Bioactive gel for accelerating wound healing and preparation method thereof

Abstract

The invention discloses bioactive gel for accelerating wound healing. The gel is prepared from the following raw materials in percentage by weight: 0.8%-1% of bomer 980, 5%-5.5% of glycerol, 15%-20% of white vaseline, 51.7%-55% of phosphate buffer, 1.8%-2% of nano royal jelly acid, 2%-3% of recombinant III type human collagen, 0.4%-0.5% of sodium hyaluronate, 1%-3% of sodium alginate, 20%-25% of polyethylene glycol-400, 1.5%-3% of dopamine-methacrylamide copolymer, 0.5%-0.8% of nano silver and 0.3%-0.5% of phenoxyethanol. The nano royal jelly acid and the nano silver have a synergistic antibacterial effect, so that the inhibition effect on drug-resistant bacteria such as MRSA and pseudomonas aeruginosa is remarkably improved, and meanwhile, the wound epithelization can be accelerated and scar formation can be reduced through the dual effects of the recombinant III type collagen and the acetylated sodium hyaluronate. The defects that a traditional dressing is narrow in antibacterial spectrum, single in repairing effect, poor in environmental adaptability and the like are overcome, and the dressing is particularly suitable for complicated wounds such as diabetic foot ulcer and second-degree burn.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to a biological active gel for accelerating wound healing and a preparation method thereof. BACKGROUND

[0002] The biological active gel is a kind of polymer network material with biological responsiveness and the function of simulating extracellular matrix, which can interact with biological systems through physical or chemical action, promote tissue repair, drug delivery or disease treatment.

[0003] It mainly absorbs a large amount of water to form a biomimetic microenvironment through a three-dimensional polymer network structure such as natural polysaccharide or synthetic polymer, and simultaneously loads bioactive molecules, and realizes controllable release by using the dynamic responsiveness of physical / chemical crosslinking points; for example, in wound repair, the gel can not only maintain a humid environment to promote cell migration, but also inhibit infection and activate regenerative signaling pathways, and finally complete the treatment function by synchronizing degradation metabolism and tissue remodeling;

[0004] However, the existing biological active gel has certain defects. First, the antibacterial property has limitations. For example, chitosan gel is only effective against gram-positive bacteria, and has low antibacterial rate against drug-resistant bacteria such as pseudomonas aeruginosa; and the activity decays quickly, with high loss of collagen activity after 6 months of storage at room temperature. The adhesion of traditional carbomer gel decreases within 12 hours under the flushing of wound exudate, resulting in unbalanced mechanical properties. Silver-containing dressings have antibacterial property but inhibit cell migration, growth factor gels promote repair but have no antibacterial property, and the interference of the microenvironment also leads to the immersion of high-moisture gels on the wound, and dry gels hinder epithelial crawling.

[0005] Therefore, the biological active gel for accelerating wound healing and the preparation method thereof are proposed to solve the problem. SUMMARY

[0006] The present application aims to provide a biological active gel for accelerating wound healing, which solves the problems of narrow antibacterial spectrum, easy inactivation of active ingredients, poor environmental adaptability and single repair function of the current biological active gel.

[0007] To achieve the above-mentioned purpose, the present application provides the following technical scheme: a biological active gel for accelerating wound healing, the raw material ratio is as follows:

[0008] Carbomer 980: 0.8%-1%, glycerin: 5-5.5%, white petrolatum: 15%-20%, phosphate buffer: 51.7%-55%, nano royal jelly acid: 1.8%-2%, recombinant human collagen type III: 2%-3%, sodium hyaluronate: 0.4%-0.5%, sodium alginate: 1%-3%, polyethylene glycol-400: 20%-25%, dopamine-methacrylamide copolymer: 1.5%-3%, nano-silver: 0.5%-0.8%, phenoxyethanol: 0.3%-0.5%.

[0009] Preferably, the carbomer 980, glycerin, white petrolatum, and phosphate buffer are gel bases.

[0010] Preferably, the nano royal jelly acid, recombinant human collagen type III, sodium hyaluronate, sodium alginate, and polyethylene glycol-400 are bioactive components, the nano royal jelly acid is a complex nanoparticle formed by 10-HDA@CDs, and the sodium hyaluronate needs to be acetylated.

[0011] Preferably, the dopamine-methacrylamide copolymer, nano-silver, and phenoxyethanol are functional strengthening components, and the nano-silver particle size is <20 nm.

[0012] The preparation method of the bioactive gel for accelerating wound healing comprises the following steps:

[0013] Step 1: Nano royal jelly acid preparation: Dissolve royal jelly acid and ammonium citrate in water at a molar ratio of 1:3, hydrothermal reaction at 180°C for 6 hours, then centrifugal purification to obtain fluorescent carbon quantum dot complex with particle size <5 nm, and adjust to 20% w / v water dispersion for standby;

[0014] Step 2: Dissolution of recombinant collagen: Take the freeze-dried collagen protein powder, and dissolve it in pre-cooled 0.01mHCl solution at 4°C to 10% w / v to avoid aggregation;

[0015] Step 3: Gel base preparation (A phase-water phase): Heat the pH 7.0 phosphate buffer to 70°C and stir at 300 rpm, then add glycerin and phosphate buffer to completely dissolve, and slowly sieve in carbomer 980, homogenize the solution at 5000 rpm x 10 min to transparent colloid, finally cool the transparent colloid to 40°C and maintain stirring to prevent solidification;

[0016] Step 4: Mixing of oil phase with active ingredients (B phase): By melting white petrolatum using 60°C water bath, polyethylene glycol-400 was added into petrolatum and mixed evenly, sodium acetylated hyaluronic acid and sodium alginate were swelled in 5% glycerol water solution before adding into the mixture, then dopamine-methacrylamide copolymer was added after the ethanol wetting concentration was less than 0.5%, and the mixture was kept at 40°C to avoid the solidification of petrolatum;

[0017] Step 5: Loading of active ingredients (C phase-low temperature phase): The nanosilver suspension was ultrasonically treated at 25 kHz for 5 min to avoid aggregation, then collagen solution and 10-HDA@CDs nanosuspension were added under ice bath environment, and finally phenoxyethanol was added to prevent volatilization at high temperature, while the temperature of C phase was ≤10°C to prevent collagen denaturation and nanoparticle aggregation;

[0018] Step 6: Gradient mixing and defoaming filling: A phase at 40°C was slowly poured into B phase at 40°C, and the homogenizer was operated at 8000 rpm for 15 min to form the initial emulsion; then the initial emulsion was cooled to 15°C, C phase was added dropwise under nitrogen protection, and paddle stirring was performed at 200 rpm for 20 min; then it was transferred to a vacuum homogenization tank at -0.08 MPa, 30°C for 10 min to remove air bubbles; finally, online filtration was performed using a 0.22 μm filter, and the product was filled into an aluminum tube / injector under nitrogen protection and sealed.

[0019] Preferably, in step 1, the core component of royal jelly, 10-hydroxy-2-decenoic acid, is combined with carbon quantum dots to form a complex nanoparticle, i.e., nanoscale royal jelly acid, through nanotechnology.

[0020] Preferably, in step 3, the materials can be added together in the mixing device, or one material can be added first, and the carbomer dispersion is prohibited from being directly added to the water phase → it must be pre-mixed with glycerol into a paste, and then slowly sieved into 70°C phosphate buffer. If "fish eye glue" appears during stirring, immediately add 5 times the amount of glycerol and heat to 80°C to rescue.

[0021] Preferably, in step 4, if sodium alginate is not swelled enough, it needs to be pre-swelled for 30 min in a 5% glycerol water solution at a solid-liquid ratio of 1:20 to reduce the sandiness caused by unswept particles; when the acrylamide residue exceeds the standard, cold ethanol is used for purification.

[0022] Preferably, in step 6, hydrophilic carbomer 980 and oil-repellent white petrolatum are prone to direct mixing and layering, so a glycerol / PEG-400 transitional phase is needed. If the viscosity decreases after sterilization, add 0.1% calcium alginate for secondary crosslinking.

[0023] Preferably, in step 6, after preparation is completed, sterilization is performed by gamma rays, thereby avoiding thermal damage to the nanostructure, the vacuum degree during vacuum degassing is ≥-0.08 MPa, and the time is ≤10 min, to prevent the gel structure from being damaged due to excessive negative pressure, and to prevent the filter membrane from adsorbing the active ingredients of collagen, a low-protein adsorption PVDF filter membrane can be used for filtration and filling.

[0024] Compared with the prior art, the beneficial effects of the present application are:

[0025] The patent application significantly improves the inhibition effect on MRSA and drug-resistant bacteria such as Pseudomonas aeruginosa through the synergistic antibacterial effect of nano royal jelly acid and nano silver, and accelerates epithelialization and reduces scar formation on the wound surface through the dual action of recombinant type III collagen and acetylated sodium hyaluronate.

[0026] The patent application can dynamically adapt to the wound environment through its intelligent responsive gel matrix (pH-sensitive swelling, humidity balance), and the strong adhesion of dopamine copolymer ensures the stability of the physical barrier. In addition, gamma ray sterilization effectively protects the stability of the active ingredients, overcoming the defects of traditional dressings such as narrow antibacterial spectrum, single repair effect, and poor environmental adaptability, and is especially suitable for complex wounds such as diabetic foot ulcers and II degree burns. DETAILED DESCRIPTION

[0027] The application will be described in more detail below by way of examples, which are merely illustrative and do not limit the scope of the application.

[0028] In one aspect of the application, the application provides a technical solution: a bioactive gel for accelerating wound healing, the raw material ratio is as follows:

[0029] Carbopol 980: 0.8%-1%, glycerol: 5-5.5%, white petrolatum: 15%-20%, phosphate buffer: 51.7%-55%, nano royal jelly acid: 1.8%-2%, recombinant type III human collagen: 2%-3%, sodium hyaluronate: 0.4%-0.5%, sodium alginate: 1%-3%, polyethylene glycol-400: 20%-25%, dopamine-methacrylamide copolymer: 1.5%-3%, nano silver: 0.5%-0.8%, phenoxyethanol: 0.3%-0.5%.

[0030] According to the embodiment of the present application, Carbopol 980, glycerin, white petrolatum, phosphate buffer are the gel matrix. According to the embodiment of the present application: where Carbopol 980 as a pH-sensitive gelling agent can provide thixotropy, glycerin as a moisturizing plasticizer can reduce the sticky feeling of white petrolatum, white petrolatum can be used for wound sealing and moisturizing, reducing water evaporation, and phosphate buffer can be used to maintain pH 6.5-7.0 and neutralize Carbopol and stabilize protein activity;

[0031] According to the embodiment of the present application, nano royal jelly acid, recombinant human collagen type III, sodium hyaluronate, sodium alginate, and polyethylene glycol-400 are bioactive components. Nano royal jelly acid is a complex nanoparticle formed by 10-HDA@CDs binding, and sodium hyaluronate needs to be acetylated. According to the embodiment of the present application: nano royal jelly acid as a carbon quantum dot carrier improves water solubility and antibacterial properties, recombinant human collagen type III can promote wound epithelial crawling and dermal reconstruction, sodium hyaluronate with a molecular weight of <10 kDa can penetrate the stratum corneum, synergize with white petrolatum to moisturize, sodium alginate can assist in liquid absorption and gelation, provide a calcium ion controlled release channel, and polyethylene glycol-400 is a bioactive component used to replace part of the petrolatum to enhance the penetration of active ingredients.

[0032] According to the embodiment of the present application, dopamine-methacrylamide copolymer, nano-silver, and phenoxyethanol are functional strengthening components, and the particle size of nano-silver is <20 nm. According to the embodiment of the present application: dopamine-methacrylamide copolymer is a highly transparent tissue adhesive that can avoid the side effects of free dopamine, nano-silver is a broad-spectrum antibacterial membrane with a dose below the cytotoxicity threshold, and phenoxyethanol can make the gel meet the required preservative properties.

[0033] Through the synergistic antibacterial effect of nano royal jelly acid and nano silver, the inhibitory effect on drug-resistant bacteria such as MRSA and Pseudomonas aeruginosa is significantly improved. At the same time, through the dual action of recombinant collagen type III and acetylated sodium hyaluronate, the epithelialization of the wound can be accelerated and the formation of scars can be reduced.

[0034] Method for preparation

[0035] In the second aspect of the present application, the present application provides a method for preparing the bioactive gel described above, which comprises: first, preparing nano royal jelly acid: dissolving royal jelly acid and ammonium citrate in water at a molar ratio of 1:3, hydrothermal reaction at 180°C for 6 hours, then centrifugal purification to obtain fluorescent carbon quantum dot complex with particle size <5nm, and adjusting to 20% w / v water dispersion for standby; then, dissolving recombinant collagen: taking freeze-dried collagen powder, dissolving in pre-cooled 0°C 0.01mHCl solution to 10% w / v, avoiding aggregation; then, preparing gel matrix (A phase-water phase): heating pH 7.0 phosphate buffer to 70°C and stirring at 300 rpm, then adding glycerol and phosphate buffer to completely dissolve, and slowly screening into carbomer 980, homogenizing the solution at 5000 rpm x 10 min to transparent colloid, finally cooling the transparent colloid to 40°C and maintaining stirring to prevent solidification; then, mixing oil phase with active ingredients (B phase): melting white vaseline using 60°C water bath, adding polyethylene glycol-400 into vaseline and mixing evenly, swelling acetylated sodium hyaluronate and sodium alginate in 5% glycerol water solution, then adding dopamine-methacrylamide copolymer after ethanol wetting concentration <0.5%, and keeping it at 40°C to prevent vaseline solidification; then, loading active components (C phase-low temperature phase): ultrasonic treatment of silver nano-suspension at 25kHz x 5 min to prevent aggregation, then adding collagen solution and 10-HDA@CDs nano liquid in ice bath environment, finally adding phenoxyethanol to prevent high temperature volatilization, while C phase temperature ≤10°C, preventing collagen denaturation and nano-particle aggregation; finally, gradient mixing and defoaming filling: slowly pouring A phase at 40°C into B phase at 40°C, homogenizing machine 8000 rpm x 15 min to form colostrum; then, cooling the colostrum to 15°C, adding C phase drop by drop under nitrogen protection, and paddle stirring at 200 rpm x 20 min; then, transferring to vacuum homogenizing tank-0.08 MPa, 30°C x 10 min to remove bubbles; finally, 0.22μm filter cartridge online filtration, nitrogen filling into aluminum tube / syringe and sealing;

[0036] The patent application improves the inhibition effect on MRSA and drug-resistant bacteria such as pseudomonas aeruginosa through the synergistic antibiosis of nano royal jelly acid and nano silver, and accelerates the epithelialization of a wound and reduces scar formation through the double action of recombinant type III collagen and acetylated sodium hyaluronate. The intelligent responsive gel matrix (pH sensitive swelling, humidity balance) can dynamically adapt to the wound environment, and the strong adhesion of dopamine copolymer ensures the stability of the physical barrier. In addition, gamma ray sterilization effectively protects the stability of active ingredients, overcomes the defects of traditional dressings such as narrow antibacterial spectrum, single repair effect and poor environmental adaptability, and is especially suitable for complex wounds such as diabetic foot ulcers and II degree burns.

[0037] In the above embodiment, the following process is added:

[0038] According to the embodiment of the present application, in step 1, the complex nanoparticles formed by combining the royal jelly core component 10-hydroxy-2-decenoic acid with carbon quantum dots through nanotechnology, i.e. nano royal jelly acid.

[0039] According to the embodiment of the present application, in step 3, the materials can be mixed together in the mixing device, or one material can be added first, and the carbomer dispersion is prohibited from directly adding water phase -> must be pre-mixed with glycerol into a paste, and then slowly screened into 70℃ phosphate buffer. If "fish eye glue" appears during stirring, immediately add 5 times glycerol and heat to 80℃ to rescue.

[0040] According to the embodiment of the present application, in step 4, when the sodium alginate is not swollen enough, 5% glycerol aqueous solution is used for pre-swelling for 30min with a solid-liquid ratio of 1:20 to reduce the sandiness caused by unsolvated particles; cold ethanol is used for washing and purifying when acrylamide residue exceeds the standard.

[0041] First, the preparation of nano royal jelly acid: the royal jelly acid and ammonium citrate were dissolved in water at a molar ratio of 1:3, and then hydrothermally reacted at 180°C for 6 hours, followed by centrifugal purification to obtain fluorescent carbon quantum dot complex with a particle size of <5 nm, and then adjusted to 20% w / v aqueous dispersion for standby; the 10-hydroxy-2-decenoic acid, a core component of royal jelly, was combined with carbon quantum dots by nanotechnology to form a composite nanoparticle, namely nano royal jelly acid. Then, the reconstituted collagen was dissolved: the freeze-dried collagen powder was dissolved in pre-cooled 0.01mHCl solution at 4°C to 10% w / v to avoid aggregation; then the gel matrix was prepared (A phase-water phase): the pH 7.0 phosphate buffer was heated to 70°C and stirred at 300 rpm, then glycerol and phosphate buffer were added to completely dissolve them, and carboxymethylcellulose 980 was slowly sieved in, the solution was homogenized at 5000 rpm for 10 min to form a transparent gel, and finally the transparent gel was cooled to 40°C and stirred to prevent solidification; the materials can be mixed together in a mixing device, or one material can be added first, and carboxymethylcellulose should not be directly added to the water phase when dispersed → it must be pre-mixed with glycerol into a slurry, and then slowly sieved into the 70°C phosphate buffer. If "fish eye gel" appears during stirring, immediately add 5 times the amount of glycerol and heat to 80°C to rescue. Then the oil phase and active ingredient are mixed (B phase): white vaseline is melted by using a 60°C water bath, polyethylene glycol-400 is added to the vaseline and mixed evenly, acetylated hyaluronic acid sodium and sodium alginate are swelled in 5% glycerol water solution before being added to the inside, then dopamine-methacrylamide copolymer is added after ethanol wetting concentration <0.5%, and it is kept at 40°C to prevent vaseline from solidifying; when sodium alginate is not swelled enough, it needs to be pre-swelled for 30 min with 5% glycerol water solution at a solid-liquid ratio of 1:20 to reduce the grit feeling of the final product caused by unswept particles; when acrylamide residue exceeds the standard, it is purified by washing with cold ethanol. Then the active component is loaded (C phase-low temperature phase): the nano silver suspension is ultrasonically treated at 25 kHz for 5 min to prevent its aggregation, then the collagen solution and 10-HDA@CDs nano liquid are added in an ice bath environment, and finally phenoxyethanol is added to prevent high-temperature volatilization, while the temperature of C phase is ≤10°C to prevent collagen denaturation and nanoparticle aggregation; finally, gradient mixing and defoaming are performed: A phase at 40°C is slowly poured into B phase at 40°C, and the homogenizer is operated at 8000 rpm for 15 min to form the initial milk; then the initial milk is cooled to 15°C, C phase is added dropwise under nitrogen protection, and paddle stirring is performed at 200 rpm for 20 min; then it is transferred to a vacuum homogenizing tank at -0.08 MPa, 30°C for 10 min to remove air bubbles; finally, 0.22μm filter cartridge online filtration is performed, and it is filled into an aluminum tube / injector after nitrogen filling and sealed.

[0042] In the above embodiments, the following process is additionally included:

[0043] According to the embodiment of the present application, in step 6, the hydrophilic carbomer 980 is directly mixed with the oleophobic white vaseline, which is easy to stratify, so it needs to be compatible through the glycerol / PEG-400 transition phase. If the viscosity decreases after sterilization, add 0.1% calcium alginate for secondary cross-linking. After the preparation is completed, sterilization is carried out by γ-rays, so as to avoid thermal damage to the nanostructure, the vacuum degree during vacuum degassing is ≥-0.08MPa, and the time is ≤10min, so as to prevent the gel structure from being damaged due to too high negative pressure. In order to prevent the filter membrane from adsorbing the active ingredients of collagen, a low-protein adsorption PVDF filter membrane can be used for filtration and filling.

[0044] First, the preparation of nano royal jelly acid: the royal jelly acid and ammonium citrate were dissolved in water at a molar ratio of 1:3, hydrothermal reaction at 180℃ for 6 hours, then centrifugal purification to get fluorescent carbon quantum dots complex with particle size <5nm, and adjusted to 20% w / v water dispersion for standby; the core component of royal jelly 10-hydroxy-2-decenoic acid combined with carbon quantum dots through nanotechnology to form a composite nanoparticle, namely nano royal jelly acid. Then the reconstituted collagen is dissolved: take the freeze-dried collagen powder, and dissolve it in pre-cooled 0.01mHCl solution to 10% w / v to avoid aggregation; then the gel matrix preparation (A phase-water phase): heat the pH 7.0 phosphate buffer to 70℃ and stir at 300 rpm, then add glycerol and phosphate buffer to completely dissolve, and slowly sieve into carbomer 980, homogenize the solution at 5000 rpm x 10 min to transparent gel, finally cool the transparent gel to 40℃ and maintain stirring to prevent solidification; materials can be added together in the mixing device for mixing, or one material can be added first, and carbomer dispersion is prohibited to be directly added to the water phase → must be pre-mixed with glycerol into a slurry, and then slowly sieved into 70℃ phosphate buffer. If "fish eye gel" appears during stirring, immediately add 5 times glycerol and heat to 80℃ to rescue. Then the oil phase and active ingredient mixing (B phase): melt white vaseline using 60℃ water bath, add polyethylene glycol-400 into vaseline and mix evenly, acetylated hyaluronic acid sodium and sodium alginate are swelled in 5% glycerol water solution before adding into the inside, then add dopamine-methacrylamide copolymer after ethanol wetting concentration <0.5%, and keep it at 40℃ to avoid vaseline solidification; if sodium alginate is not swelled enough, it needs to be pre-swelled for 30min with 5% glycerol water solution at solid-liquid ratio 1:20 to reduce the sand feeling of the final product caused by unsolved particles; when acrylamide residue exceeds the standard, use cold ethanol to wash and purify. Then the active component loading (C phase-low temperature phase): ultrasonic treatment of nano silver suspension at 25kHz x 5min to avoid aggregation, then add collagen solution and nano royal jelly acid solution in ice bath environment, finally add phenoxyethanol to prevent high temperature volatilization, while the temperature of C phase is ≤10℃ to prevent collagen denaturation and nanoparticle aggregation; finally, gradient mixing and defoaming filling: slowly pour A phase at 40℃ into B phase at 40℃, homogenizer 8000 rpm x 15 min to form colostrum; then cool the colostrum to 15℃, add C phase drop by drop under nitrogen protection, and paddle stirring at 200 rpm x 20 min; then transfer to vacuum homogenization tank-0.08 MPa, 30℃ x 10 min to remove bubbles; finally, 0.22μm filter cartridge online filtration, nitrogen filling into aluminum tube / syringe and sealing. The hydrophilic carbomer 980 and oil-repellent white vaseline are prone to separate layers when mixed directly, so glycerol / PEG-400 transition phase is needed for compatibility.If the viscosity decreases after sterilization, add 0.1% calcium alginate for secondary crosslinking. After preparation, sterilize with gamma rays to prevent thermal damage to the nanostructure. Vacuum degassing should be performed at a pressure of ≥ -0.08 MPa for ≤ 10 minutes to prevent excessive negative pressure from damaging the gel structure. To prevent the membrane from adsorbing the active collagen component, use a low-protein adsorption PVDF membrane for filtration and filling.

Claims

1. A bioactive gel for accelerating wound healing, characterized in that the raw materials are mixed in the following proportions: Carbopol 980: 0.8-1%, glycerin: 5-5.5%, white petrolatum: 15-20%, phosphate buffer: 51.7-55%, nano royal jelly acid: 1.8-2%, recombinant human collagen type III: 2-3%, sodium hyaluronate: 0.4-0.5%, sodium alginate: 1-3%, polyethylene glycol-400: 20-25%, dopamine-methacrylamide copolymer: 1.5%-3%, nano-silver: 0.5%-0.8%, phenoxyethanol: 0.3%-0.5%.

2. The bioactive gel for accelerating wound healing according to claim 1, characterized in that: The carbomer 980, glycerol, white petrolatum, phosphate buffer are gel matrix.

3. The bioactive gel for accelerating wound healing according to claim 1, wherein: The nano-royal jelly acid, recombinant type III human collagen, sodium hyaluronate, sodium alginate, polyethylene glycol-400 are bioactive components, the nano-royal jelly acid is a complex nanoparticle combined by 10-HDA@CDs, and the sodium hyaluronate needs to be acetylated.

4. The bioactive gel for accelerating wound healing according to claim 1, wherein: The dopamine-methacrylamide copolymer, nano-silver, phenoxyethanol are functional strengthening components, and the nano-silver particle size is <20 nm.

5. Process for the preparation of a bioactive gel for accelerating wound healing according to any one of claims 1 to 4, characterized in that: The method comprises the following steps: Step 1: Preparation of nano-royal jelly acid: dissolve royal jelly acid and ammonium citrate in water at a molar ratio of 1:3, hydrothermal reaction at 180 DEG C for 6 hours, then centrifugal purification to obtain fluorescent carbon quantum dot complex with a particle size of <5 nm, and adjust to 20% w / v water dispersion for standby; Step 2: Dissolution of recombinant collagen: take freeze-dried collagen powder, dissolve in 0.01mHCl solution pre-cooled to 4 DEG C to 10% w / v to avoid aggregation; Step 3: Preparation of gel matrix (A phase-water phase): heat the pH 7.0 phosphate buffer to 70 DEG C and stir at 300 rpm, then add glycerol and phosphate buffer to completely dissolve, and slowly screen in carbomer 980, homogenize the solution to transparent gel at 5000 rpm x 10 min, finally cool the transparent gel to 40 DEG C and maintain stirring to prevent solidification; Step 4: Mixing of oil phase and active ingredients (B phase): melt white petrolatum by using 60 DEG C water bath, add polyethylene glycol-400 into the petrolatum and mix uniformly, acetylated sodium hyaluronate and sodium alginate are swelled in 5% glycerol water solution and then added into the interior, then add dopamine-methacrylamide copolymer after ethanol wetting concentration <0.5%, and keep it at 40 DEG C to avoid petrolatum solidification; Step 5: Loading of active components (C phase-low temperature phase): ultrasonic treatment of nano-silver suspension at 25 kHz x 5 min to avoid aggregation, then add collagen solution and 10-HDA@CDs nano-liquid in ice bath environment, finally add phenoxyethanol to prevent high temperature volatilization, while the temperature of C phase is ≤10 DEG C to prevent collagen denaturation and nanoparticle aggregation; Step 6: Gradient mixing and defoaming filling: slowly pour A phase at 40 DEG C into B phase at 40 DEG C, homogenizer 8000 rpm x 15 min to form initial milk; then cool the initial milk to 15 DEG C, add C phase drop by drop under nitrogen protection, and paddle stirring at 200 rpm x 20 min; then transfer to vacuum homogenization tank-0.08 MPa, 30 DEG C x 10 min to remove bubbles; finally, online filtration with 0.22 mu m filter element, nitrogen filling into aluminum tube / syringe, and then sealing.

6. The method for preparing the bioactive gel for accelerating wound healing according to claim 5, characterized in that: In step 1, the complex nanoparticle combined by 10-hydroxy-2-decenoic acid, the core component of royal jelly, and carbon quantum dots through nanotechnology, namely nano-royal jelly acid.

7. The method for preparing the bioactive gel for accelerating wound healing according to claim 5, characterized in that: The in step 3, carbomer dispersion is prohibited to add water phase directly → must be mixed with glycerol into paste, and then slowly sieve into 70℃ phosphate buffer. If "fish eye glue" appears in stirring, immediately add 5 times glycerol and increase temperature to 80℃ to rescue.

8. The method for preparing the bioactive gel for accelerating wound healing according to claim 5, characterized in that: The in step 4, when sodium alginate is not swollen enough, it needs to be pre-swollen for 30min with 5% glycerol solution at solid-liquid ratio of 1:20 to reduce the grit feeling of final product caused by unsolved particles; when acrylamide residue exceeds the standard, use cold ethanol to wash and purify.

9. The method for preparing the bioactive gel for accelerating wound healing according to claim 5, characterized in that: The in step 6, hydrophilic carbomer 980 and oleophobic white vaseline are easy to separate when mixed directly, so it needs to be compatible through glycerol / PEG-400 transition phase. If viscosity decreases after sterilization, add 0.1% calcium alginate for secondary crosslinking.

10. The method for preparing the bioactive gel for accelerating wound healing according to claim 5, characterized in that: The in step 6, after preparation, sterilization is carried out through γ-ray, so as to avoid thermal damage to nanostructure; the vacuum degree during vacuum defoaming is ≥-0.08MPa, and the time is ≤10min, to prevent gel structure damage caused by too high negative pressure; in order to prevent filter membrane from adsorbing collagen active ingredients, low protein adsorption PVDF filter membrane can be used for filtration and filling.