Hyaluronic acid gel microneedle as well as preparation method and application thereof

Abstract

The invention provides a hyaluronic acid gel microneedle as well as a preparation method and application thereof.The hyaluronic acid gel microneedle is prepared from sodium hyaluronate as a raw material, the sodium hyaluronate is dissolved to obtain HA hydrogel, and the HA hydrogel is soaked in an organic solvent and dried after being circularly frozen and unfrozen to obtain the hyaluronic acid gel microneedle. The natural polymer sodium hyaluronate is used as a main raw material, so that the hydrogel is good in biocompatibility, free of toxic and harmful effects on the environment and biodegradable. The micro-needle body and the backing layer are both hyaluronic acid, have biocompatibility and degradability, have great application prospects in the biological medicine fields of drug delivery, microsampling, biosensing, local disease treatment, medical cosmetology and the like, and can be used for preparing carriers for loading drugs.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological medicine, and particularly relates to a hyaluronic acid gel microneedle and a preparation method and application thereof. BACKGROUND

[0002] As a new type of transdermal drug delivery system and biological detection platform, microneedles can penetrate the skin barrier painlessly or with low pain, and can enhance drug delivery efficiency and reduce side effects, thus having great potential in precision medicine and customized treatment. The main materials of microneedles have evolved from metal materials to biodegradable polymers (such as polylactic acid and polyglycolic acid), hydrogels, ceramics and bioactive materials. With the aid of advanced micro-nano manufacturing technologies such as 3D printing and photolithography, microneedle arrays with complex structures and different functions are manufactured to carry and release drugs on demand according to different application scenarios, thus meeting different drug delivery needs. At present, microneedle technology has made substantial progress in vaccine inoculation, diabetes management (such as insulin delivery), pain management (such as fentanyl patches), cosmetic and skin care (such as hyaluronic acid injection), disease diagnosis (such as continuous blood glucose monitoring), and has also been explored for tumor targeted therapy, anti-aging product development and non-invasive collection of biomarkers. However, some microneedle materials may have potential biological safety and immunogenicity problems, such as triggering immune response or chronic inflammation, and further efforts are needed to evaluate the biocompatibility and toxicity of long-term use of the materials. It is of great significance to develop new microneedle materials with good biocompatibility, good degradability and strong drug loading capacity, and more sophisticated and efficient micro-nano manufacturing technologies in complex and diversified biomedical application fields.

[0003] Hyaluronic acid (HA) is a water-soluble polysaccharide of biological origin, which widely exists in human cytoplasm and plays a role in lubricating joints and cell metabolism sites. It is also a natural moisturizing agent and has certain antibacterial and anti-inflammatory effects and promotes wound healing. It exhibits excellent biocompatibility and degradability and has achieved a large number of commercialization in the field of biological medicine.

[0004] Chinese Patent CN116869916A discloses a microneedle with a needle body made of a mixture of hyaluronic acid and polyvinylpyrrolidone. Chinese Patent CN115554277A discloses a microneedle with a needle body made of a hyaluronic acid aqueous solution containing zinc / cerium composite nanomaterials. Chinese Patent CN115475325A discloses a microneedle with a needle body made of a mixture of tranexamic acid and hyaluronic acid. The use of single hyaluronic acid material as the only component of the needle body and backing layer, and the preparation of hyaluronic acid gel microneedles from hyaluronic acid hydrogel without chemical cross-linking agent, has never been disclosed worldwide. SUMMARY

[0005] The technical problem solved by the present application is to overcome the deficiencies in the prior art, in particular the insufficient strength of the dry gel and aerogel obtained after drying the hyaluronic acid gel, and to provide a hyaluronic acid gel microneedle material, a preparation method therefor, and applications thereof.

[0006] The present application provides a hyaluronic acid gel microneedle, which is prepared by dissolving sodium hyaluronate to obtain a HA hydrogel, and then soaking the HA hydrogel in an organic solvent after cyclic freezing and thawing, and drying to obtain a hyaluronic acid gel microneedle.

[0007] Based on a general technical concept, the present application further provides a preparation method of the hyaluronic acid gel microneedle, which comprises the following steps:

[0008] S1, adding sodium hyaluronate to water to obtain a solution;

[0009] S2, adjusting the solution to be acidic to obtain a HA hydrogel;

[0010] S3, subjecting the HA hydrogel to a cyclic freezing-thawing operation;

[0011] S4, soaking the thawed HA hydrogel in an organic solution and drying to obtain a hyaluronic acid gel microneedle.

[0012] The preparation method described above, further, the water in the S1 is deionized water. The deionized water does not contain metal ion impurities and bacteria, and has better effect. Ordinary water can also be prepared.

[0013] The preparation method described above, further, the concentration of sodium hyaluronate in the solution in the S1 is 20 mg / mL to 500 mg / mL.

[0014] The preparation method described above, further, the solution in the S2 is adjusted to a pH of 2 to 3.

[0015] The preparation method described above, further, the cyclic freezing-thawing operation in the S3 is specifically: freezing the HA hydrogel at-20℃ in a refrigerator for 12 hours, thawing at room temperature of 25℃ for 12 hours, and repeating the freezing and thawing operation for 5 times. Freezing and thawing can enhance the strength of the gel, and the more times the better the effect.

[0016] The preparation method described above, further, the type of organic solvent is not limited, and the purpose of the organic solvent is to enhance the strength of the gel.

[0017] Based on a general technical concept, the present application provides a hyaluronic acid gel microneedle prepared by the preparation method, for use in a carrier for loading drugs.

[0018] Compared with the prior art, the present application has the advantages of:

[0019] (1) The present application provides a hyaluronic acid gel microneedle, which is mainly made of sodium hyaluronate as a natural polymer. The raw material has good biocompatibility and no toxic and harmful effects on the environment, and is biodegradable. The microneedle body and the backing layer are both hyaluronic acid, which has biocompatibility and biodegradability, and has great application prospects in the fields of drug delivery, microsampling, biosensing, local disease treatment, medical cosmetology and other biomedical fields.

[0020] (2) The present application provides a preparation method of a hyaluronic acid gel microneedle. The preparation process is simple and reliable. The addition of acid can make the sodium hyaluronate solution form a hyaluronic acid hydrogel, and the mechanical properties of the gel can be further enhanced after multiple freeze-thaw cycles. The strength of the prepared hyaluronic acid dry gel microneedle is sufficient to penetrate the skin of ordinary humans. BRIEF DESCRIPTION OF DRAWINGS

[0021] In order to make the purpose, technical scheme and advantages of the embodiments of the present application clearer, the technical scheme in the embodiments of the present application will be described clearly and completely below in combination with the drawings in the embodiments of the present application.

[0022] Figure 1 Figure 1 is a picture of the appearance of the hyaluronic acid gel microneedle in the present application.

[0023] Figure 2 Figure 2 is the scanning electron microscope result of the hyaluronic acid gel microneedle in the present application.

[0024] Figure 3 Figure 3 is the infrared spectrum detection spectrum of the hyaluronic acid gel microneedle in the present application.

[0025] Figure 4 Figure 4 is the aluminum foil puncture experiment result of the hyaluronic acid microneedle in the present application.

[0026] Figure 5 Figure 5 is the cling film puncture experiment result of the hyaluronic acid microneedle in the present application.

[0027] Figure 6 Figure 6 is the autolysis experiment result of the hyaluronic acid microneedle in the present application. DETAILED DESCRIPTION

[0028] The present application will be further described below in combination with specific preferred embodiments, but the protection scope of the present application is not limited thereby.

[0029] The materials, reagents and instruments used in the following examples can be obtained from commercial channels. The experimental methods in the following examples are conventional methods in the art, unless otherwise specified.

[0030] Example 1

[0031] A hyaluronic acid gel microneedle of the present application, the preparation method thereof comprising the following steps:

[0032] Add 300 mg of sodium hyaluronate into 10 mL of deionized water to obtain a uniform transparent solution. Add an acid to the above solution to obtain a HA hydrogel. Then freeze the hydrogel in a refrigerator at -20°C for 12 h, thaw at room temperature for 12 h, and cycle 5 times. Then immerse the freeze-thawed hydrogel in an organic solvent. Dry the obtained organic gel (ordinary freeze-drying can be used), and the final microneedle sample (HA-1) is obtained.

[0033] Example 2

[0034] A hyaluronic acid gel microneedle of the present application, the preparation method thereof comprising the following steps:

[0035] Add 1000 mg of sodium hyaluronate into 10 mL of deionized water to obtain a uniform transparent solution. Add an acid to the above solution to obtain a HA hydrogel. Then freeze the hydrogel in a refrigerator, thaw at room temperature, and cycle 3 times. Then immerse the freeze-thawed hydrogel in an organic solvent. Dry the obtained organic gel, and the final microneedle sample (HA-2) is obtained.

[0036] Example 3

[0037] A hyaluronic acid gel microneedle of the present application, the preparation method thereof comprising the following steps:

[0038] Add 1500 mg of sodium hyaluronate into 10 mL of deionized water to obtain a uniform transparent solution. Add an acid to the above solution to obtain a HA hydrogel. Then freeze the hydrogel in a refrigerator, thaw at room temperature, and cycle multiple times. Then immerse the freeze-thawed hydrogel in an organic solvent. Dry the obtained organic gel, and the final microneedle sample (HA-3) is obtained.

[0039] Figure 1 Figure is the appearance of the hyaluronic acid gel microneedle of Example 1 of the present application.

[0040] Experiment 1: The microneedle sample is subjected to scanning electron microscope test.

[0041] The hyaluronic acid gel microneedle obtained in the above example is subjected to electron microscope scanning, and the electron microscope scanning result is shown in Figure 2 .

[0042] As can be seen from the figure, the hyaluronic acid gel microneedle material can be obtained by using the method.

[0043] Experiment 2: Infrared spectrum detection.

[0044] The infrared spectrum of the hydrogel sample before drying was characterized, and the results are shown in Figure 3 .

[0045] Figure 3 The results show that the sample exhibits typical peaks characteristic of hyaluronic acid, a wide peak of hydroxyl absorption at 3600-3200 cm -1 , a methyl stretching vibration peak at 2900 cm -1 , a carboxyl stretching vibration peak at 1800-1500 cm -1 , a strong and sharp absorption peak at 1640 cm -1 , which is attributed to free carboxyl groups, multiple weak absorption peaks in the 1450-1200 cm -1 region mainly from the bending vibration of -CH2 and -CH3 on the hyaluronic acid backbone, and the stretching vibration of the C-OH bond on the sugar ring, and a sharp peak of the C-O bond stretching vibration of alcohol at 1028 cm -1 .

[0046] Experiment three: puncture experiment.

[0047] The hyaluronic acid microneedle was subjected to aluminum foil puncture experiment, and the results are shown in Figure 4 . The results show that the hyaluronic acid microneedle can easily penetrate the aluminum foil and leave a clear small hole, indicating that the microneedle has a certain mechanical strength.

[0048] The hyaluronic acid microneedle was subjected to cling film puncture experiment, and the results are shown in Figure 5 . The results show that the sample can penetrate the skin simulated by two layers of cling film, indicating that the hyaluronic acid microneedle has a certain mechanical strength and the ability to penetrate the simulated skin material with a certain elasticity.

[0049] Experiment four: self-dissolution test.

[0050] The hyaluronic acid microneedle was immersed in ethanol aqueous solution with a volume fraction of 0%, 20%, 40%, 60%, and 80%, respectively, to investigate the dissolution degree of the hyaluronic acid microneedle.

[0051] The results are shown in Figure 6As shown: the hyaluronic acid microneedle does not dissolve in 99.5% volume fraction of ethanol aqueous solution, and can be stored. The time of complete dissolution in 80% volume fraction of ethanol aqueous solution is 4 days, in 60% volume fraction of ethanol aqueous solution is 600s, in 40% volume fraction of ethanol aqueous solution is 90s, in 20% volume fraction of ethanol aqueous solution is 30s, and in water, the time of complete dissolution is within 10s. The water content of the skin varies from 20% to 70% at different depths, and the water content gradually increases from the outside to the inside. The water content of the stratum corneum in human skin is between 20% and 30%, and the water content of the deeper skin granular layer is as high as 70%. Therefore, it is proved that the hyaluronic acid microneedle prepared by the method can self-dissolve under the epidermis of the skin.

[0052] In summary, the microneedle prepared by the present application has a great application prospect in the field of biological medicine.

[0053] The above is only a preferred embodiment of the present application, and does not limit the present application in any form. Although the present application has been disclosed as above with the preferred embodiment, it is not intended to limit the present application. Any skilled person in the art can make many possible changes and modifications to the technical solutions of the present application, or modify equivalent embodiments with equivalent changes, without departing from the spirit and technical solutions of the present application. Therefore, any simple modification, equivalent replacement, equivalent change and modification made to the above embodiments according to the technical essence of the present application, which does not depart from the technical solutions of the present application, still belongs to the protection scope of the technical solutions of the present application.

Claims

1. A hyaluronic acid gel microneedle, characterized by, The hyaluronic acid gel microneedle is prepared from sodium hyaluronate, the sodium hyaluronate is dissolved to obtain a HA hydrogel, the HA hydrogel is soaked in an organic solvent after being subjected to a cyclic freezing-thawing operation, and is dried to obtain the hyaluronic acid gel microneedle.

2. A method for preparing the hyaluronic acid gel microneedles according to claim 1, characterized in that, The preparation method comprises the following steps: S1, adding sodium hyaluronate into water to obtain a solution; S2, adjusting the solution to be acidic to obtain a HA hydrogel; S3, subjecting the HA hydrogel to a cyclic freezing-thawing operation; S4, soaking the HA hydrogel after thawing in an organic solution, and drying to obtain the hyaluronic acid gel microneedle.

3. The production method according to claim 2, characterized by, The water in the S1 is deionized water.

4. The preparation method according to claim 2, characterized in that, The concentration of the sodium hyaluronate in the solution in the S1 is 20 mg / mL-500 mg / mL.

5. The preparation method according to claim 2, characterized in that, The solution is adjusted to have a pH of 2-3 in the S2.

6. The preparation method according to claim 2, characterized in that, The cyclic freezing-thawing operation in the S3 is specifically freezing the HA hydrogel at-20 ℃ in a refrigerator for 12 h, thawing at room temperature of 25 ℃ for 12 h, and repeating the freezing and thawing operation for 5 times.

7. Use of the hyaluronic acid gel microneedle prepared by the preparation method in any one of claims 2-6 in preparation of a carrier for loading drugs.

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