Preparation method of a composite protein-coated functional microsphere and its application in nasal mucosa repair

Abstract

The present invention relates to the technical field of nasal mucosa repair, specifically to a preparation method of functional microspheres wrapped with composite protein and its application in nasal mucosa repair. The present invention uses silk fibroin, sodium chloride, ectoine, sorbitol, resveratrol, borneol, curcumin, volatile oil of Magnolia officinalis, volatile oil of Xanthium sibiricum, volatile oil of Centipeda minima, bioactive glass powder, and mussel adhesive protein as raw materials. Utilizing the negatively charged property of the nasal mucosa, functional microspheres that can have a strong electrostatic adsorption effect with the nasal mucosa are prepared. A biofilm is formed on the surface of the nasal mucosa by the functional microspheres, which can prevent and reduce the contact between pathogenic microorganisms and other particulate allergic substances in the external environment and the nasal mucosa. At the same time, electrostatic adsorption can also prolong the action time of functional components on the nasal mucosa. The functional components play roles from different aspects successively, enhancing their action effects on the nasal mucosa. The functional microspheres obtained by the preparation method of the present invention can effectively treat various rhinitis, improve the symptoms of sneezing, rhinorrhea, nasal congestion, etc. in patients, and have a long-lasting effect, are not easy to relapse, have little irritation, and high safety.

Description

Technical Field

[0001] The present invention relates to the technical field of nasal mucosa repair, and specifically provides a preparation method of functional microspheres wrapped with composite protein and its application in nasal mucosa repair. Background Art

[0002] There are two types of nasal allergies. One is vasomotor rhinitis, which is mainly due to the imbalance of autonomic nerve function, resulting in the overactivity of the parasympathetic nerve and causing various symptoms similar to allergic rhinitis. It is not caused by specific antigens, but by emotional stress, increased pressure or various physical changes. The other is allergic rhinitis, also known as allergic rhinitis, which refers to a non-infectious inflammatory disease of the nasal mucosa mainly mediated by IgE (mainly histamine) release after a specific individual contacts an allergen, and involves a variety of immune active cells and cytokines. Its symptoms include watery nasal discharge, nasal congestion, sneezing, and even nasal root soreness, itching in the nose, eyes and ears, dizziness, inattention, etc. In addition, it may also induce allergic conjunctivitis, allergic dermatitis, chronic sinusitis, nasal polyps, secretory otitis media, and may even induce asthma and endanger life in severe cases. Statistics show that in recent years, the prevalence of allergic rhinitis has been increasing globally, and the number of allergic rhinitis patients in China is also increasing year by year. This not only has a negative impact on people's physical and mental health and quality of life, but also causes a certain burden on social economy, and has increasingly become a health problem concerned by the general public.

[0003] At present, sprays and nasal drops are generally used more frequently for the treatment of rhinitis. However, due to the clearance function of numerous cilia on the surface of the nasal mucosa, the residence time of liquid in the nasal cavity is only about 15 minutes, while the residence time of powder or particles in the nasal cavity is about 20 - 30 minutes. This affects the absorption and efficacy of drugs to a certain extent, so long-term use is required. Some sprays and nasal drops contain hormonal drugs. If used frequently for a long time, there will be some local side effects, such as nasal dryness, nosebleed, and even possible systemic side effects of hormones. Summary of the Invention

[0004] The purpose of the present invention is to provide a preparation method of functional microspheres wrapped with composite protein and its application in nasal mucosa repair. The functional microspheres are safe and have no side effects, can prolong the drug action time, and effectively improve the effect on nasal mucosa repair.

[0005] To achieve the above purpose, the present invention provides the following technical solution: A preparation method of functional microspheres wrapped with composite protein, which includes the following preparation steps:

[0006] Step 1: Dissolve 8-14 parts by weight of silk fibroin, 1-5 parts by weight of sodium chloride, 3-5 parts by weight of ectoin, and 2-5 parts by weight of sorbitol in pure water, and stir to obtain a silk fibroin aqueous solution with a weight percentage of 8%-14%. In the present invention, the concentration of silk fibroin is controlled at 8%-14%. If the concentration of the silk fibroin aqueous solution is too low, insufficient silk fibroin cannot be deposited on the surface of the volatile oil. If the concentration is too high, the solution viscosity is too large to prepare microspheres.

[0007] Step 2: Dissolve 0.02-0.5 parts by weight of resveratrol, 0.02-0.5 parts by weight of borneol, and 0.02-0.5 parts by weight of curcumin in 2-5 parts by weight of absolute ethanol, and mix and stir evenly with the 8%-14% silk fibroin aqueous solution in Step 1 to obtain a mixed solution.

[0008] Step 3: Add 0.05-0.5 parts by weight of bioactive glass powder to 0.2-2 parts by weight of magnolia flower volatile oil, 0.2-2 parts by weight of xanthium sibiricum volatile oil, and 0.2-2 parts by weight of centipeda minima volatile oil, quickly stir and mix, and then add it to the mixed solution in Step 2, and stir to form a white emulsion.

[0009] Step 4: Dissolve 3-9 parts by weight of mussel adhesive protein in pure water, stir to obtain a mussel adhesive protein aqueous solution with a weight percentage of 3%-9%, and mix the mussel adhesive protein aqueous solution with the white emulsion in Step 3, and stir and mix evenly.

[0010] Step 5: Add 13-33 parts by weight of absolute ethanol to the emulsion after mixing in Step 4, stir, and then centrifuge, wash, and dry to obtain a functional microsphere wrapped with a composite protein.

[0011] The stirring speed during the preparation process is controlled at 1000-5000 r / min, and the centrifugation speed is controlled at 5000-20000 r / min.

[0012] The particle size of the functional microsphere in Step (5) is 120-160 μm.

[0013] In Step (5), it is washed with absolute ethanol.

[0014] In Step (5), drying is carried out under vacuum conditions at 45-55 °C for 12 h.

[0015] The bioactive glass powder is nano-bioactive glass powder.

[0016] Apply a functional microsphere wrapped with a composite protein to nasal mucosa repair.

[0017] The magnolia flower volatile oil, xanthium sibiricum volatile oil, and centipeda minima volatile oil of the present invention are volatile components and have anti-inflammatory, anti-allergic, and nasal orifice unblocking effects.

[0018] The bioactive glass of the present invention has the functions of antibacterial and anti-inflammatory, and repairing damaged cells.

[0019] The curcumin of the present invention has the efficacy of anti-inflammatory and bactericidal.

[0020] The resveratrol of the present invention has the efficacy of antioxidant and immune regulation.

[0021] The borneol of the present invention has the efficacy of anti-inflammatory and bactericidal, and promoting the absorption of drugs by mucosa.

[0022] The ectoine of the present invention has the efficacy of a humectant and can relieve rhinitis.

[0023] The mussel adhesive protein of the present invention is a bioadhesive with characteristics such as high strength, high toughness, waterproofness, good biocompatibility, biodegradability and low toxicity. It is composed of 70 - 90 repeated polypeptide fragments, and each polypeptide segment consists of 10 amino acids, among which 2 are lysines, making it one of the few basic proteins. When at the normal physiological pH of the human body, it carries a large amount of positive charges.

[0024] The silk fibroin of the present invention is a natural protein extracted from silk, with an amphiphilic structure, containing both hydrophilic and hydrophobic groups in the molecule.

[0025] The present invention adds bioactive glass powder to the volatile oils of magnolia flower, xanthium fruit and centipeda minima, forming functional oil droplets with solid in oil. When silk fibroin encounters the functional oil droplets, it will wrap around the surface of the functional oil droplets to form the first shell layer of microspheres. In addition, since silk fibroin has a negatively charged surface in neutral aqueous solution and mussel adhesive protein has a positive charge, when the positively charged mussel adhesive protein is added, the mussel adhesive protein will automatically deposit on the surface of silk fibroin to form the second shell layer of microspheres, strengthening the first shell layer of microspheres, and finally forming functional microspheres coated with composite protein. The addition of sodium chloride and sorbitol in the present invention makes the microsphere system formed during the preparation process more stable.

[0026] When the functional microspheres of the present invention act on the nasal cavity, the positively charged mussel adhesive protein can produce a strong electrostatic interaction with the negatively charged nasal mucosa, thereby enabling the functional microspheres to adhere to the nasal mucosa and form a positively charged nano - scale biological network membrane. In addition, the mussel adhesive protein molecule contains a large amount of 3,4 - dihydroxyphenylalanine (DOPA). Oxidized DOPA and unoxidized DOPA self - oxidize and cross - link in an oxygen - containing environment to form a microscopic discontinuous network biological membrane, strengthening the adhesion effect.

[0027] Due to the electrostatic adsorption of mussel adhesive protein and nasal mucosa, the formed biofilm can prevent and reduce the contact between pathogenic microorganisms and other particulate allergic substances in the outside world and the nasal mucosa. Electrostatic adsorption can also extend the action time of functional components on the nasal mucosa, enhance the penetration ability into the mucosa, and thus enhance their action effect on the nasal mucosa. Among the functional components of the present invention, curcumin, resveratrol, borneol, and ectoin play a role first. At the same time, with the volatilization of the volatile oils of magnolia flower, xanthium fruit, and centipeda minima, when the volatile oils of magnolia flower, xanthium fruit, and centipeda minima in the functional oil droplets of the present invention are completely volatilized, the bioactive glass powder loses its oily coating and can enter the nasal mucosa to play the role of antibacterial and anti-inflammatory and repairing damaged cells. The preparation method of the present invention enables the functional microspheres to play a role in multiple dimensions. It should be noted that when the bioactive glass powder enters the nasal mucosa to play a role, the biofilm formed in the nasal cavity has formed a stable wet environment, which can overcome the irritation and instant sharp pain caused by the rapid increase in local pH when the bioactive glass powder directly contacts the nasal mucosa. This is also the reason why the bioactive glass powder is wrapped in the volatile oil in the present invention.

[0028] Compared with the prior art, the beneficial effects of the present invention are as follows: The present invention uses silk fibroin, sodium chloride, ectoin, sorbitol, resveratrol, borneol, curcumin, volatile oil of magnolia flower, volatile oil of xanthium fruit, volatile oil of centipeda minima, bioactive glass powder, and mussel adhesive protein as raw materials, and utilizes the negatively charged characteristic of the nasal mucosa to prepare functional microspheres that can have a strong electrostatic adsorption effect with the nasal mucosa. A biofilm is formed on the surface of the nasal mucosa of the functional microspheres, which can prevent and reduce the contact between pathogenic microorganisms and other particulate allergic substances in the outside world and the nasal mucosa. At the same time, electrostatic adsorption can also extend the action time of functional components on the nasal mucosa. The functional components play a role from different aspects in sequence, enhancing their action effect on the nasal mucosa. The functional microspheres obtained by the preparation method of the present invention can effectively treat various rhinitis, improve the symptoms of sneezing, rhinorrhea, nasal congestion, etc. of patients, have a lasting effect, are not easy to relapse, have little irritation, and are highly safe. Specific Embodiments

[0029] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0030] Example 1

[0031] A preparation method of functional microspheres wrapped with composite protein, which comprises the following preparation steps:

[0032] Step 1: Dissolve 8 parts by weight of silk fibroin, 1 part by weight of sodium chloride, 3 parts by weight of ectoin, and 2 - 5 parts by weight of sorbitol in pure water, and stir to obtain an 8% silk fibroin aqueous solution by weight percentage;

[0033] Step 2: Dissolve 0.02 parts by weight of resveratrol, 0.02 parts by weight of borneol, and 0.02 parts by weight of curcumin in 2 parts by weight of absolute ethanol, mix and stir evenly with the 8% silk fibroin aqueous solution in Step 1 to obtain a mixed solution;

[0034] Step 3: Add 0.05 parts by weight of bioactive glass powder to 0.2 parts by weight of volatile oil of Magnolia biondii, 0.2 parts by weight of volatile oil of Xanthium sibiricum, and 0.2 parts by weight of volatile oil of Centipeda minima, quickly stir and mix, and then add it to the mixed solution in Step 2, and stir to form a white emulsion;

[0035] Step 4: Dissolve 3 parts by weight of mussel adhesive protein in pure water, stir to obtain a 3% mussel adhesive protein aqueous solution by weight percentage, and mix the mussel adhesive protein aqueous solution with the white emulsion in Step 3, and stir to mix evenly;

[0036] Step 5: Add 13 parts by weight of absolute ethanol to the emulsion after mixing in Step 4, stir, and then centrifuge, wash, and dry to obtain a functional microsphere wrapped with a composite protein.

[0037] During the preparation process, the stirring speed is controlled at 1000 - 5000 r / min, and the centrifugation speed is controlled at 5000 - 20000 r / min.

[0038] The particle size of the functional microsphere in Step (5) is 120 μm.

[0039] In Step (5), it is washed with absolute ethanol.

[0040] In Step (5), drying is carried out under vacuum conditions at 45 °C for 12 h.

[0041] The bioactive glass powder is nano - bioactive glass powder.

[0042] Apply a functional microsphere wrapped with a composite protein to nasal mucosa repair.

[0043] Example 2

[0044] A preparation method of a functional microsphere wrapped with a composite protein, which includes the following preparation steps:

[0045] Step 1: Dissolve 10 parts by weight of silk fibroin, 3 parts by weight of sodium chloride, 4 parts by weight of ectoin, and 3 parts by weight of sorbitol in pure water, and stir to obtain a 10% silk fibroin aqueous solution by weight percentage;

[0046] Step 2: Dissolve 0.2 parts by weight of resveratrol, 0.2 parts by weight of borneol, and 0.2 parts by weight of curcumin in 4 parts by weight of absolute ethanol, and mix and stir evenly with the 10% aqueous silk fibroin solution in Step 1 to obtain a mixed solution;

[0047] Step 3: Add 0.2 parts by weight of bioactive glass powder to 1 part by weight of volatile oil of Magnolia biondii, 1 part by weight of volatile oil of Xanthium sibiricum, and 1 part by weight of volatile oil of Centipeda minima, quickly stir and mix, and then add it to the mixed solution in Step 2, and stir to form a white emulsion;

[0048] Step 4: Dissolve 6 parts by weight of mussel adhesive protein in pure water, stir to obtain a 6% aqueous mussel adhesive protein solution by weight percentage, and mix the aqueous mussel adhesive protein solution with the white emulsion in Step 3, and stir and mix evenly;

[0049] Step 5: Add 25 parts by weight of absolute ethanol to the emulsion after mixing in Step 4, stir, and then centrifuge, wash, and dry to obtain a functional microsphere wrapped with composite protein.

[0050] The stirring speed during the preparation process is controlled at 1000 - 5000 r / min, and the centrifugation speed is controlled at 5000 - 20000 r / min.

[0051] The particle size of the functional microsphere in Step (5) is 140 μm.

[0052] In Step (5), it is washed with absolute ethanol.

[0053] In Step (5), drying is carried out under vacuum conditions at 50 °C for 12 h.

[0054] The bioactive glass powder is nano-bioactive glass powder.

[0055] Apply a functional microsphere wrapped with composite protein to nasal mucosa repair.

[0056] Example 3

[0057] A preparation method of a functional microsphere wrapped with composite protein, which includes the following preparation steps:

[0058] Step 1: Dissolve 14 parts by weight of silk fibroin, 5 parts by weight of sodium chloride, 5 parts by weight of ectoine, and 5 parts by weight of sorbitol in pure water, and stir to obtain a 14% aqueous silk fibroin solution by weight percentage;

[0059] Step 2: Dissolve 0.5 parts by weight of resveratrol, 0.5 parts by weight of borneol, and 0.5 parts by weight of curcumin in 5 parts by weight of absolute ethanol, and mix and stir evenly with the 15% aqueous silk fibroin solution in Step 1 to obtain a mixed solution;

[0060] Step 3: Add 0.5 parts by weight of bioactive glass powder into 2 parts by weight of volatile oil of Magnolia biondii, 2 parts by weight of volatile oil of Xanthium sibiricum, and 2 parts by weight of volatile oil of Centipeda minima, and stir quickly to mix. Then add it to the mixed solution in Step 2 and stir to form a white emulsion.

[0061] Step 4: Dissolve 9 parts by weight of mussel adhesive protein in pure water, and stir to obtain a 9% by weight aqueous solution of mussel adhesive protein. Mix the aqueous solution of mussel adhesive protein with the white emulsion in Step 3 and stir evenly.

[0062] Step 5: Add 33 parts by weight of absolute ethanol to the emulsion after mixing in Step 4, stir, and then centrifuge, wash, and dry to obtain a functional microsphere encapsulated with composite protein.

[0063] The stirring speed during the preparation process is controlled at 1000 - 5000 r / min, and the centrifugation speed is controlled at 5000 - 20000 r / min.

[0064] The particle size of the functional microsphere in Step (5) is 160 μm.

[0065] In Step (5), it is washed with absolute ethanol.

[0066] In Step (5), the drying is carried out under vacuum conditions at 55 °C for 12 h.

[0067] The bioactive glass powder is nano-bioactive glass powder.

[0068] Apply a functional microsphere encapsulated with composite protein to nasal mucosa repair.

[0069] Comparative Example 1

[0070] The difference between the present invention and Example 2 is that in this example, bioactive glass powder is not used in Step 3: directly add 1 part by weight of volatile oil of Magnolia biondii, 1 part by weight of volatile oil of Xanthium sibiricum, and 1 part by weight of volatile oil of Centipeda minima, stir quickly to mix, then add it to the mixed solution in Step 2, and stir to form a white emulsion. The rest are the same, and finally a functional microsphere encapsulated with composite protein is also obtained.

[0071] Comparative Example 2

[0072] The difference between the present invention and Example 2 is that in Step 3: add 0.2 parts by weight of bioactive glass powder into 3 parts by weight of volatile oil of Magnolia biondii, 4 parts by weight of volatile oil of Xanthium sibiricum, and 3 parts by weight of volatile oil of Centipeda minima, stir quickly to mix, then add it to the mixed solution in Step 2, and stir to form a white emulsion. The rest are the same.

[0073] No microspheres were observed. Reason analysis: In this comparative example, the proportion of volatile oil is too high, resulting in too large microspheres and causing damage in the subsequent process.

[0074] Comparative Example 3

[0075] The difference between the present invention and Example 2 is as follows: In this example, in Step 4, 20 parts by weight of mussel adhesive protein is dissolved in pure water, and stirred to obtain a 20% by weight aqueous solution of mussel adhesive protein. Then, this aqueous solution of mussel adhesive protein is mixed with the white emulsion in Step 3 and stirred evenly. The rest are the same.

[0076] The microspheres are adhered and agglomerated. Reason analysis: In this comparative example, the concentration of mussel adhesive protein is too high, resulting in too high viscosity of the solution, and the formed microspheres are adhered and agglomerated with each other, and the dispersion is uneven.

[0077] Experimental part

[0078] 1. Animal experiment:

[0079] The products prepared in Examples 1-3 and Comparative Examples 1-3, 10 mg, are mixed with 100 ml of normal saline to prepare a suspension of 0.1 mg / ml.

[0080] Treatment plan: Ninety SD rats are randomly divided into 9 groups, with 10 rats in each group, namely ① blank control group, ② allergic rhinitis model group, ③ treatment group of Example 1, ④ treatment group of Example 2, ⑤ treatment group of Example 3, ⑥ treatment group of Comparative Example 1, ⑦ treatment group of Comparative Example 2, ⑧ treatment group of Comparative Example 3, ⑨ dexamethasone treatment group (positive drug control group).

[0081] The rats in Group do not undergo sensitization and are not given drugs. 、 、 、 、 、 、 、 The rats in Group are continuously intranasally instilled with a 10% by volume toluene-2,4-diisocyanate (TDI) olive oil solution for 7 days to sensitize the rats, and the dose is 0.1 mL / d. After the successful establishment of the model, the allergic rhinitis model group does not receive treatment. 、 、 、 、 The rats are intranasally instilled with 0.2 ml of the suspension every day, and the dexamethasone treatment group is given dexamethasone by nasal drip, twice a day. Observe continuously for 7 days.

[0082] Observation indexes of allergic rhinitis symptoms: 30 minutes after the last nasal drip, observe the severity of nose scratching, sneezing, and clear nasal discharge in the rats and score them. Score each item and calculate the total score using the superposition method.

[0083] (1)Nose scratching (nasal itching): 1 point (1 - 5 times), 2 points (6 - 15 times), 3 points (more than 15 times);

[0084] (2)Sneezing: 1 point (1 - 3 times), 2 points (4 - 10 times), 3 points (more than 11 times);

[0085] (3)Clear nasal discharge: 1 point (flows to the anterior naris), 2 points (exceeds the anterior naris), 3 points (flows over the face).

[0086] Statistical analysis: All data are expressed as ±S. ANOVA and Student - Newman - Keuls multiple comparison methods are used for comparing differences between groups; S - test analysis is completed by SPSS 11.5 statistical software. A two - sided P < 0.05 indicates significant differences.

[0087] .

[0088] Compared with the allergic rhinitis group, *P < 0.05; compared with the blank control group, #P < 0.05; n = 10.

[0089] Result analysis: Except for the blank control group, different degrees of nasal itching, sneezing, and nasal discharge symptoms occurred in other groups starting from the administration of TDI olive oil solution. The allergic symptoms worsened with the increase in the number of administrations. The rats with successful model replication had typical nasal itching, sneezing, and nasal discharge symptoms. The dexamethasone group alleviated the allergic rhinitis symptoms, showing significant differences from the allergic rhinitis group. The treatment groups of Examples 1 - 3 had a therapeutic effect on rhinitis, showing significant differences from the allergic rhinitis group (P < 0.05), and the scores after treatment were slightly better than those of the dexamethasone group. Although the treatment groups of Comparative Examples 1 - 3 had a therapeutic effect, the scores after treatment were significantly lower than those of the treatment groups of Examples 1 - 3 and the dexamethasone group, indicating that both the microspheres and the bioactive glass powder had obvious therapeutic effects on allergic rhinitis.

[0090] 2. Cytotoxicity experiment

[0091] This test was carried out in accordance with "GB∕T 16886.5 - 2017 Biological evaluation of medical devices - Part 5: In vitro cytotoxicity test" for the extraction solution test. The microspheres obtained from the previous Examples 1 - 3 were used as the test groups. During the experiment, only a small number of cells in the three sample groups showed morphological changes such as cell shrinkage, loose adherence to the wall, and no cytoplasmic granules, and the number of cells with morphological changes was less than 20%. The test samples showed a tendency of slight toxicity, with a toxicity grade of 1, and passed the toxicity test. The specific results are shown in the following table.

[0092] .

[0093] The foregoing has shown and described the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments, and the above embodiments and the descriptions in the specification are only preferred examples of the present invention and are not used to limit the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements fall within the scope of the present invention claimed. The scope of the present invention claimed is defined by the appended claims and their equivalents.

Claims

1. A method for preparing functional microspheres encapsulated with composite protein, characterized in that, It includes the following preparation steps: Step 1: Dissolve 8 - 14 parts by weight of silk fibroin, 1 - 5 parts by weight of sodium chloride, 3 - 5 parts by weight of ectoin, and 2 - 5 parts by weight of sorbitol in pure water, and stir to obtain an 8% - 14% aqueous silk fibroin solution by weight percentage; Step 2: Dissolve 0.02 - 0.5 parts by weight of resveratrol, 0.02 - 0.5 parts by weight of borneol, and 0.02 - 0.5 parts by weight of curcumin in 2 - 5 parts by weight of absolute ethanol, mix and stir evenly with the 8% - 14% aqueous silk fibroin solution in Step 1 to obtain a mixed solution; Step 3: Add 0.05 - 0.5 parts by weight of bioactive glass powder to 0.2 - 2 parts by weight of volatile oil of Magnolia liliflora Desr., 0.2 - 2 parts by weight of volatile oil of Xanthium sibiricum Patrin ex Widd., and 0.2 - 2 parts by weight of volatile oil of Centipeda minima (L.) A. Br. et Aschers., quickly stir and mix, and then add it to the mixed solution in Step 2, and stir to form a white emulsion; Step 4: Dissolve 3 - 9 parts by weight of mussel adhesive protein in pure water, stir to obtain a 3% - 9% aqueous mussel adhesive protein solution by weight percentage, and mix the aqueous mussel adhesive protein solution with the white emulsion in Step 3, and stir to mix evenly; Step 5: Add 13 - 33 parts by weight of absolute ethanol to the emulsion after mixing in Step 4, stir, and then centrifuge, wash, and dry to obtain functional microspheres wrapped with a composite protein; 2. The preparation method of a functional microsphere encapsulated with a composite protein according to claim 1, characterized in that: The stirring speed during the preparation process is controlled at 1000 - 5000 r / min, and the centrifugation speed is controlled at 5000 - 20000 r / min.

3. The preparation method of a functional microsphere encapsulated with a composite protein according to claim 1, wherein: The particle size of the functional microspheres in Step 5 is 120 - 160 μm.

4. The preparation method of a functional microsphere encapsulated with a composite protein according to claim 1, characterized in that: In Step 5, the washing is carried out with absolute ethanol.

5. The preparation method of a functional microsphere encapsulated with a composite protein according to claim 1, characterized in that: The drying in Step 5 is carried out under vacuum conditions at 45 - 55 °C for 12 h.

6. The preparation method of a functional microsphere encapsulated with a composite protein according to claim 1, characterized in that: The bioactive glass powder is nano - bioactive glass powder.