Mineralization repair type oral cavity and throat nursing material for middle-aged and elderly people and preparation method

Abstract

The invention discloses a mineralization repair type oral cavity and throat nursing material for middle-aged and elderly people and a preparation method thereof. The nursing material consists of a core mineralization precursor, a function relieving component and an auxiliary synergistic component, wherein the core mineralization precursor is a biocompatible mineralization matrix of modified hydroxyapatite loaded carbon nitride quantum dots, and is used for realizing mucous membrane mineralization repair; the functional soothing component is a mixture of dihydromyricetin, a licorice extract and a menthol-beta-cyclodextrin inclusion compound, and has mild soothing and anti-inflammatory effects; the auxiliary synergistic component is a mixture of bovine serum albumin and a gamma-polyglutamic acid-g-methoxypolyethylene glycol derivative, and a mineralized precursor is stabilized and mucous membrane adhesion is enhanced by constructing a protein nanocage structure. By optimizing the process, the production efficiency and stability are improved, the biological safety is enhanced, the nursing requirements of middle-aged and elderly oral cavity and throat sensitive mucous membranes are better met, and the oral cavity and throat sensitive mucous membrane repairing gel is suitable for oral cavity mucous membrane sensitive repairing and chronic pharyngitis auxiliary nursing.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of oral and throat care materials, in particular to a mineralization repair type oral and throat care material for the elderly and a preparation method of the material, which is especially suitable for preparing oral mucosa sensitive repair products and chronic pharyngitis auxiliary care mineralization materials for the elderly. BACKGROUND

[0002] With the increase of age, the physiological functions of the oral and throat of the elderly gradually decline, the thickness of the oral mucosa becomes thinner, the barrier function is weakened, and problems such as mucosa sensitivity, dryness and damage are prone to occur; at the same time, the resistance of the mucosa of the throat decreases, and the incidence of chronic pharyngitis significantly increases, which is manifested as symptoms such as itching, foreign body sensation and irritating cough of the throat, which seriously affects the quality of life.

[0003] At present, existing oral care products (such as toothpaste and mouthwash) mainly focus on cleaning and anti-cavity functions, and lack mineralization repair function for sensitive mucosa of the elderly; throat care products (such as throat lozenges and sprays) mainly focus on temporary relief, and cannot achieve long-term repair of mucosa damage, and often contain irritating additives, which are not suitable for sensitive mucosa of the elderly.

[0004] Hydroxyapatite (HAp) is the main inorganic component of human bones and teeth, and has good biocompatibility and mineralization repair ability, but its nanoparticles are easy to agglomerate, have poor stability in the oral and throat environment, and have insufficient adhesion to the mucosa, which limits the repair effect. Carbon nitride quantum dots (CNQDs) have excellent biological safety and fluorescence performance, and can be used as a bioactive carrier, but lack mineralization function when used alone. In addition, the functional components of existing care materials are mostly single systems, which are difficult to meet the comprehensive needs of mineralization repair, soothing anti-inflammatory and long-term stability, and the preparation process has problems such as complicated steps, low production efficiency and insufficient quality control precision.

[0005] Therefore, it is a technical problem to be solved to develop an oral and throat care material for the elderly, which has good biocompatibility, mineralization repair, mild soothing and system stability, and has a controllable and efficient preparation process. SUMMARY

[0006] In view of the deficiencies of the prior art, the present application aims to provide a mineralization repair type oral and throat care material for the elderly and a preparation method, which realizes mineralization repair, mild soothing and long-term stability of the oral and throat mucosa by optimizing the component design and preparation process, improves the production efficiency and product safety, and meets the special care needs of the elderly.

[0007] To achieve the above-mentioned purpose, the present application provides the following technical solutions:

[0008] The application discloses a mineralization repair type oral and throat care material for the middle-aged and the old, which comprises the following components in percentage by mass: a core mineralization precursor 85-94%, a functional soothing component 3-7% and an auxiliary synergistic component 2-10%.

[0009] The core mineralization precursor is a modified hydroxyapatite loaded with carbon nitride quantum dots.

[0010] The functional soothing component is a mixture of dihydro-myricetin, liquorice extract and menthol-beta-cyclodextrin inclusion compound.

[0011] The auxiliary synergistic component is a mixture of bovine serum albumin and a gamma-polyglutamic acid-g-polyethylene glycol monomethyl ether (gamma-PGA-g-mPEG) derivative.

[0012] The auxiliary synergistic component forms a protein nanocage structure through the synergistic effect of bovine serum albumin and the gamma-PGA-g-mPEG derivative, realizes the stable effect on the mineralization precursor and enhances the adhesion of the material to the mucosa.

[0013] Preferably, the mass ratio of the modified hydroxyapatite to the carbon nitride quantum dots in the modified hydroxyapatite loaded with carbon nitride quantum dots is 3:1.

[0014] Preferably, the mass ratio of dihydro-myricetin, liquorice extract and menthol-beta-cyclodextrin inclusion compound is 3:2:(1-1.8).

[0015] Preferably, the mass ratio of bovine serum albumin to the gamma-polyglutamic acid-g-polyethylene glycol monomethyl ether derivative is (1.5-2):1.

[0016] Preferably, the number average molecular weight of the gamma-polyglutamic acid-g-polyethylene glycol monomethyl ether derivative is 8000-15000 Da, the molecular weight of the polyethylene glycol monomethyl ether in the derivative is 2000-5000 Da, and the grafting rate is 30%-40%.

[0017] Preferably, the mass ratio of menthol to beta-cyclodextrin in the menthol-beta-cyclodextrin inclusion compound is 1:4.

[0018] Preferably, the preparation steps of the menthol-beta-cyclodextrin inclusion compound are as follows: mixing menthol and beta-cyclodextrin, stirring in water at 60 DEG C for 1 hour, and obtaining the menthol-beta-cyclodextrin inclusion compound through freeze drying.

[0019] Preferably, the product form of the mineralization repair type oral and throat care material for the middle-aged and the old is a dissolvable mineralization film or a slow-release mineralization gel.

[0020] Preferably, in the dissolvable mineralization film, the auxiliary synergistic component accounts for 2%-4% of the mass percentage of the care material.

[0021] Preferably, the auxiliary synergistic component accounts for 6%-10% of the mass percentage of the care material in the slow-release mineralized gel.

[0022] The care material of the present application is composed of a core mineralized precursor, a functional soothing component, and an auxiliary synergistic component, and the composition and role of each component are as follows:

[0023] (1) Core mineralized precursor: a biocompatible mineralized matrix of modified hydroxyapatite loaded with carbon nitride quantum dots, which is the core component for realizing mucosa mineralization repair. Through the microfluidic homogenization and ultrasonic-stirring composite dispersion process, the agglomeration problem of hydroxyapatite nanoparticles is solved; after cross-linking modification of pyrimidine derivatives, coordination active sites are formed on the surface, thereby improving the interaction with the mucosa; carbon nitride quantum dots are loaded on the modified hydroxyapatite at a mass ratio of 3:1, which not only enhances the biological activity of the mineralized matrix, but also uses its fluorescence performance to monitor the action trajectory of the material on the mucosa surface, thereby providing support for the evaluation of the repair effect.

[0024] (2) Functional soothing component: a compound of dihydromyricetin, licorice extract, and menthol-β-cyclodextrin inclusion complex at a mass ratio of 3:2:(1-1.8). The β-cyclodextrin inclusion can reduce the irritability of menthol and prolong the duration of the cooling sensation; dihydromyricetin plays an anti-inflammatory and antioxidant role to reduce the inflammatory response of the mucosa; licorice extract soothes the dryness and itching of the throat and inhibits irritating cough; menthol promotes the mucosal penetration of other functional components, and the three components work together to achieve the effects of "gentle soothing-anti-inflammatory-penetration promotion".

[0025] (3) Auxiliary synergistic component: a compound of bovine serum albumin and γ-polyglutamic acid-g-polyethylene glycol monomethyl ether derivative (γ-PGA-g-mPEG) at a mass ratio of (1.5-2):1 (the auxiliary synergistic component accounts for 2%-10% of the total mass of the care material). The γ-PGA-g-mPEG derivative has improved dispersibility after pretreatment, and forms a protein nanocage structure with bovine serum albumin, which on the one hand stabilizes the core mineralized precursor to avoid agglomeration or inactivation, and on the other hand enhances the adhesion of the material to the mucosa, wherein the soluble mineralized film is adapted to a low proportion (2%-4%) to ensure the dissolution rate, and the slow-release mineralized gel is adapted to a high proportion (6%-10%) to achieve long-term adhesion.

[0026] On the other hand, the present application also provides a method for preparing the above-mentioned mineralized repair type oral and throat care material for the middle-aged and elderly, which comprises the following steps:

[0027] (1) Preparation of core mineralized precursor

[0028] S1, preparation of modified hydroxyapatite:

[0029] a. 2,5-diamino-4,6-dihydroxypyrimidine and polyethylene glycol diglycidyl ether are dissolved in an ethanol / water mixed solvent at a molar ratio of 1:1.05 at 60-70°C for 6-8 hours until the epoxy groups are completely reacted;

[0030] b. The reaction solution is precipitated with diethyl ether, centrifuged, washed, and freeze-dried to obtain the PEGDE cross-linked pyrimidine derivative;

[0031] c. The derivative is dissolved in an ethanol / water mixed solvent and stirred to dissolve at 60-80°C;

[0032] d. 4-hydroxybenzaldehyde solution is added to the solution at 0-4°C, and the pH is adjusted to 4-5 with dilute acetic acid. After 2-4 hours of reaction, it is neutralized to pH 6.5-7.0 with triethylamine;

[0033] e. Hydroxyapatite nanoparticles are dispersed in an ethanol / water mixed solvent, treated by microfluidization and ultrasonic stirring combination, and then added to the reaction system of step d. Continue ultrasonic stirring reaction;

[0034] f. After the reaction is completed, the product is washed with ethanol / water mixed solvent containing Tween-80 and ultrapure water, and then dried to obtain modified hydroxyapatite;

[0035] S2, preparation of modified hydroxyapatite loaded with carbon nitride quantum dots:

[0036] A, preparation of modified HAp dispersion: modified hydroxyapatite is dispersed in water and treated by ultrasonic stirring combination to obtain a stable dispersion of 1 mg / mL, particle size 50-200 nm;

[0037] B, preparation of CNQDs solution: water-soluble carbon nitride quantum dots with a fluorescence quantum yield of ≥50% are prepared into a 0.5 mg / mL aqueous solution;

[0038] C, composite loading reaction: carbon nitride quantum dot solution is slowly added to the hydroxyapatite dispersion at a modified HAp to CNQDs mass ratio of 3:1 at 25±2°C, pH 6.5-7.5, and light shielding conditions. Stirring with magnetic stirring combined with intermittent ultrasonic reaction for 3 hours;

[0039] D, post-treatment and purification: after the reaction is completed, centrifugation, water washing and freeze-drying are carried out to obtain the modified hydroxyapatite loaded with carbon nitride quantum dots composite material;

[0040] (2) ingredient compounding

[0041] ① Preparation of functional soothing ingredients: mix dihydromyricetin, licorice extract, and menthol-β-cyclodextrin inclusion complex;

[0042] ②Auxiliary synergistic component pretreatment: dissolve γ-polyglutamic acid-g-polyethylene glycol monomethyl ether derivative in ultrapure water, and treat to transparency by ultrasonic treatment at 200 W for 10 min + low-speed stirring at 300 rpm for 5 min, then mix with the bovine serum albumin solution to form a premix;

[0043] ③Total material mixing: mix the core mineralization precursor, functional soothing component, and auxiliary synergistic component premix according to the preset ratio, uniformly stir at room temperature, and form a mixture;

[0044] (3) Shape forming

[0045] The mixture is made into a dissolvable mineralized film or a slow-release mineralized gel.

[0046] The preparation steps of the dissolvable mineralized film are as follows: add ammonium bicarbonate as a pore former to the mixture, uniformly stir, and then place in a vacuum drying oven for reaction for 3-4 h to obtain a dissolvable mineralized film with a porous structure.

[0047] The preparation steps of the slow-release mineralized gel are as follows: perform two-stage homogenization treatment on the mixture, adjust the system viscosity to 500-800 mPa·s by adding ultrapure water or xanthan gum, and obtain a slow-release mineralized gel.

[0048] Preferably, the preparation steps of the core mineralization precursor are as follows:

[0049] S1, modified hydroxyapatite preparation

[0050] a. Dissolve 2,5-diamino-4,6-dihydroxypyrimidine and polyethylene glycol diglycidyl ether (PEGDE, molecular weight 400) in a molar ratio of 1:1.05 in an ethanol / water mixed solvent (volume ratio 3:2), uniformly stir at room temperature, then heat to 65±5℃ and stir at constant temperature for 6-8 hours, monitor the characteristic peak of epoxy group every 1 hour by FTIR until the characteristic peak completely disappears.

[0051] b. Add diethyl ether to the reaction solution, centrifuge at 12000 rpm for 15 minutes to collect the precipitate, repeat the diethyl ether washing for 2 times, and freeze-dry to obtain the PEGDE cross-linked pyrimidine derivative.

[0052] c. Dissolve the PEGDE cross-linked pyrimidine derivative in an ethanol / water mixed solvent (volume ratio 3:2) and stir at 60~80℃ until completely dissolved.

[0053] d. Add a 4-hydroxybenzaldehyde solution dissolved in the same proportion of ethanol / water mixed solvent, slowly drop 1 mol / L dilute acetic acid under ice bath conditions at 0~4℃, maintain the system pH at 4-5, and stir for 2~4 hours; after the reaction is completed, neutralize with triethylamine to pH 6.5-7.0.

[0054] e. Hydroxyapatite nanoparticles were added to an ethanol / water mixed solvent (volume ratio 3:2), pretreated once by 15-20 MPa microfluidic homogenization, and then dispersed by a combination of "250 W low-temperature ultrasonic for 20 min + 800 rpm continuous mechanical stirring for 15 min". After DLS detected that the particle size distribution was 50-200 nm (no > 300 nm agglomerates), it was transferred to the reaction system of step d and continuously ultrasonically stirred.

[0055] f. After the reaction, a two-step composite washing method was used: first, 2 times of washing with ethanol / water mixed solvent (volume ratio 1:1) + 0.1% Tween-80, 12000 rpm x 12 min / time; second, 2 times of washing with ultrapure water + 200 W ultrasonic assistance for 5 min / time, 12000 rpm x 10 min / time; After collecting the solid, it was pre-dried at 30°C for 2 hours, and then freeze-dried for 24 hours to obtain modified hydroxyapatite.

[0056] g. Safety verification: gas chromatography detected solvent residue ≤35 μg / g; high performance liquid chromatography detected no PEGDE residue (detection limit ≤0.05 μg / g); 10 mg / mL water suspension pH was 6.5-7.5; CCK-8 method detected L929 cell survival rate ≥80%; subcutaneous injection of 2 mL / kg (experimental concentration) in rats, no toxicity reaction was observed for 7 days.

[0057] S2, modified hydroxyapatite loaded carbon nitride quantum dots

[0058] A, preparation of modified HAp dispersion liquid: take 100 mg of modified HAp powder prepared in step S1, add 100 mL of ultrapure water, treat by 250 W ultrasonic for 20 min + 800 rpm mechanical stirring for 10 min, prepare 1 mg / mL of uniform dispersion (DLS detects particle size 50-200 nm, no agglomeration).

[0059] B, preparation of CNQDs solution: select water-soluble carbon nitride quantum dots (CNQDs, fluorescence quantum yield ≥50%, surface containing -OH / -NH2 groups), prepare 0.5 mg / mL transparent solution with ultrapure water, calibrate the concentration deviation within ±5% by ultraviolet-visible spectrophotometer, and store in dark place.

[0060] C. Composite loading reaction: Mix the CNQDs solution slowly into the modified HAp dispersion liquid according to the ratio of "modified HAp: CNQDs = 3:1 (mass ratio)", and treat it in the dark at 25±2℃. Use the combination of "600 rpm magnetic stirring + 150 W intermittent ultrasound 5 min / hour" to react for 3 hours. Maintain the system pH 6.5-7.5 during the reaction.

[0061] D. Post-treatment and purification: After the reaction is completed, centrifuge the mixed solution at 12000 rpm for 15 minutes, collect the precipitate, wash the precipitate twice with ultrapure water (12000 rpm x 10 min / time), remove the unbound free CNQDs, freeze-dry for 24 hours, and obtain the core mineralization precursor.

[0062] Preferably, the preparation steps of the dissolvable mineralized membrane are as follows: add 0.5% (mass percentage) ammonium bicarbonate as a pore former to the mixture, stir uniformly, and then place it in a vacuum drying oven for crosslinking at -0.09 MPa vacuum degree and 35℃ for 4 hours to obtain a porous structure dissolvable mineralized membrane with a pore size of 3-8 μm.

[0063] Preferably, the preparation steps of the slow-release mineralized gel are as follows: perform two-stage homogenization on the mixture, first at 20 MPa for 5 minutes, and then at 30 MPa for 5 minutes, with online viscosity monitoring (detect every 2 minutes) during the homogenization process; if the viscosity is lower than 500 mPa·s, add 0.1%-0.2% xanthan gum (pre-dissolved in a small amount of ultrapure water) and stir for 5 minutes; if the viscosity is higher than 800 mPa·s, add 1%-2% ultrapure water; finally adjust the system viscosity to 500-800 mPa·s (25℃) to obtain the slow-release mineralized gel.

[0064] The above preparation method mainly involves the following key process optimization design:

[0065] (1) Modification of hydroxyapatite preparation optimization: Use "micro-jet homogenization pretreatment + low-temperature ultrasonic stirring composite dispersion" instead of traditional dispersion process, shorten the dispersion time to 35 minutes, and control the particle size distribution to 50-200 nm (no >300 nm agglomerates); innovate "two-step composite washing method", combine surfactants with ultrasonic assistance, reduce washing steps and solvent consumption, reduce solvent residue to ≤35 μg / g, and improve production efficiency by 40%.

[0066] (2) Carbon nitride quantum dot loading optimization: Introduce CNQDs solution concentration calibration step to ensure accurate loading ratio; use "constant temperature control + intermittent ultrasonic auxiliary stirring" to improve the loading efficiency to 95%, avoid CNQDs fluorescence quenching, and ensure mineralization activity and monitoring performance.

[0067] (3) Component compounding optimization: through high-precision weighing and HPLC content verification, ensure that the functional component allocation ratio deviation is within ±2%; auxiliary synergistic component premixing treatment, improve the dispersion uniformity, enhance the stability of the nanocage structure, and prolong the mucosal adhesion time by 50% (up to 6-8 hours).

[0068] (4) Morphology forming optimization: soluble mineralized membrane is added with ammonium bicarbonate porogen, combined with high vacuum drying, to form a uniform porous structure (pore size 3-8 μm), and the dissolution rate is stable; the slow-release mineralized gel adopts two-stage homogenization and online viscosity adjustment, and the viscosity deviation is controlled within ±20 mPa·s, and the unqualified rate is reduced from 8% to 2%.

[0069] The key parameters and safety guarantees of the nursing material product prepared by the application are as follows:

[0070] (1) Product form and parameters: the soluble mineralized membrane is a porous structure with a pore size of 3-8 μm, and is slowly dissolved in saliva for 6-8 minutes; the slow-release mineralized gel has a viscosity of 500-800 mPa·s at 25℃; the pH response range of the two forms of products is 6.5-8.0, which matches the physiological environment of the oral cavity and throat, and is mild and non-irritating.

[0071] (2) Safety verification: in addition to the conventional solvent residue, cell survival rate, and animal toxicity test, a 30-day long-term oral mucosa irritation test and pathological section analysis are supplemented, and new heavy metal (Pb, Cd, As≤0.1 mg / kg) and bovine serum albumin residue (≤1 μg / g) detection are added, to comprehensively ensure the safety of long-term use by the middle-aged and the elderly.

[0072] The application scenarios of the nursing material product prepared by the application are as follows:

[0073] 1. Oral mucosa sensitive repair product for the middle-aged and the elderly: the core mineralized precursor repairs the damaged mucosal barrier, the auxiliary synergistic component enhances the adhesion, the functional soothing component relieves the sensitivity and discomfort, and it is suitable for oral mucosa dryness, sensitivity, and slight damage.

[0074] 2. Chronic pharyngitis auxiliary care mineralized material: gently soothes the throat itching and foreign body sensation, mineralized repair enhances the mucosal resistance, and auxiliary improves the symptoms of chronic pharyngitis, the components are mild and non-irritating, and it is suitable for long-term use by the middle-aged and the elderly.

[0075] Compared with the prior art, the application has the following beneficial effects:

[0076] The mineralization-repairing oral and pharyngeal care material provided by this invention for middle-aged and elderly people achieves multiple beneficial effects through component design and process optimization: The core mineralization precursor is based on modified hydroxyapatite loaded with carbon nitride quantum dots, which not only solves the problem of easy aggregation of nanoparticles, but also enhances the interaction with the mucosa through coordination active sites, significantly improving the mineralization and repair capacity of the mucosal barrier, especially suitable for repairing dry, sensitive or slightly damaged oral mucosa; The functional soothing ingredient, through the synergistic compounding of dihydromyricetin, licorice extract and menthol-β-cyclodextrin inclusion complex, reduces irritation while prolonging the soothing effect, effectively relieving discomfort such as dry throat, itching, and foreign body sensation, and reducing mucosal inflammation through anti-inflammatory and antioxidant mechanisms; The auxiliary synergistic ingredient, through the compounding of bovine serum albumin and γ-polyglutamic acid-g-polyethylene glycol monomethyl ether derivative, constructs a stable nanostructure, which not only prevents the inactivation of the mineralization precursor, but also enhances the adhesion of the material to the mucosal surface, thereby achieving long-lasting sustained-release repair. Overall, this material combines targeted repair, gentle soothing, and long-lasting stability, fully meeting the safety, adaptability, and efficiency needs of middle-aged and elderly people for oral and throat care. Attached Figure Description

[0077] Figure 1 Transmission electron microscopy image of the core mineralization precursor.

[0078] Figure 2 Scanning electron microscope image of the core mineralization precursor.

[0079] Figure 3 FTIR spectra of modified hydroxyapatite and hydroxyapatite modified with supported carbon quantum dots (CNQDs). a: Modified hydroxyapatite, b: CNQD-supported hydroxyapatite.

[0080] Figure 4 This is a scanning electron microscope image of the soluble mineralized pattern.

[0081] Figure 5 This is the dissolution rate curve of the soluble mineralized film.

[0082] Figure 6 The effect of different concentrations of extracts from sustained-release mineralized gel on the expression of IL-10 and TNF-α in RAW264.7 cells, as described in Example 1.

[0083] Figure 7 Example 1: In vitro sustained-release characteristics (2 h-8 h) of the functional soothing component in the sustained-release mineralized gel. Detailed Implementation

[0084] The following embodiments are further illustrations of the present invention, but not limitations thereof.

[0085] The carbon nitride quantum dots (CNQDs) in the application are prepared by a bottom-up process. The preparation method of CNQDs can refer to the preparation method of CNQDs in “Preparation of Graphite Carbon Nitride Quantum Dot Assembly and Study on Its Photocatalytic Performance” (Fan Yu, Master's Thesis). The particle size of the obtained CNQDs is 5-10 nm, and the surface contains oxygen-containing groups (-COOH, -OH) and -NH2 groups, which can meet the requirements of water solubility, biocompatibility and fluorescence monitoring function, and adapt to the loading requirements of core mineralization precursors and the safety requirements of mucosa care scenes.

[0086] Example 1: Preparation of slow-release mineralization gel

[0087] 1. Preparation of core mineralization precursor

[0088] (1) Preparation of modified HAp

[0089] a. 2,5-diamino-4,6-dihydroxypyrimidine and polyethylene glycol diglycidyl ether (PEGDE, molecular weight 400) were dissolved in an ethanol / water mixed solvent (volume ratio 3:2) at a molar ratio of 1:1.05, stirred uniformly at room temperature, and then heated to 65°C for constant temperature stirring for 7 hours. The characteristic peak of the epoxy group was monitored by FTIR every 1 hour until the characteristic peak completely disappeared.

[0090] b. Ethyl ether was added to the reaction solution, and the precipitate was collected by centrifugation at 12000 rpm for 15 minutes. The ethyl ether washing was repeated twice, and the PEGDE cross-linked pyrimidine derivative was obtained by freeze-drying.

[0091] c. The PEGDE cross-linked pyrimidine derivative was dissolved in an ethanol / water mixed solvent (volume ratio 3:2) and stirred at 70°C until completely dissolved.

[0092] d. 4-Hydroxybenzaldehyde solution dissolved in the same proportion of ethanol / water mixed solvent was added, and 1 mol / L dilute acetic acid was slowly added under ice bath conditions at 0-4°C, the pH of the system was maintained at 4.5, and the reaction was stirred for 3 hours. After the reaction, the pH was neutralized to 6.8 with triethylamine.

[0093] e. The hydroxyapatite nanoparticles were added to the ethanol / water mixed solvent (volume ratio 3:2), pretreated once by 18 MPa microfluidic homogenization, and then dispersed by a combination of “250 W low-temperature ultrasonic 20 min + 800 rpm continuous mechanical stirring 15 min”. After the particle size distribution was detected by DLS to be 50-200 nm (no >300 nm agglomerates), it was transferred to the reaction system of step d and continuously ultrasonically stirred.

[0094] f. After the reaction, a two-step composite washing method was used: first, 2 times of washing with ethanol / water mixed solvent (volume ratio 1:1) + 0.1% Tween-80, 12000 rpm x 12 min / time; second, 2 times of washing with ultrapure water + 200 W ultrasonic assistance for 5 min / time, 12000 rpm x 10 min / time; after collecting the solid, 30°C vacuum pre-drying for 2 hours, and then freeze-drying for 24 hours, the modified hydroxyapatite was obtained.

[0095] g. Safety verification: gas chromatography detected solvent residue of 32 μg / g; high performance liquid chromatography detected no PEGDE residue (detection limit ≤ 0.05 μg / g); 10 mg / mL water suspension pH was 7.0; CCK-8 method detected L929 cell survival rate of 86%; subcutaneous injection of 2 mL / kg (experimental concentration) in rats, no toxicity reaction was observed for 7 days.

[0096] (2) CNQDs loading

[0097] a. Preparation of modified HAp dispersion: take 100 mg of modified HAp powder prepared in step (1), add 100 mL of ultrapure water, and treat with 250 W ultrasonic for 20 min + 800 rpm mechanical stirring for 10 min to prepare a uniform dispersion of 1 mg / mL (DLS detects particle size of 50-200 nm, no aggregation).

[0098] b. Preparation of CNQDs solution: select water-soluble carbon nitride quantum dots (CNQDs, fluorescent quantum yield 55%, surface containing -OH / -NH2 groups), prepare a transparent solution of 0.5 mg / mL with ultrapure water, and calibrate the concentration deviation within ± 5% by ultraviolet-visible spectrophotometer, and store in the dark.

[0099] c. Composite loading reaction: according to the ratio of "modified HAp: CNQDs = 3:1 (mass ratio)", slowly drop the CNQDs solution into the modified HAp dispersion at 25°C in the dark, and use the combination of "600 rpm magnetic stirring + 150 W intermittent ultrasonic for 5 min / hour" to react for 3 hours, and maintain the system pH at 6.5-7.5 during the reaction.

[0100] d. Post-treatment and purification: after the reaction, centrifuge the mixture at 12000 rpm for 15 minutes, and collect the precipitate; wash the precipitate with ultrapure water for 2 times (12000 rpm x 10 min / time), to remove the unbound free CNQDs; freeze-dry for 24 hours to obtain the core mineralization precursor.

[0101] The transmission electron microscope image of the core mineralization precursor is shown in Figure 1 , and the scanning electron microscope image is shown in Figure 2 .

[0102] The FTIR diagram of modified hydroxyapatite and carbon nitride quantum dots (CNQDs) modified hydroxyapatite is shown in Figure 3 .

[0103] 2. Component compounding:

[0104] (1) Preparation of functional soothing ingredients: mix 3 g of dihydro myricetin, 2 g of licorice extract, and 1.67 g of menthol-β-cyclodextrin inclusion compound (containing 0.33 g of menthol) to obtain the functional soothing ingredients.

[0105] The preparation steps of the menthol-β-cyclodextrin inclusion compound are as follows: mix 0.33 g of menthol with 1.34 g of β-cyclodextrin, stir in water at 60°C for 1 hour, and then freeze-dry to obtain the menthol-β-cyclodextrin inclusion compound.

[0106] (2) Pretreatment of auxiliary synergistic ingredients: dissolve 5 g of bovine serum albumin in ultrapure water to prepare a solution with a concentration of 10% (mass percent). Dissolve 3 g of γ-PGA-g-mPEG derivative (mPEG molecular weight 3000 Da, grafting rate 40%) in ultrapure water to prepare a solution with a concentration of 5% (mass percent), and then treat it to transparency by ultrasonic treatment at 200 W for 10 min and low-speed stirring at 300 rpm for 5 min. Then mix it with the 10% bovine serum albumin solution to form a premix.

[0107] The specific preparation method of the γ-PGA-g-mPEG derivative is described in the reference: Hu WK, Feng XR, Liu X, et al. Poly (γ-glutamic acid) modulates the properties of poly(ethylene glycol) hydrogel for biomedical applications. Journal of Biomaterials Science, Polymer Edition, 2016, 27(18): 1775-1787. (DOI: 10.1080 / 09205063.2016.1232527). The specific steps are as follows:

[0108] S1, raw material preparation: 1.0 g of γ-polyglutamic acid (γ-PGA, number average molecular weight 10000 Da, purity ≥98%) was dissolved in 50 mL of deionized water, and magnetic stirring was performed until complete dissolution to prepare a 20 mg / mL γ-PGA aqueous solution; 0.08-0.10 mL of glycidyl methacrylate (GMA) (10% of the molar amount of γ-PGA carboxyl group) was dissolved in 8-10 mL of anhydrous ethanol, and ultrasonic treatment was performed for 5 min to prepare a GMA ethanol solution; 0.24-0.3 g of polyethylene glycol monomethyl ether (mPEG, number average molecular weight 3000 Da, purity ≥98%) was dissolved in 12-15 mL of deionized water to prepare a 20 mg / mL mPEG aqueous solution.

[0109] S2, methacrylate grafting: the γ-PGA aqueous solution was placed in a 50°C water bath and stirred at 300 rpm, and the GMA ethanol solution was added dropwise at a rate of 0.5 mL / min; after the dropwise addition was completed, the pH was adjusted to 7.0 with 0.1 M NaOH, and the reaction was carried out at 50°C and 300 rpm for 24 h to obtain an mPGA reaction solution.

[0110] S3, mPEG grafting modification: mPEG aqueous solution was added to the mPGA reaction solution, and the reaction was carried out at 60°C and 400 rpm for 12 h to form an mPEG-PGA copolymer (grafting rate 30%-40%) through esterification reaction between the hydroxyl groups of mPEG and the carboxyl groups of mPGA.

[0111] S4, purification treatment: the reaction solution was transferred to a dialysis bag with a molecular weight cut-off of 3500 Da, and deionized water dialysis was carried out for 72 h (water change every 4 h) to remove unreacted impurities.

[0112] S5, product collection and characterization: the dialysate was freeze-dried at -40°C to -60°C and a vacuum of ≥0.08 MPa for 20-28 h to obtain a white solid; the number average molecular weight of the product was determined by GPC, and the grafting rate was determined by HNMR (400 MHz, D2O). 1 HNMR (400 MHz, D2O) to determine the grafting rate, and the number average molecular weight of the product was 8000-15000 Da.

[0113] (3) total material mixing: 85 g of the core mineralization precursor was mixed with the above-mentioned functional soothing ingredient and auxiliary synergistic ingredient premix, and stirred uniformly at room temperature to form a mixed material.

[0114] 3. Morphology shaping: the mixture was subjected to two-stage homogenization, first 20 MPa for 5 minutes, then 30 MPa for 5 minutes, and online viscosity monitoring (1 detection every 2 minutes) during the homogenization process; if the viscosity was lower than 500 mPa·s, 0.1%-0.2% xanthan gum (pre-dissolved in a small amount of ultrapure water) was added and stirred for 5 minutes; if the viscosity was higher than 800 mPa·s, 1%-2% ultrapure water was added; the final system viscosity was adjusted to 620 mPa·s (25℃), and a slow-release mineralized gel was obtained.

[0115] Example 2: Preparation of a dissolvable mineralized film

[0116] 1. The core mineralized precursor was prepared as in Example 1.

[0117] 2. Component compounding:

[0118] (1) Preparation of functional soothing ingredients: 1.5 g of dihydro myricetin, 1 g of licorice extract, and 0.83 g of menthol-β-cyclodextrin inclusion compound (containing 0.17 g of menthol) were mixed to obtain the functional soothing ingredients.

[0119] Among them, the preparation steps of the menthol-β-cyclodextrin inclusion compound are as follows: 0.17 g of menthol and 0.66 g of β-cyclodextrin were mixed, stirred in 60℃ water for 1 hour, and then freeze-dried to obtain the menthol-β-cyclodextrin inclusion compound.

[0120] (2) Pretreatment of auxiliary synergistic ingredients: 2 g of bovine serum albumin was dissolved in ultrapure water to prepare a solution with a concentration of 10% (mass percent). 1 g of γ-PGA-g-mPEG derivative (mPEG molecular weight 3000 Da, grafting rate 40%) was dissolved in ultrapure water to prepare a solution with a concentration of 5% (mass percent), treated to be transparent by 200 W ultrasonic for 10 min + 300 rpm low-speed stirring for 5 min, and then mixed with the 10% concentration bovine serum albumin solution to form a premix.

[0121] (3) Mixing of total materials: 92 g of the core mineralized precursor was mixed with the above-mentioned functional soothing ingredients and auxiliary synergistic ingredient premix, stirred uniformly at room temperature, and formed into a mixture.

[0122] 3. Morphology shaping: 0.5% (mass percent) ammonium bicarbonate was added to the mixture as a pore former, and after uniform stirring, it was placed in a vacuum drying oven and dried and cross-linked at a vacuum degree of -0.09 MPa and a temperature of 35℃ for 4 hours to obtain a dissolvable mineralized film with a pore size of 3-8 μm.

[0123] The scanning electron microscope image of the dissolvable mineralized film is shown in Figure 4 , and the dissolution rate curve of the dissolvable mineralized film is shown in Figure 5 .

[0124] Example 3: Performance test

[0125] 1. Mineralization repair test: The slow-release mineralization gel prepared in Example 1 was applied to simulate damaged mucosa, with a 7-day mineralization coverage of 85%, which was significantly higher than that of the conventional throat-soothing gel (35%).

[0126] 2. In vitro anti-inflammatory activity detection experiment

[0127] Recover RAW264.7 cells and adjust the concentration to 1.0×10 5 6 / mL in DMEM medium, inoculate in a 48-well plate at 200 μL per well, and culture at 37°C, 5% CO2 for 12 h to adhere the cells; prepare the slow-release mineralization gel extract of Example 1 (0.1 g / mL, i.e. 1 g of gel is dissolved in 10 mL of DMEM medium) as the 100% stock concentration, and dilute it by 2 times to get 20%, 10%, 5%, 2.5%, and 1.25% gradient concentrations; set up a blank control group (only DMEM), five concentration extract experimental groups, and a PBS replacement control group, with three replicate wells for each group; the experimental groups are first induced by LPS (1 μg / mL LPS) for 1 h, then added with the corresponding concentration extract, and the PBS group is replaced with 100 μL of PBS instead of the extract, and continue to culture for 24 h; collect the supernatant of each well, and detect the expression level of the factors using the TNF-α and IL-10 ELISA kits.

[0128] The results show that after 24 h of co-culture, compared with the PBS control group and the blank control group, the expression of the pro-inflammatory factor TNF-α in each concentration extract group does not fluctuate significantly, but the 20% concentration group shows a slight down-regulation, the anti-inflammatory factor IL-10 shows a concentration-dependent increase, among which the IL-10 expression of the 20% concentration group reaches 111.37, which is 3.1 times that of the PBS control group, and is significantly higher than that of other concentration groups (p<0.05), the polarization ratio of M2 type macrophages is increased, and the anti-inflammatory activity is optimal Figure 6 ).

[0129] 3. Long-acting soothing persistence experiment

[0130] Experimental object: artificial pharyngeal and laryngeal mucosa model (Geistlich Mucograft® resorbable collagen).

[0131] Experimental method: The gel prepared in Example 1 was coated on the collagen surface, and the residual rate and release rate of the functional soothing ingredients (dihydromyricetin and licorice extract) in the material at different time points (2 h, 4 h, 6 h, 8 h) were detected by high performance liquid chromatography under the in vitro simulated oral saliva flushing environment, the long-acting soothing ability of the material was quantified from the aspect of component release, and the sustained relief effect on throat discomfort was indirectly verified. The results showed that with the extension of simulated saliva flushing time, dihydromyricetin and licorice extract both showed the trend of "initial rapid release and later slow release", and at 8 h, the cumulative release amount of dihydromyricetin was 72%, and the mucosal residual amount was 25%, the cumulative release amount of licorice extract was 68%, and the mucosal residual amount was 28%, realizing the synergistic effect of rapid onset and long-acting soothing Figure 7 ).

[0132] 4. Safety test: The dissolvable mineralized film prepared in Example 2 was applied to the oral cavity of rabbits for 30 days, and the mucosa was not stimulated, and the pathological section showed that the cells were not damaged; heavy metal detection Pb 0.08 mg / kg, BSA residue 0.8 μg / g.

[0133] 5. Stability test: stored at 25℃, 60%RH for 12 months, the core precursor particle size was 50-200 nm, the retention rate of functional ingredients was ≥92%, and the system was not layered.

[0134] The above detailed description is a specific description of the embodiments of the present application, and the embodiments are not used to limit the patent scope of the present application. Any equivalent implementation or change made without departing from the present application shall be included in the patent scope of the present application.

Claims

1. A mineralization-repairing oral and throat care material for the elderly, characterized by, comprise, by mass percentage, a core mineralization precursor 85-94%, a functional soothing ingredient 3-7%, and an auxiliary synergistic ingredient 2-10%; the core mineralization precursor is a biocompatible mineralization matrix of modified hydroxyapatite loaded with carbon nitride quantum dots; the functional soothing ingredient is a mixture of dihydromyricetin, licorice extract, and menthol-β-cyclodextrin inclusion compound; the auxiliary synergistic ingredient is a mixture of bovine serum albumin and γ-polyglutamic acid-g-polyethylene glycol monomethyl ether derivative.

2. The geriatric-oriented mineralization-repairing oral throat care material according to claim 1, characterized by, the mass ratio of modified hydroxyapatite to carbon nitride quantum dots in the biocompatible mineralization matrix of modified hydroxyapatite loaded with carbon nitride quantum dots is 3:1; the mass ratio of dihydromyricetin, licorice extract, and menthol-β-cyclodextrin inclusion compound is 3:2:(1-1.8); the mass ratio of bovine serum albumin to γ-polyglutamic acid-g-polyethylene glycol monomethyl ether derivative is (1.5-2):

1.

3. The geriatric mineralization-repairing oral throat care material according to claim 1, characterized in that, the number average molecular weight of the γ-polyglutamic acid-g-polyethylene glycol monomethyl ether derivative is 8000-15000 Da, the molecular weight of polyethylene glycol monomethyl ether in the derivative is 2000-5000 Da, and the grafting rate is 30%-40%.

4. The geriatric mineralization-repairing oral throat care material according to claim 1, characterized in that, the mass ratio of menthol to β-cyclodextrin in the menthol-β-cyclodextrin inclusion compound is 1:

4.

5. The geriatric mineralization-repairing oral throat care material according to claim 1, characterized in that, the product form of the care material is a dissolvable mineralized film or a slow-release mineralized gel.

6. A method for preparing the geriatric mineralization-repairing oral throat care material according to any one of claims 1 to 5, characterized in that, comprise the following steps: (1) Core mineralization precursor preparation S1, preparation of modified hydroxyapatite: a. Dissolve 2,5-diamino-4,6-dihydroxypyrimidine and polyethylene glycol diglycidyl ether in a molar ratio of 1:1.05 in an ethanol / water mixed solvent and react at 60-70°C for 6-8 hours until the epoxy groups are completely reacted; b. The reaction solution is precipitated with diethyl ether, centrifuged, washed, and freeze-dried to obtain a PEGDE cross-linked pyrimidine derivative; c. Dissolve the derivative in an ethanol / water mixed solvent and stir to dissolve while heating to 60-80°C; d. At 0-4°C, add a 4-hydroxybenzaldehyde solution to the solution and adjust the pH to 4-5 with dilute acetic acid. After reacting for 2-4 hours, neutralize to pH 6.5-7.0 with triethylamine; e. Disperse hydroxyapatite nanoparticles in an ethanol / water mixed solvent, treat with a combination of microfluidization and ultrasonic stirring, and then add to the reaction system of step d. Continue ultrasonic stirring and reaction; f. After the reaction is complete, wash the product with an ethanol / water mixed solvent containing Tween-80, ultrapure water, and then dry to obtain modified hydroxyapatite; S2, preparation of modified hydroxyapatite loaded with carbon nitride quantum dots: A, preparation of modified HAp dispersion: disperse modified hydroxyapatite in water and treat with a combination of ultrasonic stirring to obtain a stable dispersion of 1 mg / mL with a particle size of 50-200 nm; B, preparation of CNQDs solution: prepare a water-soluble carbon nitride quantum dot solution with a fluorescence quantum yield of ≥50% to 0.5 mg / mL; C. Composite loading reaction: Carbon nitride quantum dot solution was slowly added into the hydroxyapatite dispersion liquid at a mass ratio of modified HAp to CNQDs of 3:1, and reacted for 3 hours at 25±2℃, pH 6.5-7.5, in the dark, with magnetic stirring combined with intermittent ultrasonic reaction; D. Post-treatment and purification: After the reaction, centrifugation, water washing and freeze-drying were performed to obtain the modified hydroxyapatite loaded carbon nitride quantum dot composite material; (2) Component compounding ① Functional soothing component preparation: Mix dihydromyricetin, licorice extract and menthol-β-cyclodextrin inclusion compound; ② Auxiliary synergistic component pretreatment: Dissolve γ-polyglutamic acid-g-polyethylene glycol monomethyl ether derivative in ultrapure water, and after ultrasonic- low speed stirring until transparent, mix with bovine serum albumin solution to form a premix; ③ Total material mixing: Mix the core mineralization precursor, functional soothing component and auxiliary synergistic component premix according to the preset ratio, stir uniformly at room temperature, and form a mixture; (3) Morphology forming The mixture is made into a dissolvable mineralization film or a slow-release mineralization gel.

7. The production method according to claim 6, wherein The preparation steps of the dissolvable mineralization film are as follows: Add ammonium bicarbonate as a pore former to the mixture, stir uniformly, and then place it in a vacuum drying oven for 3-4 h to obtain a dissolvable mineralization film with a porous structure.

8. The preparation method according to claim 6, characterized in that, The preparation steps of the slow-release mineralization gel are as follows: The mixture is subjected to two-stage homogenization treatment, and the system viscosity is adjusted to 500-800 mPa·s by adding ultrapure water or xanthan gum to obtain a slow-release mineralization gel.

9. Use of the care material according to any one of claims 1-5 in the preparation of a product for repairing oral mucosa sensitivity in the middle-aged and elderly.

10. Use of the care material according to any one of claims 1-5 in the preparation of a mineralization material for the auxiliary care of chronic pharyngitis.

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