Demineralized dentin remineralization mouthwash based on cp20k-p3 bionic polypeptide and application of demineralized dentin remineralization mouthwash

Abstract

The invention discloses demineralized dentin remineralization mouthwash based on cp20k-p3 bionic polypeptide and an application of the demineralized dentin remineralization mouthwash. The mouthwash is prepared by mixing the polypeptide cp20k-p3 (the amino acid sequence is CNQKHPCWRRHGKKHRGLHRKFHGNACNC) derived from barnacle mucin 20K, an aqueous solution of calcium salt and an aqueous solution of phosphate. The polypeptide can specifically recognize and adsorb an organic matrix exposed on the surface of demineralized dentin, guide hydroxyapatite crystals to directionally deposit on the demineralized part of the dentin, and form a compact mineralized layer similar to a natural structure on the surface of the dentin. Compared with a calcium phosphate mixed aqueous solution without cp20k-p3, the preparation method is simple and convenient to operate, can form a compact mineralized layer with the surface of the re-demineralized dentin, and has excellent stability in an oral environment.

Description

Technical Field

[0001] This invention belongs to the field of functional mouthwash technology and also to the field of dentin biomimetic repair technology, specifically involving demineralized dentin remineralization mouthwash based on cp20k-p3 biomimetic peptides and its application. Background Technology

[0002] The mineralizing solution used for remineralization of demineralized dentin comprises the following components: Calcium source: Calcium is one of the important components of dentin. Common calcium sources include calcium chloride and calcium nitrate. For example, a certain concentration of calcium chloride solution is added to some mineralizing solutions, usually around 0.13M.

[0003] Phosphate source: Phosphates are also an essential component of dentin mineralization, such as disodium hydrogen phosphate, whose concentration is usually around 0.26M. They work together with calcium sources to promote the formation of hydroxyapatite, thereby achieving remineralization of dentin.

[0004] Fluoride: Fluoride can enhance mineralization and improve the caries resistance of dentin. Some mineralizing solutions contain about 500 ppm of fluoride, which can promote the deposition of calcium and phosphorus ions and make the formed mineralized layer denser.

[0005] Buffers, such as Tris buffer, are used to maintain a relatively stable pH value in the mineralization solution. The pH value is generally adjusted to around 6.5 to simulate the oral cavity environment and ensure the smooth progress of the mineralization process.

[0006] Other ingredients: Some mineralizing solutions also contain bioactive substances, such as genipin. Genipin is a natural cross-linking agent that can bind to type I collagen in dentin, enhancing collagen stability and thus better promoting mineralization.

[0007] The mechanism of action of mineralizing solutions used for remineralization of demineralized dentin: The mineralizing solution provides the necessary ions for mineralization: calcium and phosphate ions replenish the minerals lost from demineralized dentin. Under the influence of the mineralizing solution, these ions are redeposited on the surface and interstices of the collagen fibers in the dentin, forming hydroxyapatite crystals, thereby repairing the mineralized structure of the dentin.

[0008] Promoting crystal growth: The components in the mineralizing solution can regulate the growth direction and speed of crystals. For example, fluoride can promote the growth of hydroxyapatite crystals along a specific direction, forming a more ordered and dense mineralized layer, which helps restore the hardness and strength of dentin.

[0009] Simulating physiological mineralization processes: Some mineralizing solutions, through special formulations and preparation methods, simulate the mineralization process under human physiological conditions. For example, using agarose hydrogel as a carrier, calcium and phosphate sources are encapsulated in different hydrogels, and then interact on the dentin surface, more closely resembling the mineralization mechanism of natural dentin, thereby achieving more effective remineralization.

[0010] The effects of mineralizing solutions used for remineralization of demineralized dentin: Improved dentin properties: After treatment with a mineralizing solution, the mechanical properties of demineralized dentin, such as hardness and elastic modulus, are significantly improved. Studies have shown that after two days of treatment with a specific mineralizing solution, the elastic modulus and hardness of dentin recover to levels close to those of normal dentin, indicating that its mechanical properties have been well restored.

[0011] Repairing dentin defects: Mineralizing solutions can promote mineralization on the surface and inside of dentin, forming a uniform and dense mineralized layer. In animal experiments, treatment with an agarose hydrogel mineralization system resulted in the formation of a well-oriented and tightly packed hydroxyapatite layer on the dentin surface, effectively repairing dentin defects.

[0012] Preventing tooth decay: Through remineralization, the mineralizing solution enhances the acid resistance and caries resistance of dentin. The addition of fluoride further improves the caries resistance of dentin, helping to prevent the occurrence and development of tooth decay.

[0013] However, traditional mineralizing solutions used for remineralization of demineralized dentin typically have limited therapeutic effects and cannot selectively identify and adhere to the surface of demineralized dentin.

[0014] There are already related studies, such as CN202510359951.6, a biomimetic polypeptide cp20k-p3 for inducing remineralization of demineralized dentin. This polypeptide can adhere to the surface of demineralized dentin and induce its remineralization, but cp20k-p3 has not been prepared as a biomimetic mouthwash. Summary of the Invention

[0015] To address the problems of slow mineralization speed and weak adhesion in existing remineralizing mouthwashes, this invention provides a mouthwash for remineralizing demineralized dentin based on a cp20k-p3 biomimetic peptide and its application. It is also a mouthwash for inducing remineralization of demineralized dentin. The biomimetic peptide cp20k-p3 can form mineralization products containing cp20k-p3, such as cp20k-p3 / ACP or cp20k-p3 / HAP, in a mineralization solution. This composite mineralization product can selectively adsorb onto the surface of demineralized dentin and ultimately form a mineralized layer on or inside the demineralized tooth surface. Compared to calcium phosphate mixed aqueous solutions without cp20k-p3, this invention is simple to operate, can form a dense mineralized layer with the surface of remineralized dentin, and exhibits excellent stability in the oral environment.

[0016] The technical solution of the present invention is as follows: A mouthwash for demineralizing dentin based on cp20k-p3 biomimetic peptide and its application: The mouthwash is prepared by mixing the white barnacle 20K-derived peptide cp20k-p3 (amino acid sequence CNQKHPCWRRHGKKHGLHRKFHGNACNC) with an aqueous solution of calcium salt and an aqueous solution of phosphate. This peptide can specifically recognize and adsorb onto the organic matrix exposed on the surface of demineralized dentin, guide the directional deposition of hydroxyapatite crystals at the demineralized site of dentin, and form a dense mineralized layer on the dentin surface that is similar to the natural structure.

[0017] The objective of this invention is achieved through the following technical solution: A mouthwash for demineralizing dentin and remineralizing mouthwash based on cp20k-p3 biomimetic peptides contains biomimetic peptide cp20k-p3 and mineralizing solution. The biomimetic polypeptide cp20k-p3 is derived from a 20K-derived polypeptide of white barnacle protein, and the amino acid sequence of the polypeptide is: CNQKHPCWRRHGKKHGLHRKFHGNACNC; Mineralizing solution contains Ca 2+ Or PO4 3- ; When the mineralizing solution is mixed with the biomimetic peptide cp20k-p3, a demineralized dentin remineralizing mouthwash based on the cp20k-p3 biomimetic peptide is obtained.

[0018] Furthermore, the biomimetic polypeptide cp20k-p3 is synthesized using a chemical synthesis method.

[0019] Furthermore, the concentration of the biomimetic polypeptide cp20k-p3 is 0.01 mg / mL to 1 mg / mL.

[0020] The mineralizing solution is mixed with the biomimetic polypeptide cp20k-p3 by sequentially reacting the biomimetic polypeptide cp20k-p3 with a substance containing PO4. 3- solutions containing Ca 2+ The solutions are mixed; in this mixed solution, the reaction forms cp20k-p3 / ACP (ACP stands for amorphous calcium phosphate, which is formed by the reaction of calcium ions and phosphate ions. When a polypeptide is present, a complex of cp20k-p3 / ACP is formed) or cp20k-p3 / HAP (HAP stands for hydroxyapatite crystal, which is formed by the continuous reaction of calcium ions and phosphate ions. The intermediate product ACP reacts to become HAP. When a polypeptide is present, a complex of cp20k-p3 / HAP is formed). The solution containing cp20k-p3 / ACP or cp20k-p3 / HAP is a demineralized dentin remineralizing mouthwash based on cp20k-p3 biomimetic polypeptide.

[0021] This invention also relates to the application of the above-mentioned mouthwash based on cp20k-p3 biomimetic polypeptide for remineralizing demineralized dentin. Specifically, the mouthwash is combined with demineralized dentin to carry out a remineralization reaction for remineralizing demineralized dentin. The mineralizing mouthwash containing cp20k-p3 has a stronger mineralization ability and can form a remineralization layer on or inside the demineralized dentin. It can seal dentinal tubules by forming hydroxyapatite crystals and restore the mechanical strength of demineralized dentin.

[0022] In mouthwash containing cp20k-p3, cp20k-p3 / ACP or cp20k-p3 / HAP mineralization products are formed, which can adhere to the surface of demineralized dentin.

[0023] Specifically, the application of the aforementioned mouthwash based on cp20k-p3 biomimetic peptides for demineralizing dentin and remineralizing includes the following steps: (1) Preparation of simulated body fluid m-SBF: 137.5 mM NaCl, 3.0 mM KCl, 1.0 mM K2HPO4, 1.5 mM MgCl2, 2.5 mM CaCl2, 0.5 mM Na2SO4, 10.0 mM HEPES, 0.77 mM NaF; (2) When cp20k-p3 is mixed with the simulated body fluid, cp20k-p3 in the mineralized solution can form cp20k-p3 / ACP at 15 min and cp20k-p3 / HAP on the sixth day.

[0024] Furthermore, mouthwashes containing cp20k-p3 / ACP or cp20k-p3 / HAP were prepared using cp20k-p3, and reacted with demineralized dentin. The demineralized dentin was then immersed in the mouthwash containing cp20k-p3 / ACP or cp20k-p3 / HAP to study the remineralization of the demineralized dentin surface by the composite mouthwash.

[0025] Furthermore, the cp20k-p3 mentioned in step (2) was prepared according to the report in CN202510359951.6.

[0026] Furthermore, mouthwashes containing cp20k-p3-FITC / ACP or cp20k-p3-FITC / HAP were prepared using FITC-labeled cp20k-p3-FITC. Demineralized dentin was then immersed in the mouthwashes for 5-15 minutes to study the selective adhesion of cp20k-p3 / ACP or cp20k-P3 / HAP to demineralized dentin.

[0027] Furthermore, the demineralized dentin remineralizing mouthwash based on cp20k-p3 biomimetic peptides works at room temperature or 37°C and within a pH range of 6-8.

[0028] Compared with the prior art, the present invention has the following beneficial effects: 1. The demineralized dentin remineralization mouthwash based on cp20k-p3 biomimetic polypeptide described in this invention, wherein the biomimetic polypeptide cp20k-p3 is used to induce demineralized dentin remineralization and can form cp20k-p3 / ACP and cp20k-p3 / HAP in the mineralization solution.

[0029] 2. The demineralized dentin remineralization mouthwash based on cp20k-p3 biomimetic peptide described in this invention, after incubation with mouthwash containing cp20k-p3, yields a mineralized product with faster crystallization speed and higher crystallinity; cp20k-p3 / ACP and cp20k-p3 / HAP can selectively adhere to the surface of demineralized dentin.

[0030] 3. The demineralizing dentin remineralizing mouthwash based on cp20k-p3 biomimetic peptide described in this invention contains 0.3 mg / mL cp20k-p3 and 3.3 mM Ca. 2+ The mouthwash reacts with demineralized dentin, forming a stable, adhesive mineralized layer within 45 minutes, effectively sealing dentinal tubules. The thickness of this mineralized layer is approximately 7.5 micrometers.

[0031] 4. The demineralized dentin remineralization mouthwash based on cp20k-p3 biomimetic peptides described in this invention, cp20k-p3 / ACP and cp20k-p3 / HAP can quickly form a firm remineralization layer on the surface of demineralized dentin, and can form ACP or HAP with selective adhesion ability, which has new and great application prospects in demineralized dentin repair. Attached Figure Description

[0032] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the embodiments of the invention to explain the invention and do not constitute a limitation thereof.

[0033] Figure 1 The figures show the results of alizarin red staining of mineral products in mouthwash containing cp20k-p3 in Example 1 of the present invention and mouthwash in Comparative Example 1 after incubation for 45 min and after standing for 1 min during incubation. A1 shows the appearance of alizarin red staining of mineral products remaining in a 24-well plate after incubation with mouthwash containing cp20k-p3 for 45 min; A2 shows the results of alizarin red staining of mouthwash containing cp20k-p3 after incubation (at 22.5 min) in a 24-well plate. A3 shows the appearance of alizarin red staining of mineral products in a 24-well plate after standing for 1 min in the well plate; A4 shows the appearance of alizarin red staining of mineral products in the control mouthwash after incubation for 45 min; B shows the appearance of alizarin red staining of mineral products in the control mouthwash after it was removed during incubation (at 22.5 min) and left to stand in a 24-well plate for 1 min; B shows the quantitative results of mineral products in mouthwash containing or without cp20k-p3 after incubation for 45 min, or during incubation. Figure 2The images shown are of fluorescently labeled FITC-tagged cp20k-p3 / ACP and cp20k-p3 / HAP prepared in this embodiment of the invention, and their adhesion fluorescence signals to the surface of demineralized dentin. A1 is an image of the demineralized tooth treated with FITC in m-SBF mineralization solution on the second day; A2 is an image of the demineralized tooth treated with cp20k-p3-FITC / ACP in m-SBF mineralization solution on the second day; A3 is an image of the demineralized dentin transversely treated with FITC in m-SBF mineralization solution on the second day; A4 is an image of the m-SBF mineralization solution on the second day. Image B1: Image of demineralized dentin after treatment with cp20k-p3-FITC / ACP in m-SBF mineralized solution on day 6; Image B2: Image of demineralized dentin after treatment with FITC in m-SBF mineralized solution on day 6; Image B3: Image of demineralized dentin after treatment with FITC in m-SBF mineralized solution on day 6; Image B4: Image of demineralized dentin after treatment with cp20k-p3-FITC / HAP in m-SBF mineralized solution on day 6. Figure 3 These are scanning electron microscope images of the surfaces of acid-etched dentin sections and demineralized tooth sections after incubation with mouthwash containing cp20k-p3, as shown in Example 3 of this invention. Detailed Implementation

[0034] The technical solution of the present invention will be clearly and comprehensively described below with reference to the embodiments and accompanying drawings. The embodiments described below are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0035] Example 1: A mouthwash for demineralizing dentin and remineralizing based on cp20k-p3 biomimetic peptides includes the following steps: cp20k-p3-induced mineralization in vitro, and an experiment on the alizarin red staining of the mineralization products. (1) Mix cp20k-p3 with phosphate-buffered saline (PBS) buffer to make the final CCP concentration 0.3 mg / mL, and let stand for 15 min; (2) Add 200 mM CaCl2 stock solution and 100 mM K2HPO4 stock solution to cp20k-p3 and PBS solution so that the final concentration of the mixed mouthwash is 3.3 mM CaCl2 and 1.6 mM K2HPO4. (3) Divide the mixed solution into two portions. One portion is placed directly in a petri dish and left to stand for 45 minutes. The other portion is aspirated into a petri dish at 22.5 minutes and left to stand for 1 minute. (4) At the designed time, remove the mouthwash, add alizarin red staining solution, stain at room temperature for 15 minutes, remove the alizarin red staining solution, and take a picture. (5) Add 10% hexadecylpyridine chloride to the culture dish after staining until the mineralized product is completely dissolved, and measure the absorbance of the solution at 562 nm.

[0036] Results analysis: Figure 1 The images show the results of alizarin red staining of mineral products in mouthwash containing CP20k-p3 (Example 1) and mouthwash from Comparative Example 1, after incubation for 45 min and 1 min of resting. The samples were divided into experimental and control groups. Experimental group: biomimetic polypeptide CP20k-p3 + simulated body fluid = experimental group mouthwash; Control group: simulated body fluid = control group mouthwash. The images also show the appearance and quantitative results of alizarin red staining in mouthwash containing CP20k-p3 (Example 1) after 1 min or 45 min of resting in a petri dish. The study aimed to evaluate the effect of CP20k-p3 on promoting calcium phosphate mineralization using alizarin red staining. During the 45 min incubation period, the mouthwash containing CP20k-p3 produced a large amount of calcium phosphate mineralization products. Figure 1 A) Quantitative analysis of the generated mineralized products revealed that mouthwash containing added cp20k-p3 produced significantly more mineralized products.

[0037] Example 2: A mouthwash for demineralizing dentin based on a cp20k-p3 biomimetic peptide was developed. cp20k-p3 was added to simulated body fluid to obtain cp20k-p3 / ACP and cp20k-p3 / HAP. The adhesion of these two mineralization products to demineralized dentin was investigated, including the following steps: (1) Prepare simulated body fluid with the following formula: 137.5 mM NaCl, 3.0 mM KCl, 1.0 mM K2HPO4, 1.5 mM MgCl2, 2.5 mM CaCl2, 0.5 mM Na2SO4, 10.0 mM HEPES, and 0.77 mM NaF; (2) FITC-labeled cp20k-p3-FITC was added to the simulated body fluid to make the concentration of the peptide in the simulated body fluid 0.3 mg / mL; (3) Etch the dentin slices with 37% phosphoric acid for 30 seconds to demineralize the surface of the slices and obtain demineralized dentin slices; (4) The composite simulated body fluid was incubated at 37°C. On the second day of incubation, a large amount of cp20k-p3-FITC / ACP was formed in the mouthwash, and on the sixth day, a large amount of cp20k-p3-FITC / HAP was formed in the mouthwash. Demineralized tooth sections were added to the two mouthwashes respectively, and incubated at room temperature for 5 min. Fluorescence signals were captured using a laser confocal microscope.

[0038] Results analysis: Figure 2 The fluorescently labeled FITC-labeled cp20k-p3 / ACP and cp20k-p3 / HAP prepared in Example 2 are shown in the adhesion fluorescence signal map of the demineralized dentin surface. Figure 2 In the diagram, A2 shows the adhesion of cp20k-p3-FITC / ACP to demineralized dentinal tubules, indicating that cp20k-p3-FITC / ACP can selectively adhere to demineralized dentinal material; A4 shows that cp20k-p3-FITC / ACP mainly adheres to intertubular dentinal material; B2 shows the adhesion of cp20k-p3-FITC / HAP to demineralized dentinal tubules, indicating that cp20k-p3-FITC / HAP can also selectively adhere to demineralized dentinal material; B4 shows that cp20k-p3-FITC / HAP mainly adheres to intertubular dentinal material.

[0039] Example 3: A mouthwash for remineralizing demineralized dentin based on cp20k-p3 biomimetic peptides involves immersing demineralized dentin slices in simulated body fluid containing cp20k-p3 to generate a mineralization layer to seal demineralized dentinal tubules. The method includes the following steps: (1) Prepare mouthwash containing cp20k-p3 according to steps (1) and (2) of Example 1; (2) Immerse the demineralized teeth in mouthwash containing cp20k-p3 and incubate at room temperature for 45 minutes; (3) Remove the soaked teeth, dry them, and take scanning electron microscope images to observe the surface morphology of the dentinal tubules; (4) The treated tooth was broken in the middle, and the cross-sectional morphology and the thickness of the remineralized layer were photographed by scanning electron microscopy.

[0040] Results analysis: Figure 3 These are scanning electron microscope images of the surfaces of acid-etched dentin sections and demineralized tooth sections after incubation with mouthwash containing cp20k-p3, as shown in Example 3. Figure 3In the images, B represents the demineralized dentin surface after treatment with mouthwash containing cp20k-p3. B1 and B2 show that the dentinal tubules are almost completely sealed, and B3 shows that the mineralized layer covering the dentinal tubules is approximately 7.5 micrometers thick. A1 and A2 are scanning electron microscope images of acid-etched dentin surfaces after soaking in mouthwash for 45 minutes in the control group. A3 is a scanning electron microscope image of a cross-section of acid-etched dentin after soaking in mouthwash for 45 minutes in the control group. B1 and B2 are scanning electron microscope images of acid-etched dentin surfaces after soaking in mouthwash for 45 minutes in the cp20k-p3 group. B3 is a scanning electron microscope image of a cross-section of acid-etched dentin after soaking in mouthwash for 45 minutes in the cp20k-p3 group.

[0041] Comparative Example 1: The difference between Comparative Example 1 and Example 1 is that no biomimetic polypeptide cp20k-p3 was added; other experimental conditions were the same as in Example 1.

[0042] pass Figure 1 As can be seen from the alizarin red staining appearance diagram, it was found that both the mouthwash taken out midway and the mouthwash incubated for 45 minutes contained only a very small amount of red when cp20k-p3 was not present. Figure 1 A); Quantitative analysis showed that mouthwash without cp20k-p3 had significantly fewer mineral products than mouthwash containing cp20k-p3 ( Figure 1 B).

[0043] The results showed that cp20k-p3 could significantly promote the formation of calcium phosphate crystals.

[0044] Comparative Example 2: The difference between Comparative Example 2 and Example 2 is that only the FITC test was used instead of cp20k-p3 labeled with FITC; other test conditions were the same as in Example 2. pass Figure 2 As can be seen, A1 and 2B1 indicate that FITC has no adhesion to either demineralized dentin or demineralized enamel, while A3 and B3 show that FITC has no adhesion to transversely cut demineralized dentin.

[0045] The results show that: FITC does not have the ability to adhere to the surface of demineralized dentin.

[0046] Comparative Example 3: The difference between Comparative Example 3 and Example 3 is that demineralized teeth were treated with a mouthwash that did not contain CP20k-p3. Everything else was the same as in Example 3. pass Figure 3As can be seen from B1 and B2, teeth treated with mouthwash that does not contain cp20k-p3 also formed a mineralized layer. However, this mineralized layer is loose and easily falls off, and cannot form a complete mineralized layer on the surface of demineralized dentin, thus fully exposing the dentinal tubules. B3 shows that when the dentinal tubules treated in this way are broken under physical force, the mineralized products randomly fall off and disappear.

[0047] This indicates that mouthwashes without CP20k-P3 cannot form a firmly adhering mineralized layer on the surface of demineralized teeth.

[0048] Results and Discussion: Example 1 compared mouthwash containing cp20k-p3 with mouthwash without cp20k-p3 in terms of the amount of mineralization products during the mineralization process and after 45 minutes of mineralization. The results showed that the amount of mineralization products generated by the mouthwash with added cp20k-p3 was significantly greater than that of the mouthwash without added cp20k-p3.

[0049] Example 2 and Comparative Example 2 verified the adhesion of the mineralization intermediates cp20k-p3-FITC / ACP and cp20k-p3-FITC / HAP generated during the mineralization process to demineralized dentin. This indicates that the possibility of FITC adhering to dentin is excluded, and cp20k-p3-FITC / ACP and cp20k-p3-FITC / HAP can adhere to demineralized dentin.

[0050] Example 3 and Comparative Example 3 verified that mouthwash containing cp20k-p3 can form a firmly adhered mineralized layer on the surface of demineralized dentinal tubules, the mineralized layer being approximately 7.5 micrometers thick, and the mineralized layer effectively seals the demineralized dentinal tubules.

[0051] The above experiments show that the mineralized solution with added cp20k-p3 has the ability to become a mouthwash. The mineralization intermediates cp20k-p3 / ACP and cp20k-p3 / HAP generated by cp20k-p3 in the mouthwash can selectively adhere to the surface of demineralized dentin and generate mineralized products with good binding ability on the surface. These mineralized products can seal the orifices of dentinal tubules.

[0052] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A mouthwash for demineralizing dentin and remineralizing based on cp20k-p3 biomimetic peptides, characterized in that, It contains biomimetic polypeptide cp20k-p3 and mineralization solution; The biomimetic polypeptide cp20k-p3 is derived from a 20K-derived polypeptide of white barnacle protein, and the amino acid sequence of the polypeptide is: CNQKHPCWRRHGKKHGLHRKFHGNACNC; Mineralizing solution contains Ca 2+ Or PO4 3- ; When the mineralizing solution is mixed with the biomimetic peptide cp20k-p3, a demineralized dentin remineralizing mouthwash based on the cp20k-p3 biomimetic peptide is obtained.

2. The mouthwash for demineralizing dentin and remineralizing based on cp20k-p3 biomimetic peptides according to claim 1, characterized in that, The biomimetic polypeptide cp20k-p3 was synthesized using a chemical synthesis method.

3. The mouthwash for demineralizing dentin and remineralizing according to claim 1, characterized in that, The concentration of the biomimetic polypeptide cp20k-p3 is 0.01 mg / mL to 1 mg / mL.

4. The mouthwash for demineralizing dentin and remineralizing according to claim 1, characterized in that, The mineralizing solution is mixed with the biomimetic polypeptide cp20k-p3 by sequentially reacting the biomimetic polypeptide cp20k-p3 with a substance containing PO4. 3- solutions containing Ca 2+ The solutions are mixed; in this mixed solution, the reaction forms cp20k-p3 / ACP or cp20k-P3 / HAP.

5. The application of a mouthwash for demineralizing dentin and remineralizing based on cp20k-p3 biomimetic peptides, characterized in that, The application of the demineralized dentin remineralizing mouthwash based on cp20k-p3 biomimetic polypeptide according to any one of claims 1-4, wherein the mouthwash is combined with demineralized dentin to carry out a remineralization reaction, and is used for demineralized dentin remineralization. The mineralizing mouthwash containing cp20k-p3 can form a remineralization layer on or inside the demineralized dentin, and seal the dentinal tubules by forming hydroxyapatite crystals and restore the mechanical strength of the demineralized dentin.

6. The application of the mouthwash based on cp20k-p3 biomimetic peptides for demineralizing dentin and remineralizing according to claim 5, characterized in that, Includes the following steps: (1) Preparation of simulated body fluid m-SBF: 137.5 mM NaCl, 3.0 mM KCl, 1.0 mM K2HPO4, 1.5 mM MgCl2, 2.5 mM CaCl2, 0.5 mM Na2SO4, 10.0 mM HEPES, 0.77 mM NaF; (2) When cp20k-p3 is mixed with simulated body fluid, cp20k-p3 in the mineralized solution forms cp20k-p3 / ACP at 15 min and cp20k-p3 / HAP on the sixth day.

7. The application of the mouthwash based on cp20k-p3 biomimetic polypeptide for demineralizing dentin and remineralizing according to claim 6, characterized in that, Mouthwashes containing cp20k-p3 / ACP or cp20k-p3 / HAP were prepared using cp20k-p3. These mouthwashes were then reacted with demineralized dentin. The demineralized dentin was then immersed in the mouthwashes containing cp20k-p3 / ACP or cp20k-p3 / HAP to study the remineralization of the demineralized dentin surface by the composite mouthwashes.

8. The application of the mouthwash based on cp20k-p3 biomimetic polypeptide for demineralizing dentin and remineralizing according to claim 6, characterized in that, Mouthwash containing cp20k-p3-FITC / ACP or cp20k-p3-FITC / HAP was prepared using FITC-labeled cp20k-p3-FITC. Demineralized dentin was then immersed in the mouthwash for 5-15 minutes to study the selective adhesion of cp20k-p3 / ACP or cp20k-P3 / HAP to demineralized dentin.

9. The application of the mouthwash based on cp20k-p3 biomimetic polypeptide for demineralizing dentin and remineralizing according to claim 6, characterized in that, The demineralized dentin remineralizing mouthwash based on cp20k-p3 biomimetic peptides works at room temperature or 37°C and within a pH range of 6-8.

Images