​in Example 1 of the present invention; Figure 2 These are SEM images of the remineralized enamel and dentin prepared by toothpaste in Examples 1-3 and Comparative Example 1 of this invention; Figure 3 This is an AFM image of the remineralized tooth enamel prepared by toothpaste in Example 2 of this invention; Figure 4 This is an XRD pattern of remineralized tooth enamel prepared by toothpaste in Example 2 of this invention; Figure 5 These are before-and-after comparison images of the toothpaste prepared in Example 2 and Comparative Example 2 of this invention, showing the toothpaste used before and after brushing. Figure 6 This is a macroscopic diagram showing the repair of oral ulcer wounds after brushing with the toothpaste prepared in Embodiment 2 and Comparative Example 2 of the present invention. Detailed Implementation

[0020] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below in conjunction with the embodiments of this invention. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0021] The present invention provides, in a first aspect, a zinc silicate-supported lysozyme multi-functional toothpaste, wherein the zinc silicate-supported lysozyme multi-functional toothpaste comprises the following components by mass fraction: 40–50 wt% (e.g., 40 wt%, 41 wt%, 42 wt%, 43 wt%, 44 wt%, 45 wt%, 46 wt%, 47 wt%, 48 wt%, 49 wt%, or 50 wt%) of abrasives; 18–25 wt% (e.g., 18 wt%, 19 wt%, 20 wt%, 21 wt%, 22 wt%, 23 wt%, 24 wt%, or 25 wt%) of humectants; 2.0–2.9 wt% (e.g., 2.0 wt%, 2.1 wt%, 2.2 wt%, 2.3 wt%, 2.4 wt%, 2.5 wt%, 2.6 wt%, 2.7 wt%, 2.8 wt%, or 2.9 wt%) of surfactants; 1.0–1.5 wt% (e.g., 1.0 wt%, 1.1 wt%, 1.2 wt%, 1.3 wt%, 1.4 wt%, or 1.5 wt%) of surfactants. 0.1 to 0.5 wt% (e.g., 0.1 wt%, 0.2 wt%, 0.3 wt%, 0.4 wt% or 0.5 wt%) of adhesives, 0.1 to 0.4 wt% (e.g., 0.1 wt%, 0.2 wt%, 0.3 wt% or 0.4 wt%) of preservatives, 0.1 to 0.4 wt% (e.g., 0.1 wt%, 0.2 wt%, 0.3 wt% or 0.4 wt%) of sweeteners, 1 to 20 wt% (e.g., 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt% or 20 wt%) of sweeteners. (wt%) Zinc silicate loaded with lysozyme and the balance being water; in this invention, the sum of the mass fractions of each component is 100%; in this invention, the zinc silicate loaded with lysozyme is obtained by loading lysozyme onto zinc silicate.

[0022] Lysozyme achieves highly effective bactericidal action by specifically hydrolyzing bacterial cell walls and is unlikely to induce drug resistance, while also possessing anti-inflammatory properties; zinc ions (Zn) 2+ Lysozyme exerts a broad-spectrum antibacterial effect by disrupting the bacterial membrane structure. Although the respective functional roles of lysozyme and zinc silicate (Zn2SiO4) have been studied, there are currently no publicly reported cases of lysozyme-loaded zinc silicate being used in toothpaste bases. This invention preferably uses zinc silicate with a porous structure. By utilizing the porous structure of zinc silicate, lysozyme can be uniformly loaded inside and / or on its surface, forming a stable sustained-release system. During use, the interaction between lysozyme and Zn2SiO4 can be achieved. 2+ The coordinated release of Zn 2+ Zinc silicate loaded with lysozyme can synergistically inhibit cariogenic bacterial biofilms and accelerate mucosal repair; it not only enhances the stability and sustained antibacterial properties of lysozyme, but also adsorbs calcium. 2+ / PO4 3- It promotes hydroxyapatite deposition, achieving tooth remineralization, while zinc silicate continuously induces the regeneration of hard tooth tissue, forming a triple mechanism of "antibacterial-mineralization-repair." Furthermore, zinc silicate, with its porous structure, can effectively adsorb pigment deposits in the oral cavity, thus helping to maintain the white appearance of teeth. This synergistic effect cannot be obtained by simply adding lysozyme or zinc silicate alone, nor is it the same as the effect produced by simply combining the two; it exhibits a significant synergistic effect. This technology breaks through the limitations of traditional toothpaste's single function, providing an innovative solution for the comprehensive prevention and treatment of oral diseases.

[0023] In the multi-functional toothpaste of the present invention, the preferred addition amount of the lysozyme-loaded zinc silicate is 1-20 wt%. Within this range, the lysozyme-loaded zinc silicate can be uniformly dispersed in the toothpaste matrix, ensuring sufficient lysozyme and Zn. 2+ This method releases zinc silicate to effectively inhibit cariogenic bacterial biofilms and repair oral mucosa, while maintaining the remineralization effect of zinc silicate on tooth hard tissues. It avoids excessively thick toothpaste texture or affecting daily use due to excessive dosage. When the amount of lysozyme-loaded zinc silicate is too low, the content of lysozyme-loaded zinc silicate in the toothpaste is insufficient, and the lysozyme loading and Zn... 2+ Limited release leads to a significant decrease in antibacterial and remineralization effects. Excessive dosage increases toothpaste viscosity, reduces foaming properties and user experience, and excessive zinc silicate loading may lead to the depletion of lysozyme and zinc. 2+ Excessive local concentration can cause oral mucosal irritation or ingredient waste. This invention controls the zinc silicate carrying lysozyme within the range of 1-20 wt%, ensuring toothpaste performance and user comfort while achieving highly efficient antibacterial and sustained-release effects, as well as synergistic effects of tooth remineralization and oral repair. Furthermore, its porous structure enables the adsorption of oral pigments, achieving multiple synergistic effects of antibacterial, sustained-release, remineralization, oral repair, and pigment adsorption.

[0024] According to some preferred embodiments, the preparation of the zinc silicate carrying lysozyme includes the following steps: S1. Dissolve lysozyme in an acetate-sodium acetate buffer solution to obtain a lysozyme solution; the present invention does not specifically limit the acetate-sodium acetate buffer solution, and any commercially available product or a product prepared by existing methods may be used; the present invention does not specifically limit the lysozyme, and any commercially available product may be used; S2. Add zinc silicate to the lysozyme solution and oscillate to load it, then centrifuge, wash and dry to obtain lysozyme-loaded zinc silicate (also referred to as lysozyme-loaded zinc silicate composite microspheres). In this invention, for example, after centrifuging to remove the supernatant, the precipitate is washed multiple times with deionized water (e.g., 3 times), and then vacuum dried at 37°C. This invention does not specifically limit the vacuum drying time, which can be conventionally selected by those skilled in the art. The vacuum drying time is, for example, 4~12h.

[0025] According to some preferred embodiments, the pH of the acetate-sodium acetate buffer solution is 5.0-6.0, preferably 5.0-5.5. In this invention, during loading, it is preferred to dissolve the lysozyme in an acetate-sodium acetate buffer solution with a pH of 5-6. Under these conditions, the lysozyme activity is superior, and it is beneficial for adsorption with zinc silicate, thereby achieving efficient loading. Since the zinc silicate preferred in this invention is a zinc silicate flower-shaped porous microsphere, this porous microsphere provides a rich internal surface area, allowing the lysozyme to not only adsorb onto the outer surface of the particles but also enter the pores and bind to the pore walls, achieving a higher loading capacity and a more stable binding state. If the pH deviates from 5-6, the adsorption effect is relatively weakened, and the lysozyme is less likely to penetrate deep into the pores for loading, resulting in reduced binding capacity, uneven release, or decreased stability. The oscillating loading is performed at 37°C and 100-150 rpm (e.g., 100, 110, 120, 130, 140, or 150 rpm). The solution is oscillated and loaded for 12 to 24 hours (e.g., 12, 15, 20 or 24 hours) at rpm; and / or the lysozyme solution contains a concentration of 10 to 50 mg / mL (e.g., 10, 20, 30, 40 or 50 mg / mL).

[0026] According to some specific embodiments, the zinc silicate carrying lysozyme is prepared as follows: S1. Dissolve lysozyme in an acetate-sodium acetate buffer solution with a pH of 5.0-6.0 to obtain a lysozyme solution with a concentration of 10-50 mg / mL; S2. Add zinc silicate to the lysozyme solution and shake at 37°C and 120 rpm for 12-24 hours. Then centrifuge (e.g., at 8000 rpm for 3-10 min) to remove the supernatant. Wash the precipitate three times with deionized water and dry it under vacuum at 37°C to obtain zinc silicate loaded with lysozyme.

[0027] According to some preferred embodiments, the mass ratio of zinc silicate to lysozyme in the lysozyme solution is (0.5~3):1 (e.g., 0.5:1, 1:1, 1.5:1, 2:1, 2.5:1 or 3:1), preferably 1:1. In this invention, it is preferred that the mass ratio of zinc silicate to lysozyme in the lysozyme solution is (0.5~3):1. This invention has found that within this range, the adsorption sites on the zinc silicate surface and the concentration of lysozyme molecules can maintain a reasonable match, ensuring that the lysozyme is fully bound to the zinc silicate surface and / or pores, while avoiding enzyme molecule aggregation, adsorption competition, and activity loss due to excessive lysozyme, thereby achieving high loading efficiency and excellent enzyme activity retention. When the mass ratio is lower than 0.5:1, the carrier amount is relatively insufficient, and the lysozyme... Excessive loading leads to decreased loading capacity and poor enzyme stability. When the mass ratio is higher than 3:1, there is too much carrier and insufficient lysozyme, leaving a large amount of carrier surface unused, reducing overall loading efficiency, slowing lysozyme release rate, and weakening antibacterial effect. Therefore, controlling the mass ratio of zinc silicate to lysozyme within the range of (0.5–3:1) achieves optimal loading balance and sustained-release performance, ensuring that lysozyme-loaded zinc silicate in oral applications has a comprehensive effect of high-efficiency antibacterial activity, stable sustained release, promotion of remineralization, and promotion of oral repair. In particular, the experimental results of this invention show that the loading capacity of lysozyme-loaded zinc silicate exhibits a trend of "first increasing and then decreasing" with the change of the lysozyme-zinc silicate (carrier) mass ratio; when the lysozyme-zinc silicate mass ratio is 1:1, the loading capacity reaches its maximum value (766.62 ± 19.97). (mg / g); This result indicates that there is a specific binding equilibrium and synergistic adsorption relationship between lysozyme and zinc silicate in this system, reflecting the efficient occupation of carrier surface sites by lysozyme and its optimized interaction, rather than a simple physical mixing adsorption.

[0028] According to some preferred embodiments, the zinc silicate in the lysozyme-carrying zinc silicate is a flower-shaped porous zinc silicate microsphere and / or a cluster-shaped zinc silicate microsphere; in this invention, the flower-shaped porous zinc silicate microsphere is prepared by the method disclosed in Chinese patent application 202310922612.5; specifically, the preparation of the flower-shaped porous zinc silicate microsphere includes the following steps: (a) adding a surfactant to deionized water and dissolving it completely to obtain a surfactant solution; (b) adding a silicon source to the surfactant solution, reacting completely, and then adding... (c) Add a zinc source and stir until homogeneous to obtain a first mixed solution; (d) Adjust the pH of the first mixed solution to a strongly acidic state (e.g., pH 1-3) to obtain a second mixed solution; (e) Place the second mixed solution in a hydrothermal reactor for hydrothermal reaction, and cool to room temperature after the reaction to obtain a hydrothermal product; (f) Wash and dry the hydrothermal product sequentially to obtain zinc silicate flower-shaped porous microspheres; In this invention, the cluster-shaped zinc silicate microspheres can be prepared, for example, by the method disclosed in Chinese Patent Application 202311754115.5.

[0029] In this invention, it is preferred to construct a sustained-release system using zinc silicate flower-shaped porous microspheres and / or cluster-shaped zinc silicate microspheres loaded with lysozyme. Compared with ordinary zinc silicate loaded with lysozyme, on the one hand, flower-shaped porous zinc silicate and cluster-shaped zinc silicate microspheres have a higher specific surface area and richer pore structure, providing more adsorption and immobilization sites, enabling uniform distribution and stable loading of lysozyme on their surface and within the pores; the flower-shaped porous zinc silicate and cluster-shaped zinc silicate microspheres significantly improve the loading capacity and binding strength of lysozyme. At the same time, the flower-shaped porous structure and cluster-shaped porous structure can effectively delay the release rate of lysozyme, forming a continuous and stable sustained-release effect and prolonging its antibacterial action time. On the other hand, the open structure of flower-shaped porous or cluster-shaped porous zinc silicate is beneficial to Zn 2+ The continuous release and Ca 2+ / PO4 3- The adsorption further promotes the deposition of hydroxyapatite, thereby enhancing the remineralization capacity of dental hard tissues. The composite system of flower-shaped porous zinc silicate and / or clustered spherical zinc silicate microspheres loaded with lysozyme in this invention is superior to ordinary zinc silicate carriers in terms of antibacterial efficacy, enzyme activity retention and remineralization promotion performance, demonstrating significant synergistic effects and outstanding technical advantages.

[0030] According to some preferred embodiments, the zinc silicate-loaded lysozyme multifunctional toothpaste contains 5-15 wt%, preferably 10-15 wt%, of lysozyme-loaded zinc silicate.

[0031] According to some preferred embodiments, the abrasive is at least one of calcium carbonate, calcium phosphate, and silicon dioxide; and / or the humectant is at least one of sorbitol, glycerin, and polyethylene glycol.

[0032] According to some preferred embodiments, the surfactant is at least one of sodium dodecyl sulfate, sodium lauryl sulfate, sodium lauroyl sarcosinate, and sodium dodecylbenzene sulfonate; and / or the adhesive is at least one of sodium carboxymethyl cellulose, hydroxyethyl cellulose, guar gum, xanthan gum, and carbomer.

[0033] According to some preferred embodiments, the preservative is at least one of parabens, sodium benzoate, and phenoxyethanol; and / or the sweetener is sodium saccharin and / or xylitol.

[0034] In a second aspect, the present invention provides a method for preparing the zinc silicate-loaded lysozyme multifunctional toothpaste described in the first aspect, the method comprising: (1) Add the formulated amount of humectant, surfactant and adhesive to water and stir evenly to obtain a first mixture. Add the formulated amount of sweetener and preservative to water and stir evenly to obtain a second mixture. In this invention, the water in the first mixture is the first part of water, the water in the second mixture is the second part of water, and the sum of the amounts of the first part of water and the second part of water is the formulated amount of water in the zinc silicate-loaded lysozyme multifunctional toothpaste. This invention does not impose specific limitations on the ratio of the first part of water and the second part of water, and those skilled in the art can make conventional choices. (2) Add the second mixture to the first mixture and stir well, then let it stand (e.g., stand for 1-3 hours) to obtain the third mixture; (3) Add the formulated amount of abrasive and the formulated amount of zinc silicate loaded with lysozyme to the third mixture and stir evenly to obtain zinc silicate loaded with lysozyme multifunctional toothpaste.

[0035] According to some specific embodiments, the preparation of the zinc silicate-loaded lysozyme multifunctional toothpaste is as follows: Weigh the abrasive, humectant, surfactant, adhesive, preservative, sweetener, zinc silicate loaded with lysozyme, and water according to the formula, wherein the water is, for example, deionized water; mix, wet, and stir the humectant, surfactant, and adhesive with water until uniform; then add the sweetener and preservative dissolved in water, and continue stirring until uniform, to obtain a uniform, semi-transparent, viscous adhesive; let it stand for a period of time to allow it to fully dissolve and expand; finally, add the abrasive and zinc silicate loaded with lysozyme, and stir mechanically until uniform, to obtain a paste with a certain viscosity and consistency, thus obtaining the zinc silicate-loaded lysozyme multifunctional toothpaste.

[0036] The present invention will be further described below by way of examples, but the scope of protection of the present invention is not limited to these embodiments. The present invention may have many other embodiments, and those skilled in the art can make various corresponding changes and modifications based on the present invention without departing from its spirit and essence; however, all such corresponding changes and modifications should fall within the scope of protection of the appended claims.

[0037] Example 1 The raw material composition of the zinc silicate-loaded lysozyme multifunctional toothpaste in this embodiment is shown in Table 1 below.

[0038] Table 1 In this embodiment, the preparation of the lysozyme-loaded zinc silicate is as follows: lysozyme is added to an acetate-sodium acetate buffer solution with a pH of 5.5 and dissolved thoroughly to obtain a lysozyme solution with a concentration of 25 mg / mL; zinc silicate (zinc silicate flower-shaped porous microspheres) is added to the lysozyme solution and loaded in a shaker at 37°C and 120 rpm for 24 hours; after centrifugation to remove the supernatant, the precipitate is washed three times with deionized water and vacuum dried at 37°C to obtain lysozyme-loaded zinc silicate; wherein, the mass ratio of zinc silicate to lysozyme in the lysozyme solution is 1:1, and the zinc silicate is zinc silicate flower-shaped porous microspheres, the preparation of which is as follows: a) Weigh 3 g of CTAB (hexadecyltrimethylammonium bromide) and add it to deionized water to make the mass ratio of CTAB to deionized water 1:99, and stir thoroughly to dissolve; b) Add 10 g of CTAB to the mixture obtained in step a) a) Add 10 mL of 2 mmol / L zinc nitrate aqueous solution to the mixture obtained in step b, and stir for 20 minutes to allow for complete reaction; b) Add 2 mol / L HNO3 aqueous solution to the mixture obtained in step c to obtain a viscous mixture with pH 3; c) Transfer the mixture obtained in step d to a hydrothermal reactor, react at 180℃ for 24 hours, and allow to cool naturally to obtain a hydrothermal product; d) Wash the hydrothermal product obtained in step e three times each with water and anhydrous ethanol, and then dry it in a 60℃ oven to obtain zinc silicate flower-shaped porous microspheres.

[0039] In this embodiment, the preparation of the zinc silicate-loaded lysozyme multifunctional toothpaste includes the following steps: ① Add the formula amount of (glycerol), the formula amount of sodium dodecyl sulfate and the formula amount of sodium carboxymethyl cellulose to 0.5 times the formula amount of water and stir evenly to obtain the first mixture. Add the formula amount of sodium saccharin and the formula amount of sodium benzoate to 0.5 times the formula amount of water and stir evenly to obtain the second mixture.

[0040] ② Add the second mixture to the first mixture and stir well. After standing, the third mixture is obtained.

[0041] ③ Add the prescribed amounts of calcium carbonate and zinc silicate loaded with lysozyme to the third mixture and stir evenly to obtain zinc silicate loaded with lysozyme multifunctional toothpaste.

[0042] Example 2 Example 2 is basically the same as Example 1, except that: The amount of zinc silicate carrying lysozyme added in the raw material composition is 10 wt%, and the other raw materials and preparation methods are the same.

[0043] Example 3 Example 3 is basically the same as Example 1, except that: The amount of zinc silicate carrying lysozyme added in the raw material composition is 15 wt%, and the other raw materials and preparation methods are the same.

[0044] Example 4 Example 4 is basically the same as Example 2, except that: In this embodiment, the zinc silicate used in the preparation of the lysozyme-loaded zinc silicate is ordinary zinc silicate microspheres. The preparation of the ordinary zinc silicate microspheres is as follows: a) 10 mmol of tetraethyl orthosilicate is added dropwise to 300g of water and stirred for 20 minutes to disperse it evenly, resulting in a mixed solution; b) 10mL of 2 mmol / L zinc nitrate aqueous solution is added dropwise to the mixed solution obtained in step a and stirred for 20 minutes to allow it to react fully; c) 2 mol / L HNO3 aqueous solution is added dropwise to the mixed solution obtained in step b to obtain a viscous mixed solution with a pH of 3; d) The mixed solution obtained in step c is transferred to a hydrothermal reactor and reacted at 180℃ for 24 hours, followed by natural cooling to obtain a hydrothermal product; e) The hydrothermal product obtained in step d is washed three times each with water and anhydrous ethanol, and then dried in a 60℃ oven to obtain ordinary zinc silicate microspheres.

[0045] Example 5 Example 5 is basically the same as Example 2, except that: In this embodiment, the zinc silicate used in the preparation of the lysozyme-loaded zinc silicate is a cluster of spherical zinc silicate microspheres; the cluster of spherical zinc silicate microspheres is prepared by referring to the method in Example 1 of Chinese Patent Application 202311754115.5.

[0046] Comparative Example 1 Comparative Example 1 is basically the same as Example 1, except that the use of zinc silicate carrying lysozyme is omitted in Comparative Example 1 and replaced with an equal amount of deionized water. The other raw materials and preparation methods are the same as those in Example 1. The raw material composition of the toothpaste in Comparative Example 1 is shown in Table 2 below.

[0047] Table 2 Comparative Example 2 Comparative Example 2 is basically the same as Example 2, except that the zinc silicate carrying lysozyme in the raw material composition is replaced with calcium silicate, while the other raw materials and preparation methods are the same as in Example 2.

[0048] Comparative Example 3 Comparative Example 3 is basically the same as Example 2, except that the zinc silicate carrying lysozyme in the raw material composition is replaced with lysozyme, while the other raw materials and preparation methods are the same as in Example 2.

[0049] Comparative Example 4 Comparative Example 4 is basically the same as Example 2, except that the zinc silicate carrying lysozyme in the raw material composition is replaced with zinc silicate flower-shaped porous microspheres. The other raw materials and preparation methods are the same as in Example 2. The preparation of zinc silicate flower-shaped porous microspheres is the same as in Example 1.

[0050] Comparative Example 5 Comparative Example 5 is basically the same as Example 2, except that the zinc silicate carrying lysozyme in the raw material composition is replaced with zinc silicate flower-shaped porous microspheres and lysozyme, and the mass ratio of zinc silicate flower-shaped porous microspheres to lysozyme is 1:1. The other raw materials and preparation methods are the same as in Example 2, and the preparation of zinc silicate flower-shaped porous microspheres is the same as in Example 1.

[0051] Comparative Example 6 Comparative Example 6 is basically the same as Example 2, except that the zinc silicate carrying lysozyme in the raw material composition is replaced with a hierarchical porous bioactive glass immobilized with lysozyme. The remaining raw materials and preparation methods are the same as in Example 2. Specifically, the preparation of the hierarchical porous bioactive glass immobilized with lysozyme is as follows: I. Preparation of mesoporous bioactive glass (MBG): 189 mL of anhydrous ethanol is added to a flask, followed by 10 g of triblock copolymer (P123), 18 mL of tetraethyl orthosilicate (TEOS), 1.17 g of calcium nitrate tetrahydrate, 1.71 mL of triethyl phosphate (TEP), and 5 mL of hydrochloric acid (0.5 mol / L). The mixture is then magnetically stirred at room temperature for 24 h to obtain mesoporous bioactive glass (MBG); II. Preparation of hierarchical porous bioactive glass: 5 g of pine pollen is impregnated in 50 mL of anhydrous ethanol and dispersed ultrasonically for 1 h. The pollen is then transferred to a 1:1 volume ratio mixture of ethanol and formaldehyde. After min, rinse repeatedly with deionized water, and finally carbonize with concentrated sulfuric acid. Add the immobilized pollen to 50 mL of 12 mol / L concentrated sulfuric acid and stir for 4 h in an 80℃ water bath. When the solution turns from yellow to black, filter and rinse repeatedly with deionized water until pH=7. Place the material in an 80℃ drying oven for 12 h. Impregnate the dried material with the prepared liquid MBG 15 times, dry in a 30℃ oven, and finally calcine in a resistance furnace at 700℃ for 8 h to prepare a hierarchical porous bioactive glass material; III. Preparation of hierarchical porous bioactive glass immobilized with lysozyme: Take 10 mg of the above hierarchical porous bioactive glass and 4 mL of lysozyme solution (4 The mixture (mg / L, pH=9, solvent tris(hydroxymethyl)aminomethane-hydrochloric acid buffer) was shaken at 180 rpm for 2 h at 37 °C. The adsorbed material was obtained by high-speed centrifugation and washed with a large amount of deionized water to prepare a hierarchical porous bioactive glass immobilized with lysozyme.

[0052] Experimental Example 1 The specific method for determining the lysozyme loading at different drug loading ratios is as follows: Lysozyme and zinc silicate flower-shaped porous microspheres were weighed at mass ratios of 2:1, 1:1, 1:2, and 1:3 for loading. Lysozyme was dissolved in an acetate-sodium acetate buffer solution at pH 5.5 to obtain a lysozyme solution with a concentration of 25 mg / mL. The zinc silicate flower-shaped porous microspheres were added to the lysozyme solution and loaded at 37℃ and 120 rpm for 24 h in a shaker. After loading, the mixture was centrifuged at 8000 rpm for 10 min, and the supernatant was diluted tenfold and the absorbance was measured at 280 nm. The lysozyme loading capacity of the zinc silicate flower-shaped porous microspheres was calculated based on the lysozyme standard curve. The above experiment was repeated three times, and the lysozyme loading capacity at different ratios is shown in Table 3.

[0053] Table 3 As shown in Table 3, when the mass ratio of lysozyme to carrier (zinc silicate flower-shaped porous microspheres) is 2:1, the loading is low, and when the mass ratio is 1:1, the loading is the highest, which is 766.62±19.97 mg / g. When the mass ratio of lysozyme to carrier (zinc silicate flower-shaped porous microspheres) is further reduced to 1:2 and 1:3, the loading shows a downward trend.

[0054] This invention also tested the lysozyme-loaded zinc silicate loading in Examples 4 and 5. Loading was performed with a 1:1 mass ratio of lysozyme to ordinary zinc silicate microspheres and a 1:1 mass ratio of lysozyme to clustered zinc silicate microspheres. Lysozyme was dissolved in an acetate-sodium acetate buffer solution at pH 5.5 to obtain a lysozyme solution with a concentration of 25 mg / mL. Ordinary zinc silicate microspheres (or clustered zinc silicate microspheres) were added to the lysozyme solution and loaded for 24 h at 37°C and 120 rpm in a shaker. After loading, the mixture was centrifuged at 8000 rpm for 10 min, and the supernatant was diluted tenfold and the absorbance was measured at 280 nm. The lysozyme loading capacity of ordinary zinc silicate microspheres or clustered zinc silicate microspheres was calculated based on the lysozyme standard curve. The above experiments were repeated 3 times; the loading capacity of zinc silicate carrying lysozyme in Example 4 was measured to be 62.15±13.26 mg / g, and the loading capacity of zinc silicate carrying lysozyme in Example 5 was measured to be 702.39±18.15 mg / g.

[0055] Experimental Example 2 The cumulative release rate of lysozyme was monitored by placing zinc silicate loaded with lysozyme in PBS at different pH values ​​and using a standard curve. The specific method is as follows: Weigh 10 mg of zinc silicate carrying lysozyme (prepared with lysozyme / carrier zinc silicate flower-shaped porous microspheres at a mass ratio of 1:1 and acetate-sodium acetate buffer at pH 5.5) from Example 1 and disperse it in 10 mL of PBS buffer at different pH values ​​(7.4 or 6.0). Place the centrifuge tube in a constant temperature shaker at 37°C and 100 rpm. Centrifuge the tube at 8000 rpm for 5 min at 1 h, 5 h, 8 h, 12 h, 24 h, 36 h, 48 h, and 72 h, respectively, collect 3 mL of supernatant, detect the absorbance, and add an equal volume of fresh PBS buffer at the corresponding pH. Calculate the cumulative release rate according to the lysozyme standard curve. Repeat the above experiment 3 times, and the results are as follows. Figure 1 As shown; from Figure 1 It can be seen that lysozyme can be continuously released under both pH=7.4 and pH=6.0 conditions. Under both conditions, lysozyme is released rapidly in the first 12 hours, then the release rate slows down, and reaches a plateau around 24 hours. In the initial stage of release (e.g., the first few hours), the release rate under pH=6.0 conditions is slightly higher than that under pH=7.4 conditions. By 72 hours, the cumulative release rates under pH=7.4 and pH=6.0 conditions are 44.67% and 45.68%, respectively.

[0056] In this embodiment, the zinc silicate carrying lysozyme (prepared at a mass ratio of lysozyme / carrier ordinary zinc silicate microspheres of 1:1 and an acetate-sodium acetate buffer pH=5.5) from Example 4 was also subjected to the same sustained-release test in PBS (pH 7.4 or 6.0). The results showed a burst release trend under both pH conditions, with cumulative release rates of approximately 31.0% (pH=7.4) and 35.0% (pH=6.0) over 72 hours.

[0057] Experimental Example 3 The antibacterial properties of the zinc silicate loaded with lysozyme added in Examples 1 and 4, the lysozyme added in Comparative Example 3, the zinc silicate flower-shaped porous microspheres added in Comparative Example 4, the lysozyme and zinc silicate flower-shaped porous microspheres added in Comparative Example 5, and the hierarchical porous bioactive glass with immobilized lysozyme added in Comparative Example 6 were tested using the CCK-8 method. The specific methods are as follows: Zinc silicate with lysozyme at a concentration of 5 mg / mL (phosphate buffer, pH = 7.4) was mixed with 1×10⁻⁶... 7Staphylococcus aureus, Escherichia coli, and Streptococcus mutans were co-cultured at 37°C. After culturing, 10% (v / v) CCK-8 solution was added, and the absorbance (OD 450nm) of the bacteria was measured using a microplate reader. The experiment was repeated three times, and the average OD value of each group was taken. The absorbance values ​​of bacteria at 450 nm detected by the ELISA reader are shown in Table 4. Specifically, when testing the antibacterial performance of added lysozyme, zinc silicate loaded with lysozyme was replaced with lysozyme; when testing the antibacterial performance of added zinc silicate flower-shaped porous microspheres, zinc silicate loaded with lysozyme was replaced with zinc silicate flower-shaped porous microspheres; when testing the antibacterial performance of added zinc silicate flower-shaped porous microspheres and lysozyme, zinc silicate loaded with lysozyme was replaced with a 1:1 mass ratio of zinc silicate flower-shaped porous microspheres and lysozyme; and when testing the antibacterial performance of added lysozyme-loaded hierarchical porous bioactive glass, zinc silicate loaded with lysozyme was replaced with lysozyme-loaded hierarchical porous bioactive glass.

[0058] Table 4: Absorbance results of different bacteria at OD 450 nm **** indicates that the difference is statistically significant compared with the control group (blank control group) (p<0.0001).

[0059] As can be seen from Table 4, the absorbance of the three bacteria in the zinc silicate group loaded with lysozyme in Example 1 was significantly lower than that of the control group (blank control), indicating that the zinc silicate material loaded with lysozyme has good antibacterial properties against different bacteria.

[0060] Test Example 4 The concentration of zinc silicate loaded with lysozyme was selected as 100 μg / mL as the test concentration for anti-inflammatory efficacy experiments to evaluate the anti-inflammatory effect of zinc silicate loaded with lysozyme. In this experiment, materials with a concentration of 100 μg / mL were selected (zinc silicate loaded with lysozyme or lysozyme or zinc silicate flower-shaped porous microspheres or zinc silicate flower-shaped porous microspheres with lysozyme or multi-level porous bioactive glass with immobilized lysozyme). HGF-1 cells were seeded in 24-well plates and co-cultured for 24 hours with 1 μg / mL lipopolysaccharide (LPS) and 100 μg / mL experimental material, respectively. Control groups included untreated cells (negative control) and cells treated with LPS only (positive control). Cell supernatants were collected, and interleukin-6 (IL-6) levels were measured using an ELISA kit following the manufacturer's instructions.

[0061] An in vitro inflammation model of human gingival fibroblasts (HGF-1) induced by lipopolysaccharide (LPS) at a concentration of 1 μg / mL was used. After treatment with material at a concentration of 100 μg / mL or without material for 24 hours, cell supernatant was collected, and interleukin-6 (IL-6) levels were measured. The results are shown in Table 5.

[0062] Table 5: Results of the anti-inflammatory efficacy test of zinc silicate loaded with lysozyme

[0063] #### indicates a statistically significant difference compared to the negative control group (p<0.0001); **** indicates a statistically significant difference compared to the positive control group (p<0.0001).

[0064] In the positive control group, cells were treated with LPS but without any materials. An inflammation model was constructed using human gingival fibroblasts (HGF-1) induced by LPS at a concentration of 1 μg / mL. The levels of the pro-inflammatory factor interleukin-6 (IL-6) are shown in Table 5. In the negative control group, cells were treated without LPS and without any materials. The lysozyme-loaded zinc silicate group in Example 1 was treated with LPS and the lysozyme-loaded zinc silicate group from Example 1. The lysozyme-loaded zinc silicate group in Example 4 was treated with LPS and the lysozyme-loaded zinc silicate group from Example 4. The lysozyme group in Comparative Example 3 was treated with LPS and lysozyme. The zinc silicate flower-shaped porous microsphere group in Comparative Example 4 was treated with LPS and the zinc silicate flower-shaped porous microsphere group. The zinc silicate flower-shaped porous microsphere and lysozyme (mass ratio 1:1) group in Comparative Example 5 was treated with LPS and the zinc silicate flower-shaped porous microsphere group. Compared with lysozyme (mass ratio 1:1), the hierarchical porous bioactive glass group with immobilized lysozyme in Comparative Example 6 consisted of cells plus LPS and immobilized lysozyme in a multi-level porous bioactive glass. As shown in Table 5, compared with the negative control group, the IL-6 level in the LPS-stimulated positive control group was significantly increased (369.32 pg / mL), indicating that the inflammation model was successfully established. In contrast, the IL-6 level in the experimental group treated with zinc silicate containing lysozyme from Example 1 was significantly reduced to 100.75 pg / mL, which was significantly lower than that in the positive control group, indicating that the material can effectively inhibit the secretion of LPS-induced pro-inflammatory cytokines and has good anti-inflammatory properties. It should be noted that the cells used in the evaluation of anti-inflammatory effects were human gingival fibroblasts (HGF-1).

[0065] Experimental Example 5 Toothpastes prepared in Examples 1, 2, and 3, as well as Comparative Example 1, were used to study their effects on the remineralization of tooth enamel and dentin. The preparation steps for tooth enamel and dentin samples were as follows: the crown portion was separated from the tooth at the pulp-enamel junction, and enamel and dentin blocks were cut using a low-speed water-cooled diamond saw to obtain 4mm × 4mm × 2mm samples. The samples were polished under running water with SiC paper to obtain a smooth surface. Specifically, dentin samples were progressively polished using 600, 1200, 2000, and 3000 grit sandpaper to remove surface irregularities and form a uniform, smooth surface. Samples used for the initial in vitro remineralization experiment of tooth enamel caries were finely polished using only 3000 grit sandpaper to remove minute surface irregularities while maintaining the integrity of the surface demineralized microstructure, in order to preserve the outermost original enamel structure. Before use, the samples were stored in a 0.5% thymol solution at 4°C. The samples were randomly divided into a demineralization group (no treatment after demineralization), a control group (immersed only in artificial saliva after demineralization), an example group, and a comparative group.

[0066] The demineralization and remineralization experimental steps are as follows: the tooth sample is etched with 37% phosphoric acid solution (volume fraction) for 30 seconds to produce a demineralized model, rinsed thoroughly with deionized water, and ultrasonically treated for 5 minutes to form an artificial caries.

[0067] The above-mentioned toothpaste was used to treat demineralized enamel and dentin by brushing 9 times a day, morning and evening. After treatment, the teeth were rinsed following the human brushing routine. Subsequently, tooth slices were immersed in 4 mL of artificial saliva (pH=6.8) and incubated at 37°C for 2 weeks, with the artificial saliva being replaced daily with fresh solution (freshly prepared artificial saliva (pH=6.8)). After incubation, the samples were rinsed several times with deionized water and air-dried at 25°C before examination. The surface morphology of dentin and the degree of dentinal tubule occlusion were observed using SEM; the surface morphology and roughness of the enamel in the demineralized group and Example 2 group were examined using atomic force microscopy; and the crystal orientation and mineral phase of the newly formed layer on the enamel surface in the control group and Example 2 group were analyzed by XRD. The experimental results are as follows: Figure 2 , Figure 3 , Figure 4 As shown; in Figure 2 In the images, the scales for the results of the demineralized group, control group, comparative example 1, example 1, example 2, and example 3 are the same.

[0068] SEM results are shown (see...) Figure 2In the demineralization group, control group, and Comparative Example 1, the enamel surface after phosphoric acid etching all exhibited a typical "scaly" demineralization morphology, exposing the hydroxyapatite (HAP) prismatic structure. The lysozyme-loaded zinc silicate treatment groups corresponding to Examples 1-3 showed a concentration-dependent effect: the 5% lysozyme-loaded zinc silicate group formed a porous mineralization network, while the 10% and 15% lysozyme-loaded zinc silicate groups induced continuous crystal layers accompanied by directional HAP prismatic growth, significantly smoothing the enamel surface and reducing roughness. AFM quantification results (see...) Figure 3 The results showed that 10% zinc silicate loaded with lysozyme reduced the surface roughness of tooth enamel from 57.68 nm (demineralized group) to 32.68 nm.

[0069] To address the potential destruction of internal dentinal tubules due to dentin damage, this invention further investigated its sealing effect. The control group remained open after 14 days of remineralization, while the zinc silicate treatment group with lysozyme-loaded zinc silicate induced mineral deposition inside and around the dentinal tubules. Both the 10% and 15% lysozyme-loaded zinc silicate groups generated dense, interlocking crystal layers, effectively sealing microporous defects. XRD (see...) Figure 4 The results showed that the diffraction peaks of hydroxyapatite on the enamel surface of the control group were weakened, while the crystal plane peaks of (002), (300) and (004) of the 10% zinc silicate-loaded lysozyme treatment group were significantly enhanced.

[0070] Experimental Example 6 Demineralized tooth enamel sections were immersed in colored tea for 3 days until they turned a deep yellow color, and photographs were taken for record-keeping. Subsequently, toothpaste prepared in Example 2 and Comparative Example 2 was used for brushing teeth 9 AM and 9 PM daily. The control group received no treatment. During the untreated period, tooth sections were immersed in deionized water. After 5 days, photographs were taken and analyzed for all groups of sections; the experimental results are as follows: Figure 5 As shown in the macroscopic photograph, although the toothpaste in Comparative Example 2 could partially remove tea stains from the enamel surface due to the abrasive effect, it still showed obvious yellow staining after treatment; while the toothpaste treatment group in Example 2 showed a more significant stain removal effect, and the staining on the enamel surface was significantly reduced.

[0071] Experimental Example 7 Eight-week-old SD rats (weighing 250-300 g, n=3) were selected. After the mandibular mucosa was dried with a sterile cotton ball, a 3 mm × 3 mm square filter paper was soaked in 50% glacial acetic acid for 5 seconds and attached to the gingival mucosa for 30 seconds to induce ulcer formation. Residual acetic acid was then removed with a sterile cotton ball soaked in PBS buffer. The filter paper was further soaked in a Streptococcus mutans suspension (1 × 10⁻⁶). 7After 5 seconds, the toothpaste (CFU / mL) was reapplied to the ulcer to establish a bacterial infection model. 24 hours after infection, the experimental group received toothpaste from Example 2 and the control group, which was treated with saline solution only, for 5 consecutive days. Wound images were taken daily to assess healing dynamics. The experimental results are as follows: Figure 6 As shown.

[0072] All rats showed typical inflammatory responses in their mandibular mucosa after modeling (referred to as day 1), characterized by local tissue swelling, congestion, and the formation of a pseudomembrane. On day 3 of intervention, the pseudomembrane thickness and ulcer area in group 2 (using the toothpaste in example 2) showed a decreasing trend. By day 5, the treatment effect showed significant differences between groups. The control group (treated with physiological saline) and comparative group 2 (using the toothpaste in comparative group 2) still had some pseudomembrane structures that had not sloughed off, while the ulcer area in group 2 had significantly decreased. By day 6 of intervention, the ulcer wounds in group 2 had basically completed epithelial regeneration and showed a physiological healing state, while the control group and comparative group 2 still had obvious pathological characteristics, including typical signs of infection such as whitening of the wound tissue and accumulation of purulent secretions.

[0073] In this invention, 8-week-old SD rats (weighing 250-300 g, n=3) were induced to form ulcers using the same method as described above, and a bacterial infection model was established. 24 hours after infection, the rats were treated with toothpaste from Examples 4, 5, and Comparative Examples 3-6, and brushed their teeth at 9 am and 9 pm daily for 6 consecutive days. The degree of ulcer non-healing was statistically analyzed, and the results are shown in Table 6.

[0074] Table 6: Ulcer area after 6 days / Initial ulcer area

[0075] As shown in Tables 4-6 above, although the combined addition of zinc silicate flower-shaped porous microspheres and lysozyme in Comparative Example 5 and the hierarchical porous bioactive glass with lysozyme immobilized in Comparative Example 6 also exhibited good antibacterial effects (see results in Table 4), this is because the combined addition of zinc silicate flower-shaped porous microspheres and lysozyme in Comparative Example 5 and the hierarchical porous bioactive glass with lysozyme immobilized in Comparative Example 6 had strong antibacterial properties in the early stages; however, the combined addition of zinc silicate flower-shaped porous microspheres and lysozyme in Comparative Example 5 and the hierarchical porous bioactive glass with lysozyme immobilized in Comparative Example 6... The multi-level porous bioactive glass containing immobilized lysozyme cannot provide sustained or continuous antibacterial effects. Therefore, as shown in Table 5, the anti-inflammatory performance results indicate that the zinc silicate flower-shaped porous microspheres and lysozyme group in Comparative Example 5 and the multi-level porous bioactive glass group containing immobilized lysozyme in Comparative Example 6 have relatively poor antibacterial performance due to the reduced antibacterial rate caused by burst release or degradation. Similarly, due to the inability to provide sustained or continuous antibacterial effects, the results in Table 6 show that the ulcer repair promotion effect of Comparative Example 5 and Comparative Example 6 is significantly reduced.

[0076] In summary, this invention has found that loading lysozyme with zinc silicate to obtain lysozyme-loaded zinc silicate can complement each other, synergistically enhancing the antibacterial and anti-inflammatory properties of the material. Furthermore, adding lysozyme-loaded zinc silicate to toothpaste can improve its mineralization and whitening effects. Therefore, this invention provides a toothpaste with multiple functions, including preventing and treating tooth decay and oral ulcers. In vitro antibacterial, anti-inflammatory, mineralization, whitening, and ulcer repair experiments have verified its antibacterial, mineralization, whitening, and ulcer repair functions, demonstrating its broad application prospects.

[0077] The parts of this invention not described in detail are techniques known to those skilled in the art.

[0078] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A multi-functional toothpaste containing zinc silicate-loaded lysozyme, characterized in that, The zinc silicate-loaded lysozyme multi-functional toothpaste contains the following components by mass fraction: 40-50 wt% abrasive, 18-25 wt% humectant, 2.0-2.9 wt% surfactant, 1.0-1.5 wt% adhesive, 0.1-0.5 wt% preservative, 0.1-0.4 wt% sweetener, 1-20 wt% zinc silicate carrying lysozyme and balance water.

2. The zinc silicate-loaded lysozyme multifunctional toothpaste according to claim 1, characterized in that, The preparation of the zinc silicate carrying lysozyme includes the following steps: S1. Dissolve lysozyme in an acetate-sodium acetate buffer solution to obtain a lysozyme solution; S2. Add zinc silicate to the lysozyme solution and oscillate to load it, then centrifuge, wash and dry to obtain zinc silicate loaded with lysozyme.

3. The zinc silicate-loaded lysozyme multifunctional toothpaste according to claim 2, characterized in that: The pH of the acetic acid-sodium acetate buffer solution is 5.0~6.0; The oscillation load is an oscillation load at 37℃ and 100~150 rpm for 12~24h; and / or The lysozyme solution contains lysozyme at a concentration of 10-50 mg / mL.

4. The zinc silicate-loaded lysozyme multifunctional toothpaste according to claim 2, characterized in that: The mass ratio of zinc silicate to lysozyme in the lysozyme solution is (0.5~3):

1.

5. The zinc silicate-loaded lysozyme multifunctional toothpaste according to claim 1 or 2, characterized in that: The zinc silicate in the zinc lysozyme-carrying zinc silicate is a flower-shaped porous zinc silicate microsphere and / or a cluster-shaped zinc silicate microsphere.

6. The zinc silicate-loaded lysozyme multifunctional toothpaste according to claim 1, characterized in that: The zinc silicate-loaded lysozyme multifunctional toothpaste contains 5-15 wt%, more preferably 10-15 wt%, of zinc silicate loaded with lysozyme.

7. The zinc silicate-loaded lysozyme multifunctional toothpaste according to claim 1, characterized in that: The friction agent is at least one of calcium carbonate, calcium phosphate, and silicon dioxide; and / or The moisturizer is at least one of sorbitol, glycerin, and polyethylene glycol.

8. The zinc silicate-loaded lysozyme multifunctional toothpaste according to claim 1, characterized in that: The surfactant is at least one selected from sodium dodecyl sulfate, sodium lauryl sulfate, sodium lauroyl sarcosinate, and sodium dodecylbenzene sulfonate; and / or The adhesive is at least one of sodium carboxymethyl cellulose, hydroxyethyl cellulose, guar gum, xanthan gum, and carbomer.

9. The zinc silicate-loaded lysozyme multifunctional toothpaste according to claim 1, characterized in that: The preservative is at least one of parabens, sodium benzoate, and phenoxyethanol; and / or The sweetener is sodium saccharin and / or xylitol.

10. A method for preparing a zinc silicate-loaded lysozyme multifunctional toothpaste according to any one of claims 1 to 9, characterized in that, The method includes: (1) Add the prescribed amount of humectant, the prescribed amount of surfactant and the prescribed amount of adhesive to water and stir evenly to obtain the first mixture; add the prescribed amount of sweetener and the prescribed amount of preservative to water and stir evenly to obtain the second mixture; (2) Add the second mixture to the first mixture and stir well. After standing, the third mixture is obtained. (3) Add the formulated amount of abrasive and the formulated amount of zinc silicate loaded with lysozyme to the third mixture and stir evenly to obtain zinc silicate loaded with lysozyme multifunctional toothpaste.

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