CN115006546B - Disulfide heterojunction material for promoting wound healing and preparation method thereof - Google Patents

Disulfide heterojunction material for promoting wound healing and preparation method thereof Download PDF

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CN115006546B
CN115006546B CN202210257880.5A CN202210257880A CN115006546B CN 115006546 B CN115006546 B CN 115006546B CN 202210257880 A CN202210257880 A CN 202210257880A CN 115006546 B CN115006546 B CN 115006546B
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sulfide
heterojunction
disulfide
solution
bismuth
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CN115006546A (en
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邓怡
黄义宣
梁坤能
黄依玲
谢克难
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Sichuan University
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    • A61K33/00—Medicinal preparations containing inorganic active ingredients
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    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/44—Oxidoreductases (1)
    • A61K38/443—Oxidoreductases (1) acting on CH-OH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
    • A—HUMAN NECESSITIES
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    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
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    • C01G—COMPOUNDS CONTAINING METALS NOT COVERED BY SUBCLASSES C01D OR C01F
    • C01G3/00—Compounds of copper
    • C01G3/12—Sulfides
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    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
    • C12Y101/03004—Glucose oxidase (1.1.3.4)
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    • C01P2004/00—Particle morphology
    • C01P2004/01—Particle morphology depicted by an image
    • C01P2004/03—Particle morphology depicted by an image obtained by SEM
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    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention discloses a disulfide heterojunction material for promoting wound healing and a preparation method thereof, wherein the disulfide heterojunction material comprises bismuth sulfide, copper sulfide, polydopamine and oxidase; the preparation method mainly comprises three steps of preparing bismuth sulfide, preparing bismuth sulfide copper sulfide heterojunction and loading oxidase on the bismuth sulfide copper sulfide heterojunction. The invention provides a disulfide heterojunction material, which can generate local hyperthermia under the irradiation of near infrared light, change the permeability of bacterial cell membranes and leak bacterial organelles or cytoplasmic matrixes in a short time so as to cause bacterial death, and has short action time in the process, so that the bacterial adaptation time is short and the drug resistance is not easy to generate; meanwhile, a large amount of active oxygen can be generated under the irradiation of near infrared light, pathogenic bacteria can be effectively killed, and a good antibacterial effect is achieved.

Description

Disulfide heterojunction material for promoting wound healing and preparation method thereof
Technical Field
The invention relates to the field of medical materials, in particular to a disulfide heterojunction material for promoting wound healing and a preparation method thereof.
Background
The skin is used as the largest organ of human body, has the functions of protecting, excreting, regulating body temperature and the like, and is the first defense line for preventing invasion of pathogenic microorganisms such as bacteria, viruses and the like. Once the skin has a wound, wound infection may occur and healing is difficult for a long time. At present, the clinical treatment of wound infection is mainly combined treatment of debridement and antibiotics. However, excessive use of antibiotics may cause the generation of drug-resistant bacteria, and there is a possibility that wound infection recurs, thereby causing difficulty in healing of wounds, endangering human health. Therefore, the material which can effectively kill bacteria and promote wound healing without generating drug resistance is of great significance.
Hydrogen sulfide is reported to promote angiogenesis and endothelial cell growth and migration. At present, the preparation of hydrogen sulfide mainly uses inorganic sodium sulfide and sodium hydrosulfide, but the biotoxicity of sodium sulfide and sodium hydrosulfide is strong, and the antibacterial effect is avoided. Therefore, it is important to select a hydrogen sulfide source that is biocompatible and has an antibacterial effect. Bismuth sulfide is a sea urchin-shaped metal sulfide, has more photocatalytic sites and good biocompatibility, has been applied to the fields of photocatalysis and the like, but has poor photo-thermal performance and low antibacterial capability of single bismuth sulfide. Copper sulfide is spherical metal sulfide, has good photo-thermal and photodynamic properties, can generate local hyperthermia and a large amount of active oxygen under the irradiation of near infrared light, has proved to have remarkable bactericidal effect, but the single copper sulfide has stronger biotoxicity and larger damage to normal cells.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a disulfide heterojunction material for promoting wound healing and a preparation method thereof.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
a disulfide heterojunction material for promoting wound healing, comprising bismuth sulfide, copper sulfide, polydopamine, and oxidase.
Further, the oxidase is lactate oxidase or glucose oxidase.
A method for preparing a disulfide heterojunction material for promoting wound healing, comprising the following steps:
step 1: preparation of bismuth sulfide
Mixing a certain amount of bismuth nitrate pentahydrate and L-cysteine, and adding deionized water for dissolution; placing the solution into a hydrothermal kettle for reaction, taking out and centrifuging after the reaction to obtain a precipitate, and drying to obtain bismuth sulfide;
step 2: preparation of bismuth sulfide copper sulfide heterojunction
S21: copper chloride and bismuth sulfide are mixed according to the mass ratio of bismuth element to copper element of 1:0.5 to 1, adding water for dissolution, and then adding a surfactant;
s22: adjusting the pH value of the solution to 9.0 by using sodium hydroxide solution, adding a reducing agent while stirring, and reacting for 5 minutes;
s23: slowly adding sodium sulfide solution into the solution in the step S22, performing hydrothermal reaction to obtain a turbid solution, centrifuging, and drying the precipitate to obtain bismuth sulfide copper sulfide heterojunction, namely disulfide metal heterojunction powder;
step 3: loading oxidase onto bismuth sulfide copper sulfide heterojunction
S31: weighing a certain amount of dopamine hydrochloride to be dissolved in a Tris solution (pH=8.6);
s32: immersing the disulfide metal heterojunction in the solution prepared in the step S31, placing the solution in a constant-temperature shaking table for reaction, centrifuging to remove supernatant after the reaction is finished, and reserving precipitate;
s33: dripping oxidase into the precipitate obtained in the step S32, and then freeze-drying to obtain the disulfide heterojunction material; the oxidase is lactate oxidase or glucose oxidase.
Further, the centrifugation time in the step 1 and the step 2 is 10 minutes, and the rotating speed is 8000rpm.
Further, 480mg of bismuth nitrate pentahydrate, 280mg of L-cysteine and 10mL of deionized water were added in the step 1, and the hydrothermal reaction kettle temperature was 150 ℃ for 12h.
Further, the copper chloride in the step 2 is used in an amount of 0.5mM, and the water consumption for dissolution is 25mL; the sodium hydroxide was used in an amount of 1mM and the sodium sulfide solution was used in an amount of 0.5mM.
Further, the surfactant in the step 2 is PVP, and the addition amount is 0.24g; the reducing agent is hydrazine hydrate, and the adding amount is 64 mu L.
Further, the temperature of the hydrothermal reaction in the step 2 is 75 ℃, and the reaction is carried out for 2 hours.
Further, the pH value of the Tris solution in the step 3 is 8.6, the concentration of the dopamine hydrochloride solution is 2mg/mL, and the concentration of the dropwise added oxidase is 1mg/mL.
Further, the temperature of the constant temperature shaking table in the step 3 is controlled to be 37 ℃, and the reaction time is 24 hours.
In the preparation process of the bismuth sulfide copper sulfide heterojunction, the pH value is regulated, hydrazine hydrate is added to firstly generate cuprous oxide microspheres on the surface of bismuth sulfide, then sodium sulfide is slowly added, and oxygen ions overflow the microsphere structure when the sulfur ions attack copper ions, so that the oxygen ions overflow at a higher speed in the process, and a hollow nanometer microsphere structure is generated. Copper sulfide and cuprous sulfide microspheres with different mass ratios can be generated on the surface of bismuth sulfide by controlling the reaction time, and the photo-thermal and photodynamic performances are enhanced due to the simple substance knot formed between the copper sulfide and the cuprous sulfide. In addition, copper sulfide and cuprous sulfide on the surface of bismuth sulfide are of microsphere structures, so that the bismuth sulfide copper sulfide heterojunction has larger specific surface area, and the catalytic activity is enhanced.
According to the invention, a one-pot method is adopted to synthesize the bismuth sulfide copper sulfide heterojunction in situ, heterojunction powder is soaked in polydopamine solution to improve the biocompatibility of the heterojunction and change the surface potential, then oxidase (lactate oxidase or glucose oxidase) solution is dripped into the heterojunction wrapping dopamine, and oxidase is loaded on the surface of the heterojunction through electrostatic interaction, so that the disulfide heterojunction material with obvious antibacterial effect and obvious wound healing promoting effect is obtained.
Compared with the prior art, the invention has the following beneficial effects:
(1) The invention provides a disulfide heterojunction material, which can generate local hyperthermia under the irradiation of near infrared light, change the permeability of bacterial cell membranes and leak bacterial organelles or cytoplasmic matrixes in a short time so as to cause bacterial death, and has short action time in the process, so that the bacterial adaptation time is short and the drug resistance is not easy to generate; meanwhile, a large amount of active oxygen can be generated under the irradiation of near infrared light, pathogenic bacteria can be effectively killed, and a good antibacterial effect is achieved.
(2) When the disulfide heterojunction material prepared by the invention is used, hydrogen sulfide can be generated in an infected microenvironment, so that the generation of blood vessels at a wound and the proliferation and migration of endothelial cells are promoted, and the rapid healing of the wound is effectively promoted.
(3) The bismuth sulfide copper sulfide heterojunction is loaded with oxidase (lactate oxidase or glucose oxidase), so that hydrogen peroxide can be promoted to generate a large amount of active oxygen under the catalysis of the disulfide heterojunction, and bacteria can be killed more effectively.
(4) The preparation method adopts a one-pot method, avoids the separation process in the post-treatment process and the process of purifying the intermediate compound, greatly improves the reaction efficiency, saves the preparation time and ensures that the preparation operation is simpler.
Drawings
FIG. 1 is an SEM characterization of bismuth sulfide, copper sulfide, and bismuth sulfide copper sulfide heterojunction of the present invention;
FIG. 2 is a photo-thermal image of a disulfide heterojunction material of the present invention;
FIG. 3 is a graph of photo-thermal stability analysis of a bismuth sulfide copper sulfide heterojunction according to the present invention;
FIG. 4 is a graph of the photodynamic performance of the disulfide heterojunction material of the present invention;
FIG. 5 is a graph of the antimicrobial properties of the disulfide heterojunction material of the present invention;
FIG. 6 is an electron microscopy scan of bacterial morphology after treatment with the disulfide heterojunction material of the present invention;
fig. 7 is a graph of wound healing promoting properties of the disulfide heterojunction material of the present invention.
Detailed Description
The invention will be further illustrated by the following description and examples, which include but are not limited to the following examples.
The invention provides a disulfide heterojunction material for promoting wound healing, which comprises bismuth sulfide, copper sulfide, polydopamine solution and loaded enzyme, wherein the loaded enzyme is lactate oxidase or glucose oxidase.
Example 1
The embodiment provides a preparation method of a disulfide heterojunction material for promoting wound healing, which comprises the following steps:
step 1: preparation of bismuth sulfide
Mixing 480mg of bismuth nitrate pentahydrate and 280mg of L-cysteine, adding 10mL of deionized water for dissolution, transferring the solution into a hydrothermal kettle at 150 ℃ for reaction for 12 hours, taking out the bismuth sulfide after the reaction is completed, centrifuging at 8000rpm for 10 minutes, retaining precipitate, and drying to obtain bismuth sulfide.
Step 2: preparation of bismuth sulfide copper sulfide heterojunction
S21: 0.5mM copper chloride and bismuth sulfide are mixed according to the mass ratio of bismuth element to copper element of 1:1, adding 25mL of water for dissolution, and then adding 0.24g of surfactant PVP;
s22: adjusting the pH to 9.0 with 1mM sodium hydroxide solution, adding 64 mu L of hydrazine hydrate while stirring, and reacting for 5 minutes;
s23: slowly adding 0.5mM sodium sulfide solution into the solution in the step S22, reacting for 2 hours at 75 ℃ to enable copper sulfide to be generated on the surface of bismuth sulfide in situ, obtaining a turbid solution, centrifuging for 10 minutes at 8000rpm, obtaining precipitate, and drying to obtain bismuth sulfide copper sulfide heterojunction powder. (As shown in FIG. 1, wherein A is bismuth sulfide, B is copper sulfide, and C is SEM characterization of a disulfide metal heterojunction.)
Step 3: preparation of a Dithiobiological heterojunction Material
S31: weighing a certain amount of dopamine hydrochloride, and dissolving the dopamine hydrochloride in a Tris solution with the pH of 8.6 to obtain a solution with the concentration of 2mg/mL of dopamine hydrochloride;
s32: immersing a proper amount of bismuth sulfide and copper sulfide heterojunction powder into the solution prepared in the step S31, placing the solution into a shaking table with constant temperature of 37 ℃ for reaction for 24 hours, centrifuging after the reaction is finished, retaining the precipitate, dripping 1mg/mL of lactic acid oxidase solution into the solution, and freeze-drying to obtain the disulfide heterojunction material capable of promoting wound healing.
And carrying out a photo-thermal performance experiment, a photodynamic performance experiment, a bacteriostasis experiment, a bacterial morphology test and an animal experiment on the prepared disulfide heterojunction material.
Comparative example: preparing copper sulfide: 0.5mM CuCl was taken 2 ·2H 2 After O was dissolved by adding 25mL of water, 0.24g of PVP was added as a surfactant, then the pH was adjusted to 9.0 with 1mM sodium hydroxide solution, 64. Mu.L of hydrazine hydrate was added as a reducing agent while stirring, a sodium sulfide solution (0.5 mM) was slowly added after 5 minutes of reaction and reacted at 75℃for 2 hours, so that copper sulfide was formed in situ on the surface of bismuth sulfide, the resulting cloudy solution was centrifuged (800 rpm,10 minutes), and the resulting precipitate was dried to obtain copper sulfide.
(1) Photo-thermal performance experiment
Experiment 1: the bismuth copper sulfide heterojunction prepared in this example, the bismuth copper sulfide heterojunction loaded with lactate oxidase and the copper sulfide in the comparative example were taken in 48-well plates, phosphate Buffer Solution (PBS) without adding material was additionally taken as a control group, 500. Mu.L of PBS was added to each sample well plate, and 808nm near infrared light (1.5W/cm 2 ) Each sample well plate was irradiated and the temperature change was captured every 30 seconds using FLIR infrared detector. The results are shown in FIG. 2, where PBS represents phosphate buffer, BC represents bismuth sulfide copper sulfide heterojunction, and BCPL represents lactate oxidase-loaded disulfide heterojunction.
Experiment 2: the bismuth copper sulfide heterojunction prepared in this example was taken in a 48-well plate, 500 μl of PBS was added to the sample well plate, the sample well plate was irradiated with 808nm near infrared light for 15 minutes, then the infrared irradiation lamp was turned off for cooling for 15 minutes, the temperature change was captured with a FIRT infrared detector every 30 seconds during the period, the operation was cycled three times, the data was recorded and the photo-thermal stability of the sample was analyzed, and the result was shown in fig. 3.
Summarizing: from fig. 2, it can be known that copper sulfide, bismuth sulfide and a disulfide heterojunction loaded with lactate oxidase can realize local hyperthermia under the irradiation of near infrared light, wherein the copper sulfide has a higher temperature rise in a shorter time and has the risk of burning skin, the bismuth sulfide has a lower temperature rise, and the disulfide heterojunction loaded with lactate oxidase has a temperature rise speed between the two, so that the temperature is easier to control; as can be seen from fig. 3, the bismuth sulfide copper sulfide heterojunction has good photo-thermal cycling capability, provides recycling possibility, and is beneficial to the stability of photo-thermal antibiosis while promoting environmental friendliness.
(2) Photodynamic performance experiment
Bismuth sulfide, bismuth sulfide copper sulfide heterojunction prepared in this example and disulfide heterojunction carrying lactate oxidase and copper sulfide in comparative example were taken in a 48-well plate, wherein Phosphate Buffer Solution (PBS) without adding material was used as a control group, 0.5mL of 100. Mu.g/mL Methylene Blue (MB) was used as an active oxygen (. OH) marker, and each group was placed in near infrared light (1.5W/cm) under a dark condition 2 ) The sample was irradiated for 15 minutes, 100. Mu.L of the solution was measured for absorbance in a microplate reader every 5 minutes, and the test results are shown in FIG. 4.
Summarizing: the disulfide heterojunction material loaded with the lactic acid oxidase degrades the most methylene blue, has the strongest capacity of combining oxygen-containing substances to generate hydroxyl free radicals, and can achieve excellent antibacterial effect.
(3) Bacteriostasis experiment
Experiment 1: bismuth sulfide, bismuth sulfide copper sulfide heterojunction prepared in this example and disulfide heterojunction carrying lactate oxidase and copper sulfide in comparative example were taken in 48-well plates, wherein Phosphate Buffer Solution (PBS) without adding material was used as a control group, 100 μl of liquid medium and 100 μl of 106CFU/mL staphylococcus aureus (s.aureus) were added to each sample well, and 3 parallel experimental groups were established. After each group was cultured under 808nm laser irradiation and under dark conditions for 20 minutes, the bacterial liquid was uniformly coated on a solid medium, and after culturing at 37℃for 24 hours, the sterilizing effect of each material was observed.
Experiment 2: 106CFU/mL Staphylococcus aureus (S.aureus) in experiment 1 above was replaced with 10 6 CFU/mL E.coli (E.coli) experiment 1 was repeated.
The results of experiments 1 and 2 are shown in FIG. 5, where NIR (+) is shown to indicate near infrared light irradiation and NIR (-) is shown to indicate no near infrared light irradiation.
Summarizing: the disulfide heterojunction loaded with the lactic acid oxidase kills most staphylococcus aureus and escherichia coli under the irradiation of near infrared light, so that the problem of wound infection can be effectively solved.
(4) Bacterial morphology testing
Experiment 1: bismuth sulfide, bismuth sulfide copper sulfide heterojunction prepared in this example and bismuth sulfide copper sulfide heterojunction carrying lactate oxidase and copper sulfide in comparative example were taken in 48-well plates, wherein Phosphate Buffer Solution (PBS) without adding material was used as a control group, 100 μl of liquid medium and 100 μl of 106CFU/mL staphylococcus aureus (s.aureus) were added to each sample well, and 3 parallel experimental groups were established. After culturing each group under 808nm laser irradiation and dark conditions for 20 minutes, the samples were centrifuged and taken out, and placed in 500. Mu.L of 2.5% glutaraldehyde, and stored at 4℃for 12 hours, then dehydrated with gradient ethanol (30%, 50%, 70%, 90% and 100% v/v), each gradient dehydrated for 10 minutes, and finally the samples were naturally air-dried and observed for bacterial morphology with a scanning electron microscope.
Experiment 2: 106CFU/mL Staphylococcus aureus (S.aureus) in experiment 1 above was replaced with 10 6 CFU/mL E.coli (E.coli) experiment 1 was repeated.
Summarizing: as shown in fig. 6, the disulfide heterojunction loaded with the lactic acid oxidase can damage the integrity of a bacterial membrane, so that bacterial organelles or cytoplasm matrixes leak, bacterial death is caused, the action time is short, the action effect is obvious, and therefore, the adaptation time of the bacteria is short, and drug resistance is not easy to generate.
(5) Animal experiment
Establishing a wound infection model: grouping and numbering mice, removing hair on the back, injecting 5% chloral hydrate (0.1 mL/10 g) into abdominal cavity for anesthesia, creating a full-thickness skin wound with a diameter of about 1 cm at 2cm outside the spinal column of the mice, and dripping 10 μl of Staphylococcus aureus bacterial liquid (1×10) by using a pipette 8 CFU/mL), and finally applying a special-component-free adhesive bandage or wrapping with sterile gauze to prevent the mice from licking or scratching wounds.
Wound treatment analysis: a photograph of the wound was taken 24 hours after the infection model was established, and the length and width of the wound were measured with a ruler. Placing materials at the wound site, respectively (vulcanizationBismuth, bismuth sulfide copper sulfide heterojunction, lactate oxidase-loaded disulfide heterojunction, and PBS), all light groups received an intensity of 1.5W/cm 2 Is irradiated for 10 minutes with near infrared light of 808 nm. The wound is exposed to air after illumination. The wounds were recorded every two days in the following eight days,
the results are shown in FIG. 7, in which Bi 2 S 3 + represents bismuth sulfide plus near-infrared light irradiation, bc+ represents bismuth sulfide copper sulfide heterojunction plus near-infrared light irradiation, bcpl+ represents lactate oxidase-loaded disulfide heterojunction plus near-infrared light irradiation; bi (Bi) 2 S 3 BC, BCPL indicate that no near infrared light irradiation was performed.
Summarizing: the wound recovery speed is the fastest and the wound area is the smallest after the disulfide heterojunction material loaded with the lactic acid oxidase is subjected to the irradiation treatment. In comparison, in the sham group treated with PBS, the wound of the mice had pus exudation and inflammation was evident. Under the synergistic effect of photo-heat and photodynamic, the material prepared by the embodiment has remarkable antibacterial effect, reduces wound inflammatory reaction, and simultaneously promotes angiogenesis and endothelial cell proliferation and migration by the generated hydrogen sulfide so as to promote wound healing rapidly.
Example 2
Based on the preparation method of the biological heterojunction material for promoting wound healing provided in the embodiment 1, the mass ratio of 0.5mM copper chloride and bismuth sulfide in the step 2 can be further as follows: 0.75, and then 25mL of water was added for dissolution.
Example 3
Based on the preparation method of the biological heterojunction material for promoting wound healing provided in the embodiment 1, the mass ratio of 0.5mM copper chloride and bismuth sulfide in the step 2 can be further as follows: 0.5, and then 25mL of water was added for dissolution.
The above embodiment is only one of the preferred embodiments of the present invention, and should not be used to limit the scope of the present invention, but all the insubstantial modifications or color changes made in the main design concept and spirit of the present invention are still consistent with the present invention, and all the technical problems to be solved are included in the scope of the present invention.

Claims (8)

1. A disulfide heterojunction material for promoting wound healing, which is characterized by comprising bismuth sulfide, copper sulfide, polydopamine and oxidase; the oxidase is lactate oxidase or glucose oxidase;
the preparation method of the disulfide heterojunction material comprises the following steps:
step 1: preparation of bismuth sulfide
Mixing a certain amount of bismuth nitrate pentahydrate and L-cysteine, and adding deionized water for dissolution; placing the solution into a hydrothermal kettle for reaction, taking out and centrifuging after the reaction to obtain a precipitate, and drying to obtain bismuth sulfide;
step 2: preparation of bismuth sulfide copper sulfide heterojunction
S21: copper chloride and bismuth sulfide are mixed according to the mass ratio of bismuth element to copper element of 1:0.5 to 1, adding water for dissolution, and then adding a surfactant;
s22: adjusting the pH value of the solution to 9.0 by using sodium hydroxide solution, adding a reducing agent while stirring, and reacting for 5 minutes;
s23: slowly adding sodium sulfide solution into the solution in the step S22, performing hydrothermal reaction to obtain a turbid solution, centrifuging, and drying the precipitate to obtain bismuth sulfide copper sulfide heterojunction, namely disulfide metal heterojunction powder;
step 3: loading oxidase onto bismuth sulfide copper sulfide heterojunction
S31: weighing a certain amount of dopamine hydrochloride and dissolving the dopamine hydrochloride into a Tris solution;
s32: immersing the disulfide metal heterojunction in the solution prepared in the step S31, placing the solution in a constant-temperature shaking table for reaction, centrifuging to remove supernatant after the reaction is finished, and reserving precipitate;
s33: and (3) dripping oxidase into the precipitate obtained in the step (S32), and then freeze-drying to obtain the disulfide heterojunction material.
2. The disulfide heterojunction material as claimed in claim 1, wherein the centrifugation time in said step 1 and said step 2 is 10 minutes, and the rotation speed is 8000rpm.
3. The disulfide heterojunction material for promoting wound healing as claimed in claim 2, wherein in said step 1, 480mg of bismuth nitrate pentahydrate, 280mg of l-cysteine are taken, the volume of deionized water added is 10mL, and the temperature of the hydrothermal reaction kettle is 150 ℃, and the reaction time is 12 hours.
4. A disulfide heterojunction material as claimed in claim 3, wherein the copper chloride in said step 2 is used in an amount of 0.5mM and the water for dissolution is 25mL; the sodium hydroxide was used in an amount of 1mM and the sodium sulfide solution was used in an amount of 0.5mM.
5. The disulfide heterojunction material as claimed in claim 4, wherein the surfactant in said step 2 is PVP and is added in an amount of 0.24g; the reducing agent is hydrazine hydrate, and the adding amount is 64 mu L.
6. The disulfide heterojunction material as claimed in claim 5, wherein the temperature of the hydrothermal reaction in said step 2 is 75 ℃, and the reaction is 2 hours.
7. The disulfide heterojunction material as claimed in claim 6, wherein the pH of the Tris solution in the step 3 is 8.6, the concentration of the dopamine hydrochloride solution is 2mg/mL, and the concentration of the oxidase is 1mg/mL.
8. The disulfide heterojunction material for promoting wound healing as claimed in claim 7, wherein the temperature of the constant temperature cradle in said step 3 is controlled to be 37 ℃ and the reaction time is 24 hours.
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