Oral ulcer repairing spray and preparation method thereof
By combining aloe exosomes, silk fibroin and mussel mucin and combining with traditional Chinese medicine ingredients, an oral ulcer repair spray was prepared, which solved the problem that the existing spray was not long-lasting and the possible scar formation, achieving lasting therapeutic effect and scar-free healing.
Patent Information
- Application Number
- CN202411993908.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-31
- Publication Date
- 2025-03-28
AI Technical Summary
The existing oral ulcer spray does not last long after use, is irritating, has poor treatment effect, and may form scars after healing, making it difficult to recover the local mucosa.
A combination of aloe vera exosomes, silk fibroin and mussel mucin is used to wrap aloe vera exosomes through modified silk fibroin nanocapsules to form stable exosome microspheres, and combined with traditional Chinese medicine ingredients such as bacillol extract and chamomile extract, a kind of oral ulcer repair spray is prepared.
This spray lasts for a long time in the oral cavity, has significant therapeutic effect, is gentle and comfortable when used, and has no scars after healing, which promotes the recovery of local mucosa.
Smart Images
Figure SMS_1
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of oral care, and in particular to an oral ulcer repairing spray and a preparation method thereof. Background Art
[0002] In recent years, oral problems have received more and more attention. Common oral problems include oral ulcers, dental calculus, caries, gum problems, oral sensitivity, bad breath problems, tooth stains and dental plaque. Oral ulcers refer to diseases in which the normal color, appearance, integrity and function of the mucosa in a certain part of the mouth are changed. There are many types of lesions, which can be combined into complex and diverse lesions. Recurrent oral ulcers are roughly divided into three types: small mouth ulcers, large mouth ulcers, and herpetiform ulcers. Small mouth ulcers are the most common and have a high recurrence rate. Clinically, the treatment of small mouth ulcers can be divided into: (1) symptomatic supportive treatment; (2) special treatment; (3) preventive treatment. Standard local treatment options usually use some drugs to relieve symptoms such as pain, including analgesics, antimicrobial agents, anti-inflammatory agents, steroids, vitamin B2, etc., and a large number of spray products have appeared.
[0003] Currently available sprays are very irritating to the skin and the effect is not long-lasting after use, so the effect is often not ideal; and larger and deeper oral ulcers will form local scars after healing. As time goes by, the scars formed after the ulcer heals will gradually soften, absorb, and shrink, but the local mucosa is difficult to restore to its original state. Summary of the invention
[0004] The purpose of the present invention is to address the deficiencies in the prior art. The present invention provides an oral ulcer spray and a preparation method thereof. The oral ulcer spray provided by the present invention has a long-lasting effect in the oral cavity, a good therapeutic effect, a mild and comfortable feeling when used, and no scars after healing.
[0005] The purpose of the present invention is achieved through the following technical solutions: A mouth ulcer repair spray, comprising a component A and a component B in a volume ratio of 1:1. Component A is made of aloe exosomes, silk fibroin solution, and mussel mucin solution; Component B is composed of the following raw materials in weight percentage: 1.0-1.5wt% of hyacinthus annuus alcohol extract Chamomile extract 1.5-2.0wt% Trehalose 2.0-3.0wt% Sorbitol 2.5-3.5wt% Erythritol 0.5-1.0wt% Polysorbate-800 1.5-2.5wt% Water balance.
[0006] The above-mentioned method for preparing the oral ulcer repair spray comprises the following preparation steps: Step (1): preparing the aloe exosomes, silk fibroin solution and mussel mucin solution in component A respectively; Step (2): 1 part by weight of aloe exosomes and 10 parts by weight of silk fibroin solution are mixed, stirred evenly and allowed to stand for 2-3 hours; Step (3): adding 10-15 parts by weight of mussel mucin solution to the substance obtained in step (2), stirring evenly and then standing for 2-3 hours to obtain liquid A containing exosome microspheres; Step (4): dissolving the chamomile extract, chamomile extract, trehalose, sorbitol, erythritol and polysorbate 800 in water according to the formula amount of component B, stirring evenly, and removing bubbles under vacuum to obtain liquid B; Step (5): Mix the liquid A obtained in step (3) and the liquid B obtained in step (4) in a volume ratio of 1:1, stir evenly, and finally dispense into a spray bottle to obtain an oral ulcer repair spray.
[0007] Preferably, before adding the mussel mucin solution in step (3), the pH is adjusted to 7.0 using a sodium bicarbonate solution with a mass percentage concentration of 5%, so that the mussel mucin can be deposited on the surface of the silk fibroin.
[0008] Wherein, the aloe exosomes in step (1) are prepared by the following method: Step (1.1): put 10 kg of aloe vera into a homogenizer, beat it into a pulp, and then filter it to remove obvious fibers and impurities, and collect the filtrate; Step (1.2): The filtrate is centrifuged at 500 g for 10-15 min at 4°C, the supernatant is collected and centrifuged at 2000 g for 15-20 min at 4°C, the supernatant is collected again and centrifuged at 15000 g for 30-40 min at 4°C, and finally filtered through a 0.22 μm filter membrane to collect the filtrate; Step (1.3): The filtrate is centrifuged at 120,000 g for 60-90 min at 4°C, and the supernatant is discarded; the precipitate is resuspended with PBS, and the precipitate is further centrifuged at 120,000 g for 60-90 min at 4°C, the supernatant is discarded, and the precipitate is resuspended with 50 ml of PBS to obtain aloe exosomes.
[0009] Preferably, the silk fibroin solution in step (1) is prepared by the following method: dissolving the silk fibroin in a PBS buffer solution, and using acetic acid dropwise to adjust the pH of the silk fibroin solution to 4.0, and fixing the volume to a concentration of 1.0-1.5% (w / v) to obtain a silk fibroin solution.
[0010] Preferably, after obtaining the silk fibroin solution in step (1), the method further comprises: adding 1-3 parts by weight of polyethylene glycol diglycidyl ether and 1-3 parts by weight of 1-ethyl-3-(3-methylaminopropyl) carbodiimide (EDC) to 100 parts by weight of the silk fibroin solution, reacting for 30-60 minutes, and dropping 1,2-hexanediol embedding of SOD during the reaction. Ethylene glycol diglycidyl ether and 1-ethyl-3-(3-methylaminopropyl) carbodiimide introduce new functional groups on the silk fibroin, which can be used for further biological modification and attachment of specific targeting molecules or drugs. When SOD is fused to the silk fibroin microspheres, it is beneficial to the stability of SOD, and a more stable system is formed after the subsequent addition of mussel mucin and aloe exosomes.
[0011] Preferably, the mussel mucin solution in step (1) is prepared by the following method: dissolving the mussel mucin in a PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise using acetic acid, and fixing the volume to a final concentration of 0.5-1.2% (w / v).
[0012] Preferably, the concentration of PBS buffer is 0.01M.
[0013] As a natural nanovesicle structure, the aloe exosomes of the present invention have good biocompatibility, can upregulate the expression of antioxidant enzymes to remove free radicals, help eliminate the inflammatory response in the wound repair process, and restore the normal redox level of the skin. Silk protein is a natural polymer protein that can promote cell growth, inhibit bacterial activity, and has good moisture retention. It also has good biocompatibility and biodegradability. The present invention encapsulates aloe exosomes by modified silk protein nanocapsules to improve the stability of aloe exosomes. Mussel mucin (MAP) contains lysine and dopa, which can form a microscopic biological scaffold with positive charge in water isolation and adhesion, attracting negatively charged cells, such as epidermal cells, fibroblasts, endothelial cells, nerve cells, etc., to quickly adhere to the wall, crawl, replace and grow, and promote wound healing. After adding mussel mucin, mussel mucin will automatically deposit on the surface of silk protein, and its dopa group will self-crosslink to form a mesh structure, which will reinforce the silk protein shell and make the microspheres more stable.
[0014] The present invention selects two kinds of traditional Chinese medicine components to cooperate with aloe exosomes-silk fibroin-mussel mucin. The alcohol extract of chamomile flower contains a variety of aromatic alcohol compounds such as aromatic alcohol and geraniol. The use of chamomile flower alcohol extract has anti-inflammatory, antipruritic, moisturizing and repairing effects. Chamomile extract Chamomile extract mainly contains active ingredients such as flavonoids, saponins, polysaccharides, volatile oils, etc., which can inhibit the growth of microorganisms such as Staphylococcus aureus, Streptococcus and Candida albicans, and has anti-inflammatory and sedative effects.
[0015] Trehalose is a safe and reliable natural sugar. It is a non-reducing sugar composed of two glucose molecules with an α, α-1, 1-glycosidic bond and has very stable properties. Trehalose forms a unique protective film on the cell surface under harsh environments such as high temperature, high cold, high osmotic pressure and drying and dehydration, effectively protecting the two protein molecules of silk fibroin and mussel mucin of the present invention from denaturation and inactivation, thereby protecting the stability of the entire system.
[0016] Beneficial effects of the present invention: the present invention uses an effective combination of aloe exosomes-silk fibroin-mussel mucin and traditional Chinese medicine ingredients in the oral ulcer repair spray. Aloe exosomes can effectively promote healing. Silk fibroin cooperates with mussel mucin to wrap the aloe exosomes to form exosome microspheres, thereby improving the stability of the aloe exosomes, and then improving the stability of the entire system, which is conducive to the stable and long-lasting effect of various drug effects. The oral ulcer spray of the present invention has a long-lasting effect in the oral cavity, good therapeutic effect, is gentle and comfortable when used, and leaves no scars after healing. DETAILED DESCRIPTION
[0017] The present invention is further described in conjunction with the following examples.
[0018] Example 1
[0019] A mouth ulcer repair spray, comprising a component A and a component B in a volume ratio of 1:1. Component A is made of aloe exosomes, silk fibroin solution, and mussel mucin solution; Component B is composed of the following raw materials in weight percentage: 1.0wt% of hyacinthus annuus alcohol extract Chamomile extract 1.5wt% Trehalose 2.0wt% Sorbitol 2.5wt% Erythritol 0.5wt% Polysorbate-800 1.5wt% Water balance.
[0020] The above-mentioned method for preparing the oral ulcer repair spray comprises the following preparation steps: Step (1): preparing the aloe exosomes, silk fibroin solution and mussel mucin solution in component A respectively; Step (2): 1 part by weight of aloe exosomes and 10 parts by weight of silk fibroin solution were mixed, stirred evenly and allowed to stand for 2 hours; Step (3): adding 10 parts by weight of the mussel mucin solution to the substance obtained in step (2), stirring evenly and then standing for 2 hours to obtain a liquid A containing exosome microspheres, wherein the pH is adjusted to 7.0 with a sodium bicarbonate solution having a mass percentage concentration of 5% before adding the mussel mucin solution; Step (4): dissolving the chamomile extract, chamomile extract, trehalose, sorbitol, erythritol and polysorbate 800 in water according to the formula amount of component B, stirring evenly, and removing bubbles under vacuum to obtain liquid B; Step (5): Mix the liquid A obtained in step (3) and the liquid B obtained in step (4) in a volume ratio of 1:1, stir evenly, and finally dispense into a spray bottle to obtain an oral ulcer repair spray.
[0021] Wherein, the aloe exosomes in step (1) are prepared by the following method: Step (1.1): put 10 kg of aloe vera into a homogenizer, beat it into a pulp, and then filter it to remove obvious fibers and impurities, and collect the filtrate; Step (1.2): The filtrate was centrifuged at 500 g for 10 min at 4°C, the supernatant was collected and centrifuged at 2000 g for 15 min at 4°C, the supernatant was collected again and centrifuged at 15000 g for 30 min at 4°C, and finally filtered through a 0.22 μm filter membrane to collect the filtrate; Step (1.3): The filtrate was centrifuged at 120,000 g for 60 min at 4°C, and the supernatant was discarded; the precipitate was resuspended with PBS, and the precipitate was further centrifuged at 120,000 g for 60 min at 4°C, the supernatant was discarded, and the precipitate was resuspended with 50 ml of PBS to obtain aloe exosomes.
[0022] The silk fibroin solution in step (1) is prepared by the following method: dissolving the silk fibroin in a PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise using acetic acid, and fixing the volume to a concentration of 1.0% (w / v) to obtain a silk fibroin solution.
[0023] Wherein, after obtaining the silk fibroin solution in step (1), the method further comprises: adding 1 part by weight of polyethylene glycol diglycidyl ether and 1 part by weight of 1-ethyl-3-(3-methylaminopropyl)carbodiimide (EDC) to 100 parts by weight of the silk fibroin solution, reacting for 30 minutes, and dropping 1 part by weight of SOD 1,2-hexanediol embedding material during the reaction. The 1,2-hexanediol embedding material of SOD is obtained by drying 4 parts by weight of 1,2-hexanediol in a forced air drying oven at 65°C to remove moisture, adding 1 part by weight of SOD, and mixing in a shaker at -20°C until condensation.
[0024] The mussel mucin solution in step (1) is prepared by the following method: dissolving the mussel mucin in a PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise using acetic acid, and fixing the volume to a final concentration of 0.5% (w / v).
[0025] Example 2
[0026] A mouth ulcer repair spray, comprising a component A and a component B in a volume ratio of 1:1. Component A is made of aloe exosomes, silk fibroin solution, and mussel mucin solution; Component B is composed of the following raw materials in weight percentage: 1.2wt% of hyacinthus annuus alcohol extract Chamomile extract 1.8wt% Trehalose 2.5wt% Sorbitol 3.0wt% Erythritol 0.8wt% Polysorbate-800 2.0wt% Water balance.
[0027] The above-mentioned method for preparing the oral ulcer repair spray comprises the following preparation steps: Step (1): preparing the aloe exosomes, silk fibroin solution and mussel mucin solution in component A respectively; Step (2): 1 part by weight of aloe exosomes and 10 parts by weight of silk fibroin solution were mixed, stirred evenly and allowed to stand for 3 hours; Step (3): add 12 parts by weight of mussel mucin solution to the substance obtained in step (2), stir evenly and let stand for 3 hours to obtain liquid A containing exosome microspheres, and adjust the pH to 7.0 with a sodium bicarbonate solution with a mass percentage concentration of 5% before adding the mussel mucin solution; Step (4): dissolving the chamomile extract, chamomile extract, trehalose, sorbitol, erythritol and polysorbate 800 in water according to the formula amount of component B, stirring evenly, and removing bubbles under vacuum to obtain liquid B; Step (5): Mix the liquid A obtained in step (3) and the liquid B obtained in step (4) in a volume ratio of 1:1, stir evenly, and finally dispense into a spray bottle to obtain an oral ulcer repair spray.
[0028] Wherein, the aloe exosomes in step (1) are prepared by the following method: Step (1.1): put 10 kg of aloe vera into a homogenizer, beat it into a pulp, and then filter it to remove obvious fibers and impurities, and collect the filtrate; Step (1.2): The filtrate was centrifuged at 500 g for 12 min at 4 °C, the supernatant was collected and centrifuged at 2000 g for 18 min at 4 °C, the supernatant was collected again and centrifuged at 15000 g for 35 min at 4 °C, and finally filtered through a 0.22 μm filter membrane to collect the filtrate; Step (1.3): The filtrate was centrifuged at 120,000 g for 70 min at 4°C, and the supernatant was discarded; the precipitate was resuspended with PBS, and the precipitate was further centrifuged at 120,000 g for 70 min at 4°C, the supernatant was discarded, and the precipitate was resuspended with 50 ml of PBS to obtain aloe exosomes.
[0029] The silk fibroin solution in step (1) is prepared by the following method: dissolving the silk fibroin in a PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise using acetic acid, and fixing the volume to a concentration of 1.3% (w / v) to obtain a silk fibroin solution.
[0030] Wherein, after obtaining the silk fibroin solution in step (1), the method further comprises: adding 2 parts by weight of polyethylene glycol diglycidyl ether and 2 parts by weight of 1-ethyl-3-(3-methylaminopropyl)carbodiimide (EDC) to 100 parts by weight of the silk fibroin solution, reacting for 50 minutes, and dropping 2 parts by weight of SOD 1,2-hexanediol embedding material during the reaction. The SOD 1,2-hexanediol embedding material is obtained by drying 4 parts by weight of 1,2-hexanediol in a forced air drying oven at 65°C to remove moisture, adding 1 part by weight of SOD, and mixing in a shaker at -20°C until condensation.
[0031] The mussel mucin solution in step (1) is prepared by the following method: dissolving the mussel mucin in a PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise using acetic acid, and fixing the volume to a final concentration of 0.8% (w / v).
[0032] Example 3
[0033] A mouth ulcer repair spray, comprising a component A and a component B in a volume ratio of 1:1. Component A is made of aloe exosomes, silk fibroin solution, and mussel mucin solution; Component B is composed of the following raw materials in weight percentage: 1.5wt% of hyacinthus annuus alcohol extract Chamomile extract 2.0wt% Trehalose 3.0wt% Sorbitol 3.5wt% Erythritol 1.0wt% Polysorbate-800 2.5wt% Water balance.
[0034] The above-mentioned method for preparing the oral ulcer repair spray comprises the following preparation steps: Step (1): preparing the aloe exosomes, silk fibroin solution and mussel mucin solution in component A respectively; Step (2): 1 part by weight of aloe exosomes and 10 parts by weight of silk fibroin solution were mixed, stirred evenly and allowed to stand for 3 hours; Step (3): adding 15 parts by weight of mussel mucin solution to the substance obtained in step (2), stirring evenly and then standing for 3 hours to obtain liquid A containing exosome microspheres; Step (4): dissolving the chamomile extract, chamomile extract, trehalose, sorbitol, erythritol and polysorbate 800 in water according to the formula amount of component B, stirring evenly, and removing bubbles under vacuum to obtain liquid B; Step (5): Mix the liquid A obtained in step (3) and the liquid B obtained in step (4) in a volume ratio of 1:1, stir evenly, and finally dispense into a spray bottle to obtain an oral ulcer repair spray.
[0035] Wherein, the aloe exosomes in step (1) are prepared by the following method: Step (1.1): put 10 kg of aloe vera into a homogenizer, beat it into a pulp, and then filter it to remove obvious fibers and impurities, and collect the filtrate; Step (1.2): The filtrate was centrifuged at 500 g for 15 min at 4°C, the supernatant was collected and centrifuged at 2000 g for 20 min at 4°C, the supernatant was collected again and centrifuged at 15000 g for 40 min at 4°C, and finally filtered through a 0.22 μm filter membrane to collect the filtrate; Step (1.3): The filtrate was centrifuged at 120,000 g for 90 min at 4°C, and the supernatant was discarded; the precipitate was resuspended with PBS, and the precipitate was further centrifuged at 120,000 g for 90 min at 4°C, and the supernatant was discarded. The precipitate was resuspended with 50 ml of PBS to obtain aloe exosomes.
[0036] The silk fibroin solution in step (1) is prepared by the following method: dissolving the silk fibroin in a PBS buffer solution, and using acetic acid to adjust the pH of the silk fibroin solution to 4.0 dropwise, and fixing the volume to a concentration of 1.5% (w / v) to obtain a silk fibroin solution.
[0037] Wherein, after obtaining the silk fibroin solution in step (1), the method further comprises: adding 3 parts by weight of polyethylene glycol diglycidyl ether and 3 parts by weight of 1-ethyl-3-(3-methylaminopropyl)carbodiimide (EDC) to 100 parts by weight of the silk fibroin solution, reacting for 60 minutes, and dropping 3 parts by weight of SOD 1,2-hexanediol embedding material during the reaction. The SOD 1,2-hexanediol embedding material is obtained by drying 4 parts by weight of 1,2-hexanediol in a forced air drying oven at 65°C to remove moisture, adding 1 part by weight of SOD, and mixing in a shaker at -20°C until condensation.
[0038] The mussel mucin solution in step (1) is prepared by the following method: dissolving the mussel mucin in a PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise using acetic acid, and fixing the volume to a final concentration of 1.2% (w / v).
[0039] Comparative Example 1
[0040] The difference between this comparative example and Example 2 is that the silk fibroin is not modified in this example.
[0041] A mouth ulcer repair spray, comprising a component A and a component B in a volume ratio of 1:1. Component A is made of aloe exosomes, silk fibroin solution, and mussel mucin solution; Component B is composed of the following raw materials in weight percentage: 1.2wt% of hyacinthus annuus alcohol extract Chamomile extract 1.8wt% Trehalose 2.5wt% Sorbitol 3.0wt% Erythritol 0.8wt% Polysorbate-800 2.0wt% Water balance.
[0042] The above-mentioned method for preparing the oral ulcer repair spray comprises the following preparation steps: Step (1): preparing the aloe exosomes, silk fibroin solution and mussel mucin solution in component A respectively; Step (2): 1 part by weight of aloe exosomes and 10 parts by weight of silk fibroin solution were mixed, stirred evenly and allowed to stand for 3 hours; Step (3): add 12 parts by weight of mussel mucin solution to the substance obtained in step (2), stir evenly and let stand for 3 hours to obtain liquid A containing exosome microspheres, and adjust the pH to 7.0 with a sodium bicarbonate solution with a mass percentage concentration of 5% before adding the mussel mucin solution; Step (4): dissolving the chamomile extract, chamomile extract, trehalose, sorbitol, erythritol and polysorbate 800 in water according to the formula amount of component B, stirring evenly, and removing bubbles under vacuum to obtain liquid B; Step (5): Mix the liquid A obtained in step (3) and the liquid B obtained in step (4) in a volume ratio of 1:1, stir evenly, and finally dispense into a spray bottle to obtain an oral ulcer repair spray.
[0043] Wherein, the aloe exosomes in step (1) are prepared by the following method: Step (1.1): put 10 kg of aloe vera into a homogenizer, beat it into a pulp, and then filter it to remove obvious fibers and impurities, and collect the filtrate; Step (1.2): The filtrate was centrifuged at 500 g for 12 min at 4 °C, the supernatant was collected and centrifuged at 2000 g for 18 min at 4 °C, the supernatant was collected again and centrifuged at 15000 g for 35 min at 4 °C, and finally filtered through a 0.22 μm filter membrane to collect the filtrate; Step (1.3): The filtrate was centrifuged at 120,000 g for 70 min at 4°C, and the supernatant was discarded; the precipitate was resuspended with PBS, and the precipitate was further centrifuged at 120,000 g for 70 min at 4°C, the supernatant was discarded, and the precipitate was resuspended with 50 ml of PBS to obtain aloe exosomes.
[0044] The silk fibroin solution in step (1) is prepared by the following method: dissolving the silk fibroin in a PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise using acetic acid, and fixing the volume to a concentration of 1.3% (w / v) to obtain a silk fibroin solution.
[0045] The mussel mucin solution in step (1) is prepared by the following method: dissolving the mussel mucin in a PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise using acetic acid, and fixing the volume to a final concentration of 0.8% (w / v).
[0046] Comparative Example 2
[0047] The difference between this comparative example and Example 2 is that this example does not contain mussel mucin.
[0048] A mouth ulcer repair spray, comprising a component A and a component B in a volume ratio of 1:1. Component A is made of aloe exosomes and silk fibroin solution; Component B is composed of the following raw materials in weight percentage: 1.2wt% of hyacinthus annuus alcohol extract Chamomile extract 1.8wt% Trehalose 2.5wt% Sorbitol 3.0wt% Erythritol 0.8wt% Polysorbate-800 2.0wt% Water balance.
[0049] The above-mentioned method for preparing the oral ulcer repair spray comprises the following preparation steps: Step (1): preparing aloe exosomes and silk fibroin solutions in component A respectively; Step (2): 1 part by weight of aloe exosomes and 10 parts by weight of silk fibroin solution are mixed, stirred evenly and allowed to stand for 3 hours to obtain liquid A containing exosome microspheres; Step (3): dissolving the chamomile extract, chamomile extract, trehalose, sorbitol, erythritol and polysorbate 800 in water according to the formula amount of component B, stirring evenly, and removing bubbles under vacuum to obtain liquid B; Step (5): Mix the liquid A obtained in step (2) and the liquid B obtained in step (3) in a volume ratio of 1:1, stir evenly, and finally dispense into a spray bottle to obtain an oral ulcer repair spray.
[0050] Wherein, the aloe exosomes in step (1) are prepared by the following method: Step (1.1): put 10 kg of aloe vera into a homogenizer, beat it into a pulp, and then filter it to remove obvious fibers and impurities, and collect the filtrate; Step (1.2): The filtrate was centrifuged at 500 g for 12 min at 4 °C, the supernatant was collected and centrifuged at 2000 g for 18 min at 4 °C, the supernatant was collected again and centrifuged at 15000 g for 35 min at 4 °C, and finally filtered through a 0.22 μm filter membrane to collect the filtrate; Step (1.3): The filtrate was centrifuged at 120,000 g for 70 min at 4°C, and the supernatant was discarded; the precipitate was resuspended with PBS, and the precipitate was further centrifuged at 120,000 g for 70 min at 4°C, the supernatant was discarded, and the precipitate was resuspended with 50 ml of PBS to obtain aloe exosomes.
[0051] The silk fibroin solution in step (1) is prepared by the following method: dissolving the silk fibroin in a PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise using acetic acid, and fixing the volume to a concentration of 1.3% (w / v) to obtain a silk fibroin solution.
[0052] Wherein, after obtaining the silk fibroin solution in step (1), the method further comprises: adding 2 parts by weight of polyethylene glycol diglycidyl ether and 2 parts by weight of 1-ethyl-3-(3-methylaminopropyl)carbodiimide (EDC) to 100 parts by weight of the silk fibroin solution, reacting for 50 minutes, and dropping 2 parts by weight of SOD 1,2-hexanediol embedding material during the reaction. The SOD 1,2-hexanediol embedding material is obtained by drying 4 parts by weight of 1,2-hexanediol in a forced air drying oven at 65°C to remove moisture, adding 1 part by weight of SOD, and mixing in a shaker at -20°C until condensation.
[0053] Experimental part:
[0054] 1. Stability test: The products prepared in Examples 1-3 and Comparative Examples 1-2 were subjected to periodic placement tests. The results are shown in Table 1.
[0055] Table 1: Stability test
[0056] The results of the stability test showed that comparative example 1 also had precipitation and stratification at room temperature, indicating that aloe exosomes had better stability, and the system stability was greatly reduced due to the lack of silk fibroin modification; comparative example 2 had precipitation and stratification at 48°C and 4°C, and its performance was better than comparative example 1, but worse than that of examples 1-3, indicating that the system stability was also reduced due to the lack of mussel mucin.
[0057] 2. Application of oral ulcer repair spray in skin damage repair Male BALB / c mice aged 6-8 weeks were randomly divided into 6 groups, a blank group (saline group), and 10 mice in each group. On day 0, the mice were anesthetized with 5% chloral hydrate, and two circular wounds with a diameter of 10 mm were created on the back of the mice with a skin puncher. The corresponding drug formulation was applied to the wound once a day, 100 microliters per wound, covering the entire wound, for 7 consecutive days, and then the drug was stopped and observed for 7 days. During the experiment, the wound changes were recorded every 2 days, and the wound healing rate was finally calculated. The wound healing rate (%) = (original wound area - unhealed wound area) / original wound area × 100%, and the results are shown in Table 2.
[0058] Table 2: Wound healing rate
[0059] Number of days Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Blank group Day 1 0 0 0 0 0 0 Day 3 23% 24% 23% 15% 18% 14% Day 7 62% 63% 61% 50% 54% 47% Day 10 91% 92% 91% 82% 87% 77% Day 14 98% 98% 98% 90% 95% 86% The experimental data results show that the wound healing rate of the rats treated with the oral ulcer repair spray modified with silk fibroin in Example 1-3 was significantly higher than that of the control group 1, and the wound healing rate of the rats treated with the oral ulcer repair spray modified with mussel mucin in Example 1-3 was significantly higher than that of the control group 2.
[0060] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.
Claims
1. An oral ulcer repair spray, characterized in that: It includes component A and component B in a volume ratio of 1:
1. Component A is made of aloe exosomes, silk fibroin solution, and mussel mucin solution; Component B is composed of the following raw materials in weight percentage: 1.0-1.5wt% of hyacinthus annuus alcohol extract Chamomile extract 1.5-2.0wt% Trehalose 2.0-3.0wt% Sorbitol 2.5-3.5wt% Erythritol 0.5-1.0wt% Polysorbate-800 1.5-2.5wt% Water balance.
2. A method for preparing an oral ulcer repair spray as claimed in claim 1, characterized in that: It includes the following preparation steps: Step (1): preparing the aloe exosomes, silk fibroin solution and mussel mucin solution in component A respectively; Step (2): 1 part by weight of aloe exosomes and 10 parts by weight of silk fibroin solution are mixed, stirred evenly and allowed to stand for 2-3 hours; Step (3): adding 10-15 parts by weight of mussel mucin solution to the substance obtained in step (2), stirring evenly and then standing for 2-3 hours to obtain liquid A containing exosome microspheres; Step (4): dissolving the chamomile extract, chamomile extract, trehalose, sorbitol, erythritol and polysorbate 800 in water according to the formula amount of component B, stirring evenly, and removing bubbles under vacuum to obtain liquid B; Step (5): Mix the liquid A obtained in step (3) and the liquid B obtained in step (4) in a volume ratio of 1:1, stir evenly, and finally dispense into a spray bottle to obtain an oral ulcer repair spray.
3. The method for preparing an oral ulcer repair spray according to claim 1, characterized in that: The aloe exosomes in step (1) are prepared by the following method: Step (1.1): put 10 kg of aloe vera into a homogenizer, beat it into a pulp, and then filter it to remove obvious fibers and impurities, and collect the filtrate; Step (1.2): The filtrate is centrifuged at 500 g for 10-15 min at 4°C, the supernatant is collected and centrifuged at 2000 g for 15-20 min at 4°C, the supernatant is collected again and centrifuged at 15000 g for 30-40 min at 4°C, and finally filtered through a 0.22 μm filter membrane to collect the filtrate; Step (1.3): The filtrate is centrifuged at 120,000 g for 60-90 min at 4°C, and the supernatant is discarded; the precipitate is resuspended with PBS, and the precipitate is further centrifuged at 120,000 g for 60-90 min at 4°C, the supernatant is discarded, and the precipitate is resuspended with 50 ml of PBS to obtain aloe exosomes.
4. The method for preparing an oral ulcer repair spray according to claim 1, characterized in that: The silk fibroin solution in step (1) is prepared by the following method: dissolving silk fibroin in PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise with acetic acid, and fixing the volume to a concentration of 1.0-1.5% (w / v) to obtain a silk fibroin solution.
5. The method for preparing an oral ulcer repair spray according to claim 4, characterized in that: After obtaining the silk fibroin solution in step (1), the method further comprises: adding 1-3 parts by weight of polyethylene glycol diglycidyl ether and 1-3 parts by weight of 1-ethyl-3-(3-methylaminopropyl)carbodiimide (EDC) to 100 parts by weight of the silk fibroin solution, reacting for 30-60 minutes, and adding dropwise 1,2-hexanediol-embedded SOD during the reaction.
6. The method for preparing an oral ulcer repair spray according to claim 1, characterized in that: The mussel mucin solution in step (1) is prepared by the following method: dissolving the mussel mucin in a PBS buffer solution, and adjusting the pH of the silk fibroin solution to 4.0 dropwise using acetic acid to a final concentration of 0.5-1.2% (w / v).