Silk fibroin hydrogel liposome with transdermal delivery effect and preparation method and application thereof

Abstract

The application belongs to the technical field of biological materials, and particularly relates to a silk fibroin-loaded hydrogel liposome with transdermal delivery effect and a preparation method and application thereof. The preparation method comprises the following steps: dissolving degummed silk in a neutral salt solution to obtain a silk fibroin solution, performing dialysis filtration to obtain a silk fibroin aqueous solution; then performing a diazo coupling reaction on the silk fibroin aqueous solution to obtain an azide-functionalized silk fibroin aqueous solution; dissolving phospholipid and cholesterol in a solvent to obtain a mixed solution; adding the azide-functionalized silk fibroin aqueous solution into the mixed solution to perform ultrasonic emulsification to obtain an emulsion; sequentially performing reverse evaporation treatment on the emulsion to obtain a gelled silk fibroin-loaded liposome; and performing freeze-drying on the gelled silk fibroin-loaded liposome to obtain the silk fibroin-loaded hydrogel liposome with transdermal delivery effect.

Description

Technical Field

[0001] This invention belongs to the field of biomaterials technology, specifically relating to a transdermal delivery method for a silk fibroin hydrogel liposome, its preparation method, and its application. Background Technology

[0002] The skin, the largest and most accessible organ in the human body, plays a vital role in maintaining homeostasis, defending against invading microorganisms, and protecting against environmental attacks such as high temperatures, chemicals, and toxins. Transdermal drug delivery delivers bioactive agents through the skin to achieve local or systemic effects; it is non-invasive and allows for self-administration. Transdermal delivery systems have become a cutting-edge research area for delivering small molecule drugs, proteins, small peptides, and vaccines, avoiding the "first-pass effect" of the liver and gastrointestinal tract, maintaining stable and sustained plasma drug concentrations, reducing dosing frequency, and improving drug bioavailability.

[0003] Chinese patent (publication number CN119280079A) discloses a method for preparing liposomes containing hair growth active ingredients. This invention utilizes active substances to regulate hormone levels, improve scalp blood circulation, and stimulate anti-hair loss factors, thereby promoting hair growth in multiple ways. Furthermore, it uses lecithin and oil-soluble active ingredients as raw materials to form liposomes that integrally encapsulate the active substances, overcoming the drawback of weak transdermal absorption due to significant differences in water solubility between various active substances. This improves the stability and bioavailability of the active substances, significantly enhancing the hair growth effect of the prepared liposomes containing hair growth active ingredients. However, there is still a lack of sufficient research in the prior art on preparing hydrogel liposomes using silk fibroin and applying liposomes to transdermal delivery systems.

[0004] Therefore, how to use silk fibroin as a raw material, functionalize it, and use it in combination with phospholipids and cholesterol to prepare silk fibroin hydrogel liposomes, and successfully apply them to transdermal delivery systems, has become a direction that needs to be studied. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the present invention aims to provide a silk fibroin hydrogel liposome with transdermal delivery capability, its preparation method, and its application. Using silk fibroin as raw material, it undergoes diazo coupling treatment to obtain azide-functionalized silk fibroin, which is then used in conjunction with phospholipids and cholesterol. Through processes such as reverse evaporation and ultrasonic treatment, the silk fibroin hydrogel liposome is prepared and successfully applied to a transdermal delivery system.

[0006] The technical solution adopted by the present invention to solve the above-mentioned technical problems is as follows:

[0007] A first aspect of the present invention provides a method for preparing a silk fibroin hydrogel liposome with transdermal delivery capability, characterized by comprising the following steps:

[0008] S1: Dissolve the degummed silk in a neutral salt solution to obtain a silk fibroin solution, dialyze and filter to obtain an aqueous solution of silk fibroin; then perform a diazo coupling reaction on the aqueous solution of silk fibroin to obtain an aqueous solution of azide-functionalized silk fibroin.

[0009] S2: Dissolve phospholipids and cholesterol in a solvent to obtain a mixed solution;

[0010] S3: The aqueous solution of azide-functionalized silk fibroin is added to the mixed solution and ultrasonically emulsified to obtain an emulsion;

[0011] S4: The emulsion is subjected to counter-evaporation treatment in sequence to obtain gelled silk fibroin liposomes;

[0012] S5: Freeze-dry the gelled fibroin liposomes to obtain fibroin hydrogel liposomes with transdermal delivery effect.

[0013] The following are preferred technical solutions of the present invention, but are not intended to limit the technical solutions provided by the present invention. The purpose and beneficial effects of the present invention can be better achieved and realized through the following preferred technical solutions.

[0014] As a preferred embodiment of the present invention, the steps of the diazo coupling reaction include: dissolving 0.6-0.8 parts of 4-azidoaniline hydrochloride in a mixture of 4-6 parts of acetonitrile and 4-6 parts of deionized water, then adding 4-6 parts of 0.01-0.02 mol / L hydrochloric acid aqueous solution and mixing evenly, placing it in an ice bath and controlling the temperature at 0-5°C, then slowly adding 4-6 parts of 0.06-0.08 mol / L sodium nitrite aqueous solution and stirring for 10-20 min to obtain a diazonium salt solution; adjusting the pH of 90-100 parts of silk fibroin aqueous solution to 7-8 with borate buffer, then slowly adding 20-24 parts of the diazonium salt solution under light-protected and ice bath conditions and stirring for 2-4 h, terminating the reaction and dialysis purification.

[0015] The nanofiber structure of azidolated silk fibroin mimics the skin's natural channels. Combined with the lipid bilayer of liposomes, it can reduce the lipid order of the stratum corneum and improve transdermal delivery efficiency. At the same time, azidolated silk fibroin can be chemically anchored to the surface of liposomes to form a "protein-phospholipid bilayer" hybrid membrane, which enhances the penetration of liposomes into the skin barrier.

[0016] As a preferred embodiment of the present invention, the neutral salt solution in step S1 is a lithium bromide / water binary system or a calcium chloride / ethanol / water ternary system.

[0017] As a preferred embodiment of the present invention, the molar ratio of lithium bromide to water in the lithium bromide / water binary system is 1:5; and the molar ratio of calcium chloride to ethanol to water in the calcium chloride / ethanol / water ternary system is 1:2:8.

[0018] As a preferred embodiment of the present invention, the dissolution temperature in step S1 is 50~70℃ and the dissolution time is 4~6h.

[0019] As a preferred embodiment of the present invention, the mass concentration fraction of the silk fibroin aqueous solution in step S1 is 2-4%.

[0020] The neutral salt solution of this invention is a lithium bromide / water binary system or a calcium chloride / ethanol / water ternary system. The lithium bromide / water binary system achieves rapid dissolution by disrupting the β-sheet structure of silk fibroin, transforming it into a soluble random coil conformation. In the calcium chloride / ethanol / water ternary system, calcium ions can disrupt the ionic and hydrogen bonds of silk fibroin and attack the crystalline region. Ethanol reduces the polarity of water, enhances the penetration into hydrophobic regions, and inhibits protein oxidative degradation during dissolution. The ternary system can achieve efficient dissolution under mild conditions.

[0021] As a preferred embodiment of the present invention, the phospholipid in step S2 is selected from any one or more of egg yolk lecithin, soybean lecithin, dipalmitoylphosphatidylcholine, and distearate phosphatidylcholine.

[0022] As a preferred embodiment of the present invention, the concentration of phospholipids in the mixed solution in step S2 is 1~20 mg / mL, and the concentration of cholesterol in the mixed solution is 0.2~2 mg / mL.

[0023] As a preferred embodiment of the present invention, the organic solvent in step S2 is chloroform or dichloromethane.

[0024] Phospholipid-cholesterol complex membranes are similar to stratum corneum lipids, which can promote the fusion of liposomes with the skin barrier, making it easier for silk fibroin to diffuse into the deeper layers of the skin; the presence of cholesterol can enhance the moisturizing ability of liposomes, soften the stratum corneum through hydration, expand the intercellular spaces, and promote the penetration of active ingredients.

[0025] As a preferred embodiment of the present invention, the volume ratio of the azide-functionalized silk fibroin aqueous solution and the mixed solution in step S3 is 1:(5~10).

[0026] As a preferred embodiment of the present invention, the ultrasound time in step S3 is 10-30 seconds and the number of ultrasound cycles is 1-3.

[0027] As a preferred technical solution of the present invention, the reverse evaporation process in step S4 includes: transferring the emulsion to a rotary evaporator and evaporating it at 50-80 rpm for 6-24 hours at 30-60°C to obtain a silk fibroin liposome film; adding 4-6 parts of phosphate buffer solution to the silk fibroin liposome film and incubating it at 25-40°C for 2-8 hours, and then inducing silk fibroin gelation by ultrasonic treatment.

[0028] As a preferred embodiment of the present invention, the ultrasonic treatment time is 10-30 minutes and the number of ultrasonic treatments is 1-3 times.

[0029] As a preferred embodiment of the present invention, the freeze-drying conditions in step S5 include: a temperature of -20 to -40°C and a time of 36 to 48 hours.

[0030] As a preferred embodiment of the present invention, the particle size of the transdermal delivery fibroin hydrogel liposome is 90~100nm and the zeta potential is -9.4~-9.6mV.

[0031] A second aspect of the present invention provides a fibroblast protein hydrogel liposome with transdermal delivery effect prepared by the method described in the first aspect.

[0032] A third aspect of the present invention provides the application of a transdermal delivery fibroin hydrogel liposome prepared by the method described in the first aspect in skin care products.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] (1) The silk fibroin raw material of the present invention has good compatibility, is degradable, has excellent mechanical properties and film-forming properties, and can enhance the stability of liposomes; silk fibroin liposomes can be prepared simply and efficiently by reverse evaporation, ultrasonic treatment and other methods, and the liposomes can effectively penetrate the stratum corneum of the skin, solving the problem that large molecular silk fibroin is difficult to penetrate the stratum corneum of the skin.

[0035] (2) The neutral salt solution of the present invention is a lithium bromide / water binary system or a calcium chloride / ethanol / water ternary system. The lithium bromide / water binary system achieves rapid dissolution by destroying the β-sheet structure of silk fibroin, which transforms it into a soluble random coil conformation. The calcium ions in the calcium chloride / ethanol / water ternary system can destroy the ionic bonds and hydrogen bonds of silk fibroin and attack the crystallization region. Ethanol reduces the polarity of water and enhances the penetration of hydrophobic regions, while inhibiting the oxidative degradation of protein during dissolution. The ternary system can achieve efficient dissolution under mild conditions.

[0036] (3) The nanofiber structure of azide-modified silk fibroin mimics the natural channels of the skin. Combined with the lipid bilayer of liposomes, it can reduce the lipid order of the stratum corneum and improve the transdermal delivery efficiency. At the same time, azide-modified silk fibroin can be chemically anchored to the surface of liposomes to form a "protein-phospholipid bilayer" mixed membrane, which enhances the penetration of liposomes into the skin barrier. Detailed Implementation

[0037] To facilitate understanding of the present invention, the following embodiments are provided. Those skilled in the art should understand that these embodiments are merely illustrative and should not be construed as limiting the scope of the invention.

[0038] The sources of some components in the examples and comparative examples are as follows:

[0039] Lithium bromide, CAS No. 7550-35-8, was purchased from Shanghai Maclean Biochemical Technology Co., Ltd.

[0040] Calcium chloride, CAS No. 10043-52-4, was purchased from Shanghai Maclean Biochemical Technology Co., Ltd.

[0041] 4-Azide aniline hydrochloride, CAS No. 91159-79-4, was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0042] p-Toluenesulfonic acid, CAS No. 104-15-4, purchased from Sinopharm Chemical Reagent Co., Ltd.

[0043] Sodium nitrite, CAS No. 7632-00-0, was purchased from Sinopharm Chemical Reagent Co., Ltd.

[0044] Borate buffer, product number B407195, was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0045] Egg yolk lecithin, product number L305002, was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0046] Soy lecithin, CAS No. 8002-43-5, was purchased from Shaanxi Xintianyu Biotechnology Co., Ltd.

[0047] Dipalmitoylphosphatidylcholine, product number S26236, was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0048] Distearate phosphatidylcholine, product number S50960, was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0049] Cholesterol, CAS No. 57-88-5, purchased from Sinopharm Chemical Reagent Co., Ltd.

[0050] Example 1

[0051] This embodiment provides a method for preparing fibroin hydrogel liposomes with transdermal delivery effect, characterized by comprising the following steps:

[0052] S1: Dissolve 10 parts of degummed silk (at 65℃ for 4 hours) in a 40-part lithium bromide / water binary system (lithium bromide:water molar ratio of 1:5) to obtain a silk fibroin solution. After dialyzing for 3 days, filter to obtain a 3% (w / w) silk fibroin aqueous solution. Dissolve 0.8 parts of 4-azidoaniline hydrochloride in a mixture of 6 parts acetonitrile and 6 parts deionized water, then add 6 parts of 0.01~0.02 mol / L hydrochloric acid aqueous solution and mix well. Place in an ice bath and control the temperature at 0℃. Then slowly add 6 parts of 0.08 mol / L sodium nitrite aqueous solution and stir for 20 minutes to obtain a diazonium salt solution. Adjust the pH of 100 parts of the silk fibroin aqueous solution to 8 with borate buffer. Then slowly add 24 parts of the diazonium salt solution under light-protected and ice bath conditions and stir for 4 hours. After terminating the reaction, dialyze to purify and obtain an azide-functionalized silk fibroin aqueous solution.

[0053] S2: Dissolve egg yolk lecithin and cholesterol in chloroform to obtain a mixed solution (the concentration of egg yolk lecithin in the mixed solution is 20 mg / mL, and the concentration of cholesterol in the mixed solution is 2 mg / mL).

[0054] S3: Add 1 part of the azide-functionalized silk fibroin aqueous solution to 5 parts of the mixed solution and perform ultrasonic emulsification (15s for 2 times) to obtain an emulsion;

[0055] S4: Transfer the emulsion to a rotary evaporator and evaporate it at 50°C and 60 rpm for 8 hours to obtain a silk fibroin liposome film; add 5 parts of phosphate buffer solution to the silk fibroin liposome film and incubate it at 40°C for 2 hours, then sonicate it (for 10 minutes, 3 times) to induce silk fibroin gelation to obtain gelled silk fibroin liposomes.

[0056] S5: The gelled fibroin liposomes are freeze-dried (temperature -20℃, time 48h) to obtain fibroin hydrogel liposomes with transdermal delivery effect (particle size 100nm, zeta potential -9.6mV).

[0057] Example 2

[0058] This embodiment provides a method for preparing fibroin hydrogel liposomes with transdermal delivery effect, characterized by comprising the following steps:

[0059] S1: Dissolve 10 parts of degummed silk in a calcium chloride / ethanol / water ternary system (calcium chloride:ethanol:water molar ratio of 1:2:8) to obtain a silk fibroin solution. Dialyze and filter to obtain a 2% (w / w) silk fibroin aqueous solution. Dissolve 0.6 parts of 4-azidoaniline hydrochloride in a mixture of 4 parts acetonitrile and 4 parts deionized water. Then add 4 parts of 0.01 mol / L hydrochloric acid aqueous solution and mix well. Place the solution in an ice bath at 5°C and slowly add 4 parts of 0.06 mol / L sodium nitrite aqueous solution while stirring for 10 min to obtain a diazonium salt solution. Adjust the pH of 90 parts of the silk fibroin aqueous solution to 7 with borate buffer. Then slowly add 20 parts of the diazonium salt solution under light-protected and ice bath conditions while stirring for 2 h. After terminating the reaction, dialyze and purify to obtain an azide-functionalized silk fibroin aqueous solution.

[0060] S2: Dissolve soybean lecithin and cholesterol in dichloromethane to obtain a mixed solution (the concentration of soybean lecithin in the mixed solution is 1 mg / mL, and the concentration of cholesterol in the mixed solution is 0.2 mg / mL).

[0061] S3: Add 1 part of the azide-functionalized silk fibroin aqueous solution to 10 parts of the mixed solution and perform ultrasonic emulsification (time 20s, number of times 2) to obtain an emulsion;

[0062] S4: Transfer the emulsion to a rotary evaporator and evaporate it at 50 rpm for 6 hours at 60°C to obtain a silk fibroin liposome film; add 4 parts of phosphate buffer solution to the silk fibroin liposome film and incubate it at 25°C for 8 hours, then sonicate it (30 min for 1 time) to induce silk fibroin gelation to obtain gelled silk fibroin liposomes.

[0063] S5: The gelled fibroin liposomes are freeze-dried (temperature -40℃, time 36h) to obtain fibroin hydrogel liposomes with transdermal delivery effect (particle size 90nm, zeta potential -9.4mV).

[0064] Example 3

[0065] This embodiment provides a method for preparing fibroin hydrogel liposomes with transdermal delivery effect, characterized by comprising the following steps:

[0066] S1: Dissolve 10 parts of degummed silk in a 50-part lithium bromide / water binary system (lithium bromide:water molar ratio of 1:5) to obtain a silk fibroin solution. Dialyze and filter to obtain a 4% (w / w) silk fibroin aqueous solution. Dissolve 0.7 parts of 4-azidoaniline hydrochloride in a mixture of 5 parts acetonitrile and 5 parts deionized water. Then add 5 parts of 0.015 mol / L hydrochloric acid aqueous solution and mix well. Place the solution in an ice bath at 4°C and slowly add 5 parts of 0.07 mol / L sodium nitrite aqueous solution while stirring for 15 min to obtain a diazonium salt solution. Adjust the pH of 95 parts of the silk fibroin aqueous solution to 7.4 with borate buffer. Then slowly add 22 parts of the diazonium salt solution under light-protected and ice bath conditions while stirring for 3 h. After terminating the reaction, dialyze and purify to obtain an azide-functionalized silk fibroin aqueous solution.

[0067] S2: Dissolve phospholipids (egg yolk lecithin, soybean lecithin, dipalmitoylphosphatidylcholine, distearate phosphatidylcholine) and cholesterol in chloroform to obtain a mixed solution (the concentration of phospholipids in the mixed solution is 1 mg / mL, and the concentration of cholesterol in the mixed solution is 0.2 mg / mL).

[0068] S3: Add 3 parts of the azide-functionalized silk fibroin aqueous solution to 10 parts of the mixed solution and perform ultrasonic emulsification (30s, 1 time) to obtain an emulsion;

[0069] S4: The emulsion is transferred to a rotary evaporator and evaporated at 80 rpm for 24 h at 30 °C to obtain a silk fibroin liposome film; 4 parts of phosphate buffer solution are added to the silk fibroin liposome film and incubated at 30 °C for 6 h, and then ultrasonically treated (for 10 min, 3 times) to induce silk fibroin gelation to obtain gelled silk fibroin liposomes;

[0070] S5: The gelled fibroin liposomes are freeze-dried (at -30°C for 40 hours) to obtain fibroin hydrogel liposomes with transdermal delivery effect (particle size 95 nm, zeta potential -9.5 mV).

[0071] Comparative Example 1

[0072] This embodiment provides a method for preparing hydrogel liposomes, characterized by comprising the following steps:

[0073] S1: Dissolve 10 parts of degummed silk (at 65℃ for 4 hours) in 40 parts of lithium bromide / water binary system (the molar ratio of lithium bromide to water is 1:5) to obtain a silk fibroin solution. After dialyzing for 3 days, filter to obtain a silk fibroin aqueous solution with a mass concentration of 3%.

[0074] S2: Dissolve egg yolk lecithin and cholesterol in chloroform to obtain a mixed solution (the concentration of egg yolk lecithin in the mixed solution is 20 mg / mL, and the concentration of cholesterol in the mixed solution is 2 mg / mL).

[0075] S3: Add 1 part of the silk fibroin aqueous solution to 5 parts of the mixed solution and perform ultrasonic emulsification (15s for 2 times) to obtain an emulsion;

[0076] S4: Transfer the emulsion to a rotary evaporator and evaporate it at 50°C and 60 rpm for 8 hours to obtain a silk fibroin liposome film; add 5 parts of phosphate buffer solution to the silk fibroin liposome film and incubate it at 40°C for 2 hours, then sonicate it (for 10 minutes, 3 times) to induce silk fibroin gelation to obtain gelled silk fibroin liposomes.

[0077] S5: The gelled fibroin liposomes were freeze-dried (at -20°C for 48 hours) to obtain hydrogel liposomes (particle size 180 nm, zeta potential -10.6 mV).

[0078] The transdermal delivery performance of the hydrogel liposomes provided in the above examples and comparative examples was tested using the following methods: In vitro transdermal delivery was tested using a Franz diffusion cell. Before the experiment, pig ear skin was thawed naturally in physiological saline and cleaned, then the skin surface was blotted dry with filter paper. The skin was fixed, and 6.5 mL of PBS and 0.5 mL of bispecific antibody were added to the receiving cell. Air bubbles were removed to allow the solution to contact the skin. The receiving solution was tested under constant temperature magnetic stirring conditions at 37°C and 350 rpm. 2 mL of hydrogel liposomes was placed in the supply chamber as the experimental group. At predetermined time intervals (1, 3, 6, 9, 12, and 24 hours), 2 mL of liquid from the receiving chamber was collected, and an equal amount of PBS was added. Air bubbles were removed to allow the liquid to contact the skin. After collecting the liquid from the receiving chamber, it was filtered through a 0.22 μm filter membrane into a liquid chromatography vial and analyzed by HPLC. The cumulative permeation per unit volume was calculated.

[0079] The performance test data above are shown in Table 1.

[0080] Table 1 Performance Test Results

[0081]

[0082] As can be seen from the above, this invention uses silk fibroin as raw material and performs diazo coupling treatment to obtain azide-functionalized silk fibroin. It is then used in conjunction with phospholipids and cholesterol, and through processes such as reverse evaporation and ultrasonic treatment, silk fibroin-loaded hydrogel liposomes are prepared and successfully applied to transdermal delivery systems.

Claims

1. A method for preparing a transdermal delivery method for silk fibroin hydrogel liposomes, characterized in that, Includes the following steps: S1: Dissolve the degummed silk in a neutral salt solution to obtain a silk fibroin solution, dialyze and filter to obtain an aqueous solution of silk fibroin; then perform a diazo coupling reaction on the aqueous solution of silk fibroin to obtain an aqueous solution of azide-functionalized silk fibroin. S2: Dissolve phospholipids and cholesterol in a solvent to obtain a mixed solution; S3: The aqueous solution of azide-functionalized silk fibroin is added to the mixed solution and ultrasonically emulsified to obtain an emulsion; S4: The emulsion is subjected to counter-evaporation treatment in sequence to obtain gelled silk fibroin liposomes; S5: Freeze-dry the gelled silk fibroin liposomes to obtain silk fibroin hydrogel liposomes with transdermal delivery effect. The steps of the diazo coupling reaction include: dissolving 0.6-0.8 parts of 4-azidoaniline hydrochloride in a mixture of 4-6 parts of acetonitrile and 4-6 parts of deionized water, then adding 4-6 parts of 0.01-0.02 mol / L hydrochloric acid aqueous solution and mixing thoroughly. The mixture is placed in an ice bath at a controlled temperature of 0-5°C, and then 4-6 parts of 0.06-0.08 mol / L sodium nitrite aqueous solution are slowly added dropwise and stirred for 10-20 min to obtain a diazonium salt solution. 90-100 parts of silk fibroin aqueous solution are adjusted to pH 7-8 using borate buffer, and then 20-24 parts of the diazonium salt solution are slowly added under light-protected and ice bath conditions and stirred for 2-4 h. After terminating the reaction, the solution is purified by dialysis. The neutral salt solution in step S1 is either a lithium bromide / water binary system or a calcium chloride / ethanol / water ternary system. In the lithium bromide / water binary system, the molar ratio of lithium bromide to water is 1:5; in the calcium chloride / ethanol / water ternary system, the molar ratio of calcium chloride to ethanol to water is 1:2:

8. In step S2, the phospholipid is selected from any one or more of egg yolk lecithin, soybean lecithin, dipalmitoylphosphatidylcholine, and distearate phosphatidylcholine. The concentration of phospholipids in the mixed solution in step S2 is 1~20 mg / mL, and the concentration of cholesterol in the mixed solution is 0.2~2 mg / mL; The solvent in step S2 is chloroform or dichloromethane; In step S3, the volume ratio of the azide-functionalized silk fibroin aqueous solution to the mixed solution is 1:(5~10). In step S3, the ultrasound duration is 10-30 seconds, and the number of ultrasound cycles is 1-3. The reverse evaporation process in step S4 includes: transferring the emulsion to a rotary evaporator and evaporating it at 50-80 rpm for 6-24 hours at 30-60°C to obtain a silk fibroin liposome film; adding 4-6 parts of phosphate buffer solution to the silk fibroin liposome film and incubating it at 25-40°C for 2-8 hours, followed by ultrasonic treatment to induce silk fibroin gelation. The freeze-drying conditions in step S5 include: a temperature of -20 to -40°C and a time of 36 to 48 hours.

2. The method for preparing a transdermal delivery hydrogel liposome according to claim 1, characterized in that, The transdermal delivery-enabled fibroin hydrogel liposomes have a particle size of 90-100 nm and a zeta potential of -9.4 to -9.6 mV.

3. A transdermal delivery hydrogel liposome, characterized in that, Obtained by the preparation method as described in any one of claims 1-2.

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