Preparation method of antibacterial and antioxidant hemostatic hydrogel dressing capable of releasing phycocyanin

Abstract

The present application reports a preparation method of an antibacterial and antioxidant hemostatic hydrogel dressing which can release phycocyanin. Put kaolin into a water solution of lysozyme to adsorb lysozyme, then put it into a mixed water solution of γ-methacryloxypropyltrimethoxysilane and γ-aminopropyltriethoxysilane, and polysiloxane is generated on the surface of kaolin. Mix the lysozyme-adsorbed kaolin treated with silane with a water solution of spirulina and sodium alginate, and get a casting solution after standing and defoaming. After scraping the casting solution into a film, spray a water solution containing calcium chloride and lysozyme to the surface of the film to make the film solidify and crosslink, and get an antibacterial and antioxidant hemostatic hydrogel dressing which can release phycocyanin. Lysozyme embedded in the hydrogel and polysiloxane is released near spirulina to destroy the cell wall of spirulina, thereby releasing phycocyanin. Lysozyme, phycocyanin and kaolin respectively endow the hydrogel dressing of the present application with antibacterial, antioxidant and hemostatic functions.

Description

TECHNICAL FIELD

[0001] The application is a preparation method of an antibacterial and antioxidant hemostatic hydrogel dressing with sustained release of phycocyanin, belonging to the field of biomaterials and medicines. BACKGROUND

[0002] At present, the main hemostatic components of military hemostatic materials at home and abroad are chitosan or clay particles (mainly kaolin and zeolite), and the carriers are traditional gauze or high molecular dressing. Historically, a variety of minerals have been used for wound hemostasis, such as zeolite, montmorillonite, etc., which have gradually withdrawn due to too many side effects. The exothermic reaction of zeolite during hemostasis can cause local burns, and montmorillonite particles are easy to fall off, causing damage to the inner wall of blood vessels and organ embolism, etc. Kaolin is mainly composed of small flaky, tubular, and laminar kaolinite mineral clusters with a particle size of less than 2 microns, which belongs to silicate inert minerals, and has abundant natural reserves in China. Kaolin contains a large amount of Al2O3, SiO2, and a small amount of Fe2O3, TiO2, as well as trace amounts of K2O, Na2O, CaO, and MgO. When kaolin comes into contact with blood, it can directly stimulate coagulation factor XII to start the endogenous coagulation pathway, and then activate coagulation factor XI to form fibrin monomers from fibrinogen, which combine into fibrin polymers to form water-insoluble blood fibers. In the absence of coagulation factor XII, kaolin can directly activate coagulation factor XI to achieve hemostasis. Another hemostatic mechanism of kaolin is physical hemostasis. When kaolin comes into contact with blood flowing out of a damaged wound, it can quickly absorb water molecules in the blood, concentrate blood platelets and thrombin, and make coagulation factors and platelets aggregate and deposit to achieve hemostasis

Military Medicine, 2017, 41(2): 141-145

[0003] Wound dressing or artificial skin can prevent bacteria from invading the wound bed, effectively maintain the moisture of the wound, absorb wound exudates, and have good biocompatibility. In commercial wound dressings, most are prepared from natural biological polymers. Gelatin or collagen crosslinked with glutaraldehyde can reduce immunogenicity and is suitable for the treatment of full-thickness skin wounds. Non-woven fabric of chitin fiber is used for burn treatment and has shown good results in clinical studies. Poly(L-leucine) sponge is also used as a dressing for full-thickness skin wounds in clinical trials. Matsuda et al. embedded microspheres of poly-L-lactic acid containing the antibiotic Tobramycin in the silicone layer on the upper layer of the double-layer artificial skin. As the microspheres continue to degrade, the antibiotic in them can be released slowly, preventing external bacterial infection

Biomaterials, 1992, 13:119

[0004] Alginate is a natural polysaccharide isolated from kelp, sargassum and other brown seaweed. It is widely available and inexpensive. As early as 1951, Blaine et al. had explored the possibility of calcium alginate as a hemostatic material. British Winter found that the surface of the wound in a humid environment heals faster than in a dry environment. The humid environment speeds up the migration of epidermal cells from healthy skin to the wound, thus speeding up the healing of the wound. Under the guidance of the principle of "wet therapy", alginate-based medical dressings, gauze, bandages and other products have been widely used due to their superior moisture retention and overall easy removal.

Knitting Industry, 2004, 32(5): 60-63

Ann R Coll Surg Engl., 1986, 68(1): 27-28

[0005] Both kaolin and alginate lack antibacterial and bacteriostatic properties. Nanosilver-loaded calcium alginate dressing has good antibacterial properties, but the high concentration of silver ions can easily cause heavy metal ion toxicity, and the silver ions are easily lost, causing the antibacterial properties to decrease. Lysozyme, also known as muramidase, is a non-toxic, harmless and highly safe protein that kills microorganisms by hydrolyzing the mucopolysaccharide of microorganisms, causing the cell wall to rupture and the contents to escape. It has no side effects on the human body, does not remain in the body, and has certain health functions. The role of lysozyme in food preservation has attracted widespread attention from society, and it has been widely used in the preservation of aquatic products and meat products. It can be used as an antibacterial agent in wound dressing.

[0006] Phycocyanin is a deep blue powder isolated from Spirulina, which is one of the rare pigment proteins in nature. It is not only bright in color, but also a nutrient-rich protein with complete amino acid composition and high essential amino acid content. Phycocyanin has the functions of antioxidant, anti-infection, anti-allergy, anti-cancer, promoting blood cell regeneration, and promoting the synthesis of elastin in the human body. In Europe, the United States, Japan and other countries, phycocyanin is widely used as a high-grade natural pigment for food and cosmetics, and is made into biochemical drugs. Due to its good color characteristics and nutritional effects, phycocyanin is widely developed and applied as a natural blue pigment and nutritional health ingredient in food, cosmetics, diagnosis and treatment, rehabilitation health products, and other fields. As the source of phycocyanin, Spirulina has also received more and more attention in recent years. Spirulina is a filamentous blue-green algae rich in various proteins, vitamins and trace elements, which is green, safe, easy to obtain, non-toxic and side effect, and has high nutritional value and disease prevention and health value. It has also been found to have many pharmacological effects and can be used as a biological active supplement with physiological effects of promoting cell vitality or survival, so it has attracted much attention in the biomedical industry. Spirulina is becoming increasingly important in the fields of health products, food and medicine. Moreover, there is no report on the toxic and side effects of Spirulina products on humans. It can be used to resist viruses, oxidation, improve human immunity and assist in the treatment of diseases. Choi et al. also used the electrospinning method and prepared a alginate / PCL nanofiber wound dressing containing Spirulina extract by CaCl2 crosslinking, and the results showed that the dressing can accelerate wound healing by improving the moisture retention, biological safety and effectiveness of the wounded skin. The impregnation of alginate increases the adhesion and humidity of the skin, accelerates wound healing, and does not cause cytotoxicity

[0007]

Biotechnology and Bioprocess Engineering, 2017, 22(6) :679-685

[0008] In view of potential harm of heavy metal ion antibacterial agent in dressing, drug resistance of bacteria caused by antibiotic abuse, lack of antibacterial and antioxidant function of traditional dressing, easy loss and high price of directly added phycocyanin, etc., the application reports a preparation method of antibacterial and antioxidant hemostatic hydrogel dressing capable of releasing phycocyanin. Kaolin is put into a lysozyme aqueous solution to adsorb lysozyme, and then put into a mixed aqueous solution of gamma-methacryloxypropyltrimethoxysilane and gamma-aminopropyltriethoxysilane, and polysiloxane is formed on the surface of kaolin. The lysozyme-adsorbed kaolin treated with silane is mixed with an aqueous solution of spirulina and sodium alginate, and a casting solution is obtained after standing and defoaming. The casting solution is scraped into a film, and an aqueous solution containing calcium chloride and lysozyme is sprayed onto the surface of the film to make the film solidify and crosslink, thereby obtaining an antibacterial and antioxidant hemostatic hydrogel dressing capable of releasing phycocyanin. The lysozyme embedded in the hydrogel and polysiloxane is released near the spirulina to destroy the cell wall of the spirulina, thereby releasing the phycocyanin. The lysozyme, phycocyanin and kaolin respectively endow the hydrogel dressing of the application with antibacterial, antioxidant and hemostatic functions. SUMMARY

[0009] In view of the deficiencies of the prior art, the technical problem to be solved by the application is potential harm of heavy metal ion antibacterial agent in dressing, drug resistance of bacteria caused by antibiotic abuse, lack of antibacterial and antioxidant function of traditional dressing, easy loss and high price of directly added phycocyanin, etc. The technical solution of the application to solve the aforementioned problems is to provide a preparation method of antibacterial and antioxidant hemostatic hydrogel dressing capable of releasing phycocyanin.

[0010] The application provides a preparation method of antibacterial and antioxidant hemostatic hydrogel dressing capable of releasing phycocyanin, characterized by comprising the following steps:

[0011] a) a lysozyme aqueous solution with a mass percentage concentration of 0.1% to 5% is prepared with deionized water, kaolin is dispersed into the lysozyme aqueous solution under stirring to obtain a kaolin mixed aqueous solution with a mass percentage concentration of 0.5% to 5% of kaolin, the dispersibility of the kaolin is improved by ultrasonic treatment at low power, the pH of the mixed aqueous solution is adjusted to 4-5, and the lysozyme is fully adsorbed onto the kaolin under stirring, the lysozyme-adsorbed kaolin is separated by filtration with filter paper, and then put into a mixed aqueous solution of gamma-methacryloxypropyltrimethoxysilane and gamma-aminopropyltriethoxysilane with a mass percentage concentration of 0.2% to 2% respectively, the components are uniformly dispersed by ultrasonic treatment at low power, and the mixture is left to stand for 12-36 hours, the precipitate is obtained by filtration, and the unreacted substances are washed away with deionized water to obtain lysozyme-adsorbed kaolin wrapped with a layer of polysiloxane, which is freeze-dried for later use;

[0012] b) dispersing the lysozyme-adsorbed and polysiloxane-coated kaolin obtained in step a) in a mass percentage concentration of 0.2% to 5% into deionized water under stirring, then adding sodium alginate in a mass percentage concentration of 1.0% to 3% and continuing to stir to dissolve the sodium alginate, and obtaining a casting solution after standing and defoaming;

[0013] c) preparing a water-soluble solution of lysozyme in a mass percentage concentration of 0.1% to 5% and a water-soluble solution of soluble calcium salt in a mass percentage concentration of 1.0% to 20% as a crosslinking agent water solution, and pouring the crosslinking agent water solution into a spray bottle for standby;

[0014] d) pouring the casting solution obtained in step b) onto a clean and flat glass plate, scraping a liquid film with a film scraper, and immediately spraying the crosslinking agent water solution in the spray bottle in step c) onto the surface of the liquid film, so that calcium ions in the soluble calcium salt crosslink with sodium alginate to form a calcium alginate hydrogel film, and the calcium alginate hydrogel film is taken out after preliminary shaping and immediately sprayed with the crosslinking agent water solution on the other side of the film, and the hydrogel film is obtained after sufficient crosslinking, which is an antibacterial and antioxidant hemostatic hydrogel dressing capable of releasing phycocyanin; the hydrogel dressing is cut into the required shape and size with a mold for standby;

[0015] e) since the lysozyme bacteria are adsorbed and embedded by polysiloxane, and lysozyme is easily adsorbed by alginate, the lysozyme in the hydrogel dressing has a slow-release performance, the slow-released lysozyme meets the spirulina to destroy the cell wall of the spirulina, so as to achieve the purpose of slow-releasing phycocyanin; the slow-release performance of phycocyanin in the hydrogel dressing obtained in step d) is tested in physiological saline, and the hydrogel dressing has a longer ability to release phycocyanin than the dressing directly added with phycocyanin and spirulina, and can continuously release phycocyanin for 24 to 60 hours;

[0016] f) the antibacterial rate of the hydrogel dressing on Staphylococcus aureus reaches 65% to 100%;

[0017] g) the clearance rate of the phycocyanin released by the hydrogel dressing on 2,2'-azobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt reaches 60% to 96% after 7 hours.

[0018] The soluble calcium salt in the application is any one or a mixture of two or more of calcium chloride, calcium phosphate, and calcium nitrate, and the antibacterial and antioxidant hemostatic hydrogel dressing has a hemostatic time of 1 to 5 minutes for a rat liver lobe resection bleeding. DETAILED DESCRIPTION

[0019] The specific embodiments of the application are described below, but the application is not limited by the embodiments.

[0020] Example 1.

[0021] a) A 0.1% by mass lysozyme aqueous solution was prepared using deionized water, and kaolin was dispersed into the lysozyme aqueous solution under stirring to obtain a 0.5% by mass kaolin mixed aqueous solution. The dispersion of the kaolin was improved by ultrasonic treatment at low power, the pH of the mixed aqueous solution was adjusted to 4, and the lysozyme was fully adsorbed onto the kaolin under stirring. The kaolin with adsorbed lysozyme was separated by filtration using filter paper, and then was put into a 0.2% by mass mixed aqueous solution of γ-methacryloxypropyltrimethoxysilane and γ-aminopropyltriethoxysilane to uniformly disperse the components by ultrasonic treatment at low power. The mixture was allowed to stand for 12 hours, and the precipitate was collected by filtration and washed with deionized water to remove unreacted substances, thereby obtaining lysozyme-adsorbed kaolin wrapped with a layer of polysiloxane. The product was freeze-dried for later use;

[0022] b) The freeze-dried lysozyme-adsorbed kaolin wrapped with a layer of polysiloxane obtained in step a) was dispersed into deionized water under stirring to obtain a 0.2% by mass aqueous solution. Then, 0.5% by mass spirulina and 1.0% by mass sodium alginate were added to the aqueous solution, and the sodium alginate was fully dissolved by further stirring. After standing to remove bubbles, a casting solution was obtained for later use;

[0023] c) A mixed aqueous solution of 0.1% by mass lysozyme and 1.0% by mass calcium chloride was prepared as a crosslinking agent aqueous solution, and was poured into a spray bottle for later use;

[0024] d) The casting solution obtained in step b) was poured onto a clean and flat glass plate, and a doctor blade was used to scrape the solution into a liquid film with uniform thickness. Then, the crosslinking agent aqueous solution in the spray bottle of step c) was immediately sprayed onto the surface of the liquid film. The calcium ions in the soluble calcium salt were crosslinked with the sodium alginate to form a calcium alginate hydrogel film. After the preliminary shaping of the calcium alginate hydrogel film, the other side of the film was immediately sprayed with the crosslinking agent aqueous solution. After sufficient crosslinking, an antibacterial and antioxidant hemostatic hydrogel dressing capable of releasing phycocyanin was obtained. The hydrogel dressing was cut into the desired shape and size using a mold for later use;

[0025] e) Since the lysozyme bacteria were adsorbed and embedded with polysiloxane, and the lysozyme was easily adsorbed by alginate, the lysozyme inside the hydrogel dressing had a slow-release property. The slow-released lysozyme could destroy the cell wall of spirulina, thereby achieving the purpose of slow-releasing phycocyanin. The slow-release property of phycocyanin in the hydrogel dressing obtained in step d) was tested in physiological saline. The hydrogel dressing had a longer-lasting ability to release phycocyanin than the dressing directly added with phycocyanin and spirulina, and could continuously release phycocyanin for 24 hours;

[0026] f) the bacteriostatic rate of the hydrogel dressing against Staphylococcus aureus reaches 65%;

[0027] g) the clearance rate of the phycocyanin released by the hydrogel dressing against 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt reaches 60% after 7 hours, and the hemostatic time for a rat liver lobectomy hemorrhage is 5 minutes.

[0028] Example 2.

[0029] a) prepare a 5% (mass percentage) lysozyme aqueous solution with deionized water, disperse kaolin into the lysozyme aqueous solution under stirring to obtain a 5% (mass percentage) kaolin mixed aqueous solution, improve the dispersibility of the kaolin under low-power ultrasonic, adjust the pH of the mixed aqueous solution to 5, and fully adsorb the lysozyme onto the kaolin under stirring, separate the lysozyme-adsorbed kaolin by filtration with filter paper, and then put it into a 2% (mass percentage) mixture aqueous solution of γ-methacryloxypropyltrimethoxysilane and γ-aminopropyltriethoxysilane to uniformly disperse the components under low-power ultrasonic, stand for 36 hours of reaction, filter and take the precipitate, and wash away the unreacted substances with deionized water to obtain lysozyme-adsorbed kaolin wrapped with a layer of polysiloxane, which is ready for use after freeze-drying;

[0030] b) disperse 5% (mass percentage) freeze-dried lysozyme-adsorbed kaolin wrapped with a layer of polysiloxane obtained in step a) and 2% (mass percentage) spirulina into deionized water under stirring, then add 3% (mass percentage) sodium alginate, continue to stir to fully dissolve the sodium alginate, and obtain a casting solution after standing and defoaming;

[0031] c) prepare a mixed aqueous solution of 5% (mass percentage) lysozyme and 20% (mass percentage) calcium nitrate as a crosslinker aqueous solution, and pour the crosslinker aqueous solution into a spray bottle for standby;

[0032] d) pour the casting solution obtained in step b) onto a clean and flat glass plate, scrape it into a liquid film with a film scraper, then immediately spray the crosslinker aqueous solution in the spray bottle in step c) onto the surface of the liquid film, crosslink the calcium alginate hydrogel film generated by the crosslinking of calcium ions in the soluble calcium salt and sodium alginate, take out the preliminarily shaped calcium alginate hydrogel film immediately, spray the crosslinker aqueous solution onto the other side of the film for crosslinking, and obtain an antibacterial, antioxidant and hemostatic hydrogel dressing capable of releasing phycocyanin after sufficient crosslinking; the hydrogel dressing is carved into the required shape and size with a mold for standby;

[0033] e) due to the adsorption of lysozyme and the embedding of polysiloxane, and the adsorption of lysozyme by alginate, the lysozyme inside the hydrogel dressing has a slow-release performance, the slow-released lysozyme meets the spirulina and destroys the cell wall of spirulina, so as to achieve the purpose of slow release of phycocyanin; the slow-release performance of phycocyanin in the hydrogel dressing obtained in step d) is tested in physiological saline, and the hydrogel dressing has a longer ability to release phycocyanin than the dressing directly added with phycocyanin and spirulina, and can continuously release phycocyanin for 60 hours;

[0034] f) the bacteriostatic rate of the hydrogel dressing to Staphylococcus aureus reaches 100%;

[0035] g) the clearance rate of the released phycocyanin of the hydrogel dressing to 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt reaches 96% after 7 hours, and the hemostatic time of the liver resection bleeding of rats is 1 minute.

[0036] Example 3.

[0037] a) prepare a 0.5% (mass percentage) lysozyme aqueous solution with deionized water, disperse kaolin into the lysozyme aqueous solution under stirring to obtain a 2.5% (mass percentage) kaolin mixed aqueous solution, improve the dispersibility of kaolin by ultrasonic under low power, adjust the pH of the mixed aqueous solution to 4.5, and make lysozyme fully adsorbed onto kaolin under stirring, then separate the lysozyme-adsorbed kaolin by filtration with filter paper, and then put it into a 1.0% (mass percentage) mixed aqueous solution of γ-methacryloxypropyltrimethoxysilane and γ-aminopropyltriethoxysilane, uniformly disperse each component by ultrasonic under low power, and stand for 24 hours of reaction, then take the precipitate by filtration, and wash away unreacted substances with deionized water to obtain kaolin wrapped with a layer of polysiloxane and adsorbed with lysozyme, which is ready for use after freeze-drying;

[0038] b) disperse 3% (mass percentage) of the freeze-dried kaolin wrapped with a layer of polysiloxane and adsorbed with lysozyme obtained in step a) and 1.5% (mass percentage) of spirulina into deionized water under stirring, then add 2.5% (mass percentage) of sodium alginate, continue to stir to make sodium alginate fully dissolved, and then stand for defoaming to obtain a casting solution ready for use;

[0039] c) prepare a mixed aqueous solution of 1.0% (mass percentage) lysozyme and 2.5% (mass percentage) calcium dihydrogen phosphate as a crosslinking agent aqueous solution, and pour the crosslinking agent aqueous solution into a spray bottle ready for use;

[0040] d) Pour the casting solution obtained in step b) on a clean flat glass plate, and use a doctor blade to form a liquid film with uniform thickness, then immediately spray the crosslinking agent aqueous solution in the spray bottle of step c) onto the surface of the liquid film, the calcium ions in the soluble calcium salt crosslink with sodium alginate to form a calcium alginate hydrogel film, after the calcium alginate hydrogel film is initially shaped, it is immediately sprayed with a crosslinking agent aqueous solution to crosslink the other side of the film, and after sufficient crosslinking, an antibacterial and antioxidant hemostatic hydrogel dressing capable of releasing phycocyanin is obtained; the hydrogel dressing is carved into the required shape and size using a mold for standby;

[0041] e) Because the lysozyme is adsorbed and embedded by polysiloxane, and the lysozyme is easily adsorbed by alginate, the lysozyme inside the hydrogel dressing has a slow-release performance, and the slow-released lysozyme encounters spirulina to destroy the cell wall of spirulina, thereby achieving the purpose of slow-releasing phycocyanin; the slow-release performance of phycocyanin in the hydrogel dressing obtained in step d) is tested in physiological saline, and the hydrogel dressing has a longer ability to release phycocyanin than the dressing directly added with phycocyanin and spirulina, and can continuously release phycocyanin for 48 hours;

[0042] f) The antibacterial rate of the hydrogel dressing to Staphylococcus aureus reaches 90%;

[0043] g) The clearance rate of the phycocyanin released by the hydrogel dressing to 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt reaches 90% after 7 hours, and the hemostatic time for the liver lobe resection bleeding of rats is 2 minutes.

[0044] Example 4.

[0045] a) Prepare a lysozyme aqueous solution with a mass percentage concentration of 1.0% by using deionized water, disperse kaolin into the lysozyme aqueous solution under stirring to obtain a kaolin mixed aqueous solution with a mass percentage concentration of 2.0% of kaolin, improve the dispersibility of kaolin under low power ultrasonic, adjust the pH of the mixed aqueous solution to 4.2, and adsorb lysozyme to kaolin under stirring, then filter the adsorbed lysozyme on kaolin, and then put it into a mixture aqueous solution of 2.0% mass percentage concentration of γ-methacryloxypropyltrimethoxysilane and γ-aminopropyltriethoxysilane, ultrasonic dispersion under low power, stand for 24 hours, filter the precipitate, and wash away the unreacted substances with deionized water to obtain adsorbed lysozyme kaolin wrapped with a layer of polysiloxane, which is freeze-dried for standby;

[0046] b) dispersing the freeze-dried lysosome-adsorbed kaolin coated with polysiloxane obtained in step a) in deionized water under stirring to obtain a casting solution, then adding sodium alginate with a mass percentage concentration of 2.8%, and continuing to stir to fully dissolve the sodium alginate, and then standing to remove bubbles to obtain a casting solution ready for use;

[0047] c) preparing a mixed aqueous solution of lysosome with a mass percentage concentration of 2.0% and calcium chloride with a mass percentage concentration of 5.0% as a crosslinking agent aqueous solution, and pouring the crosslinking agent aqueous solution into a spray bottle for standby;

[0048] d) pouring the casting solution obtained in step b) onto a clean and flat glass plate, using a doctor blade to scrape a liquid film with uniform thickness, and then immediately spraying the crosslinking agent aqueous solution in the spray bottle of step c) onto the surface of the liquid film, so that the calcium ions in the soluble calcium salt crosslink with the sodium alginate to form a calcium alginate hydrogel film, and after the calcium alginate hydrogel film is initially formed, it is removed immediately to spray the crosslinking agent aqueous solution onto the other side of the crosslinked film, and after sufficient crosslinking, a hydrogel dressing capable of releasing phycocyanin is obtained, which is used to cut into the required shape and size with a mold for standby;

[0049] e) Since the lysozyme bacteria are adsorbed and embedded with polysiloxane, and the lysozyme is easily adsorbed by alginate, the lysozyme inside the hydrogel dressing has a slow-release performance, and the slow-released lysozyme destroys the cell wall of the spirulina when it meets the spirulina, thereby achieving the purpose of slow-releasing phycocyanin; the slow-release performance of phycocyanin in the hydrogel dressing obtained in step d) is tested in physiological saline, and the hydrogel dressing of the present application has a longer release of phycocyanin than the dressing directly added with phycocyanin and spirulina, and can continuously release phycocyanin for 48 hours;

[0050] f) The hydrogel dressing of the present application has a bacteriostatic rate of 99.5% on Staphylococcus aureus;

[0051] g) The phycocyanin released by the hydrogel dressing of the present application has a clearance rate of 2,2'-azobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt of 94% after 7 hours, and a hemostatic time of 1.5 minutes for a liver lobe resection hemorrhage in a rat.

[0052] Example 5.

[0053] a) prepare a 1.0% by mass lysozyme aqueous solution with deionized water, disperse kaolin into the lysozyme aqueous solution under stirring to obtain a 3% by mass kaolin mixed aqueous solution, improve the dispersibility of kaolin by ultrasonic treatment at low power, adjust the pH of the mixed aqueous solution to 4.8, and fully adsorb lysozyme onto kaolin under stirring, then separate the lysozyme-adsorbed kaolin by filtration with filter paper, and then put the lysozyme-adsorbed kaolin into a 1.5% by mass mixed aqueous solution of γ-methacryloxypropyltrimethoxysilane and γ-aminopropyltriethoxysilane, uniformly disperse the components by ultrasonic treatment at low power, and stand for reaction for 32 hours, then take the precipitate by filtration, and wash away unreacted substances with deionized water to obtain lysozyme-adsorbed kaolin wrapped with a layer of polysiloxane, and freeze-dry the lysozyme-adsorbed kaolin for later use;

[0054] b) disperse 4% by mass lysozyme-adsorbed kaolin wrapped with a layer of polysiloxane obtained in step a) and 1% by mass spirulina into deionized water under stirring, then add 2% by mass sodium alginate, continue to stir to fully dissolve the sodium alginate, and obtain a casting solution after standing and defoaming for later use;

[0055] c) prepare a 0.5% by mass mixed aqueous solution of lysozyme and 2% by mass calcium chloride as a crosslinking agent aqueous solution, and pour the crosslinking agent aqueous solution into a spray bottle for later use;

[0056] d) pour the casting solution obtained in step b) onto a clean and flat glass plate, scrape the liquid film into a uniform thickness with a film scraper, then immediately spray the crosslinking agent aqueous solution in the spray bottle in step c) onto the surface of the liquid film, crosslink calcium ions in the soluble calcium salt with sodium alginate to form a calcium alginate hydrogel film, take the preliminarily shaped calcium alginate hydrogel film immediately, spray the crosslinking agent aqueous solution onto the other side of the film, and fully crosslink to obtain an antibacterial and antioxidant hemostatic hydrogel dressing capable of releasing phycocyanin; and use a mold to carve the hydrogel dressing into a desired shape and size for later use;

[0057] e) since the lysozyme bacteria are adsorbed and embedded with polysiloxane, and lysozyme is easily adsorbed by alginate, the lysozyme in the hydrogel dressing has a slow-release performance, the slow-released lysozyme destroys the cell wall of spirulina when meeting the spirulina, so as to achieve the purpose of slow-releasing phycocyanin; test the slow-release performance of phycocyanin in the hydrogel dressing obtained in step d) in physiological saline, and the hydrogel dressing of the application has a longer ability to release phycocyanin than the dressing directly added with phycocyanin and spirulina, and can continuously release phycocyanin for 52 hours;

[0058] f) the antibacterial rate of the hydrogel dressing of the application on Staphylococcus aureus reaches 92%;

[0059] The phycocyanin released by the hydrogel dressing of the present invention achieved an 86% clearance rate of 2,2'-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt after 7 hours, and the hemostasis time for bleeding in rat middle lobe resection was 2.5 minutes.

Claims

1. A method for preparing an antibacterial and antioxidant hemostatic hydrogel dressing with sustained release of phycocyanin, characterized by The method comprises the following steps: a) preparing a lysozyme aqueous solution with a mass percentage of 0.1-5%, dispersing kaolin into the lysozyme aqueous solution under stirring to obtain a kaolin mixed aqueous solution with a mass percentage of 0.5-5%, and adjusting the pH of the mixed aqueous solution to 4-5 under ultrasonic treatment at low power to adsorb lysozyme onto kaolin under stirring, separating the kaolin with adsorbed lysozyme by filtration with filter paper, and then placing the kaolin with adsorbed lysozyme into a mixed aqueous solution of γ-methacryloxypropyltrimethoxysilane and γ-aminopropyltriethoxysilane with a mass percentage of 0.2-2% respectively, uniformly dispersing the components under ultrasonic treatment at low power, and standing for 12-36 hours for reaction, filtering the precipitate, and washing the unreacted substances with deionized water to obtain kaolin with adsorbed lysozyme wrapped with a layer of polysiloxane, which is freeze-dried for later use; b) dispersing the freeze-dried kaolin with adsorbed lysozyme wrapped with a layer of polysiloxane obtained in step a) with a mass percentage of 0.2-5% into deionized water under stirring, and then adding sodium alginate with a mass percentage of 1.0-3% to fully dissolve the sodium alginate under continuous stirring, and standing for defoaming to obtain a casting solution for later use; c) preparing a mixed aqueous solution of lysozyme with a mass percentage of 0.1-5% and soluble calcium salt with a mass percentage of 1.0-20% as a crosslinking agent aqueous solution, and pouring the crosslinking agent aqueous solution into a spray bottle for later use; d) pouring the casting solution obtained in step b) onto a clean and flat glass plate, scraping the liquid film with a film scraper to have a uniform thickness, and then immediately spraying the crosslinking agent aqueous solution in the spray bottle in step c) onto the surface of the liquid film, crosslinking calcium ions in the soluble calcium salt with sodium alginate to form a calcium alginate hydrogel film, taking the preliminarily shaped calcium alginate hydrogel film immediately, spraying the crosslinking agent aqueous solution onto the other side of the film for crosslinking, and obtaining an antibacterial and antioxidant hemostatic hydrogel dressing capable of releasing phycocyanin after sufficient crosslinking; and cutting the hydrogel dressing into a desired shape and size with a mold for later use; e) the lysozyme is adsorbed and embedded by polysiloxane, and the lysozyme is easily adsorbed by alginate, so that the lysozyme in the hydrogel dressing has a slow-release performance, the slow-released lysozyme destroys the cell wall of spirulina to achieve the purpose of slow release of phycocyanin; the slow-release performance of phycocyanin in the hydrogel dressing obtained in step d) is tested in physiological saline, and the hydrogel dressing has a longer ability to release phycocyanin than the dressing directly added with phycocyanin and spirulina, and can continuously release phycocyanin for 24-60 hours; f) the antibacterial rate of the hydrogel dressing to Staphylococcus aureus reaches 65-100%; g) the clearance rate of the released phycocyanin in the hydrogel dressing to 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt reaches 60-96% after 7 hours. The soluble calcium salt is any one or a mixture of two or more of calcium chloride, calcium phosphate, and calcium nitrate.

2. A process for the preparation of an alginate-releasing antibacterial, antioxidant, hemostatic hydrogel dressing according to claim 1, characterized in that ​ 3. The method for preparing a sustained-release phycocyanin antibacterial, antioxidant, and hemostatic hydrogel dressing as described in claim 1, characterized in that... The antibacterial and antioxidant hemostatic hydrogel dressing has a hemostatic time of 1-5 minutes for liver lobe resection hemorrhage of rats.