Exosome oral spray and preparation method thereof

Abstract

The invention relates to an exosome oral spray and a preparation method thereof, and relates to the technical field of oral sprays.The exosome oral spray is prepared from, by weight, 0.2-0.7 part of human umbilical cord mesenchymal stem cell exosome, 1-5 parts of natural extract, 8-15 parts of moisturizing solubilizer, 0.1-0.5 part of preservative, 0.1-0.5 part of essence and 78-90 parts of deionized water; the natural extract is obtained by performing water extraction on raw materials of clinacanthus nutans and xanthoceras sorbifolia bunge kernels in a weight ratio of (14-20): (5-11). According to the invention, the human umbilical cord mesenchymal stem cell exosome and the natural extract with specific composition are taken as functional components, and the exosome and the natural extract cooperate with each other and act together, so that the repairing effect of the exosome oral spray on skin injury is remarkably improved, the treatment of common oral diseases such as oral ulcer and oral mucositis is facilitated, and the application is wide.

Description

Technical Field

[0001] The present invention relates to the technical field of oral sprays, and in particular to an exosome oral spray and a preparation method thereof. Background Art

[0002] In the field of oral health, oral diseases such as oral ulcers and oral mucositis are common and prone to recurrence, causing a lot of inconvenience and pain to patients. The occurrence of these diseases is often related to factors such as imbalance in the oral environment, decreased immunity, and bacterial or viral infections. Traditional treatment methods mostly use local medications, such as anti-inflammatory drugs and analgesics, which can temporarily relieve symptoms but often cannot cure them, and long-term use may cause side effects.

[0003] In recent years, with the continuous development of biotechnology, the application of exosomes in the medical field has gradually attracted attention. Exosomes are tiny vesicles secreted by cells, rich in a variety of bioactive substances, such as growth factors, cytokines, etc., and have multiple functions such as promoting cell proliferation, differentiation, tissue repair and immune regulation. Therefore, the application of exosomes in the treatment of oral diseases is expected to promote the repair and regeneration of oral mucosa from the root, improve oral immunity, and thus achieve the purpose of treating oral diseases. However, there are relatively few exosome oral products on the market, and there are some problems, such as poor repair effect on damaged skin and mucosa, which need to be further improved to meet the use needs of oral care products. Summary of the invention

[0004] In order to solve the above problems, the present invention provides an exosome oral spray and a preparation method thereof.

[0005] In a first aspect, the present invention provides an exosome oral spray, which comprises the following components in parts by weight: 0.2-0.7 parts of human umbilical cord mesenchymal stem cell exosomes, 1-5 parts of natural extracts, 8-15 parts of moisturizing solubilizers, 0.1-0.5 parts of preservatives, 0.1-0.5 parts of flavors, and 78-90 parts of deionized water; The natural extract is obtained by water extraction using alligator mouth flower and Xanthoceras sorbifolia seed kernel as raw materials in a weight ratio of (14-20):(5-11).

[0006] Furthermore, the exosome oral spray comprises the following components in parts by weight: 0.4 parts of human umbilical cord mesenchymal stem cell exosomes, 3.5 parts of natural extracts, 12 parts of moisturizing solubilizers, 0.2 parts of preservatives, 0.4 parts of flavors and 83.5 parts of deionized water.

[0007] Furthermore, the weight ratio of the alligator mouth flower to the Xanthoceras sorbifolia seed kernel is 16:9.

[0008] Furthermore, the water extraction method is reduced pressure reflux extraction.

[0009] Furthermore, the working condition parameters of the reduced pressure reflux extraction include: temperature of 60-68°C, pressure of 16-23 kPa, time of 1-2 hours, and number of times of 1-3 times.

[0010] Furthermore, the moisturizing solubilizer is composed of betaine and 1,2-propylene glycol in a weight ratio of (4-7):(1-3).

[0011] Furthermore, the preservative includes sodium benzoate, and the flavor includes blueberry flavor.

[0012] In a second aspect, the present invention provides a method for preparing the exosome oral spray according to any one of the first aspects, the preparation method comprising the following steps: Ultrasonic stirring is performed to mix human umbilical cord mesenchymal stem cell exosomes, natural extracts, moisturizing solubilizers and water to obtain a first mixture; Adding preservatives and flavors to the first mixed material and stirring and mixing to obtain a second mixed material; The second mixed material is homogenized to obtain the exosome oral spray.

[0013] Furthermore, the working condition parameters of the ultrasonic stirring and mixing include: ultrasonic power of 200~400W, stirring speed of 100~200rpm, and time of 5~15 minutes; the working condition parameters of the homogenization treatment include: homogenization pressure of 15~20Mpa, and time of 5~10 minutes.

[0014] The above technical solution provided by the embodiment of the present invention has at least the following advantages compared with the prior art: The embodiment of the present invention provides an exosome oral spray, which uses human umbilical cord mesenchymal stem cell exosomes and a natural extract of a specific composition as effective ingredients. The two cooperate with each other and significantly improve the repair effect of the exosome oral spray on skin damage under the joint action, which is beneficial for the treatment of common oral diseases such as oral ulcers and oral mucositis, and has a wide range of uses. Specifically: 1) The present invention can form a complementary effect by combining exosomes of human umbilical cord mesenchymal stem cells with extracts of alligator mouth flower and Xanthoceras sorbifolia seed kernel. On the one hand, the growth factors and cytokines provided by exosomes can promote cell proliferation and differentiation, and combined with the anti-inflammatory and antioxidant components in natural extracts, enhance the repair effect and can repair damaged oral mucosa faster. On the other hand, the immunomodulatory effect of exosomes and the pharmacological activity of natural extracts complement each other, can more comprehensively regulate the oral microenvironment, enhance the resistance of oral mucosa, and improve immune regulation ability, which can comprehensively improve oral health and prevent the recurrence of diseases such as oral ulcers and oral mucositis.

[0015] 2) The present invention combines the moisturizing and protective effects of natural extracts with the repairing effects of exosomes, and the raw materials are green, natural and safe, making the oral spray more comfortable during use, reducing pain and discomfort, and helping patients to persist in using it, thereby improving the treatment effect. DETAILED DESCRIPTION

[0016] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0017] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0018] In a first aspect, the present invention provides an exosome oral spray, which comprises the following components in parts by weight: 0.2-0.7 parts of human umbilical cord mesenchymal stem cell exosomes, 1-5 parts of natural extracts, 8-15 parts of moisturizing solubilizers, 0.1-0.5 parts of preservatives, 0.1-0.5 parts of flavors, and 78-90 parts of deionized water; The natural extract is obtained by water extraction using alligator mouth flower and Xanthoceras sorbifolia seed kernel as raw materials in a weight ratio of (14-20):(5-11).

[0019] The embodiment of the present invention provides an exosome oral spray, which uses human umbilical cord mesenchymal stem cell exosomes and a natural extract of a specific composition as effective ingredients. The two cooperate with each other and significantly improve the repair effect of the exosome oral spray on skin damage under the joint action, which is beneficial for the treatment of common oral diseases such as oral ulcers and oral mucositis, and has a wide range of uses. Specifically: 1) The present invention can form a complementary effect by combining exosomes of human umbilical cord mesenchymal stem cells with extracts of alligator mouth flower and Xanthoceras sorbifolia seed kernel. On the one hand, the growth factors and cytokines provided by exosomes can promote cell proliferation and differentiation, and combined with the anti-inflammatory and antioxidant components in natural extracts, enhance the repair effect and can repair damaged oral mucosa faster. On the other hand, the immunomodulatory effect of exosomes and the pharmacological activity of natural extracts complement each other, can more comprehensively regulate the oral microenvironment, enhance the resistance of oral mucosa, and improve immune regulation ability, which can comprehensively improve oral health and prevent the recurrence of diseases such as oral ulcers and oral mucositis.

[0020] 2) The present invention combines the moisturizing and protective effects of natural extracts with the repairing effects of exosomes, and the raw materials are green, natural and safe, making the oral spray more comfortable during use, reducing pain and discomfort, and helping patients to persist in using it, thereby improving the treatment effect.

[0021] In some specific embodiments, the human umbilical cord mesenchymal stem cell exosomes in the present invention can be directly used, such as Shanghai Fuyu Biotechnology Co., Ltd., product number WM-23GK0032 (mass concentration of 1 mg / 1L, BCA protein concentration of 0.5~1.0 mg / mL) and other commercially available human umbilical cord mesenchymal stem cell exosomes; or they can be made according to the human umbilical cord mesenchymal stem cell exosome preparation process disclosed in the prior art (such as high-speed centrifugation, density gradient centrifugation, etc.), and the following methods can be used specifically: 1. Umbilical cord collection and initial processing: Under sterile conditions, fresh umbilical cords are collected from healthy pregnant women, and the Wharton's jelly tissue in the umbilical cord is carefully separated to avoid contamination; the separated Wharton's jelly tissue is cut into small pieces and evenly attached to the wall of the culture bottle to ensure that the cells can fully adhere to the wall and grow; complete culture medium is added and placed under appropriate culture conditions to allow umbilical cord mesenchymal stem cells to naturally crawl out and proliferate.

[0022] 2. Cell expansion and culture supernatant collection: After the cells grow to a certain density, they are passaged and expanded to obtain a sufficient number of umbilical cord mesenchymal stem cells; the culture medium supernatant of the fifth generation cells is selected for collection, at which time the cell state is stable and the exosome secretion amount is high.

[0023] 3. Preliminary centrifugation to remove residual cells: The collected culture medium supernatant was dispensed into centrifuge tubes and centrifuged at 300G for 10 minutes to remove the remaining live cells in the supernatant; the first supernatant after centrifugation was carefully collected to avoid inhaling the cell pellet.

[0024] 4. Further centrifugation to remove dead cells: The first supernatant was again dispensed into centrifuge tubes and centrifuged at 3000G for 30 minutes to remove dead cells and larger cell debris; the second supernatant after centrifugation was collected to ensure the purity of the exosomes.

[0025] 5. Ultracentrifugation to remove cell debris: Transfer the second supernatant to a clean ultracentrifuge tube and centrifuge at 10,000G for 30 minutes to completely remove cell debris and other impurities; carefully collect the third supernatant after centrifugation, which mainly contains exosomes and other small molecules.

[0026] 6. Final ultracentrifugation to obtain exosomes: The third supernatant was again dispensed into clean ultracentrifuge tubes and ultracentrifuged at 100,000G for 90 minutes to precipitate the exosomes. After the centrifugation, the supernatant was discarded and an appropriate amount of physiological saline was added to each tube to resuspend the precipitate to obtain an exosome resuspension.

[0027] 7. Filtration, sterilization and finished product preparation: The exosome resuspension was transferred to a new centrifuge tube and filtered and sterilized using a 0.22 μm filter to ensure the sterility of the exosomes; the filtered liquid was human umbilical cord mesenchymal stem cell exosomes.

[0028] The crocodile mouth flower in the present invention, scientific name Clinacanthus nutans (Burm. f.) Lindau, is a herbaceous plant of the genus Clinacanthus of the Acanthaceae family, and belongs to a plant with medicinal and edible properties. Modern scientific research has found that the crocodile mouth flower is rich in flavonoids, lupeol, betulin, sitosterol, polypeptides, and a variety of trace elements and vitamins, and these components give the crocodile mouth flower a variety of pharmacological effects, such as improving immunity, anti-tumor, anti-inflammatory and antibacterial, and anti-oxidation. At the same time, the crocodile mouth flower can also lower blood pressure, lower blood lipids, and promote intestinal peristalsis, and has a positive effect on the prevention and treatment of a variety of diseases.

[0029] The Xanthoceras sorbifolia seed kernels of the present invention are made from seeds of Xanthoceras sorbifolia of the Sapindaceae family, through processes such as drying, magnetic separation, shelling, and screening, and are natural plant products that are both medicinal and edible. According to traditional medical records and modern scientific research, Xanthoceras sorbifolia seed kernels have a variety of medicinal effects. For example, Xanthoceras sorbifolia saponins, an extract from Xanthoceras sorbifolia seed kernels, are very important drugs for improving brain function. At the same time, a variety of active ingredients in Xanthoceras sorbifolia seed kernels, such as flavonoids, coumarin compounds, etc., have a variety of pharmacological effects such as antioxidant, anti-tumor, and anti-inflammatory. In addition, Xanthoceras sorbifolia seed kernels are also rich in unsaturated fatty acids and essential fats for the human body, which help to lower cholesterol, soften blood vessels, and prevent cardiovascular diseases.

[0030] In some specific embodiments, the exosome oral spray comprises the following components in parts by weight: 0.4 parts of human umbilical cord mesenchymal stem cell exosomes, 3.5 parts of natural extracts, 12 parts of moisturizing solubilizers, 0.2 parts of preservatives, 0.4 parts of flavors and 83.5 parts of deionized water.

[0031] In some specific embodiments, the weight ratio of the alligator mouth flower to the Xanthoceras sorbifolia seed kernel is 16:9.

[0032] In some specific embodiments, the water extraction method is reduced pressure reflux extraction.

[0033] In some specific embodiments, the working condition parameters of the reduced pressure reflux extraction include: temperature of 60-68°C, pressure of 16-23 kPa, time of 1-2 hours, and number of times of 1-3 times. Preferably, the temperature of the reduced pressure reflux extraction is 65°C, and the pressure of the reduced pressure reflux extraction is 20 kPa.

[0034] In some specific embodiments, the moisturizing solubilizing agent is composed of betaine and 1,2-propylene glycol in a weight ratio of (4-7):(1-3). Preferably, the moisturizing solubilizing agent is composed of betaine and 1,2-propylene glycol in a weight ratio of 5:2.

[0035] In some specific embodiments, the preservative comprises sodium benzoate, and the flavor comprises blueberry flavor.

[0036] In a second aspect, based on the same inventive concept, the present invention provides a method for preparing the exosome oral spray according to any one of the first aspects, the preparation method comprising the following steps: Ultrasonic stirring is performed to mix human umbilical cord mesenchymal stem cell exosomes, natural extracts, moisturizing solubilizers and water to obtain a first mixture; Adding preservatives and flavors to the first mixed material and stirring and mixing to obtain a second mixed material; The second mixed material is homogenized to obtain the exosome oral spray.

[0037] The preparation method of the exosome oral spray provided by the present invention is simple to operate, does not need to rely on any additional special equipment, greatly reduces the production threshold and cost, and is suitable for industrial mass production; the prepared exosome oral spray can seamlessly connect to the existing filling process while maintaining its high activity, and easily achieves efficient filling into the spray bottle, ensuring that the final form of the product is both easy to carry and easy to use. At the same time, the preparation method is based on the exosome oral spray described in any one of the first aspects, and therefore has at least the beneficial effects of the exosome oral spray described in any one of the first aspects, which will not be repeated here.

[0038] In some specific embodiments, the working condition parameters of the ultrasonic stirring and mixing include: ultrasonic power of 200~400W, stirring speed of 100~200rpm, and time of 5~15 minutes; the working condition parameters of the homogenization treatment include: homogenization pressure of 15~20Mpa, and time of 5~10 minutes.

[0039] It should be noted that the components and raw materials involved in the exosome oral spray and the preparation method thereof provided in the embodiments of the present invention, unless otherwise specified or specifically described, can be directly commercially available products or homemade using existing public preparation methods; at the same time, the steps and parameters involved, unless otherwise specified or specifically described, can be carried out according to the existing processing technology of oral sprays or directly using existing equipment, and the present invention document will not repeat them one by one.

[0040] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples that do not specify specific conditions are usually measured according to national standards. If there is no corresponding national standard, then the conditions recommended by the manufacturer are followed.

[0041] Example 1 This example provides an exosome oral spray, which includes the following components in parts by weight: 0.4 parts of human umbilical cord mesenchymal stem cell exosomes, 3.5 parts of natural extracts, 12 parts of moisturizing solubilizers, 0.2 parts of preservatives, 0.4 parts of flavors and 83.5 parts of deionized water; The human umbilical cord mesenchymal stem cell exosomes are selected from Shanghai Fuyu Biotechnology Co., Ltd., with the product number WM-23GK0032; The natural extract is obtained by water extraction of crocodile mouth flower and Xanthoceras sorbifolia seed kernels in a weight ratio of 16:9 as raw materials, and the preparation method thereof comprises the following steps: mixing and crushing the crocodile mouth flower and Xanthoceras sorbifolia seed kernels, and then adding water accounting for 8 times the total weight of the crocodile mouth flower and Xanthoceras sorbifolia seed kernels for vacuum reflux extraction, wherein the vacuum reflux extraction temperature is 65° C., the vacuum reflux extraction pressure is 20 kPa, the vacuum reflux extraction time is 1.5 hours, the vacuum reflux extraction is performed twice, filtering, combining the obtained water extracts, and concentrating under reduced pressure and drying to obtain the natural extract; The moisturizing solubilizing agent is composed of betaine and 1,2-propylene glycol in a weight ratio of 5:2; The preservative is sodium benzoate, and the flavor is blueberry flavor.

[0042] The preparation method of the above-mentioned exosome oral spray comprises the following steps: Ultrasonic stirring is performed to mix human umbilical cord mesenchymal stem cell exosomes, natural extracts, moisturizing solubilizers and water to obtain a first mixture; Adding preservatives and flavors to the first mixed material and stirring and mixing to obtain a second mixed material; homogenizing the second mixed material to obtain the exosome oral spray; The working condition parameters of the ultrasonic stirring and mixing include: ultrasonic power of 300W, stirring speed of 160rpm, and time of 10 minutes; the working condition parameters of the homogenization treatment include: homogenization pressure of 18Mpa, and time of 7 minutes.

[0043] Example 2 This example provides an exosome oral spray, which includes the following components in parts by weight: 0.2 parts of human umbilical cord mesenchymal stem cell exosomes, 1 part of natural extract, 8 parts of moisturizing solubilizer, 0.2 parts of preservative, 0.1 parts of flavor and 78 parts of deionized water; The human umbilical cord mesenchymal stem cell exosomes are selected from Shanghai Fuyu Biotechnology Co., Ltd., with the product number WM-23GK0032; The natural extract is obtained by water extraction of crocodile mouth flower and Xanthoceras sorbifolia seed kernels in a weight ratio of 20:11 as raw materials, and the preparation method thereof comprises the following steps: mixing and crushing the crocodile mouth flower and Xanthoceras sorbifolia seed kernels, and then adding water accounting for 8 times the total weight of the crocodile mouth flower and Xanthoceras sorbifolia seed kernels for vacuum reflux extraction, wherein the vacuum reflux extraction temperature is 60° C., the vacuum reflux extraction pressure is 16 kPa, the vacuum reflux extraction time is 1.5 hours, the vacuum reflux extraction is performed twice, filtering, combining the obtained water extracts, and concentrating under reduced pressure and drying to obtain the natural extract; The moisturizing solubilizing agent is composed of betaine and 1,2-propylene glycol in a weight ratio of 7:3; The preservative is sodium benzoate, and the flavor is blueberry flavor.

[0044] The preparation method of the above-mentioned exosome oral spray comprises the following steps: Ultrasonic stirring is performed to mix human umbilical cord mesenchymal stem cell exosomes, natural extracts, moisturizing solubilizers and water to obtain a first mixture; Adding preservatives and flavors to the first mixed material and stirring and mixing to obtain a second mixed material; homogenizing the second mixed material to obtain the exosome oral spray; The working condition parameters of the ultrasonic stirring and mixing include: ultrasonic power of 400W, stirring speed of 200rpm, and time of 5 minutes; the working condition parameters of the homogenization treatment include: homogenization pressure of 15Mpa, and time of 10 minutes.

[0045] Example 3 This example provides an exosome oral spray, which includes the following components in parts by weight: 0.7 parts of human umbilical cord mesenchymal stem cell exosomes, 5 parts of natural extracts, 15 parts of moisturizing solubilizers, 0.5 parts of preservatives, 0.5 parts of flavors and 90 parts of deionized water; The human umbilical cord mesenchymal stem cell exosomes are selected from Shanghai Fuyu Biotechnology Co., Ltd., with the product number WM-23GK0032; The natural extract is obtained by water extraction of crocodile mouth flower and Xanthoceras sorbifolia seed kernels in a weight ratio of 14:5 as raw materials, and the preparation method thereof comprises the following steps: mixing and crushing the crocodile mouth flower and Xanthoceras sorbifolia seed kernels, and then adding water accounting for 8 times the total weight of the crocodile mouth flower and Xanthoceras sorbifolia seed kernels for vacuum reflux extraction, wherein the vacuum reflux extraction temperature is 68° C., the vacuum reflux extraction pressure is 23 kPa, the vacuum reflux extraction time is 1.5 hours, the vacuum reflux extraction is performed twice, filtering, combining the obtained water extracts, and concentrating under reduced pressure and drying to obtain the natural extract; The moisturizing solubilizing agent is composed of betaine and 1,2-propylene glycol in a weight ratio of 4:1; The preservative is sodium benzoate, and the flavor is blueberry flavor.

[0046] The preparation method of the above-mentioned exosome oral spray comprises the following steps: Ultrasonic stirring is performed to mix human umbilical cord mesenchymal stem cell exosomes, natural extracts, moisturizing solubilizers and water to obtain a first mixture; Adding preservatives and flavors to the first mixed material and stirring and mixing to obtain a second mixed material; homogenizing the second mixed material to obtain the exosome oral spray; The working condition parameters of the ultrasonic stirring and mixing include: ultrasonic power of 200W, stirring speed of 100rpm, and time of 15 minutes; the working condition parameters of the homogenization treatment include: homogenization pressure of 20Mpa, and time of 5 minutes.

[0047] Example 4 This example provides an exosome oral spray and a preparation method thereof, which is different from Example 1 only in that: in the preparation method of the natural extract, the temperature of the reduced pressure reflux extraction is adjusted to 86° C., and the pressure of the reduced pressure reflux extraction is adjusted to 55 kPa; the remaining steps and parameters are the same.

[0048] Comparative Example 1 This example provides an exosome oral spray and a preparation method thereof, which is different from Example 1 only in that: the amount of natural extract added is 0 parts (that is, no natural extract is added and the preparation process of adding natural extract is omitted); the remaining steps and parameters are the same.

[0049] Comparative Example 2 This example provides an exosome oral spray and a preparation method thereof, which is different from Example 1 only in that the natural extract is obtained by water extraction using alligator mouth flower as raw material (i.e., without adding Xanthoceras sorbifolia seed kernel); the remaining steps and parameters are the same.

[0050] Comparative Example 3 This example provides an exosome oral spray and a preparation method thereof, which is different from Example 1 only in that the natural extract is obtained by water extraction using Xanthoceras sorbifolia seed kernels as raw materials (i.e., without adding Alligator Mouth Flower); the remaining steps and parameters are the same.

[0051] Comparative Example 4 This example provides an exosome oral spray and a preparation method thereof, which is different from Example 1 only in that the natural extract is obtained by water extraction using alligator mouth flower and Xanthoceras sorbifolia seed kernels in a weight ratio of 9:25 as raw materials; the remaining steps and parameters are the same.

[0052] Test Case In this example, the exosome oral sprays obtained in the above Examples 1 to 4 and Comparative Examples 1 to 4 were tested for their damaged skin repair effects.

[0053] The test method is as follows: 1. Experimental animals: 6-8 week-old healthy male BALB / c mice were selected as experimental subjects and were first subjected to adaptive feeding for 2 weeks to ensure that the mice were in a stable physiological state.

[0054] 2. Grouping and sample size: The mice were randomly divided into 9 groups, 5 mice in each group, for a total of 45 mice. Among them, 1 group was used as a blank control group (treated with normal saline only, marked as group C), and the remaining 8 groups were experimental groups, which were respectively administered with the exosome oral sprays obtained in Examples 1 to 4 (respectively marked as groups E1 to E4) and Comparative Examples 1 to 4 (respectively marked as groups P1 to P4).

[0055] 3. Wound model establishment: On day 0 of the experiment, mice were anesthetized with 5% chloral hydrate, and then two circular full-thickness skin wounds with a diameter of approximately 10 mm were created symmetrically on both sides of the back of the mice using a skin punch to ensure that the wound conditions of each group of mice were consistent.

[0056] 4. Sample application: From the day the wound is created, apply the corresponding test sample to the mouse wound once a day, applying 100 microliters to each wound each time, ensuring that the sample evenly covers the entire wound area. This process is continued for 3 consecutive days to observe the initial treatment effect.

[0057] 5. Data collection and evaluation: Wound photography record: On the 4th day after the application of the sample, high-definition photography was performed on the back wounds of the mice to record the changes in wound morphology. Wound healing rate calculation: The wound healing rate was calculated using the formula as follows: Wound healing rate (%) = (S1-S2) / S1×100%, where S1 is the initial wound area and S2 is the wound area after the experimental test.

[0058] The test results are shown in Table 1.

[0059] Table 1 As shown in Table 1, compared with the control groups of comparative example 1 (group P1), comparative example 2 (group P2) and comparative example 3 (group P3), the exosome oral sprays developed in the four embodiments of the present invention, namely, embodiment 1 (group E1), embodiment 2 (group E2), embodiment 3 (group E3) and embodiment 4 (group E4), showed higher therapeutic efficiency and better therapeutic effect in promoting wound healing of damaged skin in a short period of time, indicating that the present invention uses human umbilical cord mesenchymal stem cell exosomes and natural extracts of specific composition as effective ingredients, and the two cooperate with each other, thereby greatly improving the treatment and repair ability of the oral spray for skin damage, and can be applied to the treatment of common oral diseases such as oral ulcers and oral mucositis, providing patients with a new and more effective treatment option. Further, by comparing comparative example 4 (group P4) with embodiment 1 (group E1), the skin repair effect of embodiment 1 (group E1) of the present invention is more significant, indicating that the amount of Xanthoceras chinensis seeds in the natural extract should not be too much.

[0060] In addition, in this example, an oral mucosal irritation test was performed on the composition provided in the embodiment of the present invention with reference to the YY / T0127.13-2009 pharmaceutical industry standard. The test results show that the composition provided in the embodiment of the present invention has an oral mucosal tissue reaction grade of 0, that is, there is no tissue irritation reaction, which can meet the use requirements of oral care products.

[0061] In summary, the embodiments of the present invention provide an exosome oral spray and a preparation method thereof, which uses human umbilical cord mesenchymal stem cell exosomes and a natural extract of a specific composition as effective ingredients. The two work synergistically with each other, and the combined effect significantly improves the repair effect of the obtained exosome oral spray on skin damage, which is beneficial for the treatment of common oral diseases such as oral ulcers and oral mucositis, and has a wide range of uses.

[0062] Various embodiments of the present invention may be presented in the form of a range; it should be understood that the description in the form of a range is only for convenience and brevity, and should not be understood as a rigid limitation on the scope of the present invention; therefore, the range description should be considered to have specifically disclosed all possible sub-ranges and single numerical values ​​within the range. For example, the range description from 1 to 6 should be considered to have specifically disclosed sub-ranges, such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as single numbers within the range, such as 1, 2, 3, 4, 5 and 6, which apply regardless of the range. In addition, whenever a numerical range is indicated herein, it is meant to include any cited number (fractional or integer) within the indicated range.

[0063] The foregoing is merely a specific embodiment of the present invention, which enables those skilled in the art to understand or implement the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features claimed herein.

Claims

1. An exosome oral spray, characterized in that: The exosome oral spray comprises the following components in parts by weight: 0.2-0.7 parts of human umbilical cord mesenchymal stem cell exosomes, 1-5 parts of natural extracts, 8-15 parts of moisturizing solubilizers, 0.1-0.5 parts of preservatives, 0.1-0.5 parts of flavors, and 78-90 parts of deionized water; The natural extract is obtained by water extraction using alligator mouth flower and Xanthoceras sorbifolia seed kernel as raw materials in a weight ratio of (14-20):(5-11).

2. The exosome oral spray according to claim 1, characterized in that The exosome oral spray comprises the following components in parts by weight: 0.4 parts of human umbilical cord mesenchymal stem cell exosomes, 3.5 parts of natural extracts, 12 parts of moisturizing solubilizers, 0.2 parts of preservatives, 0.4 parts of flavors and 83.5 parts of deionized water.

3. The exosome oral spray according to claim 1, characterized in that The weight ratio of the alligator mouth flower to the Xanthoceras sorbifolia seed kernel is 16:

9.

4. The exosome oral spray according to claim 1, characterized in that The water extraction method is reduced pressure reflux extraction.

5. The exosome oral spray according to claim 4, characterized in that The working condition parameters of the reduced pressure reflux extraction include: temperature of 60-68° C., pressure of 16-23 kPa, time of 1-2 hours, and number of times of 1-3 times.

6. The exosome oral spray according to claim 1, characterized in that The moisturizing solubilizing agent is composed of betaine and 1,2-propylene glycol in a weight ratio of (4-7):(1-3).

7. The exosome oral spray according to claim 1, characterized in that The preservative includes sodium benzoate, and the flavor includes blueberry flavor.

8. A method for preparing the exosome oral spray according to any one of claims 1 to 7, characterized in that: The preparation method comprises the following steps: Ultrasonic stirring is performed to mix human umbilical cord mesenchymal stem cell exosomes, natural extracts, moisturizing solubilizers and water to obtain a first mixture; Adding preservatives and flavors to the first mixed material and stirring and mixing to obtain a second mixed material; The second mixed material is homogenized to obtain the exosome oral spray.

9. The method for preparing the exosome oral spray according to claim 8, characterized in that: The working condition parameters of the ultrasonic stirring and mixing include: ultrasonic power of 200-400W, stirring speed of 100-200rpm, and time of 5-15 minutes; the working condition parameters of the homogenization treatment include: homogenization pressure of 15-20Mpa, and time of 5-10 minutes.

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