Gel for repairing endometrial injury and preparation method thereof

Abstract

The invention discloses an endometrial injury repairing gel and a preparation method thereof, and relates to the field of gels, chitosan and acetic acid are mixed and stirred to obtain a chitosan solution, hyaluronic acid and poloxamer are added, then thermal response particles and a traditional Chinese medicine compound extract containing tea polyphenol are added, and standing is performed to obtain the endometrial injury repairing gel, the tea polyphenol is added and introduced into the traditional Chinese medicine compound extract, phenolic hydroxyl groups of the tea polyphenol interact with hydroxyl groups and carboxyl groups in hyaluronic acid, and nanofibers are combined, so that a three-dimensional network structure of the gel becomes more compact, the mechanical property is improved, steric hindrance is increased, and the enzymolysis resistance of the gel is improved. Besides, thermal response particles with specific structures, which are formed by fibers and nanoparticles, are added, so that the hydrogel has temperature-sensitive response, the hydrogel has a more stable gel structure at the in-vivo temperature of 35-40 DEG C, and the adhesion of the hydrogel to an affected part is greatly improved, so that the medicinal effect of active substances in the traditional Chinese medicine extract is optimized.

Description

Technical Field

[0001] The invention relates to the field of gels, and in particular to a gel for repairing endometrial damage and a preparation method thereof. Background Art

[0002] Endometrial damage refers to the damage to the base of the endometrium due to external factors, which causes the endometrium to lose its repair function. At this time, even if the ovarian function is normal, women will not have menstruation, which seriously affects pregnancy.

[0003] Causes of endometrial damage:

[0004] 1. Severe endocrine disorders will lead to hormone imbalance in the body, disrupt the cyclical changes of the endometrium, and cause endometrial damage.

[0005] 2. Inflammatory infection, such as inflammation in the uterine cavity. Long-term inflammatory damage can damage the endometrium and ultimately affect women's pregnancy.

[0006] 3. Artificial abortion is the main factor of endometrial damage. Multiple medical abortions and surgical abortions may cause endometrial damage. Some patients even stop menstruating after abortion, which shows that abortion seriously damages the endometrium. The main methods for repairing damaged endometrium include general treatment, drug treatment, surgical treatment and other methods.

[0007] Among them, endometritis is one of the common clinical diseases in obstetrics, which often affects the health of patients. The pathogens of endometritis are mainly Escherichia coli, Streptococcus, Pseudomonas aeruginosa, and Campylobacter. It can also be followed by Burkholderia, Salmonella and other infectious diseases that invade the reproductive system. If not treated in time, it can lead to uterine pyometra, hydrops, adhesion of the uterus and surrounding tissues, inflammation of the fallopian tubes and other diseases.

[0008] At present, vaginal suppositories, lavages, and oral tablets have been reported as endometrial repair preparations, but their efficacy is poor. The main reason is that these existing preparations cannot ensure that the therapeutic drug is in close contact with the endometrial damage site and maintain an effective concentration for a long time. In addition, due to the rapid secretion and renewal of endometrial mucus, the therapeutic drug in the damaged uterine cavity is quickly lost, and it is impossible to achieve an effective drug treatment concentration. These factors have become bottlenecks that hinder the application of current preparations in endometrial damage repair. Summary of the invention

[0009] The purpose of the present invention is to solve at least one of the technical problems existing in the prior art and to provide a gel for repairing endometrial damage and a preparation method thereof.

[0010] The technical solution of the present invention is as follows:

[0011] The present invention discloses a method for preparing a gel for repairing endometrial damage.

[0012] Chitosan and acetic acid are mixed and stirred to obtain a chitosan solution, hyaluronic acid and poloxamer are added, and then the thermal response particles and the traditional Chinese medicine composite extract containing tea polyphenols are added, and the mixture is allowed to stand to obtain;

[0013] The traditional Chinese medicine composite extract comprises the following raw materials in parts by weight: 1-10 parts of saffron, 10-20 parts of seabuckthorn seeds, 5-8 parts of cnidium monnieri, 5-15 parts of fennel, 5-16 parts of dried ginger, 3-15 parts of cinnamon, 10-22 parts of angelica, 5-17 parts of chuanxiong, 3-13 parts of red peony root, 7-24 parts of myrrh, 8-21 parts of corydalis, 8-21 parts of lindera, 7-21 parts of pollen and 5-20 parts of tea.

[0014] As a preferred embodiment of the present invention, the preparation method of the thermally responsive particles is as follows:

[0015] The plant fiber and calcium hydroxide powder are mixed, water is added and stirred, a cationic surfactant is added at the same time, and then carbon dioxide is introduced at 1-5Mpa for reaction to obtain the product.

[0016] As a preferred embodiment of the present invention, the cationic surfactant is added in an amount of 1-5 wt % of the total mass of the mixture of plant fiber and calcium hydroxide powder.

[0017] As a preferred embodiment of the present invention, the amount of hyaluronic acid added is 3-5wt% of the chitosan solution.

[0018] As a preferred embodiment of the present invention, the added amount of the poloxamer accounts for 10-15wt% of the chitosan solution.

[0019] As a preferred embodiment of the present invention, the aspect ratio of the plant fiber is 5-8:1.

[0020] As a preferred embodiment of the present invention, the preparation method of the Chinese medicine composite extract is as follows:

[0021] The raw material is crushed, refluxed or impregnated with 6-12 times water or 50%-95% ethanol or other acceptable solvents for 1-3 times, each time for 0.5-2h, the filtrate is combined, passed through a 100-600 mesh filter cloth, and concentrated at 55℃-85℃ to a thick paste with a relative density of 1.05-1.2.

[0022] The present invention also discloses a gel for repairing endometrial damage, which is prepared by any of the above preparation methods.

[0023] The beneficial effects of the present invention are:

[0024] (1) The present invention introduces tea polyphenols into the composite extract of traditional Chinese medicine. The phenolic hydroxyl groups of tea polyphenols interact with the hydroxyl groups and carboxyl groups in hyaluronic acid and combine with nanofibers to make the three-dimensional network structure of the gel more compact and improve the mechanical properties. On the other hand, the steric hindrance is increased, the enzymatic reaction is reduced, and the enzymatic resistance of the gel is improved, thereby optimizing the medicinal effects of the active substances in the traditional Chinese medicine extract and improving the repair ability of the endometrium.

[0025] (2) The presence of nanoparticles can change the microenvironment of plant fibers, affect the attachment and growth of bacteria and fungi, inhibit the activity of microorganisms, release ions that promote microbial proliferation, and enhance fiber degradation. It can also enhance the stability of the gel. In addition, the specific structure formed by the fibers and nanoparticles has a temperature-sensitive response to the hydrogel. By adjusting the amount added, it can have a more stable gel structure at a body temperature of 35-40°C, greatly improving its attachment to the affected area and the medicinal properties of the active substances, thereby improving the repair ability of the endometrium.

[0026] (3) The present invention effectively controls the mechanical strength and degradation rate of the hydrogel for repairing endometrial damage by changing the composition ratio of the Chinese herbal extract and the thermal response particles. The hydrogel for repairing endometrial damage has low immunogenicity and has multiple advantages of rapid repair of the endometrium. It provides the best physical, chemical and biological microenvironment for endometrial repair, effectively guides the directional arrangement of cells, has good mechanical properties, and achieves rapid repair of endometrial damage. Therefore, the present invention adopts pure Chinese herbal extracts, combines thermal response particles, and selects a gel prepared by a suitable composition ratio, which can effectively improve the abnormal symptoms of the endometrium, increase the HPV conversion rate and shorten the conversion time, reduce the level of inflammatory cytokines, effectively improve the microenvironment of the affected area, and improve the repair ability of the endometrium. DETAILED DESCRIPTION

[0027] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. If no specific techniques or conditions are specified in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used that do not specify the manufacturer are all conventional products that can be obtained commercially.

[0028] The preparation method of the cationic surfactant in the embodiment is as follows:

[0029] Take 30g of anhydrous glucose, 44g of 3-chloro-1,2-propylene glycol and 0.3g of p-toluenesulfonic acid. React for 50min at a pressure of 200mmHg and a temperature of 90°C to obtain a light yellow transparent viscous liquid, which is neutralized to pH≈7 with a saturated NaOH aqueous solution. Then add the neutralized solution to a molecular distiller, control the temperature to 100°C, and the pressure to 1mba to remove excess 3-chloro-1,2-propylene glycol. Obtain solid 3-chloro-2-hydroxypropyl glycoside. Dissolve the chloroglycoside in water and prepare a 50wt% aqueous solution. Add 65g of the above chloroglycoside solution, stir and heat. When the reaction temperature is 50°C, slowly add a mixed solution of 15.5g of dodecyl dimethyl tertiary amine and 32g of ethanol to the reactor through a feed funnel, and control the addition to be completed within 0.5 hours. At a temperature of 60°C, react for 12 hours to obtain a dodecyl hydroxypropyl glycoside cationic surfactant.

[0030] Example 1

[0031] Chitosan and acetic acid are mixed and stirred to obtain a chitosan solution with a mass concentration of 5wt%, hyaluronic acid and poloxamer are added, and then the thermal response particles and the traditional Chinese medicine composite extract containing tea polyphenols are added, and the mixture is allowed to stand to obtain;

[0032] The Chinese medicine composite extract comprises the following raw materials in parts by weight: 8 parts of saffron, 14 parts of seabuckthorn seeds, 7 parts of cnidium monnieri, 10 parts of fennel, 12 parts of dried ginger, 10 parts of cinnamon bark, 18 parts of angelica sinensis, 15 parts of chuanxiong, 10 parts of red peony root, 20 parts of myrrh, 15 parts of yanhusuo, 14 parts of lindera odorata, 15 parts of pollen pollen and 15 parts of tea.

[0033] The preparation method of the thermal response particles is as follows:

[0034] The plant fiber and calcium hydroxide powder are mixed, water is added and stirred, a cationic surfactant is added at the same time, and then carbon dioxide is introduced at 3Mpa for reaction to obtain the product.

[0035] The added amount of the cationic surfactant is 3wt% of the total mass of the mixture of the plant fiber and the calcium hydroxide powder.

[0036] The added amount of the hyaluronic acid accounts for 3wt% of the chitosan solution.

[0037] The added amount of the poloxamer accounts for 10 wt % of the chitosan solution.

[0038] The plant fiber has an aspect ratio of 8:1 and a particle size of 10 μm in the length direction.

[0039] The thermal response particles and the traditional Chinese medicine composite extract account for 5wt% and 50wt% of the chitosan solution respectively.

[0040] The preparation method of the Chinese medicine composite extract is as follows:

[0041] The raw material was crushed, extracted with 10 times water reflux for 3 times, each time for 1 hour, the filtrates were combined, filtered through a 300-mesh filter cloth, and concentrated at 65°C to a thick paste with a relative density of 1.15.

[0042] Example 2

[0043] Chitosan and acetic acid are mixed and stirred to obtain a chitosan solution with a mass concentration of 5wt%, hyaluronic acid and poloxamer are added, and then the thermal response particles and the traditional Chinese medicine composite extract containing tea polyphenols are added, and the mixture is allowed to stand to obtain;

[0044] The Chinese medicine composite extract comprises the following raw materials in parts by weight: 8 parts of saffron, 14 parts of seabuckthorn seeds, 7 parts of cnidium monnieri, 10 parts of fennel, 12 parts of dried ginger, 10 parts of cinnamon bark, 18 parts of angelica sinensis, 15 parts of chuanxiong, 10 parts of red peony root, 20 parts of myrrh, 15 parts of yanhusuo, 14 parts of lindera odorata, 15 parts of pollen pollen and 15 parts of tea.

[0045] The preparation method of the thermal response particles is as follows:

[0046] The plant fiber and calcium hydroxide powder are mixed, water is added and stirred, a cationic surfactant is added at the same time, and then carbon dioxide is introduced at 3Mpa for reaction to obtain the product.

[0047] The added amount of the cationic surfactant is 3wt% of the total mass of the mixture of the plant fiber and the calcium hydroxide powder.

[0048] The added amount of the hyaluronic acid accounts for 4wt% of the chitosan solution.

[0049] The added amount of the poloxamer accounts for 12 wt % of the chitosan solution.

[0050] The plant fiber has an aspect ratio of 8:1 and a particle size of 10 μm in the length direction.

[0051] The thermal response particles and the traditional Chinese medicine composite extract account for 5wt% and 50wt% of the chitosan solution respectively.

[0052] The preparation method of the Chinese medicine composite extract is as follows:

[0053] The raw material was crushed, refluxed and extracted with 12 times water for 3 times, each time for 1 hour, the filtrates were combined, filtered through a 300-mesh filter cloth, and concentrated at 55°C to a thick paste with a relative density of 1.2.

[0054] Example 3

[0055] Chitosan and acetic acid are mixed and stirred to obtain a chitosan solution with a mass concentration of 5wt%, hyaluronic acid and poloxamer are added, and then the thermal response particles and the traditional Chinese medicine composite extract containing tea polyphenols are added, and the mixture is allowed to stand to obtain;

[0056] The Chinese medicine composite extract comprises the following raw materials in parts by weight: 8 parts of saffron, 14 parts of seabuckthorn seeds, 7 parts of cnidium monnieri, 10 parts of fennel, 12 parts of dried ginger, 10 parts of cinnamon bark, 18 parts of angelica sinensis, 15 parts of chuanxiong, 10 parts of red peony root, 20 parts of myrrh, 15 parts of yanhusuo, 14 parts of lindera odorata, 15 parts of pollen pollen and 15 parts of tea.

[0057] The preparation method of the thermal response particles is as follows:

[0058] The plant fiber and calcium hydroxide powder are mixed, water is added and stirred, a cationic surfactant is added at the same time, and then carbon dioxide is introduced at 4Mpa for reaction to obtain the product.

[0059] The added amount of the cationic surfactant is 3wt% of the total mass of the mixture of the plant fiber and the calcium hydroxide powder.

[0060] The added amount of the hyaluronic acid accounts for 4wt% of the chitosan solution.

[0061] The added amount of the poloxamer accounts for 12 wt % of the chitosan solution.

[0062] The plant fiber has an aspect ratio of 7:1 and a particle size of 10 μm in the length direction.

[0063] The thermal response particles and the traditional Chinese medicine composite extract account for 5wt% and 50wt% of the chitosan solution respectively.

[0064] The preparation method of the Chinese medicine composite extract is as follows:

[0065] The raw material was crushed, extracted with 10 times water reflux for 3 times, each time for 1 hour, the filtrates were combined, filtered through a 300-mesh filter cloth, and concentrated at 65°C to a thick paste with a relative density of 1.15.

[0066] Example 4

[0067] Different from Example 1, the aspect ratio of the plant fiber is 7:1, and the particle size in the length direction is 12 μm.

[0068] Example 5

[0069] Different from Example 1, the aspect ratio of the plant fiber is 5:1, and the particle size in the length direction is 12 μm.

[0070] Comparative Example 1

[0071] Different from Example 1, no thermal responsive particles are added.

[0072] Comparative Example 2

[0073] Different from Example 1, no tea leaves were added.

[0074] The above embodiments and comparative examples were subjected to performance tests, and the test results are shown in Table 1.

[0075] Sample Gel time at 37℃(s) Gel-like for a long time Example 1 45.1 27h Example 2 45.8 25h Example 3 45.7 26h Example 4 45.5 24d Example 5 45.2 25d Comparative Example 1 62.5 16h Comparative Example 2 48.1 14h

[0076] As can be seen from the above table, the performance of the embodiment is better than that of the comparative example. The main reason may be that in the embodiment, tea polyphenols are introduced into the composite extract of traditional Chinese medicine, and the phenolic hydroxyl groups of tea polyphenols interact with the hydroxyl and carboxyl groups in hyaluronic acid, and the nanofibers are combined to make the three-dimensional network structure of the gel more compact, improve the mechanical properties, and on the other hand, increase the steric hindrance, reduce the enzymatic reaction, and improve the enzymatic resistance of the gel, thereby optimizing the medicinal properties of the active substances in the traditional Chinese medicine extract. Furthermore, the presence of nanoparticles can change the plant fiber microenvironment, affect the attachment and growth of bacteria and fungi, inhibit the activity of microorganisms, release ions that promote microbial proliferation, and enhance fiber degradation. And it can enhance the stability of the gel. In addition, the specific structure formed by the fiber and the nanoparticles has a temperature-sensitive response to the hydrogel. By adjusting the amount of addition, it can have a more stable gel structure at a body temperature of 35-40°C, greatly improving its attachment to the affected area, and greatly improving the medicinal properties of the active substances. Furthermore, the present invention can also control the mechanical strength and degradation rate of the hydrogel for repairing endometrial damage by changing the component ratio. The hydrogel for repairing endometrial damage has low immunogenicity and possesses multiple advantages for rapid repair of the endometrium. It provides an optimal physical, chemical and biological microenvironment for endometrial repair, effectively guides directional cell alignment, has good mechanical properties, and achieves rapid repair of endometrial damage.

[0077] The above-mentioned embodiments only express the preferred implementation modes of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the present invention. It should be pointed out that, for ordinary technicians in this field, various other corresponding changes and deformations can be made according to the technical solutions and concepts described above, and all these changes and deformations should fall within the scope of protection of the claims of the present invention.

Claims

1. A method for preparing a gel for repairing endometrial damage, characterized in that: Chitosan and acetic acid are mixed and stirred to obtain a chitosan solution, hyaluronic acid and poloxamer are added, and then the thermal response particles and the traditional Chinese medicine composite extract containing tea polyphenols are added, and the mixture is allowed to stand to obtain; The preparation method of the thermal response particles is as follows: The plant fiber and calcium hydroxide powder are mixed, water is added and stirred, a cationic surfactant is added at the same time, and then carbon dioxide is introduced at 1-5Mpa for reaction to obtain the product.

2. The method for preparing a gel for repairing endometrial damage according to claim 1, characterized in that: The aspect ratio of the plant fiber is 5-8:

1.

3. The method for preparing a gel for repairing endometrial damage according to claim 1, characterized in that: The added amount of the cationic surfactant is 1-5wt% of the total mass of the mixture of the plant fiber and the calcium hydroxide powder.

4. The method for preparing a gel for repairing endometrial damage according to claim 1, characterized in that: The added amount of the hyaluronic acid accounts for 3-5wt% of the chitosan solution.

5. The method for preparing a gel for repairing endometrial damage according to claim 1, characterized in that: The added amount of the poloxamer is 10-15 wt % of the chitosan solution.

6. The method for preparing a gel for repairing endometrial damage according to claim 1, characterized in that: The thermal response particles and the traditional Chinese medicine composite extract account for 3-10wt% and 40-60wt% of the chitosan solution respectively.

7. The method for preparing a gel for repairing endometrial damage according to claim 1, characterized in that: The traditional Chinese medicine composite extract comprises the following raw materials in parts by weight: 1-10 parts of saffron, 10-20 parts of seabuckthorn seeds, 5-8 parts of cnidium monnieri, 5-15 parts of fennel, 5-16 parts of dried ginger, 3-15 parts of cinnamon, 10-22 parts of angelica, 5-17 parts of chuanxiong, 3-13 parts of red peony root, 7-24 parts of myrrh, 8-21 parts of corydalis, 8-21 parts of lindera, 7-21 parts of pollen and 5-20 parts of tea.

8. The method for preparing a gel for repairing endometrial damage according to claim 7, characterized in that: The preparation method of the Chinese medicine composite extract is as follows: The raw material is crushed, refluxed or impregnated with 6-12 times water or 50%-95% ethanol or other acceptable solvents for 1-3 times, each time for 0.5-2h, the filtrate is combined, passed through a 100-600 mesh filter cloth, and concentrated at 55℃-85℃ to a thick paste with a relative density of 1.05-1.

2.

9. A gel for repairing endometrial damage, characterized in that: The method is prepared by any one of claims 1 to 8.