Silk fibroin liquid band-aid as well as preparation method and application thereof

Abstract

The invention discloses a composition which comprises silk fibroin and a film-forming material, and the composition can be used for liquid dressings, particularly liquid band-aids. The invention also provides a preparation method and application of the composition.

Description

Technical Field

[0001] This invention belongs to the field of medical technology, and specifically discloses a silk fibroin liquid bandage, its preparation method and application. Background Technology

[0002] Acute wounds refer to injuries caused by physical, thermal, or electrical factors that disrupt the normal structure and function of tissues, such as abrasions, incisions, and tears. As one of the largest organs in the human body, skin wounds are common in daily life. If not treated promptly, they can lead to secondary damage, bacterial infection, and scar formation. Therefore, timely protection and repair of wounds are crucial for ensuring rapid and orderly healing. The wound healing process includes four stages: hemostasis, inflammation, proliferation, and remodeling. This is a complex, dynamic, and orderly biological process. An ideal healing process should include rapid hemostasis, appropriate inflammatory response (sterilization, removal of tissue debris, etc.), continuous angiogenesis (skin regeneration), and appropriate collagen synthesis and deposition (reconstruction of damaged skin). Problems at any stage can delay wound healing or affect the final outcome. For example, a prolonged inflammatory phase leading to chronic ulcers will significantly prolong healing time, while excessive development in the proliferation and remodeling phases can lead to pyogenic granulomas and keloids, affecting aesthetics.

[0003] Bandage-type adhesive bandages, as a traditional treatment method, are one of the most commonly used first-aid medical supplies in people's lives. They consist of a long strip of adhesive tape with gauze in the middle. They are simple and convenient to use, but they have the following shortcomings: when changing them, the gauze part sticks to the wound, and tearing it off may cause secondary damage; they have poor breathability and waterproofness, and are not effective in protecting hands and bathing; they cannot provide effective protection for moving parts such as joints, and while they may feel restrictive, the edges may also lift up and expose the wound, affecting the healing speed.

[0004] Liquid bandages are a novel form of protection, consisting of a film-forming substance dissolved in a suitable solvent. They are applied to the wound via spraying or smearing to form a protective film. Compared to traditional bandages, liquid bandages offer advantages such as wider coverage, better breathability and waterproofing, and consistent wound coverage, even on irregularly shaped or mobile wounds. Therefore, liquid bandages are of great significance for treating skin wounds caused in daily life or military training. However, currently available liquid bandages suffer from drawbacks such as poor hemostasis, poor biocompatibility, lack of immunomodulatory effects, weak antioxidant capacity, and insufficient collagen production promotion, leading to slow wound healing. Furthermore, the use of large amounts of organic solvents makes them prone to cracking and produces a pungent odor. Therefore, there is an urgent need to develop a safe, effective, and mechanically controllable liquid bandage product. Summary of the Invention

[0005] The purpose of this invention is to provide a silk fibroin liquid bandage that has a short film-forming time, forms a flexible and transparent film, is conducive to wound adhesion, and is easy to peel off and remove.

[0006] This invention provides the following technical solutions:

[0007] In a first aspect, the present invention provides a pharmaceutical composition comprising silk fibroin (SF) and a film-forming material.

[0008] In a preferred embodiment, the film-forming material is a water-soluble film-forming material and / or a fat-soluble film-forming material;

[0009] Preferably, the water-soluble film-forming material is selected from one or more of polyvinyl alcohol (PVA), carbomer, methyl cellulose (MC), hydroxyethyl cellulose, hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose, chitosan, sodium alginate, and sodium carboxymethyl cellulose.

[0010] Preferably, the lipid-soluble film-forming material is selected from one or more of nitrocellulose (NC), polyvinyl butyral (PVB), acrylates, ethyl cellulose (EC), cellulose acetate (CA), and polyurethane.

[0011] In a preferred embodiment, the silk fibroin content is 0.5wt%-20wt%, preferably 1wt%-10wt%, more preferably 1wt%-5wt%, even more preferably 1wt%-3wt%, and most preferably 2wt%-3wt%, based on the total mass of the composition.

[0012] In a preferred embodiment, the content of the film-forming material is 8 wt%-25 wt%, preferably 10 wt%-22 wt%, more preferably 10 wt%-20 wt%, and even more preferably 16 wt%-20 wt%, based on the total mass of the composition.

[0013] In a preferred embodiment, the composition further comprises a solvent, which is an aqueous phase and / or an organic phase. Preferably, the solvent is a mixture of an aqueous phase and an organic phase. More preferably, the volume percentage of the aqueous phase is 0%-40% based on the total volume of the composition, preferably 20%-40%.

[0014] In a preferred embodiment, the silk fibroin has a molecular weight of 8-100 kDa; preferably 8-50 kDa or 50-100 kDa.

[0015] In a preferred embodiment, the film-forming material contains polyvinyl butyral (PVB), and preferably, the viscosity of the PVB does not exceed 2800 mPa·s.

[0016] In a preferred embodiment, the composition further comprises an additive;

[0017] Preferably, the additive is selected from one or more of self-adhesives, plasticizers, and cosolvents. More preferably, the additive is selected from one or more of polyols (including polyethylene glycol, glycerin, propylene glycol, sorbitol, xylitol), dibutyl phthalate, tributyl acetyl citrate, sucrose isobutyrate, sucrose benzoic acid, acetyl citrate, mineral oil, hyaluronic acid (HA), and polyvinylpyrrolidone (PVP).

[0018] In a preferred embodiment, the organic phase is selected from one or more of alcohols, low molecular weight alkanes and cycloalkanes, alkanes, ethers, siloxanes, volatile fluorocarbons, polysorbates, and dehydrated sorbitan fatty acid esters;

[0019] Preferably, the alcohols are selected from one or more of methanol, ethanol, butanol, benzyl alcohol, phenylene glycol, tert-butanol, and isopropanol; the cycloalkanes are selected from one or more of n-pentane, hexane, heptane, cyclohexane, and isooctane; the alkyl esters are selected from one or more of ethyl acetate, butyl acetate, n-propyl acetate, isobutyl acetate, hexyl acetate, isopropyl myristate, castor oil, and isobutyl isobutyrate; the ethers are selected from one or more of hexamethyldisiloxane and diethyl ether; the siloxanes are selected from one or more of hexamethyldisiloxane, decamethylcyclopentanesiloxane, octamethyltrisiloxane, and polydimethylsiloxane; and the volatile fluorocarbons are selected from one or more of pentafluoropropane and perfluoroheptane.

[0020] In a preferred embodiment, the composition further contains a drug, preferably selected from one or more of antibiotics, metal ions, anti-inflammatory agents, and active ingredients of traditional Chinese medicine.

[0021] In a preferred embodiment, the composition is a solution, gel, semi-solid, spray, or suspension; preferably, the composition is a liquid dressing; more preferably, the composition is a liquid bandage; and even more preferably, the film-forming time of the composition is less than 3 minutes.

[0022] In a preferred embodiment, the composition comprises the following components:

[0023] - Silk fibroin, based on the total mass of the composition, wherein the content of the silk fibroin is 1wt%-3wt%, and the molecular weight of the silk fibroin is 8–50kDa or 50–100kDa;

[0024] -PVB, based on the total mass of the composition, the content of PVB is 16 wt%;

[0025] - Water, with a volume percentage of 25% based on the total volume of the composition;

[0026] - Ethanol, wherein the volume percentage of ethanol is 75% based on the total volume of the composition. In a second aspect, the present invention provides a method for preparing the aforementioned composition, comprising the following steps:

[0027] (1) Dissolve SF in the aqueous phase to obtain an SF solution; dissolve the film-forming material in the aqueous or organic phase to obtain a film-forming material solution;

[0028] (2) Mix the above SF solution and film-forming material solution;

[0029] Preferably, the mixing method is mechanical stirring, high-speed shearing, high-pressure homogenization, or molecular diffusion during static settling.

[0030] In a third aspect, the present invention provides the use of the foregoing composition or the composition prepared according to the foregoing method in the preparation of a medicament for the prevention and / or treatment of lesions and / or inflammatory diseases;

[0031] Preferably, the injury is selected from superficial mechanical damage or burns to the skin, deep skin damage, or internal organ damage;

[0032] Preferably, the inflammatory disease includes superficial skin inflammation, deep skin inflammation, or visceral inflammation.

[0033] Beneficial effects:

[0034] When SF is used alone as a film-forming material, the film-forming time is as long as several hours, and the film is not easy to peel off and is easily broken, making it difficult to be an effective liquid bandage. When used as a liquid dressing, SF has poor mechanical properties, does not adhere firmly, and is easy to fall off the wound surface when rubbed, while also staining the clothing worn, making it difficult to quickly and effectively protect the wound.

[0035] This invention combines SF with a specific film-forming material, resulting in a short film-forming time and a flexible, transparent film that is easy to peel off and remove, thus avoiding secondary damage to the wound caused by mechanical peeling.

[0036] The SF liquid bandage of this invention forms a film with a rough surface and a large number of three-dimensional mesh pores inside, which has a good effect on skin cell adhesion, proliferation and migration, good hemostasis effect, and can accelerate the wound healing process.

[0037] The SF liquid bandage of this invention increases breathability and water retention, improves the metabolic process of substances in the application area, and thus accelerates wound healing.

[0038] The SF liquid bandage of this invention has good antioxidant and anti-inflammatory effects, can scavenge free radicals, is beneficial to the regulation of the immune microenvironment at the wound site, increases M2 macrophages, and has significant effects on promoting wound healing, reducing scar hyperplasia, and promoting the regeneration of hair follicles of skin appendages.

[0039] The present invention provides a method for uniformly dissolving and / or dispersing SF in an aqueous solution and / or organic solvent containing a film-forming material. The preparation process is simple and easy to implement, and has good prospects for widespread application. Brief description of the attached diagram

[0040] Figure 1 The film appearance morphology formed by the SF liquid bandage of the present invention is shown;

[0041] Figure 2 This is a scanning electron microscope (SEM) image of the film formed by the SF liquid bandage of the present invention;

[0042] Figure 3A This is an atomic force microscope (AFM) image of the film formed by the SF liquid bandage of this invention. Figure 3B for Figure 3A The arithmetic mean height Ra, root mean square Ra Rq, and height values ​​of the AFM measurement results relative to the reference plane were statistically analyzed.

[0043] Figure 4 The Fourier transform infrared (FT-IR) spectra of the SF liquid bandage solution and the film formed therefrom, and the secondary structure of SF in different compositions are shown.

[0044] Figure 5 The rheological curve of the SF liquid bandage of the present invention is shown below;

[0045] Figure 6 The invention demonstrates that the SF liquid bandage can be applied in 10 seconds. -1 Viscosity at shear rate (compared to PVB group, * indicates P<0.05, ** indicates P<0.01, **** indicates P<0.0001);

[0046] Figure 7 The air permeability-time curve of the film formed by the SF liquid bandage of the present invention;

[0047] Figure 8 The results of air permeability of the film formed by the SF liquid bandage of the present invention at different time points (compared with the PVB group, * indicates P<0.05, ** indicates P<0.01);

[0048] Figure 9 The waterproofing results of the film formed by the SF liquid bandage of the present invention (compared with the PVB group, * indicates P<0.05, ** indicates P<0.01);

[0049] Figure 10 This is the result of the water absorption of the film formed by the SF liquid bandage of the present invention;

[0050] Figure 11 The mechanical properties of films formed from liquid bandages with different SF content are shown, including tensile strength and elongation.

[0051] Figure 12 The mechanical properties (Young's modulus, tensile strength, and elongation) of the film formed by the SF liquid bandage of the present invention are shown.

[0052] Figure 13 The invention demonstrates the removability of the SF liquid bandage, wherein the red portion represents the components of the SF liquid bandage, showing that the SF liquid bandage can be completely dissolved in 70% ethanol in 10 minutes;

[0053] Figure 14 The results of the blood compatibility of the SF liquid bandage of the present invention are shown, wherein the upper figure shows a photograph of the sample after centrifugation, and the lower figure shows the OD value of the supernatant of different samples at 570 nm after centrifugation.

[0054] Figure 15 The effects of the SF liquid bandage of the present invention on cell viability are shown (compared with the TCP group, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, **** indicates P<0.0001; compared with the PVB group, # indicates P<0.05, ## indicates P<0.01, ### indicates P<0.001, #### indicates P<0.0001).

[0055] Figure 16 The effects of the SF liquid bandage of the present invention on different cell morphologies are shown. The upper figure shows the effect of different liquid bandages on the morphology of mouse fibroblasts (L929), the middle figure shows the effect of different liquid bandages on the morphology of mouse mononuclear macrophage leukemia cells (Raw 264.7), and the lower figure shows the effect of different liquid bandages on the morphology of human umbilical vein endothelial cells (HUVEC).

[0056] Figure 17 This invention demonstrates the effects of the SF liquid bandage of the present invention on free radicals DPPH and ABTS. + Its ability to clear;

[0057] Figure 18 The in vitro antioxidant activity of the SF liquid bandage of the present invention against different cells (L929 and Raw264.7) was demonstrated (compared with the positive control PC group, *** indicates P<0.001, **** indicates P<0.0001);

[0058] Figure 19This demonstrates the effect of the SF liquid bandage of the present invention on macrophage polarization;

[0059] Figure 20 The in vitro anti-inflammatory effect of the SF liquid bandage of the present invention was demonstrated, namely the effect of different liquid bandages on the expression levels of pro-inflammatory factors TNF-α and IL-6;

[0060] Figure 21A and Figure 21B The SF liquid bandage of the present invention demonstrates its ability to promote cell migration in vitro, wherein... Figure 21A Images and migration rate results of L929 cells at the scratch site observed under a microscope at 0h, 6h, 12h, and 24h are shown. Figure 21B Images and migration rate results of HUVEC cells at the scratch site observed under a microscope at 0h, 6h, 12h, and 24h are shown.

[0061] Figure 22 The results show the hemostatic effect of the SF liquid bandage of the present invention in vivo (compared with the blank group, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001);

[0062] Figure 23 The in vivo healing effect of the SF liquid bandage of the present invention is shown (compared with the blank group, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, **** indicates P<0.0001);

[0063] Figure 24 The in vivo anti-inflammatory effect of the SF liquid bandage of the present invention on day 3 was demonstrated, namely the effect of different liquid bandages on the expression levels of different pro-inflammatory factors;

[0064] Figure 25 The in vivo anti-inflammatory effect of the SF liquid bandage of the present invention on day 7 was demonstrated, namely the effect of different liquid bandages on the expression levels of different pro-inflammatory factors;

[0065] Figure 26 The images show the in vivo healing effect of the SF liquid bandage of the present invention, namely H&E staining images and wound length data;

[0066] Figure 27 The in vivo healing effect of the SF liquid bandage of the present invention is shown in Masson staining images and collagen deposition fraction;

[0067] Figure 28 The in vivo healing-promoting effect of the SF liquid bandage of the present invention is shown in the Sirius red staining image and immunohistochemical results.

[0068] Figures 29A to 29C The skin irritation of the SF liquid bandage of the present invention in rats was demonstrated, wherein... Figure 29A This indicates the test sites for different liquid bandages. Figure 29B Photos of the test sites taken at 0h, 1h, 24h, 48h, 72h, and 96h after the liquid bandage was removed. Figure 29C This indicates the primary irritation index of different liquid bandages. Invention Details

[0070] definition

[0071] As used herein, the term "silk fibroin" includes silk fibroin, insect or spider silk fibroin, or recombinant silk fibroin. In one embodiment, the silk fibroin is obtained from domestic silkworms.

[0072] As used in this article, the term "liquid bandage," also known as liquid adhesive bandage or liquid plaster, is a new type of skin wound protection product. A liquid bandage is a liquid or semi-solid substance applied or sprayed onto a wound that quickly forms a protective film on the wound surface, isolating bacteria, keeping the wound moist, and promoting wound healing. Compared to traditional bandages, liquid bandages offer better adhesion, breathability, and moisture permeability, while also reducing the likelihood of allergic reactions and being easier to apply.

[0073] As used herein, the term "dressing" refers to various medical materials used to cover wounds / injuries. Its function is to temporarily act as a partial barrier for the skin during wound healing, providing protection or support for wound re-epithelialization or transition to permanent reconstruction. Dressings may include, but are not limited to, gauze made of natural cotton or synthetic fibers, bandages, sponge dressings consisting of a hydrophobic outer layer and an inner layer of hydrophilic foam, and hydrocolloid dressings that are permeable to water vapor but can block bacteria, and also absorb wound exudate and remove some necrotic tissue.

[0074] As used herein, the term "film-forming material" refers to a material that, in a liquid or semi-solid state, can form a uniform, continuous film with a certain strength and durability on a coated surface through drying or other curing processes. Commonly used film-forming materials include, but are not limited to: polyethylene materials, such as polyvinyl alcohol, polyvinylpyrrolidone, and ethylene-vinyl acetate copolymer; polypropylene materials, such as methyl polypropylene and methacrylic acid-methyl acrylate copolymer; cellulose materials, such as carboxymethyl cellulose, methyl cellulose, and ethyl cellulose; and natural polymer materials, such as gelatin, guar gum, corn gum, and sodium alginate.

[0075] As used herein, the term "gel" refers to a thick liquid or semi-solid formulation in the form of a solution, suspension, or emulsion. Gel matrices are single-phase dispersion systems and can be aqueous or oil-based. Aqueous gel matrices are generally composed of water, glycerol or propylene glycol with cellulose derivatives, carbomer and alginate, tragacanth gum, gelatin, starch, etc.; oil-based gel matrices are composed of liquid paraffin with polyethylene or fatty oils with colloidal silica or aluminum soaps, zinc soaps, etc.

[0076] As used herein, the term "salt" includes, for example, salts of inorganic acids and salts of organic acids. Examples of salts may include hydrochlorides, phosphates, pyrophosphates, hydrobroms, sulfates, sulfinates, nitrates, malates, maleates, fumarates, tartrates, succinates, citrates, acetates, lactates, methanesulfonates, p-toluenesulfonates, 2-hydroxyethylsulfonates, benzoates, salicylates, stearates, and alkylates (e.g., acetates, HOOC-(CH2)). n -COOH, where n is 0-4). Furthermore, if the compound herein is obtained as an acid addition salt, the free base can be obtained by alkalizing a solution of the acid salt. Conversely, if the compound herein is a free base, the addition salt (particularly a pharmaceutically acceptable addition salt) can be prepared by dissolving the free base in a suitable organic solvent and treating the solution with acid, according to conventional procedures for preparing acid addition salts from base compounds. Those skilled in the art will understand the various synthetic methods that can be used to prepare non-toxic, pharmaceutically acceptable addition salts. Detailed Implementation Plan

[0077] Silk fibroin (SF) is mainly derived from silkworms and contains 18 amino acids. Its microstructure includes crystalline and amorphous regions. The crystalline regions form stable β-sheets, providing toughness and tensile strength; the amorphous regions provide ductility and elasticity. Studies have shown that SF has many advantages, such as its natural origin, good biocompatibility and biodegradability, and controllable mechanical properties.

[0078] Based on the aforementioned characteristics of SF, it is used in this invention as a liquid bandage or dressing. In one embodiment, this invention provides a silk fibroin liquid bandage, which has the characteristics of short film-forming time, flexible and transparent film, which is conducive to wound adhesion, and easy to peel off and remove.

[0079] Example

[0080] The technical solution of the present invention will be described in detail below with reference to the embodiments. However, those skilled in the art should understand that the following embodiments are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention.

[0081] The main materials and their sources are as follows:

[0082] Silk fibroin, SF-S with a molecular weight of 8-50kDa (batch number SPSDPK1T010) and SF-M with a molecular weight of 50-100kDa (batch number SLPSXK1N005), was provided by Shenzhen Huasi Biotechnology Co., Ltd.

[0083] Comparative sample 1 was prepared according to the method of Example 2 in the specification of the patent (application number: 201510856810.1);

[0084] Comparative sample 2 is a commercially available product, Kobayashi Pharmaceutical Liquid Bandage, with a specification of 10g.

[0085] The performance of the samples in the following examples and similar products (comparison samples 1 and 2) was evaluated using the following methods:

[0086] 1. Irritating

[0087] The irritancy was evaluated using the nasal inhalation method. Specific evaluation criteria are as follows:

[0088] Stimulating evaluation Fraction No pungent odor 0 Slightly pungent odor (weak irritation, can be masked) 1 A strong, pungent odor (highly irritating, but can be masked). 2 Strong, pungent odor (highly irritating and cannot be masked). 3

[0089] 2. Film formation time and film state

[0090] Membrane preparation: Take 0.05 mL of liquid bandage sample and inject it onto a dry glass plate, ensuring a consistent spread area (1 cm × 1 cm). Immediately start timing with a timer. Stop timing when the film is no longer sticky to the touch. Record the film formation time and observe whether a film can be formed, as well as the flexibility and transparency of the formed film.

[0091] Example 1: Liquid Bandages with Different Components

[0092] Liquid bandages with different components were prepared. The composition of the components is shown in Table 1. The appearance of each sample was recorded, and the irritation, film-forming time and film state were evaluated.

[0093] Table 1 Liquid Band-Aids with Different Components

[0094]

[0095] Preparation method:

[0096] The preparation steps for samples 1-2 are as follows:

[0097] Step (1): Weigh the amount of SF (SF-S molecular weight is 8-50kDa; SF-M molecular weight is 50-100kDa) according to the prescription in Table 1, and add it in portions to a beaker containing the amount of water in Table 1, stirring until completely dissolved;

[0098] Step (2): Weigh PVB into a stoppered glass bottle according to the prescription amount in Table 1, add anhydrous ethanol according to the prescription amount in Table 1, and stir at 500 rpm until completely dissolved.

[0099] Step (3): Add SF aqueous solution to PVB ethanol solution and stir at 500 rpm until completely mixed to obtain liquid bandage composition.

[0100] Sample 3: Add glycerol to the mixed solution of PVB and SF in step (3) above, and mix well to obtain the sample.

[0101] Sample 4: Prepared according to step (1) above.

[0102] Sample 5: Prepare an SF aqueous solution according to step (1) above, then add the prescribed amount of anhydrous ethanol and mix well to obtain the final product.

[0103] Sample 6: Weigh SF according to the prescription amount, and add it in portions to a beaker containing the prescription amount of anhydrous ethanol under stirring conditions, allowing it to be completely dispersed.

[0104] Sample 7: Prepare a PVB ethanol solution according to step (2) above, then add the prescribed amount of water and stir to mix it completely.

[0105] As shown in Table 1, samples 1-3 can form good films with a film formation time of less than 3 minutes and are less irritating than control samples 1 and 2. Samples 4 and 5 contain only SF and no film-forming material, resulting in a longer film formation time, making the film difficult to peel off and exhibiting poor water resistance. Sample 6 cannot form a film. Sample 7 contains only film-forming material and no SF, resulting in a longer film formation time.

[0106] Example 2: Liquid Bandages with Different Film-Forming Materials and Their Doses

[0107] Liquid bandages were prepared by combining SF with different film-forming materials and their amounts. The composition is shown in Table 2. The appearance of each sample was recorded, and the irritation, film-forming time and film state were evaluated.

[0108] Table 2 Liquid Bandages with Different Film-Forming Materials

[0109]

[0110]

[0111] The preparation methods for samples 8-10 are the same as those for samples 1-2 in Example 1.

[0112] The preparation steps for sample 11 are as follows:

[0113] Step (1): Weigh SF according to the prescription amount, add it in portions to a beaker containing the prescription amount of water under stirring, and let it dissolve completely. Then add the film-forming material PVA and stir until it is completely dissolved.

[0114] Step (2): Add the aqueous solution containing SF and PVA to the prescribed amount of ethanol solution and stir until completely mixed.

[0115] The preparation steps for sample 12 are as follows:

[0116] Step (1): Weigh SF according to the prescription amount, add it in portions to a beaker containing the prescription amount of water under stirring, and let it dissolve completely. Then add the film-forming material PVA and stir until it is completely dissolved.

[0117] Step (2): Weigh the film-forming material PVB into a stoppered glass bottle according to the prescription amount, add the prescription amount of anhydrous ethanol, and stir until completely dissolved;

[0118] Step (3): Add the aqueous solution containing SF and PVA to the ethanol solution containing PVB and stir until completely mixed.

[0119] The preparation method of sample 13 is the same as that of samples 1-2 in Example 1, except that PVB is replaced with EC in the prescription amount in Table 2.

[0120] The preparation method of samples 14-15 is the same as that of sample 11, except that PVA is replaced with the amount of MC or HPC prescribed in Table 2.

[0121] The above results indicate that when the mass percentage of the film-forming material is greater than 8%, the film-forming time of the liquid bandage is less than 3 minutes.

[0122] Example 3: Liquid Bandages with Different SF Doses

[0123] Liquid bandages with different SF dosages were prepared. The composition is shown in Table 3. The appearance of each sample was recorded, and the irritation, film-forming time, and film state were evaluated.

[0124] Table 3 Liquid Bandages with Different SF Doses

[0125]

[0126] The preparation method is the same as that for samples 1-2 in Example 1.

[0127] The results showed that liquid bandages with a SF mass percentage of 1-3% formed a film in less than 3 minutes.

[0128] Example 4: Liquid Bandages with Different Additives and Doses

[0129] Liquid adhesive bandages with different additives and dosages were prepared. The composition is shown in Table 4. The appearance of each sample was recorded, and the irritation, film-forming time and film state were evaluated.

[0130] Table 4 Liquid Bandages with Different Additives and Doses

[0131]

[0132]

[0133] The preparation methods for samples 21-24 are the same as those for sample 3 in Example 1;

[0134] The preparation method of sample 25 is the same as that of sample 12 in Example 2.

[0135] The results showed that the addition of additives improved the film-forming properties of the liquid bandage.

[0136] Example 5: Liquid Band-Aids with Different Proportions of Organic and Aqueous Phases

[0137] Liquid bandages with different ratios of organic and aqueous phases were prepared. The composition is shown in Table 5. The appearance of each sample was recorded, and the irritation, film-forming time and film state were evaluated.

[0138] Table 5. Liquid Band-Aids with Different Proportions of Organic and Aqueous Phases

[0139]

[0140] The preparation method is the same as that for samples 1-2 in Examples.

[0141] As can be seen from the table above, liquid bandages can be prepared when the total volume of the aqueous and organic phases is 100% and the volume percentage of the aqueous phase is 0-40%. Among them, liquid bandages with an aqueous phase volume percentage of 20%-40% show better film formation and stability.

[0142] Example 6: Comparison of film-forming ability of film-forming materials with different viscosity grades

[0143] Film-forming ability was investigated using PVB of different viscosity grades as film-forming materials. The composition is shown in Table 6.

[0144] Table 6 Comparison of film-forming capabilities of film-forming materials with different viscosity grades

[0145]

[0146]

[0147] Preparation method: Weigh the film-forming material PVB into a stoppered glass bottle according to the prescription amount, add the prescription amount of anhydrous ethanol, and stir until completely dissolved to obtain the final product.

[0148] Table 6 shows that PVB with a viscosity greater than 2800 mPa·s is not easy to apply.

[0149] The following describes the characterization, performance testing, and in vitro and in vivo efficacy evaluation of the SF liquid bandage of the present invention. Samples 1-2, 7, and 16-22 are designated as S-1%, M-1%, PVB, S-0.5%, S-2%, S-3%, M-2%, M-3%, PEG 200, and PEG 400, respectively. Comparative samples 1 and 2 are designated as PVP-PVB and Xiaolin, respectively.

[0150] Example 7: Morphological Characterization

[0151] Take 0.8 ml of each of samples 1-2, 7, and 16-20 and inject them into 6-well plates, ensuring consistent development area. Allow the plates to stand for about 12 hours to allow complete film formation. Remove the films and perform morphological characterization.

[0152] 1. Membrane appearance

[0153] The appearance of the films formed from samples 7, 18, and 19 is as follows: Figure 1 As shown.

[0154] The results showed that the film formed by the PVB group (sample 7) was bright, flexible and transparent; while the films formed by the S-3% group (sample 18) and the M-2% group (sample 19) were rough, flexible and relatively transparent.

[0155] 2. Membrane micromorphology

[0156] The thin films formed from samples 7, 1, and 16-18 were observed on their outer surfaces using a scanning electron microscope (SEM). Figure 2 First row) and internal cross-sectional micromorphology ( Figure 2 (second line), the result is as follows Figure 2 As shown.

[0157] The results showed that, compared with the PVB group (sample 7), the films formed by samples 1 and 16-18 had a porous network structure, and the SF liquid bandage of the present invention is beneficial for the absorption of wound exudate.

[0158] 3. Membrane surface roughness

[0159] The surface roughness of the films formed from samples 1-2, 7, and 17-20 was observed using atomic force microscopy (AFM). The results are shown in the figure. Figure 3A The arithmetic mean height Ra, root mean square height Rq, and height value relative to the reference plane of the AFM measurement results were statistically analyzed. The results are shown in […]. Figure 3B .

[0160] The results showed that, compared with the PVB group (sample 7), the film surfaces formed by samples 1-2 and 17-20 were rougher, and the SF liquid bandage of the present invention is beneficial to cell adhesion and proliferation.

[0161] 4. SF secondary structure

[0162] The thin films formed from samples 7, 18, and 19 were analyzed using Fourier transform infrared (FT-IR) to determine the relative proportions of α-helices and β-sheets in the SF secondary structure. The results are shown in [Figure number missing]. Figure 4 .

[0163] The results showed that, compared with the FT-IR spectrum of sample 7, the FT-IR spectra of samples 18 and 19 showed new peaks, indicating that SF and PVB were successfully mixed. In addition, SF in samples 18 and 19 mainly existed in the form of α-helices in the liquid. During the film formation process, the α-helices quickly transformed into β-sheet structures, which improved the stability of the film and increased its water resistance, which is beneficial for practical use.

[0164] Example 8: Performance Testing

[0165] 1. Rheological properties

[0166] The rheological curves of samples 7, 18, and 19 were measured using a rotational rheometer. Measurement conditions: Shear rate: 0.1 s⁻¹. -1 -100s -1 Temperature: 37℃, Sample loading volume: 0.32ml. Results are shown below. Figure 5 .

[0167] The results show that the SF liquid bandage of the present invention exhibits shear-thinning behavior, has better stability during static storage, and decreases viscosity during application, which is beneficial for uniform coating. After coating, it immediately becomes viscous upon standing, making it less prone to flow loss.

[0168] The rotational rheometer was used to detect the samples 1-2, 7, 17-19, and control sample 2 at 10 s. -1 Viscosity values ​​at shear rates, the results are shown in [reference needed]. Figure 6 .

[0169] The results showed that, compared with the PVB group (sample 7), the addition of SF could significantly increase the viscosity of the film-forming material, and the present invention had better viscosity than Xiaolin (comparative sample 2), which is beneficial for adhesion to the wound during application.

[0170] 2. Breathability

[0171] Take 0.5 ml of each of sample 7, samples 17-19 and control sample 2, inject them into a 12-well plate, and let it stand for about 12 hours to allow the film to form completely. Then peel off the film and test the air permeability.

[0172] Add a certain volume of distilled water to a vial (φ: 1.4cm) until the water level is 5mm from the mouth of the vial. Cover the mouth of the vial with the sample membrane and seal the mouth and membrane edges with sealing film. Weigh the initial weight M0 of each sample membrane and place it in a 37℃ incubator, weighing it at different time points. The formula for calculating water vapor transmission rate is as follows: WVTR (mg / cm³) 2 / h)=(M0-Mx) / s·t, where M0 is the initial weight (mg), Mx is the weight at different time points (mg), and s is the surface area of ​​the vial (cm²). 2 ), where t is the placement time (h).

[0173] The air permeability curves of samples 7, 18, and 19 are shown below. Figure 7 The results showed that the WVTR of samples 18 and 19 was higher than that of the PVB group (sample 7) from 3h to 72h, indicating that the SF liquid bandage of the present invention has good breathability.

[0174] The air permeability results of samples 7, 17-19 and control sample 2 at 12h, 24h and 48h are shown in the figure. Figure 8 The results showed that, compared with PVB (sample 7), the SF liquid bandage of the present invention has better breathability and is superior to Xiaolin (comparative sample 2).

[0175] 3. Water resistance

[0176] Take 0.5 ml of each of samples 1-2, 7, 17-19 and control sample 2, inject them into a 12-well plate, and let it stand for about 12 hours to allow the film to form completely. Then peel off the film to test the water resistance.

[0177] A certain volume of distilled water was added to each vial (φ: 1.4cm). The film formed by each sample was then placed over the vial opening, and the opening and the edge of the film were sealed with sealing film. The initial weight M0 was measured, and the vial was placed upside down on a petri dish at room temperature for 24 hours. The weight of the inverted vial was then measured and recorded as M24h. Water resistance was characterized by water loss, calculated using the following formula: Water loss (%) = (M0 - M24h) / Mw × 100, where M0 is the initial weight (g), M24h is the weight after 24 hours (g), and Mw is the initial mass of distilled water (g). Results are shown below. Figure 9 .

[0178] The results show that, compared with the PVB group, the SF liquid bandage of the present invention has a lower water loss rate and better waterproof performance.

[0179] 4. Water absorption

[0180] Take 0.5 ml of each of samples 1-2, 7, and 17-19 and inject them into 12-well plates. Allow the plates to stand for about 12 hours to allow the membranes to form completely. Then peel off the membranes and test their water absorption.

[0181] The membrane was pre-weighed (M0) and then added to a centrifuge tube containing 10 mL of PBS. The tube was placed in a 37°C constant-temperature shaking water bath and removed at different time points (12h, 24h, 36h, 48h, 60h, 72h). The surface solution was wiped off with filter paper, and the membrane was weighed again. The water absorption rate was calculated using the following formula: Water absorption rate (%) = (Mx - M0) / M0 × 100%, where Mx is the weight (g) at different time points, and M0 is the initial weight (g). Results are shown below. Figure 10 .

[0182] The results showed that the absorbency of SF liquid bandages in samples 1-2 and 17-19 was higher than that in the PVB group (sample 7), indicating that the introduction of SF has the ability to increase the absorption of wound tissue exudate.

[0183] 5. Mechanical properties

[0184] Take 0.8 ml of samples 1-2, 7, 16-19, 21-22 and control samples 1-2, and inject them into 6-well plates respectively. Let them stand for about 12 hours to allow the film to form completely, then peel off the film and test its mechanical properties.

[0185] The tensile strength and elongation at break of films formed from liquid bandages with different SF-S dosages (i.e., samples 1, 7, 16-18, and 21-22) were determined at room temperature using a universal testing machine (AGS-X-10N, Japan). Specifically, rectangular films cut into 10mm × 10mm pieces were tested at a tensile speed of 5mm / min. The tensile strength and elongation at break were calculated based on the corresponding tensile stress-strain curves. The formula for calculating elongation at break is as follows: Elongation at break (%) = (L... break -L original ) / L original ×100, where L break L is the film length (mm) under maximum stress. original The initial length of the film (mm) is shown in the results. Figure 11 .

[0186] The results showed that the tensile strength and elongation at break of the PVB group (sample 7) were lower than those of samples 1, 17-19 and 21-22, while the films formed by the addition of PEG 200 and PEG 400 (i.e. samples 21-22) did not significantly improve the tensile strength and elongation at break.

[0187] The films formed from samples 1-2, 7, 16-18, and control sample 1-2 were tested using a universal testing machine (Instron 5967, USA) at room temperature. The specific testing method was as follows: rectangular films cut into 10 mm × 5 mm sections were tested at a tensile speed of 5 mm / min. Young's modulus, tensile strength, and elongation at break were obtained from the corresponding tensile stress-strain curves. The results are shown below. Figure 12 .

[0188] The results showed that SF liquid bandage has suitable rigidity (Young's modulus and tensile strength) and toughness (elongation at break), falling between the two marketed products, and provides good comfort.

[0189] 6. Removal properties

[0190] 0.5 ml of sample 19 was pipetted into a 24-well plate and allowed to stand for approximately 12 hours to allow complete film formation. The film was then peeled off and added to 10%, 30%, 50%, 70%, and 90% ethanol solutions, respectively. The elimination of the film was observed at 10 min, 30 min, 1 h, 1.5 h, 2 h, 2.5 h, 3 h, 4 h, 5 h, and 12 h. Results are shown below. Figure 13 .

[0191] The results showed that SF liquid bandage could be completely dissolved in 70% ethanol in 10 minutes, indicating that it has good removability. It can be removed without mechanical peeling, simply by rinsing and soaking, thus avoiding secondary damage to the healing wound.

[0192] 7. Blood compatibility

[0193] Take 0.05 ml of each of the following samples: Sample 1-2, Sample 17-19, and Control Sample 1-2, and inject them onto a dry glass plate. Ensure the spread area is consistent and allow the plate to stand for about 12 hours to allow complete film formation. Remove the film and perform a blood compatibility test.

[0194] The specific method was as follows: Fresh blood was collected from the abdominal aorta of rats using an anticoagulant tube containing an anticoagulant. The blood was thoroughly mixed, centrifuged at 3000 rpm for 15 min to obtain erythrocyte pellet. The supernatant was discarded, and the pellet was washed three times with PBS solution. The pellet was then resuspended in PBS (2%, v / v) for later use. 250 μL of the 2% erythrocyte suspension was thoroughly mixed with 250 μL of PBS, 250 μL of Triton X-100 (1%, v / v), and 250 μL of PBS containing the sample, respectively, and incubated at 37°C for 1 h. After incubation, the pellet was centrifuged at 3000 rpm for 15 min, and the OD value at 570 nm was measured using the supernatant. Three biological replicates were set up for each group. Results are shown below. Figure 14 .

[0195] The results show that the SF liquid bandage of the present invention has good blood compatibility (<5%) and good safety.

[0196] 8. Cell compatibility

[0197] Take 0.05 ml of each of the following samples: Sample 1-2, Sample 7, Sample 17-19, and Control Sample 1-2, and inject them onto a dry glass plate, ensuring a consistent spreading area. Allow the plate to stand for approximately 12 hours to allow complete film formation. Remove the film and add it to a certain volume of basal culture medium (10 mg / ml). Incubate at 37°C for 24 hours. Then remove the film and filter the remaining culture medium through a 0.2 μm sterile microporous membrane to obtain the corresponding extract, which will be used as the test sample.

[0198] Cell viability:

[0199] L929 cells (mouse fibroblasts) in the logarithmic growth phase were injected with 1×10 5 Cells / wells were seeded in 96-well plates. After cell adhesion, the culture medium was removed, and the cells were cultured with the above-mentioned extract for 72 hours. The extract was then removed, and culture medium containing 10% CCK8 reagent was added to each well. After incubation in an incubator for a period of time, the OD value at 450 nm was measured. The results are shown below. Figure 15 .

[0200] The results showed that, compared with the blank culture medium group (TCP) without any added samples, the cell viability of control sample 2 was significantly reduced; the SF liquid bandages of each group of the present invention can significantly promote cell growth and are beneficial to the wound healing process.

[0201] Cell morphology:

[0202] L929 cells in logarithmic growth phase were fed a 1×10⁻⁶ dose. 5 Cells / wells were seeded in 24-well plates. After cell adhesion, the culture medium was removed, and the cells were cultured with the test extract for 24 h and 48 h. The extract was then removed, and AM / PI reagent was added to each well. The plates were then placed in an incubator, and after 30 min, the cells were removed and observed for morphology under a fluorescence microscope. Raw and HUVECs cells were tested using the same method. Results are shown below. Figure 16 .

[0203] The results showed that, compared with the blank culture medium group (TCP) without any samples, the number of cells in control sample 2 was significantly reduced; the SF liquid bandages of samples 1-2 and 17-19 significantly promoted cell growth, which was better than control sample 1 and was beneficial to the wound healing process.

[0204] 9. Free radical scavenging ability – antioxidant properties

[0205] The SF liquid bandage of this invention is effective against DPPH and ABTS. + The antioxidant capacity is evaluated by its scavenging ability, and the procedure is as follows:

[0206] DPPH: Take 0.4 ml of each of the following samples: Sample 1-2, Sample 7, Sample 17-20, and Control Sample 1-2, and add them to 1.2 ml of anhydrous ethanol solution containing 100 μM DPPH. After mixing completely, let stand at 37°C in the dark for 30 min, centrifuge at 10000 rpm for 10 min, and take the supernatant to measure the absorbance at 517 nm.

[0207] ABTS + Add 0.1 ml of samples 1-2, 7, 17-20, and control samples 1-2 to 0.3 ml of ABTS. + After thoroughly mixing the working solution, incubate at 37°C in the dark for 30 min, centrifuge at 10000 rpm for 10 min, and measure the OD value at 734 nm using the supernatant. Results are shown below. Figure 17 .

[0208] The results show that the present invention is effective against DPPH and ABTS. + The removal rates were all high, and their antioxidant effects were similar to those of control sample 2 and superior to those of control sample 1.

[0209] Example 9: In vitro efficacy evaluation

[0210] The regulatory effect of this invention on the immune microenvironment was evaluated by its effects on macrophage polarization, anti-inflammation, and antioxidant activity.

[0211] The prescribed amounts of SF and PVB from samples 7, 17-20, and control sample 1 were added to a certain volume of basal culture medium. After being fully dissolved, the solutions were filtered through a 0.2 μm sterile microporous membrane to obtain the corresponding extracts.

[0212] 1. Antioxidant properties

[0213] L929 cells in logarithmic growth phase were fed a 1×10⁻⁶ dose. 5 Cells / wells were seeded in 24-well plates. After cell adhesion, the culture medium was removed, and the above-mentioned extract (containing 0.1 mg / ml of reactive oxygen species Rousp) was added to each well. After incubation for 30 min, the extract was removed, and basal medium containing 10 μM DCFH-DA was added to each well. The plates were then incubated for another 30 min. The culture medium was removed, and the cells were washed with PBS. Basal medium containing 1X Hoechst was added to each well, and the plates were incubated for 10 min. Green fluorescence was observed under a fluorescence microscope, and the average fluorescence intensity was analyzed using ImageJ. Raw cells were tested using the same method. Results are shown below. Figure 18 .

[0214] The results showed that, compared with the positive control group (PC group) which only added Rousp, all groups of SF liquid bandage had antioxidant capacity, which could significantly alleviate the oxidative stress induced by Rousp and promote the healing process.

[0215] 2. Cell polarization

[0216] Raw 264.7 cells in logarithmic growth phase were fed at a rate of 2 × 10⁻⁶. 5 Cells / wells were seeded in 24-well plates. After cell adhesion, the original culture medium was discarded, and the cells were washed with PBS. Separate extraction solutions were added, with basal medium containing 1 μg / ml LPS as a control. After 24 h of culture, the supernatant was discarded, and the cells were washed with PBS. The cells were then fixed and permeabilized with 4% paraformaldehyde / 0.5% Triton X-100 solution, washed with PBS, and each well was incubated with 100 nM FITC-labeled phalloidin working solution. The plates were incubated at room temperature in the dark for 30 min, washed with PBS, and finally mounted with a DAPI-containing anti-fluorescence quenching mounting medium. Green fluorescence was observed under a fluorescence microscope. Results are shown below. Figure 19 .

[0217] The results showed that, compared with the LPS-induced positive control (M1 Type group, cells showed pseudopodia), all groups of SF liquid bandage could reduce macrophage polarization to M1 type, and the cells still had a round shape, with better anti-polarization ability than control sample 1.

[0218] 3. LPS-induced anti-inflammatory ability

[0219] Raw cells in logarithmic growth phase were fed at a rate of 2 × 10⁻⁶. 5 Cells / wells were seeded in 12-well plates. After cell attachment, the original culture medium was discarded, and the cells were washed with PBS. Extraction solutions were added to each well, and the cells were incubated for 48 hours. Then, DMEM medium containing 1 μg / ml LPS was added for 3 hours of stimulation. The supernatant and wells were collected, and ELISA experiments were performed on TNF-α and IL-6 to detect their gene expression levels. Results are shown below. Figure 20 .

[0220] The results showed that, compared with the LPS-induced positive control (M1 Type group), sample 17-20 significantly reduced the expression levels of LPS-induced pro-inflammatory factors TNF-α and IL-6, demonstrating significant anti-inflammatory capabilities.

[0221] 4. Promotes cell migration

[0222] Fibroblasts and endothelial cells possess sufficiently strong migration capabilities to accelerate wound healing. This was evaluated using a cell scratch assay.

[0223] The prescribed amounts of SF and PVB from samples 7, 17-20, and control sample 1 were added to a certain volume of basal culture medium. After being fully dissolved, the solutions were filtered through a 0.2 μm sterile microporous membrane to obtain the corresponding extracts.

[0224] L929 cells in logarithmic growth phase were fed a 1×10⁻⁶ dose. 5 Cells / wells were seeded in 24-well plates. After cell adhesion, the cells were removed and streaked vertically along the center of each well using a 1-200 μL yellow pipette tip. The supernatant was discarded, and the cells were washed with PBS. Then, the appropriate extraction solutions were added, and cell migration at the streaks was observed under a microscope at 0 h, 6 h, 12 h, and 24 h. Results are shown below. Figure 21A The same method was used to test HUVECs cells, and the results are shown in the figure. Figure 21B .

[0225] The results showed that, compared with the TCP group without any samples, samples 17-20 all promoted the migration of L929 and HUVECs cells, and the cell migration rate increased with time, showing better results than control sample 1. This invention has the effect of reducing scratches.

[0226] Example 10: In vivo effect evaluation

[0227] 1. Hemostatic effect

[0228] The hemostatic effect of the SF liquid bandage of the present invention was evaluated using a mouse liver hemorrhage model.

[0229] SD rats were anesthetized and fixed. An abdominal incision was made to expose the liver, and the serous fluid around the liver was removed with gauze. Pre-weighed filter paper (M1), wrapped in sealing film, was placed under the liver, and hepatic bleeding was induced using a 6mm diameter perforator. After 10 seconds of free bleeding, samples 7, 18-19, and control samples 1-2 were injected into the bleeding sites, respectively. Hemostasis time was recorded, and the weight of the filter paper absorbing blood during hemostasis (M2) was measured. Blood loss was calculated using the following formula: Blood loss (mg) = M2 - M1. Results are shown below. Figure 22 .

[0230] The results showed that samples 18-19 could significantly shorten bleeding time and reduce bleeding volume, which was beneficial for rapid hemostasis and had a better hemostatic effect than control sample 1.

[0231] 2. Promotes wound healing

[0232] A full-thickness incision model was used in mice. ICR mice were anesthetized and fixed. An approximately 8 mm diameter full-thickness skin incision was made in the center of the spine of the hairless mice using a motorized punch. Samples 7, 18-19, and control samples 1-2 were applied, and the incisions were secured with gauze and bandages. The treated mice were placed in a dry, clean environment, and the dressings were changed daily. The healing of the wounds on the backs of the mice was observed on days 0, 3, 7, 10, and 14, and changes in wound area were analyzed using ImageJ. Results are shown below. Figure 23 .

[0233] The results showed that samples 18-19 significantly shortened the healing time in the early stages of wound healing (days 3, 7, and 10), which was superior to control samples 1 and 2.

[0234] 3. Anti-inflammatory

[0235] Inflammatory factors (IFN-γ, TNF-α, IL-12p70, IL-1β, IL-6, IL-17A, and IFN-β) in wound skin tissue homogenates on days 3 and 7 were detected using flow cytometry (BD FACSCelesta) and a flow cytometry kit (Biolegend). Results are shown below. Figure 24 (Day 3) and Figure 25 (Day 7)

[0236] The results showed that, compared with the blank group without any samples, the treatment with the present invention's samples 18-19 for 3 and 7 days could reduce the expression levels of pro-inflammatory factors. The present invention's SF liquid bandage can reduce the inflammatory response at the wound site, which is conducive to accelerating the transformation from the inflammatory phase to the proliferative phase, thereby promoting wound healing.

[0237] 4. Collagen deposition and regeneration of skin appendages

[0238] Skin tissue from the wound was fixed with 4% paraformaldehyde, embedded in paraffin, and sectioned. H&E, Masson's red, and Sirius red staining, as well as immunohistochemical staining for VEGF, CD31, and CK5, were performed. Positive area analysis was conducted using CaseViewer 2.1 and ImageJ. H&E staining results are shown below. Figure 26 Masson staining results are shown in [link to results]. Figure 27 Sirius red staining and immunohistochemical results are shown in [link to results]. Figure 28 .

[0239] The results showed that, compared with the blank group without any samples, samples 18-19 of the present invention could accelerate wound contraction at different healing stages and promote collagen deposition; at the same time, they increased the positive area of ​​VEGF and CD31, promoted the increase of CK5, which was beneficial to the formation of angiogenesis in the proliferative phase and the regeneration of hair follicles in the remodeling phase. Their effects were better than those of control samples 1 and 2, and they showed a trend of promoting the deposition of type I and type III collagen.

[0240] 5. Skin irritation

[0241] Safety was assessed using a rat skin irritation test.

[0242] SD rats (350 mg / kg) were anesthetized and fixed with 10% chloral hydrate. The skin on their backs was divided into sections using a marker. Samples 18-19, control samples 1-2, 1×PBS solution, and 20% SLS solution (the solution was applied to gauze at the corresponding locations) were applied to these sections. After 4 hours of incubation in a dry, clean environment, the gauze and samples were removed. The backs were washed with warm water to remove any remaining material. Photos were taken at 0h, 1h, 24h, 48h, 72h, and 96h after removal, and the erythema and edema at each time point were recorded. Scoring was performed according to GB / T 16886.10 standards to obtain the primary irritation index. Results are shown below. Figures 29A to 29C .

[0243] The results showed that the SF liquid bandage of the present invention had no irritation and good safety, while both control sample 1 and control sample 2 had mild irritation.

Claims

1. A pharmaceutical composition comprising silk fibroin (SF) and a film-forming material.

2. The composition according to claim 1, wherein the film-forming material is a water-soluble film-forming material and / or a fat-soluble film-forming material; Preferably, the water-soluble film-forming material is selected from one or more of polyvinyl alcohol (PVA), carbomer, methyl cellulose (MC), hydroxyethyl cellulose, hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose, chitosan, sodium alginate, and sodium carboxymethyl cellulose. Preferably, the lipid-soluble film-forming material is selected from one or more of nitrocellulose (NC), polyvinyl butyral (PVB), acrylates, ethyl cellulose (EC), cellulose acetate (CA), and polyurethane.

3. The composition according to claim 1 or 2, wherein the silk fibroin content is 0.5wt%-20wt%, preferably 1wt%-10wt%, more preferably 1wt%-5wt%, even more preferably 1wt%-3wt%, and most preferably 2wt%-3wt%, based on the total mass of the composition.

4. The composition according to any one of claims 1-3, wherein the content of the film-forming material is 8 wt%-25 wt%, preferably 10 wt%-22 wt%, more preferably 10 wt%-20 wt%, and even more preferably 16 wt%-20 wt%, based on the total mass of the composition.

5. The composition according to any one of claims 1-4, wherein the composition further comprises a solvent, the solvent being an aqueous phase and / or an organic phase, preferably, the solvent being a mixture of an aqueous phase and an organic phase, more preferably, the volume percentage of the aqueous phase being 0%-40% based on the total volume of the composition, preferably 20%-40%.

6. The composition according to any one of claims 1-5, characterized in that, The molecular weight of the silk fibroin is 8-100 kDa; preferably 8-50 kDa or 50-100 kDa.

7. The composition according to any one of claims 1-6, characterized in that, The film-forming material contains polyvinyl butyral (PVB), and preferably, the viscosity of the PVB does not exceed 2800 mPa·s.

8. The composition according to any one of claims 1-7, characterized in that, The composition also contains additives; Preferably, the additive is selected from one or more of self-adhesives, plasticizers, and cosolvents. More preferably, the additive is selected from one or more of polyols (including polyethylene glycol, glycerin, propylene glycol, sorbitol, xylitol), dibutyl phthalate, tributyl acetyl citrate, sucrose isobutyrate, sucrose benzoic acid, acetyl citrate, mineral oil, hyaluronic acid (HA), and polyvinylpyrrolidone (PVP).

9. The composition according to any one of claims 1-8, characterized in that, The organic phase is selected from one or more of the following: alcohols, low molecular weight alkanes and cycloalkanes, alkanes, ethers, siloxanes, volatile fluorocarbons, polysorbates, and dehydrated sorbitan fatty acid esters; Preferably, the alcohols are selected from one or more of methanol, ethanol, butanol, benzyl alcohol, phenylene glycol, tert-butanol, and isopropanol; the cycloalkanes are selected from one or more of n-pentane, hexane, heptane, cyclohexane, and isooctane; the alkyl esters are selected from one or more of ethyl acetate, butyl acetate, n-propyl acetate, isobutyl acetate, hexyl acetate, isopropyl myristate, castor oil, and isobutyl isobutyrate; the ethers are selected from one or more of hexamethyldisiloxane and diethyl ether; the siloxanes are selected from one or more of hexamethyldisiloxane, decamethylcyclopentanesiloxane, octamethyltrisiloxane, and polydimethylsiloxane; and the volatile fluorocarbons are selected from one or more of pentafluoropropane and perfluoroheptane.

10. The composition according to any one of claims 1-9, characterized in that, The composition also contains a drug, preferably selected from one or more of antibiotics, metal ions, anti-inflammatory drugs, and active ingredients of traditional Chinese medicine.

11. The composition according to any one of claims 1-10, wherein the composition is a solution, gel, semi-solid, spray, or suspension; preferably, the composition is a liquid dressing; more preferably, the composition is a liquid bandage; and even more preferably, the film-forming time of the composition is less than 3 minutes.

12. The composition according to any one of claims 1-11, comprising the following components: - Silk fibroin, based on the total mass of the composition, wherein the content of the silk fibroin is 1wt%-3wt%, and the molecular weight of the silk fibroin is 8–50kDa or 50–100kDa; -PVB, based on the total mass of the composition, the content of PVB is 16 wt%; - Water, with a volume percentage of 25% based on the total volume of the composition; - Ethanol, wherein the volume percentage of ethanol is 75% based on the total volume of the composition.

13. A method for preparing the composition according to any one of claims 1-12, comprising the following steps: (1) Dissolve SF in the aqueous phase to obtain an SF solution; The film-forming material is dissolved in an aqueous phase or an organic phase to obtain a film-forming material solution; (2) Mix the above SF solution and film-forming material solution; Preferably, the mixing method is mechanical stirring, high-speed shearing, high-pressure homogenization, or molecular diffusion during static settling.

14. Use of the composition according to any one of claims 1-12 or the composition prepared by the method according to claim 13 in the preparation of a medicament for the prevention and / or treatment of lesions and / or inflammatory diseases; Preferably, the injury is selected from superficial mechanical damage or burns to the skin, deep skin damage, or internal organ damage; Preferably, the inflammatory disease includes superficial skin inflammation, deep skin inflammation, or visceral inflammation.

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