External composition for promoting surgical incision healing and preparation method thereof

Abstract

The invention provides an external composition for promoting surgical incision healing and a preparation method thereof, and belongs to the technical field of medicines. Comprising a component A and a component B. The component A is powder and is prepared from banyan bark powder, traditional Chinese medicine powder, a decellularized material, collagen peptide and modified nano-hydroxyapatite according to the mass ratio of (2-4): (7-10): (3-5): (1-3): (2-4). And the component B is liquid and consists of carboxylated ionic liquid, chlorogenic acid / oleanolic acid-loaded chitosan, platelet-rich plasma and water in a mass ratio of (3-5): (5-8): (10-12): (200-300). The external composition for promoting surgical incision healing has good antibacterial, antioxidant and anti-inflammatory effects, can relieve pain, promote tissue regeneration, stop bleeding and restore, is safe and not prone to sensitization, achieves sustained and controlled release drug release, promotes surgical incision healing and scar repair, and has wide application prospects.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly relates to an external composition for promoting surgical incision healing and a preparation method thereof. Background Art

[0002] Surgical wounds are sharp instrument wounds, which have the characteristics of neat wounds but relatively large damage depth. Their wounds often penetrate the entire skin layer and reach the muscular layer. For parts with thick adipose tissue, the adipose tissue itself does not have the ability of self-growth and healing, and problems such as fat liquefaction will occur, which will greatly affect the wound healing. With the popularization of the application scope of surgical treatment, surgical treatment may be involved in the treatment of diseases in many disciplines such as orthopedics, oncology, neurology, oral and maxillofacial, etc.

[0003] Skin wounds rely on the self-renewal and healing of skin tissues. After the wound occurs, the body will initiate a healing cascade, starting processes such as hemostasis, self-defense, epithelial cell proliferation and coverage, and wound healing. Among them, the surgical process itself also involves operations such as hemostasis and anti-infection. Therefore, the speed of epithelial cell proliferation and coverage of the wound is the main reason affecting the healing speed of postoperative skin wounds. This process involves angiogenesis and blood supply at the surgical wound site. Clinical observations have found that most wounds with more bleeding heal well, while on the contrary, the wounds heal poorly or do not heal. Regarding the role of inflammatory response in wound healing, generally speaking, a moderate inflammatory response, as a defense mechanism, is beneficial to wound healing; while an excessive inflammatory response will delay wound healing.

[0004] Promoting surgical wound healing is one of the effective ways to relieve the pain of patients and reduce complications such as postoperative infections. Therefore, the research on drugs for promoting surgical wound healing has always been one of the hotspots.

[0005] Chinese invention patent CN105412980B discloses a medical glue for outpatient minor surgical incisions and a preparation method thereof, which includes raw materials in the following weight parts: aloe protein, yam protein, soluble ginkgo polysaccharide, polyethylene oxide aqueous solution, a-cyanoacrylate, chitosan, polyvinyl alcohol, polyaldehyde alginate, pectin, gelatin, hydrogenated camellia oil, etc. The present invention has good adhesive properties and can disinfect and inhibit wound infections. However, during the postoperative repair period, it needs to be frequently coated to meet the needs of wound repair, which is very troublesome to use and the drug consumption is large. Summary of the Invention

[0006] The purpose of the present invention is to provide an external composition for promoting surgical incision healing and a preparation method thereof, which have good antibacterial, antioxidant and anti-inflammatory effects, relieve pain and promote granulation, stop bleeding and restore vitality, and are safe and not easily sensitized, realizing sustained and controlled drug release, promoting the healing of surgical incisions and the repair of scars, and having broad application prospects.

[0007] The technical solution of the present invention is realized as follows: The present invention provides an external composition for promoting surgical incision healing, comprising component A and component B. Component A is a powder, which is composed of banyan bark powder, traditional Chinese medicine powder, acellular material, collagen peptide, and modified nano-hydroxyapatite, with a mass ratio of 2-4:7-10:3-5:1-3:2-4; Component B is a liquid, which is composed of carboxylated ionic liquid, chlorogenic acid / oleanolic acid-loaded chitosan, platelet-rich plasma, and water, with a mass ratio of 3-5:5-8:10-12:200-300.

[0008] As a further improvement of the present invention, the traditional Chinese medicine powder is prepared by washing, drying, pulverizing, and sieving lithospermum root, millettia reticulata, and achyranthes aspera to obtain the traditional Chinese medicine powder; the collagen peptide is selected from at least one of fish collagen peptide, pig collagen peptide, and bovine collagen peptide; the carboxylated ionic liquid is selected from at least one of 1-carboxymethyl-3-methylimidazolium chloride and 1-carboxymethylpyridinium bromide.

[0009] As a further improvement of the present invention, the mass ratio of lithospermum root, millettia reticulata, and achyranthes aspera is 3-5:5-8:1-3, and the mesh number of the sieve for sieving is 100-200 mesh.

[0010] As a further improvement of the present invention, the preparation method of the modified nano-hydroxyapatite is as follows: S1. Add nano-hydroxyapatite into Tris-HCl solution, add dopamine hydrochloride, and heat and stir to react to obtain a modified material; S2. Add 2-hydroxyterephthalic acid into water, add NHS and EDC, stir to activate, add the modified material, stir to react, centrifuge, add the product into a mixed solvent of N,N-dimethylformamide and acetic acid, add zirconium tetrachloride, carry out hydrothermal reaction, centrifuge, wash, and dry to obtain UiO-66-OH@modified material; S3. Dissolve inorganic salts in water, add UiO-66-OH@modified material, stir to react, centrifuge, wash, and dry to obtain Ag / Cu-deposited UiO-66-OH@modified material; S4. Add vascular endothelial growth factor-α and fibroblast growth factor into water, add NHS and EDC, stir to activate, add Ag / Cu-deposited UiO-66-OH@modified material, stir to react, centrifuge, wash, and dry to obtain modified nano-hydroxyapatite.

[0011] As a further improvement of the present invention, in step S1, the pH value of the Tris-HCl solution is 8.5 - 9.5, the mass ratio of nano-hydroxyapatite to dopamine hydrochloride is 8 - 10:2 - 3, the temperature of the heating and stirring reaction is 45 - 55 °C, and the time is 3 - 5 h; in step S2, the mass ratio of 2-hydroxyterephthalic acid, NHS, EDC to the modified material is 3 - 5:1 - 2:1 - 2:7 - 10, the mass ratio of the product to zirconium tetrachloride is 10 - 15:2 - 4, the temperature of the hydrothermal reaction is 120 - 130 °C, and the time is 20 - 24 h; in step S3, the inorganic salts include silver nitrate and copper salt, the mass ratio is 3 - 5:2 - 4, the copper salt is selected from at least one of copper chloride, copper nitrate, and copper sulfate, and the mass ratio of the inorganic salts to UiO-66-OH@modified material is 0.5 - 0.9:5 - 7; in step S4, the mass ratio of vascular endothelial growth factor-α, fibroblast growth factor, NHS, EDC to Ag / Cu-deposited UiO-66-OH@modified material is 0.2 - 0.4:0.1 - 0.3:0.5 - 1:0.5 - 1:8 - 10.

[0012] As a further improvement of the present invention, the preparation method of the chlorogenic acid / oleanolic acid-loaded chitosan is as follows: T1. Dissolve chitosan in an acid solution to obtain a chitosan solution; T2. Add chlorogenic acid and oleanolic acid to water, add NHS and EDC, stir for activation, add the chitosan solution, stir for reaction, dialyze, and dry to obtain chlorogenic acid / oleanolic acid-loaded chitosan.

[0013] As a further improvement of the present invention, in step T1, the concentration of the chitosan solution is 2 - 3 wt%, and the acid solution is a 1 - 2 wt% acetic acid or lactic acid solution; in step T2, the mass ratio of chlorogenic acid, oleanolic acid, NHS, EDC to the chitosan solution is 0.5 - 1:0.5 - 1:0.3 - 0.5:0.3 - 0.5:150 - 200.

[0014] As a further improvement of the present invention, the preparation method of the decellularized material is as follows: U1. Wash and depilate the pigskin, take the dermis layer, crush it, and wash it with hydrogen peroxide and alkali solution respectively to obtain the pretreated pigskin dermis material; U2. Add the pretreated pigskin dermis material to salt water, perform repeated freeze-thaw treatment, filter, wash, and dry to obtain the preliminary decellularized material; U3. Add the preliminary decellularized material to water, add ficin and lysozyme, perform enzymatic hydrolysis, filter, wash, and dry to obtain the decellularized material.

[0015] As a further improvement of the present invention, in step U1, the concentration of hydrogen peroxide is 28 - 32 wt%, the concentration of the alkali solution is 8 - 12 wt%, and the alkali is NaOH or KOH; in step U2, the brine is an NaCl solution with a concentration of 3 - 5 wt%, and the number of times of the repeated freeze-thaw treatment is 3 - 5 times; in step U3, the mass ratio of the preliminary decellularized material, ficin, and lysozyme is 10:1 - 2:1 - 2, the temperature of the enzymatic hydrolysis is 36 - 40 °C, and the time is 8 - 10 h.

[0016] The present invention further protects a preparation method of the above-mentioned external composition for promoting surgical incision healing, which is characterized by including the following steps: (1) Mix the banyan bark powder, traditional Chinese medicine powder, decellularized material, collagen peptide, and modified nano-hydroxyapatite evenly in proportion to obtain component A; (2) Mix the carboxylated ionic liquid, chlorogenic acid / oleanolic acid-loaded chitosan, platelet-rich plasma, and water evenly in proportion to obtain component B, and store it at 2 - 6 °C.

[0017] The present invention adopts the form of combining component A in powder form and component B in liquid form. First, sprinkle component A on the wound, and then spray component B in liquid form. The modified nano-hydroxyapatite with metal ions in component A can serve as a cross-linking center to promote the cross-linking between the collagen peptide in component A and the chitosan and carboxylated ionic liquid in component B, thereby solidifying into a hydrogel state, ensuring that the powder of component A will not spill, and prolonging the administration time.

[0018] The present invention has the following beneficial effects: The banyan bark powder in component A of the present invention is made into porous micropowder by micronization technology, which has hemostatic and antibacterial effects, can effectively adsorb exudate, keep the wound dry, and promote wound healing. The traditional Chinese medicine powder is made from a traditional Chinese medicine compound including lithospermum, millettia reticulata, achyranthes bidentata, etc., which can clear heat and detoxify, promote blood circulation to remove stasis, relieve pain and promote granulation through external application, and is extended to the application of chronic wounds such as bedsore, accelerating healing and reducing scar formation.

[0019] Collagen peptide can promote the repair of surgical wounds, and its stability is enhanced after binding with the epidermal growth factor released by modified nano-hydroxyapatite, accelerating the healing speed of surgical incisions, and at the same time having the advantages of high safety and no immunogenicity.

[0020] The modified nano-hydroxyapatite prepared by the present invention uses nano-hydroxyapatite as a carrier. After modification, the surface is provided with amino groups, and then it condenses with 2-hydroxyterephthalic acid and undergoes an in-situ hydrothermal reaction to obtain UiO-66-OH@modified material. Through its groups such as amino and hydroxyl groups, it forms complex bonds with metal ions Ag and Cu (Ag has antibacterial properties, and Cu ions can participate in the formation of enzymes and collagen), fixing the metal ions, and can condense and couple with vascular endothelial growth factor-α and fibroblast growth factor 1. The prepared nanostructured material has a high specific surface area and good permeability, can more easily penetrate the cell barrier, improve the delivery efficiency of active ingredients, can act more precisely on the wound site, and enhance the therapeutic effect. And it can slowly release vascular endothelial growth factor-α and fibroblast growth factor 1. Vascular endothelial growth factor-α (VEGF) promotes angiogenesis, providing sufficient nutrition and oxygen for wound healing; adding fibroblast growth factor (FGF) stimulates the proliferation of fibroblasts and the synthesis of collagen, accelerating the formation of granulation tissue. The synergistic effect of multiple natural ingredients promotes the healing of surgical incisions and the repair of scars.

[0021] The chitosan loaded with chlorogenic acid / oleanolic acid prepared by the present invention loads chlorogenic acid and oleanolic acid on chitosan through the condensation reaction of carboxyl and amino groups. It not only has good biocompatibility and excellent biodegradability, but also avoids the possible side effects of traditional synthetic materials. At the same time, chlorogenic acid and oleanolic acid have good antibacterial and antioxidant properties. Oleanolic acid can promote the proliferation of fibroblasts and the synthesis of collagen, and at the same time promote the synthesis and secretion of collagen, increasing the content of collagen at the wound, which helps the contraction of the wound and tissue reconstruction. Chlorogenic acid can promote the smooth transition of the wound from the inflammatory phase to the proliferation phase by inhibiting the inflammatory reaction. The two have a synergistic effect and promote wound healing.

[0022] Platelet-rich plasma contains a variety of growth factors and bioactive substances, rich in various growth factors such as platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), epidermal growth factor (EGF), vascular endothelial growth factor-α (VEGF), etc., which can bind to the receptors on the surface of target cells, activate the signal transduction pathway in cells, and promote the proliferation and differentiation of various wound-healing-related cells such as fibroblasts, endothelial cells, and keratinocytes.

[0023] In the present invention, the carboxylated ionic liquid and the chitosan loaded with chlorogenic acid / oleanolic acid are combined, and under the action of metal ions, they are crosslinked to prepare a hydrogel, which improves the drug stability and skin permeability, realizes the composite functions of anti-inflammatory, antibacterial and wound healing promotion, improves the drug solubility and release characteristics, prolongs the local action time, and has pH responsiveness. A large number of amino groups (-NH2) are contained in its molecular chain. Under acidic conditions, the amino group is easily protonated to form a positively charged ammonium ion (-NH3+ ), which increases the hydrophilicity of the polymer. At the same time, the electrostatic repulsion generated by ionization stretches the molecular chain, causing the material to swell. In a neutral or alkaline environment, the ammonium ions are deprotonated and turned back into amino groups. The electrostatic repulsion between the molecular chains is weakened, and the material shrinks. During the inflammation period of the wound, the microenvironment is acidic, and the hydrogel can release more drugs or active ingredients to exert antibacterial and anti-inflammatory effects; as the wound heals, the microenvironment tends to be neutral, and the release rate slows down, avoiding drug waste and side effects. At the same time, the imidazole group of the carboxylated ionic liquid presents a quaternary ammonium salt structure, which has good antibacterial properties and ensures that the wound is not infected.

[0024] The decellularized material prepared by the present invention provides an ideal microenvironment for cell adhesion, proliferation and differentiation. By constructing a three-dimensional structure similar to the extracellular matrix, the composition can better interact with the wound tissue, guide cell migration and tissue repair. For example, a hydrogel material is used, and its porous structure can simulate the spatial structure of the extracellular matrix, promoting cell growth and tissue reconstruction. Under the synergistic effect of high osmotic pressure and repeated freezing and thawing, the decellularized material of the present invention destroys the cell structure and promotes the release of cell components, thereby improving the decellularization effect. The hypertonic solution treats the tissue, causing the cells to swell or shrink and rupture due to osmotic pressure imbalance. Repeated washing with physiological saline combined with hypotonic solution treatment can effectively remove cell components. The freeze-thaw cycle forms ice crystals in the cells through repeated freezing and melting, destroys the cell membrane and organelles, thereby achieving the purpose of decellularization, and is commonly used to treat tissues such as skin and blood vessels. Further, under the action of ficin and lysozyme, the cell components are further removed and a good de-antigenization effect is played, making the prepared decellularized material safe and not easy to cause allergies.

[0025] The external composition for promoting surgical wound healing prepared by the present invention has good antibacterial, antioxidant and anti-inflammatory effects, relieves pain, promotes tissue regeneration, stops bleeding and restores vitality, is safe and not prone to allergies, realizes sustained and controlled release of drugs, promotes surgical wound healing and scar repair, and has broad application prospects. DETAILED DESCRIPTION

[0026] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] The particle size range of nano-hydroxyapatite is 300 - 600 nm. Ficin, 1.2 million U / g, lysozyme, 20,000 U / g. NHS, N-hydroxysuccinimide; EDC, 1-ethyl-(3-dimethylaminopropyl)carbodiimide. UiO-66-NH2 is prepared by a conventional method, and the reference can be: Duan Zhanggui, et al. Preparation of UiO-66-NH2 Supported Copper Catalyst and Its Catalytic Oxidation of Alcohols [J]. Chinese Journal of Inorganic Chemistry, 2024, 40(3): 496 - 506.

[0028] Preparation Example 1 Preparation of Traditional Chinese Medicine Powder Wash 3 g of Lithospermum erythrorhizon, 5 g of Spatholobus suberectus, and 1 g of Achyranthes aspera, dry them, pulverize them, and pass through a 100-mesh sieve to obtain the traditional Chinese medicine powder.

[0029] Preparation Example 2 Preparation of Traditional Chinese Medicine Powder Wash 5 g of Lithospermum erythrorhizon, 8 g of Spatholobus suberectus, and 3 g of Achyranthes aspera, dry them, pulverize them, and pass through a 200-mesh sieve to obtain the traditional Chinese medicine powder.

[0030] Preparation Example 3 Preparation of Traditional Chinese Medicine Powder Wash 4 g of Lithospermum erythrorhizon, 6 g of Spatholobus suberectus, and 2 g of Achyranthes aspera, dry them, pulverize them, and pass through a 150-mesh sieve to obtain the traditional Chinese medicine powder.

[0031] Preparation Example 4 Preparation of Modified Nano-Hydroxyapatite The method is as follows: S1. Add 8 g of nano-hydroxyapatite to 200 mL of Tris-HCl solution with a pH value of 8.5, add 2 g of dopamine hydrochloride, heat to 45 °C, and stir and react for 3 h to obtain the modified material; S2. Add 3 g of 2-hydroxyterephthalic acid to 200 mL of water, add 1 g of NHS and 1 g of EDC, stir and activate for 30 min, add 7 g of the modified material, stir and react for 10 h, centrifuge to obtain the product, add 10 g of the product to 4 L of a mixed solvent of N,N-dimethylformamide and acetic acid (volume ratio 39:1), add 2 g of zirconium tetrachloride, carry out hydrothermal reaction at 120 °C for 20 h, centrifuge, wash, and dry to obtain UiO-66-OH@modified material; S3. Dissolve 0.5 g of inorganic salt in 100 mL of water, add 5 g of UiO-66-OH@modified material, stir and react for 30 min, centrifuge, wash, and dry to obtain Ag / Cu-deposited UiO-66-OH@modified material; The inorganic salt includes silver nitrate and copper sulfate, and the mass ratio is 3:2; S4. Add 0.2 g of vascular endothelial growth factor-α and 0.1 g of fibroblast growth factor 1 to 100 mL of water, add 0.5 g of NHS and 0.5 g of EDC, stir and activate for 40 min, add 8 g of Ag / Cu deposited UiO-66-OH@modified material, stir and react for 12 h, centrifuge, wash, and dry to obtain modified nano-hydroxyapatite.

[0032] Preparation Example 5 Preparation of Modified Nano-Hydroxyapatite The method is as follows: S1. Add 10 g of nano-hydroxyapatite to 200 mL of Tris-HCl solution with a pH value of 9.5, add 3 g of dopamine hydrochloride, heat to 55 °C, stir and react for 5 h to obtain a modified material; S2. Add 5 g of 2-hydroxyterephthalic acid to 200 mL of water, add 2 g of NHS and 2 g of EDC, stir and activate for 30 min, add 10 g of the modified material, stir and react for 10 h, centrifuge to obtain a product. Add 15 g of the product to 4 L of a mixed solvent of N,N-dimethylformamide and acetic acid (volume ratio 39:1), add 4 g of zirconium tetrachloride, carry out hydrothermal reaction at 130 °C for 24 h, centrifuge, wash, and dry to obtain UiO-66-OH@modified material; S3. Dissolve 0.9 g of inorganic salt in 100 mL of water, add 7 g of UiO-66-OH@modified material, stir and react for 30 min, centrifuge, wash, and dry to obtain Ag / Cu deposited UiO-66-OH@modified material; The inorganic salt includes silver nitrate and copper nitrate, and the mass ratio is 5:4; S4. Add 0.4 g of vascular endothelial growth factor-α and 0.3 g of fibroblast growth factor 1 to 100 mL of water, add 1 g of NHS and 1 g of EDC, stir and activate for 40 min, add 10 g of Ag / Cu deposited UiO-66-OH@modified material, stir and react for 12 h, centrifuge, wash, and dry to obtain modified nano-hydroxyapatite.

[0033] Preparation Example 6 Preparation of Modified Nano-Hydroxyapatite The method is as follows: S1. Add 9 g of nano-hydroxyapatite to 200 mL of Tris-HCl solution with a pH value of 9, add 2.5 g of dopamine hydrochloride, heat to 50 °C, stir and react for 4 h to obtain a modified material; S2. Add 4 g of 2-hydroxyterephthalic acid to 200 mL of water, add 1.5 g of NHS and 1.5 g of EDC, stir and activate for 30 min, add 8 g of the modified material, stir and react for 10 h, centrifuge to obtain the product. Add 12 g of the product to a mixed solvent of 4 L of N,N-dimethylformamide and acetic acid (volume ratio 39:1), add 3 g of zirconium tetrachloride, carry out a hydrothermal reaction at 125 °C for 22 h, centrifuge, wash, and dry to prepare UiO-66-OH@modified material; S3. Dissolve 0.7 g of inorganic salt in 100 mL of water, add 6 g of UiO-66-OH@modified material, stir and react for 30 min, centrifuge, wash, and dry to prepare Ag / Cu-deposited UiO-66-OH@modified material; The inorganic salt includes silver nitrate and copper chloride, and the mass ratio is 4:3; S4. Add 0.3 g of vascular endothelial growth factor-α and 0.2 g of fibroblast growth factor 1 to 100 mL of water, add 0.7 g of NHS and 0.7 g of EDC, stir and activate for 40 min, add 9 g of Ag / Cu-deposited UiO-66-OH@modified material, stir and react for 12 h, centrifuge, wash, and dry to prepare the modified nano-hydroxyapatite.

[0034] Comparative Preparation Example 1 Compared with Preparation Example 6, the difference is that step S2 is not carried out.

[0035] Specifically as follows: S1. Add 9 g of nano-hydroxyapatite to 200 mL of Tris-HCl solution with a pH value of 9, add 2.5 g of dopamine hydrochloride, heat to 50 °C, and stir and react for 4 h to prepare the modified material; S2. Dissolve 0.7 g of inorganic salt in 100 mL of water, add 6 g of the modified material, stir and react for 30 min, centrifuge, wash, and dry to prepare Ag / Cu-deposited modified material; The inorganic salt includes silver nitrate and copper chloride, and the mass ratio is 4:3; S3. Add 0.3 g of vascular endothelial growth factor-α and 0.2 g of fibroblast growth factor 1 to 100 mL of water, add 0.7 g of NHS and 0.7 g of EDC, stir and activate for 40 min, add 9 g of Ag / Cu-deposited modified material, stir and react for 12 h, centrifuge, wash, and dry to prepare the modified nano-hydroxyapatite.

[0036] Comparative Preparation Example 2 Compared with Preparation Example 6, the difference is that silver nitrate is not added in step S3.

[0037] Specifically as follows: S3. Dissolve 0.7 g of copper chloride in 100 mL of water, add 6 g of UiO-66-OH@modified material, stir and react for 30 min, centrifuge, wash, and dry to obtain Ag / Cu deposited UiO-66-OH@modified material.

[0038] Comparative Preparation Example 3 Compared with Preparation Example 6, the difference lies in that copper chloride was not added in step S3.

[0039] Specifically as follows: S3. Dissolve 0.7 g of silver nitrate in 100 mL of water, add 6 g of UiO-66-OH@modified material, stir and react for 30 min, centrifuge, wash, and dry to obtain Ag / Cu deposited UiO-66-OH@modified material.

[0040] Comparative Preparation Example 4 Compared with Preparation Example 6, the difference lies in that step S3 was not carried out.

[0041] Specifically as follows: S1. Add 9 g of nano-hydroxyapatite to 200 mL of Tris-HCl solution with a pH value of 9, add 2.5 g of dopamine hydrochloride, heat to 50 °C, and stir and react for 4 h to obtain a modified material; S2. Add 4 g of 2-hydroxyterephthalic acid to 200 mL of water, add 1.5 g of NHS and 1.5 g of EDC, stir and activate for 30 min, add 8 g of the modified material, stir and react for 10 h, centrifuge to obtain a product, add 12 g of the product to 4 L of a mixed solvent of N,N-dimethylformamide and acetic acid (volume ratio 39:1), add 3 g of zirconium tetrachloride, carry out hydrothermal reaction at 125 °C for 22 h, centrifuge, wash, and dry to obtain UiO-66-OH@modified material; S3. Add 0.3 g of vascular endothelial growth factor-α and 0.2 g of fibroblast growth factor 1 to 100 mL of water, add 0.7 g of NHS and 0.7 g of EDC, stir and activate for 40 min, add 9 g of UiO-66-OH@modified material, stir and react for 12 h, centrifuge, wash, and dry to obtain modified nano-hydroxyapatite.

[0042] Comparative Preparation Example 5 Compared with Preparation Example 6, the difference lies in that step S4 was not carried out.

[0043] Specifically as follows: S1. Add 9 g of nano-hydroxyapatite to 200 mL of Tris-HCl solution with a pH value of 9, add 2.5 g of dopamine hydrochloride, heat to 50 °C, and stir and react for 4 h to obtain a modified material; S2. Add 4 g of 2-hydroxyterephthalic acid to 200 mL of water, add 1.5 g of NHS and 1.5 g of EDC, stir and activate for 30 min, add 8 g of the modified material, stir and react for 10 h, centrifuge to obtain the product. Add 12 g of the product to a mixed solvent of 4 L of N,N-dimethylformamide and acetic acid (volume ratio 39:1), add 3 g of zirconium tetrachloride, carry out hydrothermal reaction at 125 °C for 22 h, centrifuge, wash, and dry to prepare UiO-66-OH@modified material; S3. Dissolve 0.7 g of inorganic salt in 100 mL of water, add 6 g of UiO-66-OH@modified material, stir and react for 30 min, centrifuge, wash, and dry to prepare Ag / Cu-deposited UiO-66-OH@modified material, which is the modified nano-hydroxyapatite; The inorganic salt includes silver nitrate and copper chloride, and the mass ratio is 4:3.

[0044] Preparation Example 7 Preparation of Chlorogenic Acid / Oleanolic Acid-Loaded Chitosan The method is as follows: T1. Dissolve chitosan in a 1 wt% acetic acid solution to obtain a 2 wt% chitosan solution; T2. Add 0.5 g of chlorogenic acid and 0.5 g of oleanolic acid to 50 mL of water, add 0.3 g of NHS and 0.3 g of EDC, stir and activate for 30 min, add 150 g of the chitosan solution, stir and react for 8 h, dialyze, and dry to prepare chlorogenic acid / oleanolic acid-loaded chitosan.

[0045] Preparation Example 8 Preparation of Chlorogenic Acid / Oleanolic Acid-Loaded Chitosan The method is as follows: T1. Dissolve chitosan in a 2 wt% lactic acid solution to obtain a 3 wt% chitosan solution; T2. Add 1 g of chlorogenic acid and 1 g of oleanolic acid to 50 mL of water, add 0.5 g of NHS and 0.5 g of EDC, stir and activate for 30 min, add 200 g of the chitosan solution, stir and react for 8 h, dialyze, and dry to prepare chlorogenic acid / oleanolic acid-loaded chitosan.

[0046] Preparation Example 9 Preparation of Chlorogenic Acid / Oleanolic Acid-Loaded Chitosan The method is as follows: T1. Dissolve chitosan in a 1.2 wt% acetic acid solution to obtain a 2.5 wt% chitosan solution; T2. Add 0.8 g of chlorogenic acid and 0.7 g of oleanolic acid to 50 mL of water, add 0.4 g of NHS and 0.4 g of EDC, stir and activate for 30 min, add 170 g of the chitosan solution, stir and react for 8 h, dialyze, and dry to prepare chlorogenic acid / oleanolic acid-loaded chitosan.

[0047] Comparative Preparation Example 6 Compared with Preparation Example 9, the difference lies in that chlorogenic acid was not added in step T2.

[0048] Specifically as follows: T1. Dissolve chitosan in a 1.2 wt% acetic acid solution to obtain a 2.5 wt% chitosan solution; T2. Add 1.5 g of oleanolic acid to 50 mL of water, add 0.4 g of NHS and 0.4 g of EDC, stir and activate for 30 min, add 170 g of the chitosan solution, stir and react for 8 h, dialyze, and dry to obtain oleanolic acid-loaded chitosan.

[0049] Comparative Preparation Example 7 Compared with Preparation Example 9, the difference lies in that oleanolic acid was not added in step T2.

[0050] Specifically as follows: T1. Dissolve chitosan in a 1.2 wt% acetic acid solution to obtain a 2.5 wt% chitosan solution; T2. Add 1.5 g of chlorogenic acid to 50 mL of water, add 0.4 g of NHS and 0.4 g of EDC, stir and activate for 30 min, add 170 g of the chitosan solution, stir and react for 8 h, dialyze, and dry to obtain chlorogenic acid-loaded chitosan.

[0051] Comparative Preparation Example 8 Compared with Preparation Example 9, the difference lies in that step T2 was not carried out.

[0052] Specifically as follows: T1. Dissolve chitosan in a 1.2 wt% acetic acid solution to obtain a 2.5 wt% chitosan solution.

[0053] Preparation Example 10 Preparation of Composite Enzyme The method is as follows: Add 1 g of ficin and 1 g of lysozyme to 100 mL of water, add 0.5 g of NHS and 0.5 g of EDC, stir and activate for 30 min, add 5 g of UiO-66-NH2, stir and react for 12 h, then centrifuge, wash, and dry. The product is treated under high pressure of 200 MPa for 15 min to obtain the composite enzyme.

[0054] The composite enzyme prepared by the present invention dehydrates and condenses ficin and lysozyme and immobilizes them on UiO-66-NH2, greatly increasing the sites for enzymatic catalytic reactions, shortening the catalytic reaction distance, accelerating the reaction rate, and reducing the reaction activation energy, thereby enabling a reaction to be carried out at room temperature for a short time. After ultra-high pressure treatment, the hydrophobic part of the enzyme structure is exposed, the conformation changes, and no chemical reagents are added. While significantly improving the catalytic efficiency of the enzyme, it is safe and non-toxic, reducing the use and emission of chemical substances and being more environmentally friendly.

[0055] Preparation Example 11 Preparation of acellular material The method is as follows: U1. Clean and depilate the pigskin, take the dermis layer, crush it, and wash it with 28 wt% hydrogen peroxide solution and 8 wt% NaOH solution respectively to obtain the pretreated pigskin dermis material; U2. Add 10 g of the pretreated pigskin dermis material to 100 mL of a 3 wt% NaCl solution, perform freeze-thaw treatment 3 times, filter, wash, and dry to obtain the preliminary acellular material; U3. Add 10 g of the preliminary acellular material to 200 mL of water, add 1 g of ficin and 1 g of lysozyme, enzymatically hydrolyze at 36 °C for 8 h, filter, wash, and dry to obtain the acellular material.

[0056] Preparation Example 12 Preparation of acellular material The method is as follows: U1. Clean and depilate the pigskin, take the dermis layer, crush it, and wash it with 32 wt% hydrogen peroxide solution and 12 wt% KOH solution respectively to obtain the pretreated pigskin dermis material; U2. Add 10 g of the pretreated pigskin dermis material to 100 mL of a 5 wt% NaCl solution, perform freeze-thaw treatment 5 times, filter, wash, and dry to obtain the preliminary acellular material; U3. Add 10 g of the preliminary acellular material to 200 mL of water, add 2 g of ficin and 2 g of lysozyme, enzymatically hydrolyze at 40 °C for 10 h, filter, wash, and dry to obtain the acellular material.

[0057] Preparation Example 13 Preparation of acellular material The method is as follows: U1. Clean and depilate the pigskin, take the dermis layer, crush it, and wash it with 30 wt% hydrogen peroxide solution and 10 wt% NaOH solution respectively to obtain the pretreated pigskin dermis material; U2. Add 10 g of the pretreated pigskin dermis material to 100 mL of a 4 wt% NaCl solution, perform freeze-thaw treatment 4 times, filter, wash, and dry to obtain the preliminary acellular material; U3. Add 10 g of the preliminarily decellularized material to 200 mL of water, add 1.5 g of ficin and 1.5 g of lysozyme, enzymatically hydrolyze at 37 °C for 9 h, filter, wash, and dry to obtain the decellularized material.

[0058] Preparation Example 14 Preparation of Decellularized Material The method is as follows: U1. Clean and depilate the pigskin, take the dermis layer, comminute it, and wash it with 30 wt% hydrogen peroxide solution and 10 wt% NaOH solution respectively to obtain the pretreated pigskin dermis material; U2. Add 10 g of the pretreated pigskin dermis material to 100 mL of a 4 wt% NaCl solution, perform repeated freeze-thaw treatment 4 times, filter, wash, and dry to obtain the preliminarily decellularized material; U3. Add 10 g of the preliminarily decellularized material to 200 mL of water, add 3 g of the composite enzyme prepared in Preparation Example 10, enzymatically hydrolyze at room temperature for 1 h, filter, wash, and dry to obtain the decellularized material.

[0059] Comparative Preparation Example 9 Compared with Preparation Example 14, the difference is that step U2 is not carried out.

[0060] Specifically as follows: U1. Clean and depilate the pigskin, take the dermis layer, comminute it, and wash it with 30 wt% hydrogen peroxide solution and 10 wt% NaOH solution respectively to obtain the pretreated pigskin dermis material; U2. Add 10 g of the pretreated pigskin dermis material to 200 mL of water, add 3 g of the composite enzyme prepared in Preparation Example 10, enzymatically hydrolyze at room temperature for 1 h, filter, wash, and dry to obtain the decellularized material.

[0061] Comparative Preparation Example 10 Compared with Preparation Example 14, the difference is that step U3 is not carried out.

[0062] Specifically as follows: U1. Clean and depilate the pigskin, take the dermis layer, comminute it, and wash it with 30 wt% hydrogen peroxide solution and 10 wt% NaOH solution respectively to obtain the pretreated pigskin dermis material; U2. Add 10 g of the pretreated pigskin dermis material to 100 mL of a 4 wt% NaCl solution, perform repeated freeze-thaw treatment 4 times, filter, wash, and dry to obtain the preliminarily decellularized material, which is the decellularized material.

[0063] Test Example 1 Perform performance tests on the decellularized materials prepared in Preparation Examples 11 - 14 and Comparative Preparation Examples 9 - 10.

[0064] The cytotoxicity test of the material was carried out in accordance with GB / T 16886.5-2017. The results are shown in Table 1.

[0065] Table 1

[0066] As can be seen from the above table, the acellular materials prepared in Preparation Examples 11-14 of the present invention have low cytotoxicity.

[0067] Referring to the method in YY / T1465.2-2016, SPF-grade BALB / c mice were selected. The experimental animals were randomly divided into a negative control group, a positive control (BSA) group, Preparation Examples 11-14 groups, and Comparative Preparation Examples 9-10 groups, with 10 animals in each group. They were sacrificed and sampled at the 4th week after implantation.

[0068] Experimental sample group: 30 mg of the sample was implanted subcutaneously in the back of the mice in the test group.

[0069] Negative control group: The same surgical operation was performed on the animals in the test group, but the product was not implanted.

[0070] Positive control (BSA) group: BSA (bovine serum albumin) was used as the positive control. 3 mg of BSA was mixed with 9 mL of phosphate buffered saline (PBS), and then mixed with complete Freund's adjuvant (CFA) at a volume ratio of 1:1 to form an emulsion. 0.12 mL was injected subcutaneously into each animal, once a week for a total of 4 immunizations.

[0071] After 4 weeks, the mice were sacrificed, blood was collected from the heart, and after standing at room temperature for 30 min, centrifuged at 600×g for 20 min, and the serum was collected and stored at -20°C in aliquots. The contents of IgG, IgM, and complement C3a in the mouse serum were detected according to the requirements of the mouse IgG ELISA kit, mouse IgM ELISA kit, and mouse complement C3a ELISA kit, respectively.

[0072] The results are shown in Table 2.

[0073] Table 2

[0074] As can be seen from the above table, the acellular materials prepared in Preparation Examples 11-14 of the present invention have low immunogenicity. Example 1

[0075] This example provides an external composition for promoting surgical incision healing.

[0076] The preparation method includes the following steps: (1) Mix 2 g of banyan bark powder (100 mesh), 7 g of the traditional Chinese medicine powder prepared in Preparation Example 1, 3 g of the acellular material prepared in Preparation Example 11, 1 g of fish collagen peptide, and 2 g of the modified nano-hydroxyapatite prepared in Preparation Example 4 evenly to obtain Component A; Mix 3 g of 1-carboxymethyl-3-methylimidazolium chloride, 5 g of the chlorogenic acid / oleanolic acid-loaded chitosan prepared in Preparation Example 7, 10 g of platelet-rich plasma, and 200 g of water evenly to obtain Component B, and store it at 4 °C. Example 2

[0077] This example provides an external composition for promoting surgical incision healing.

[0078] The preparation method includes the following steps: (1) Mix 4 g of banyan bark powder (100 mesh), 10 g of the traditional Chinese medicine powder prepared in Preparation Example 2, 5 g of the acellular material prepared in Preparation Example 12, 3 g of fish collagen peptide, and 4 g of the modified nano-hydroxyapatite prepared in Preparation Example 5 evenly to obtain Component A; (2) Mix 5 g of 1-carboxymethyl-3-methylimidazolium chloride, 8 g of the chlorogenic acid / oleanolic acid-loaded chitosan prepared in Preparation Example 8, 12 g of platelet-rich plasma, and 300 g of water evenly to obtain Component B, and store it at 4 °C. Example 3

[0079] This example provides an external composition for promoting surgical incision healing.

[0080] The preparation method includes the following steps: (1) Mix 3 g of banyan bark powder (100 mesh), 8 g of the traditional Chinese medicine powder prepared in Preparation Example 3, 4 g of the acellular material prepared in Preparation Example 13, 2 g of fish collagen peptide, and 3 g of the modified nano-hydroxyapatite prepared in Preparation Example 6 evenly to obtain Component A; (2) Mix 4 g of 1-carboxymethyl-3-methylimidazolium chloride, 7 g of the chlorogenic acid / oleanolic acid-loaded chitosan prepared in Preparation Example 9, 11 g of platelet-rich plasma, and 250 g of water evenly to obtain Component B, and store it at 4 °C. Example 4

[0081] Compared with Example 3, the difference is that the acellular material is prepared in Preparation Example 14.

[0082] Comparative Example 1 Compared with Example 3, the difference is that the modified nano-hydroxyapatite is prepared in Comparative Preparation Example 1.

[0083] Comparative Example 2 Compared with Example 3, the difference is that the modified nano-hydroxyapatite is prepared in Comparative Preparation Example 2.

[0084] Comparative Example 3 Compared with Example 3, the difference lies in that the modified nano-hydroxyapatite is prepared from Comparative Preparation Example 3.

[0085] Comparative Example 4 Compared with Example 3, the difference lies in that the modified nano-hydroxyapatite is prepared from Comparative Preparation Example 4.

[0086] Comparative Example 5 Compared with Example 3, the difference lies in that the modified nano-hydroxyapatite is prepared from Comparative Preparation Example 5.

[0087] Comparative Example 6 Compared with Example 3, the difference lies in that the chlorogenic acid / oleanolic acid-loaded chitosan is prepared from Comparative Preparation Example 6.

[0088] Comparative Example 7 Compared with Example 3, the difference lies in that the chlorogenic acid / oleanolic acid-loaded chitosan is prepared from Comparative Preparation Example 7.

[0089] Comparative Example 8 Compared with Example 3, the difference lies in that the chlorogenic acid / oleanolic acid-loaded chitosan is prepared from Comparative Preparation Example 8.

[0090] Comparative Example 9 Compared with Example 3, the difference lies in that the acellular material is prepared from Comparative Preparation Example 9.

[0091] Comparative Example 10 Compared with Example 3, the difference lies in that the acellular material is prepared from Comparative Preparation Example 10.

[0092] Comparative Example 11 Compared with Example 3, the difference lies in that 1-carboxymethyl-3-methylimidazolium chloride is not added.

[0093] Comparative Example 12 Compared with Example 3, the difference lies in that the chlorogenic acid / oleanolic acid-loaded chitosan is not added.

[0094] Test Example 2 Take SPF-grade NIH male mice, divide them into 18 groups, use distilled water as the negative control group, use fluocinonide acetate ointment as the positive control group, and use the external compositions for promoting surgical incision healing prepared in Examples 1-4 and Comparative Examples 1-12 as Test Groups 1-4 and Comparative Groups 1-12.

[0095] Apply xylene evenly to both the inner and outer surfaces of the right auricle of each mouse for inflammation induction at a dose of 100 μL / mouse. Leave the left ear untreated as the blank control group. 30 minutes after xylene-induced inflammation, except for the negative control group, animals in each group were given the corresponding test substance on the right ear (the administration method was to sprinkle 0.05 g / mouse of component A at the test position and then evenly spray 0.5 mL / mouse of component B). When administering, note that in the initial stage of gel formation, it should be evenly applied to both the inner and outer surfaces of the right auricle. Animals in the negative control group were given distilled water on the right auricle at a dose of 0.1 mL / mouse. 1 hour after administering the test substance, the animals were sacrificed by cervical dislocation. Cut off the auricles on both sides, wash the test substance on the right auricle with normal saline, and dry the water. Overlap the bilateral auricles, punch out ear pieces from the left and right ears with a puncher with a diameter of 8 mm, weigh the ear pieces of both ears respectively, and calculate the swelling value. The smaller the swelling value, the better the anti-inflammatory effect.

[0096] Swelling value = m 右耳耳片 -m 左耳耳片 The results are shown in Table 3.

[0097] Table 3

[0098] Note: Compared with the negative control group, **P < 0.01, *P < 0.05.

[0099] As can be seen from the above table, the external composition for promoting surgical incision healing prepared in Examples 1-4 of the present invention has a very good anti-inflammatory effect.

[0100] Test Example 3 Randomly divide Balb / c mice into 17 groups, with 10 mice in each group, namely the control group (PBS solution), Examples 1-4 groups, and Comparative Examples 1-12 groups. After anesthetizing the mice intraperitoneally with 1% sodium pentobarbital, depilate the back, wash with warm normal saline, dry, disinfect with iodine tincture, and cut off the full-thickness skin with a diameter of about 1 cm on the back of the mice to prepare a wound model. Inoculate 100 μL of Escherichia coli (ATCC25922) dissolved in normal saline at a density of 2×10 6 cfu / mL on the skin wound. 1 day after infection, take the control group and each experimental group and apply the external composition for promoting surgical incision healing to the back wound. The usage method is to sprinkle 0.1 g / mouse of component A at the test position and then evenly spray 1 mL / mouse of component B. Observe and record the wound healing area and healing time on the 0th and 14th days after trauma respectively, calculate the wound healing rate according to the following formula, and evaluate the wound healing speed.

[0101] Wound healing rate = (wound area on the 0th day - area after the 14th day) / wound area on the 0th day × 100%.

[0102] The results are shown in Table 4.

[0103] Table 4

[0104] As can be seen from the above table, the external composition for promoting surgical incision healing prepared in Examples 1-4 of the present invention can significantly promote wound healing.

[0105] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the protection scope of the present invention.

Claims

1. An external composition for promoting surgical incision healing, characterized in that, It includes component A and component B. Component A is a powder, which is composed of banyan bark powder, traditional Chinese medicine powder, acellular material, collagen peptide, and modified nano-hydroxyapatite, with a mass ratio of 2-4:7-10:3-5:1-3:2-4; Component B is a liquid, which is composed of carboxylated ionic liquid, chlorogenic acid / oleanolic acid-loaded chitosan, platelet-rich plasma, and water, with a mass ratio of 3-5:5-8:10-12:200-300.

2. The external composition for promoting surgical incision healing according to claim 1, wherein The traditional Chinese medicine powder is prepared by washing, drying, pulverizing, and sieving lithospermum root, millettia reticulata, and achyranthes aspera to obtain the traditional Chinese medicine powder; The collagen peptide is selected from at least one of fish collagen peptide, pig collagen peptide, and bovine collagen peptide; The carboxylated ionic liquid is selected from at least one of 1-carboxymethyl-3-methylimidazolium chloride and 1-carboxymethylpyridinium bromide.

3. The external composition for promoting surgical incision healing according to claim 2, wherein The mass ratio of lithospermum root, millettia reticulata, and achyranthes aspera is 3-5:5-8:1-3, and the mesh number of the sieve for sieving is 100-200 mesh.

4. The external composition for promoting surgical incision healing according to claim 1, wherein The preparation method of the modified nano-hydroxyapatite is as follows: S1. Add nano-hydroxyapatite into Tris-HCl solution, add dopamine hydrochloride, heat and stir to react to obtain a modified material; S2. Add 2-hydroxyterephthalic acid into water, add NHS and EDC, stir for activation, add the modified material, stir to react, centrifuge, add the product into a mixed solvent of N,N-dimethylformamide and acetic acid, add zirconium tetrachloride, carry out hydrothermal reaction, centrifuge, wash, and dry to obtain UiO-66-OH@modified material; S3. Dissolve the inorganic salt in water, add UiO-66-OH@modified material, stir to react, centrifuge, wash, and dry to obtain Ag / Cu-deposited UiO-66-OH@modified material; S4. Add vascular endothelial growth factor-α and fibroblast growth factor into water, add NHS and EDC, stir for activation, add Ag / Cu-deposited UiO-66-OH@modified material, stir to react, centrifuge, wash, and dry to obtain the modified nano-hydroxyapatite.

5. The external composition for promoting surgical incision healing according to claim 4, wherein In step S1, the pH value of the Tris-HCl solution is 8.5 - 9.5, the mass ratio of nano-hydroxyapatite to dopamine hydrochloride is 8 - 10:2 - 3, the temperature of the heating and stirring reaction is 45 - 55 °C, and the time is 3 - 5 h; in step S2, the mass ratio of 2-hydroxyterephthalic acid, NHS, EDC to the modified material is 3 - 5:1 - 2:1 - 2:7 - 10, the mass ratio of the product to zirconium tetrachloride is 10 - 15:2 - 4, the temperature of the hydrothermal reaction is 120 - 130 °C, and the time is 20 - 24 h; in step S3, the inorganic salts include silver nitrate and copper salt, the mass ratio is 3 - 5:2 - 4, the copper salt is selected from at least one of copper chloride, copper nitrate, and copper sulfate, and the mass ratio of the inorganic salts to UiO-66-OH@modified material is 0.5 - 0.9:5 - 7; in step S4, the mass ratio of vascular endothelial growth factor-α, fibroblast growth factor, NHS, EDC to Ag / Cu-deposited UiO-66-OH@modified material is 0.2 - 0.4:0.1 - 0.3:0.5 - 1:0.5 - 1:8 - 10.

6. The external composition for promoting surgical incision healing according to claim 1, wherein The preparation method of the chitosan loaded with chlorogenic acid / oleanolic acid is as follows: T1. Dissolve chitosan in an acid solution to obtain a chitosan solution; T2. Add chlorogenic acid and oleanolic acid to water, add NHS and EDC, stir for activation, add the chitosan solution, stir for reaction, dialyze, and dry to obtain chitosan loaded with chlorogenic acid / oleanolic acid.

7. The external composition for promoting surgical incision healing according to claim 6, wherein In step T1, the concentration of the chitosan solution is 2 - 3 wt%, and the acid solution is a 1 - 2 wt% acetic acid or lactic acid solution; in step T2, the mass ratio of chlorogenic acid, oleanolic acid, NHS, EDC to the chitosan solution is 0.5 - 1:0.5 - 1:0.3 - 0.5:0.3 - 0.5:150 - 200.

8. The external composition for promoting surgical incision healing according to claim 1, wherein The preparation method of the decellularized material is as follows: U1. Wash and depilate the pigskin, take the dermis layer, crush it, and wash it with hydrogen peroxide and alkali solution respectively to obtain the pretreated pigskin dermis material; U2. Add the pretreated pigskin dermis material to salt water, perform repeated freeze-thaw treatment, filter, wash, and dry to obtain the preliminary decellularized material; U3. Add the preliminary decellularized material to water, add ficin and lysozyme, enzymatically hydrolyze, filter, wash, and dry to obtain the decellularized material.

9. The external composition for promoting surgical incision healing according to claim 8, wherein, In step U1, the concentration of the hydrogen peroxide is 28 - 32 wt%, the concentration of the alkali solution is 8 - 12 wt%, and the alkali is NaOH or KOH; in step U2, the salt water is a 3 - 5 wt% NaCl solution, and the number of repeated freeze-thaw treatments is 3 - 5 times; in step U3, the mass ratio of the preliminary decellularized material, ficin and lysozyme is 10:1 - 2:1 - 2, and the temperature of the enzymatic hydrolysis is 36 - 40 °C, and the time is 8 - 10 h.

10. A method for preparing an external composition for promoting surgical incision healing as described in claim 1, characterized in that, It includes the following steps: (1) Mix the Ficus microcarpa bark powder, traditional Chinese medicine powder, decellularized material, collagen peptide, and modified nano-hydroxyapatite in proportion to obtain component A; (2)Mix the carboxylated ionic liquid, chlorogenic acid / oleanolic acid-loaded chitosan, platelet-rich plasma, and water proportionally and uniformly to obtain Component B, and store it at 2-6 °C.

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