CN111588902A

CN111588902A - Large-area wound first-aid dressing and preparation method thereof

Info

Publication number: CN111588902A
Application number: CN202010446119.7A
Authority: CN
Inventors: 张进; 林哲
Original assignee: Ruisi Fujian Engineering Research Center Co ltd
Current assignee: Ruisi Fujian Engineering Research Center Co ltd
Priority date: 2020-05-25
Filing date: 2020-05-25
Publication date: 2020-08-28

Abstract

The invention belongs to the field of biomaterial preparation and biomedical application, and particularly relates to a large-area wound first-aid dressing and a preparation method thereof. A wound first aid dressing comprising the following components: modified chitosan, biomass protein fiber and polyphenol compounds. The modified chitosan/biomass protein fiber/polyphenol composite material is prepared by uniformly mixing modified chitosan, biomass protein fiber and polyphenol compound at room temperature. The dressing prepared by the invention has the advantages of high hemostasis efficiency, high bonding speed, wide application range, strong targeted antibacterial property, safety, no toxicity, good biocompatibility and convenience in operation in a wet environment. The raw materials have wide sources, can be used for hemostasis and healing repair of various wound surfaces, is expected to replace the traditional medical adhesive, and has wide clinical application prospect.

Description

Large-area wound first-aid dressing and preparation method thereof

Technical Field

The invention belongs to the field of biomaterial preparation and biomedical application, and particularly relates to a large-area wound first-aid dressing and a preparation method thereof.

Background

For many years, emergency treatment of sudden accidents in daily life, wound hemostasis during operation of doctors on patients and rescue and hemostasis measures of injured soldiers in war are mostly realized by physical compression, such as using bandages, gauze, rubber strips and the like, but the effect is limited. In clinical medicine, the requirements of bandaging and hemostasis of a full chain of a treatment ladder are met, particularly for acute severe wound bleeding in an emergency, due to the moisture state diversity and the dynamic complexity of an emergent bleeding environment, the existing commercially available hemostatic products cannot achieve the effects of quickly and effectively stopping bleeding and resisting bacteria, and hemorrhagic shock is very easy to occur to endanger life. The hemostatic materials in the market at present mainly comprise collagen, oxidized regenerated cellulose, alginates, zeolite, chitosan and the like.

At present, the commercial medical adhesive generally seals wounds by spraying, pressing and other modes to achieve the aim of hemostasis, does not have the function of endogenous hemostasis, and has a plurality of defects. Zeolite has limited application range and is not easily applied to the wound-down area, and the hemostatic effect is reduced by the blood flow washing away the particles. More seriously, when the zeolite absorbs moisture, a large amount of heat is emitted, the highest temperature can reach 44 ℃, and the injured tissues are seriously thermally damaged and burnt; the fibrin adhesive has the risks of high price, long preparation time, virus infection carrying and the like, and is very easy to cause serious tissue adhesion, difficult to remove, great in side effect and the like when used in an abdominal cavity; the alpha-cyanoacrylate tissue adhesive has high requirements on the surface of an adhesive tissue, the wound surface needs to be kept dry and clean and blood/digestive juice does not exist, so that great constraints exist in the practical application of the alpha-cyanoacrylate tissue adhesive, such as complex operation, long time consumption and the like. Meanwhile, the commercially available hemostatic materials release toxic substances such as formaldehyde and cyanogen when degraded, and cannot be used together with antibiotic ointment or vaseline gauze. Otherwise, it will cause inflammatory reaction in body tissues and even delay wound healing. Non-woven gauze impregnated with kaolin, which is not biodegradable, has problems in that it must be removed from the wound before surgery, etc.

Patent CN108578759A discloses a chitosan derivative colloidal fluid consisting of sodium carboxyaminodextran polysaccharide, sodium carboxymethylcellulose, niacinamide, glycerin, mannitol, microcrystalline cellulose, antioxidant, gelatin, glutaraldehyde, emulsifier and water for injection; the time required for complete degradation of the protective film formed on the surface of the wound is long, the recovery and effective cleaning rate of the prior art cannot meet the use requirement, the wound infection is caused, and the problems in actual production cannot be solved.

Patent CN109966544A discloses a composite sponge prepared by using alkyl chitosan as a matrix and compounding graphene oxide with alkyl chitosan, wherein the obtained composite sponge has excellent hemostatic property and blood absorption ability. Wherein, the hemostatic material contains lauraldehyde, benzaldehyde and other substances, has certain stimulation effect on eyes and respiratory mucosa, has no bacteriostatic property, and can cause inflammatory reaction on the surface of a wound or in vivo.

Patent CN201610006406.X discloses a biological nutrition dressing for treating skin wound, and its preparation method, wherein the adjuvants comprise nanometer silk fibroin, mineral, vitamin, oleum Menthae Dementholatum, chitosan, and polyvinyl alcohol. Experiments show that: wherein, the chitosan content is less than 2%, and the dressing has poor bacteriostatic effect and is not beneficial to the growth of wounds.

Patent CN106334209A discloses a polydopamine modified chitosan hemostatic dressing, which contains chitosan, polydopamine and at least one calcium salt, the tissue viscosity of the material is too high, secondary injury can be caused to the wound when the dressing is changed, and the polydopamine solution is obtained by adjusting the pH of the hydrochloride of dopamine to be alkaline, and the alkaline solution can be irritant to the skin and can influence the healing of the bleeding part.

Disclosure of Invention

The invention solves the technical problem of providing a dressing which is prepared by taking modified chitosan, natural biomass protein fiber and natural polyphenol compound as main raw materials, has a bacteriostatic effect and can be applied to large-area wound first aid;

correspondingly, the invention also provides a preparation method of the large-area wound emergency dressing.

In order to achieve the purpose, the invention adopts the following technical scheme:

a large area wound first aid dressing comprising the following components: modified chitosan, biomass protein fiber and polyphenol compounds.

Further, the mass ratio of the modified chitosan to the biomass protein fiber to the polyphenol compound is 1-1000: 1-1000.

Further, the biological protein fiber is one or the combination of more than two of silk fibroin, keratin, collagen and marine mussel protein.

Further, the modified chitosan is one or the combination of more than two of dodecyl chitosan, quaternary ammonium salt chitosan and carboxyl chitosan.

Further, the polyphenol compound is one or the combination of more than two of phenolic hydroxyl compound, tannin compound, phenolic acid compound and anthocyanin; preferably one or a combination of more than two of flavone, tannic acid, gallic acid, dopamine and catechol.

Further, the paint also comprises one or more than two of the following components: silver-containing compounds, antimicrobial peptides, aspirin, curcumin, celecoxib, or analgesics.

Further, the modified chitosan fiber and polyphenol compound mixed material comprises a first ingredient and a second ingredient, wherein the first ingredient is modified chitosan, and the second ingredient is a mixed material of a biological protein fiber and a polyphenol compound.

The invention also provides a preparation method of the large-area wound emergency dressing, which comprises the following steps: adding the biomass protein fiber solution and the polyphenol compound solution into the modified chitosan solution, and fully and uniformly mixing to obtain the dressing.

Further, the mixing speed is 0.01-1L/min.

Further, the modified chitosan solution is mainly prepared by dissolving modified chitosan in aluminum chloride hexahydrate;

or/and

the preparation method of the biomass protein fiber solution comprises the following steps: processing silk with Na₂CO₃Degumming and drying, dissolving by a lithium bromide solution, calcium chloride, absolute ethyl alcohol or a water ternary solution, dialyzing and filtering to obtain a biomass protein fiber solution.

(III) advantageous effects

Compared with the prior art, the invention has the advantages that,

1. the invention combines the natural biomass protein fiber net structure and the cation effect of the modified chitosan, and can quickly capture and activate red blood cells and platelets; in addition, due to self-assembly between polyphenol compounds and natural biomass protein fiber molecules and chelation between the modified chitosan and the polyphenol polymers, a double network is formed, so that high hemostasis efficiency and rapid adhesion in a wet environment are realized.

2. The modified chitosan, the biomass protein fiber and the polyphenol compound are derived from organisms, so that the modified chitosan, the biomass protein fiber and the polyphenol compound have good biocompatibility and antibacterial property.

3. The dressing prepared by the invention has wide application, can be used as a hemostatic adhesive material for tissues such as organs, joints, skin, nerves, blood vessels, muscles, mucous membranes, bones, eyes and the like, can realize rapid adhesion in a wet environment (including water, PBS buffer solution and blood environment), and has the hemostatic time within 15 s.

4. The auxiliary material obtained by the invention is verified by in vivo and in vitro experiments to have good biocompatibility of tissues and blood, and combines the functions of procoagulant, strong sealing and antibiosis and antiphlogosis. After hemostasis is carried out in time, the artificial vascular wall is formed through the powerful physical sealing effect of the material on the tissue, and the bacteria are positioned and disintegrated while the main wound is protected and the function of the vascular machine is recovered. The new generation of composite hemostatic and antibacterial material capable of replacing suture solves the problems of wound tearing bleeding, infection necrosis and other complications caused by improper postoperative care from the technical aspect, and has positive effects on the formation of new tissues and the healing of the tissues.

Drawings

FIG. 1 shows the result of cytotoxicity test of adjuvant obtained in example 7 of the present invention;

FIG. 2 is a graph showing the results of an in vitro hemostatic test on the experimental group of dressings obtained in example 7 of the present invention;

FIG. 3 shows the results of in vitro hemostasis tests on the dressing control group obtained in example 7 of the present invention.

Detailed Description

For a further understanding of the invention, reference will now be made to the preferred embodiments of the invention by way of example, and it is to be understood that the description is intended to further illustrate features and advantages of the invention, and not to limit the scope of the claims.

[ first embodiment ] to provide a toner

The invention provides a large-area wound first-aid dressing which comprises the following components: modified chitosan, biomass protein fiber and polyphenol compounds.

In the invention, modified chitosan with good biocompatibility, biomass protein fiber and polyphenol compound are selected as main raw materials, and a new hemostatic material which can realize quick adhesion and bacteriostasis in a wet environment (including water, PBS buffer solution and blood environment) is obtained through self-assembly and compounding.

Among them, modified chitosan, such as: the dodecyl chitosan is a natural mucopolysaccharide, has good biocompatibility, cationic effect and the characteristic of presenting a stereo molecular net structure in blood, and can quickly capture and activate red blood cells and platelets to form blood clots. Further, modified chitosan, such as: the chain segment of the dodecyl chitosan molecule can be positioned to a phospholipid bilayer of a bacterial cell membrane to target bacteria, and the anti-inflammatory and bactericidal effects can be further improved by polyphenol compounds or addition of other bactericidal substances, so that the bacteria are killed, and the bactericidal function is realized.

Wherein, the polyphenol compound and natural biomass protein fiber (such as silk fibroin) are self-assembled, and modified chitosan and polyphenol polymer are chelated to form a double network, thereby realizing the effect of rapid hemostasis.

On the one hand, the invention promotes blood coagulation based on the function of the modified chitosan for gathering red blood cells, and the chain segment of the modified chitosan can be positioned to a phospholipid bilayer of a bacterial cell membrane to target bacteria, so that the bacteria are killed by a bactericidal substance. On the other hand, the fast sealing of injured tissues can be realized based on the super strong wet tissue bonding capability of the functionalized bio-protein fibers, the obtained adhesive has the advantages of high hemostasis efficiency in a wet environment, high bonding speed, wide application range, strong targeted antibacterial property, safety, no toxicity, good biocompatibility and convenience in operation, and the adhesive is suitable for wide application and can overcome the aortic hemorrhage.

Experiments show that:

1. the hemostatic time of the auxiliary material in the invention for large-area wounds is within 15 s;

2. the auxiliary materials in the invention have the hemostatic time under the same hemostatic condition which is obviously shorter than that of the hemostatic material containing the modified chitosan and the biomass protein fiber but not containing the polyphenol compound;

3. the auxiliary materials in the invention have the hemostatic time under the same hemostatic condition which is obviously shorter than that of the hemostatic material containing polyphenol compounds and modified chitosan but not containing biomass protein fibers;

4. the auxiliary materials in the invention have the hemostatic time under the same hemostatic condition which is obviously shorter than that of the hemostatic material containing polyphenol compounds and biomass protein fibers but not containing modified chitosan.

The components in the invention are preferably in the following weight ratio, and the mass ratio of the modified chitosan, the biomass protein fiber and the polyphenol compound is 1-1000: 1-1000;

preferably: the mass ratio of the modified chitosan to the biomass protein fiber to the polyphenol compound is 1-15: 1-15;

preferably: the mass ratio of the modified chitosan to the biomass protein fiber to the polyphenol compound is 3-6: 1-10: 1-15;

more preferably: the mass ratio of the modified chitosan to the biomass protein fiber to the polyphenol compound is 15: 1: 15.

The biological protein fiber in the invention can be one or the combination of more than two of silk fibroin, keratin, collagen and marine mussel protein; preferably silk fibroin.

The modified chitosan in the invention can be one or the combination of more than two of dodecyl chitosan, quaternary ammonium salt chitosan and carboxyl chitosan; preferably, it is a dodecyl chitosan.

The polyphenol compound in the invention is one or the combination of more than two of phenolic hydroxyl compound, tannin compound, phenolic acid compound and anthocyanin; preferably one or a combination of more than two of flavone, tannic acid, gallic acid, dopamine and catechol.

The marine source dodecyl chitosan, the natural high polymer silk fibroin protein, the natural component polyphenol compound contained in the plant and the marine brown algae extract alginate gel are used as main raw materials, and the novel hemostatic material which can be quickly bonded and can inhibit bacteria under the wet environment (including water, PBS buffer solution and blood environment) is obtained through self-assembly and compounding.

The dodecyl chitosan is a natural mucopolysaccharide, has good biocompatibility, cationic effect and the characteristic of presenting a stereo molecular net structure in blood, and can quickly capture and activate red blood cells and platelets to form blood clots. In addition, the chain segment of the dodecyl chitosan molecule can be positioned to a phospholipid bilayer of a bacterial cell membrane to target bacteria, and further, the bacteria are killed through silver ions, so that the sterilization function is realized.

In order to improve the sterilization and pain-relieving functions of the large-area wound emergency dressing, the dressing also comprises one or more than two of the following components: silver-containing compounds, antimicrobial peptides, aspirin, curcumin, celecoxib, or analgesics.

Preferably a silver-containing compound, more preferably a silver-ion-containing compound, more preferably silver alginate; in the process of promoting blood coagulation of the dressing, the chain segment of the modified chitosan molecule can be positioned to a phospholipid bilayer of a bacterial cell membrane to target bacteria, the bacteria can be killed by silver ions in the silver alginate, and the alginate radical ions participate in the formation of a double network, so that the hemostatic function is improved.

For the convenience of the dressing, the large-area wound first-aid dressing comprises a first ingredient and a second ingredient, wherein the first ingredient is modified chitosan, and the second ingredient is a mixture of biological protein fibers and polyphenol compounds. When in use, the first ingredient and the second ingredient are mixed to prepare the dressing to realize the hemostasis function in a wet environment. Wherein the first ingredient and the second ingredient must be stored separately before use.

[ second embodiment ] to provide a medicine for treating diabetes

The embodiment provides a preparation method of a large-area wound first-aid dressing, which comprises the following steps: adding the biomass protein fiber solution and the polyphenol compound solution into the modified chitosan solution, and fully and uniformly mixing to obtain the dressing.

Preferably, the biomass protein fiber solution and the polyphenol compound solution are sequentially added into the modified chitosan solution, and the silver alginate can be further added after the polyphenol compound solution is added; by adopting the adding method, the auxiliary materials have better and more stable network structure.

Wherein the mixing speed is 0.01-1L/min, and stirring is carried out during mixing.

Wherein the concentration of the modified chitosan in the modified chitosan solution is 0.001-10 g/mL; the preferable modified chitosan solution is dodecyl chitosan solution, and the preparation method comprises the following steps:

dissolving chitosan in an acetic acid solution, adding lauraldehyde and a phase transfer catalyst sodium dodecyl sulfate to react to generate Schiff base, adjusting the Schiff base to 4-5 by using a NaOH solution, and then adding sodium dodecyl sulfate

NaBH is dripped₄Heating and stirring the solution for reduction reaction, and then adding lauraldehyde and NaBH again₄Performing secondary reduction reaction, purifying by using the solubility difference of chitosan with different alkylation substitution degrees in a chloroform solvent, specifically, washing a product obtained by secondary reduction precipitation to be neutral by using distilled water, removing excessive lauraldehyde and a reduction product lauryl alcohol by using acetone, purifying by using a Soxhlet extractor, dissolving in chloroform, adding acetone to obtain a precipitate, and filtering to obtain dodecyl chitosan; and dissolving the obtained dodecyl chitosan into an aluminum chloride hexahydrate solution, and stirring at the constant temperature of 10-60 ℃ for 0.5-6 hours to obtain a dodecyl chitosan solution.

Wherein the concentration of the biomass protein fibers in the biomass protein fiber solution is 0.001-10 g/mL;

the biomass protein fiber solution is preferably silk fibroin solution, and the preparation method comprises the following steps: heating deionized water to boil, adding Na₂CO₃Degumming silkC, processing; after drying, dissolving the mixture by using a lithium bromide solution, calcium chloride, absolute ethyl alcohol or a ternary water solution, and then dialyzing the mixture by using deionized water; centrifuging and filtering the dialyzed solution to obtain a silk fibroin solution;

wherein the concentration of the polyphenol compound in the polyphenol compound solution is 0.001-10 g/mL;

wherein the silver alginate is mainly prepared by mixing a soluble silver salt solution with the concentration of 0.001-10 g/mL and an alginate solution with the concentration of 0.001-10 g/mL.

In order to better understand the above technical solutions, exemplary embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. While exemplary embodiments of the invention are shown in the drawings, it should be understood that the invention can be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

Example 1

The large-area wound first-aid dressing is prepared from dodecyl chitosan, silk fibroin, keratin and tannic acid according to the mass ratio of 1000: 1: 1000.

Example 2

The large-area wound first-aid dressing is prepared from quaternary ammonium salt chitosan, silk fibroin, gallic acid and silver alginate according to the mass ratio of 1: 1000: 1.

Example 3

The large-area wound first-aid dressing is prepared from carboxylated chitosan, marine mussel protein, dopamine and curcumin in a mass ratio of 1: 15: 1.

Example 4

The large-area wound first-aid dressing is prepared from dodecyl chitosan, quaternary ammonium salt chitosan silk fibroin, keratin, gallic acid and silver alginate according to the mass ratio of 2: 3: 2: 1.

Example 5

The large-area wound first-aid dressing is prepared from carboxylated chitosan, marine mussel protein, polydopamine and silver alginate according to the mass ratio of 5: 10: 1: 15.

Example 6

The large-area wound first-aid dressing is prepared from dodecyl chitosan, silk fibroin, tannic acid and silver alginate according to the mass ratio of 10: 15: 1: 5: 10.

Example 7

Step S1, preparing a dodecyl chitosan solution: 4.0g of chitosan was weighed into 120mL of a 5% acetic acid solution, and dissolved by stirring. Adding 12.0g of lauraldehyde and a phase transfer catalyst sodium dodecyl sulfate, heating, stirring and reacting for 2-9 hours, adjusting the pH to 4-5 by using a NaOH solution, and slowly dropwise adding 10% NaBH with the mass concentration of 1.5 times of excessive amount₄The solution is subjected to reduction reaction, heating and stirring are continued for 2 to 3 hours, and lauraldehyde and NaBH are added again₄Performing secondary reduction reaction to obtain precipitate, washing with distilled water to neutrality, removing excessive lauraldehyde and reduction product lauryl alcohol with acetone, purifying with Soxhlet extractor for 48 hr, dissolving in chloroform to remove insoluble substance, adding acetone to obtain precipitate, and filtering to obtain dodecyl chitosan. Dissolving 15g of aluminum chloride hexahydrate in water, adding 15g of newly prepared dodecyl chitosan, heating to 40 ℃, and stirring for 6 hours to obtain a dodecyl chitosan solution with the concentration of 40%.

Step S2, preparing silk fibroin solution: heating 5L of deionized water to boil, adding 8g of sodium carbonate, fully dissolving, adding 7g of raw silkworm silk, degumming at 100 ℃, fully washing with deionized water, and drying in an oven at 80 ℃ to obtain degummed silk fibroin fiber; and (3) putting the degummed silk fibroin fiber into a LiBr solution, dissolving for 5h at 70 ℃, dialyzing for 72h in deionized water in a dialysis bag, filtering, centrifuging, and repeating twice to obtain a silk fibroin solution with the mass fraction of 10%.

Step S3, catechol solution preparation: dissolving 15g of catechol in deionized water, and stirring for 0.5h to obtain a catechol solution with the mass concentration of 50%.

Step S4, preparing silver alginate solid: mixing 15g of silver nitrate and 15g of sodium alginate solution at room temperature, and drying the obtained water-insoluble product to obtain the silver alginate solid.

Step S5, preparing a large-area wound first-aid dressing: the obtained twelve-position alkylated chitosan solution, the silk fibroin solution, the catechol solution and the silver alginate solid are prepared according to the mass ratio of 1: 5: 1. Sequentially adding the silk fibroin solution, the catechol solution and the silver alginate solid into the dodecyl alkylated chitosan solution, and fully and uniformly mixing to obtain the large-area wound first-aid dressing with the antibacterial effect and capable of realizing rapid hemostasis of the aorta.

Example 8

The other points are the same as example 7 except that, in step S5, the obtained dodecagonal alkylated chitosan solution, silk fibroin solution, catechol solution and silver alginate solid were prepared in a mass ratio of 1: 15: 1.

Example 9

The other points are the same as example 7 except that, in step S5, the obtained dodecagonal alkylated chitosan solution, silk fibroin solution, catechol solution and silver alginate solid were prepared in a mass ratio of 1: 5: 1.

Example 10

Example 11

The other points are the same as example 7 except that, in step S5, the obtained dodecagonal alkylated chitosan solution, silk fibroin solution, catechol solution and silver alginate solid were prepared in a mass ratio of 1: 5.

Example 12

The other points are the same as example 7 except that, in step S5, the obtained dodecagonal alkylated chitosan solution, silk fibroin solution, catechol solution and silver alginate solid were prepared in a mass ratio of 1: 15.

Example 13

The other points are the same as example 7 except that, in step S5, the obtained dodecagonal alkylated chitosan solution, silk fibroin solution, catechol solution and silver alginate solid were prepared in a mass ratio of 5: 1.

Example 14

The other points are the same as example 7 except that, in step S5, the obtained dodecagonal alkylated chitosan solution, silk fibroin solution, catechol solution and silver alginate solid were prepared in a mass ratio of 15: 1.

Comparative example 1

The procedure of example 7 was otherwise the same, except that in step S5, the catechol solution and the silver alginate solid were sequentially added to the twelve-position alkylated chitosan solution to obtain an adjuvant.

Comparative example 2

The other point is that, like example 7, in step S5, the silk fibroin solution and the silver alginate solid are sequentially added to the twelve-position alkylated chitosan solution to obtain an adjuvant.

Comparative example 3

The other point is that the silk fibroin solution, the catechol solution, and the silver alginate solid are mixed to prepare an auxiliary material in step S5, which is the same as example 7.

Comparative example 4

The procedure of example 7 was otherwise identical except that, in step S5, silver alginate solid was added to the twelve-position alkylated chitosan solution to obtain an adjuvant.

Comparative example 5

The other point is that, like example 7, in step S5, the silk fibroin solution, the catechol solution, and the silver alginate solid were sequentially added to the twelve-position alkylated chitosan solution to obtain an adjuvant.

Comparative example 6

Comparative example 7

The resulting dressing was prepared according to patent cn201610006406.

The following experiments prove that the specific embodiment of the invention has the following effects:

experiment one

The dressing obtained in example 7 was subjected to a cytotoxicity test, the test results of which are shown in fig. 1, while the hemostatic effect of the dressing obtained in example 7 was optimized through an in vitro coagulation test. As can be seen from FIG. 1, the dressing obtained by the present invention has good biocompatibility.

Experiment two

Performance tests were performed on the adjuvants prepared in examples 7 to 14 of the present invention, and the results shown in Table 1 were obtained.

TABLE 1 Performance testing of adjuvants prepared in inventive examples 7-14

As can be seen from Table 1, the composite material dressing prepared in the embodiments 7 to 14 of the invention has the advantages of antibacterial effect, capability of realizing rapid hemostasis of the aorta, good mechanical property, high hemostasis efficiency and excellent comprehensive performance, and is suitable for various clinical conditions. The dressing rapidly captures and activates red blood cells and platelets and accelerates the release of blood coagulation factors by the cationic effect of chitosan and the characteristic that natural alginate fibers are in a stereo molecular net structure in blood; by means of self-assembly between polyphenol compounds and silk fibroin biological macromolecules and chelation between natural mucopolysaccharide and polyphenol polymers, a double network is formed inside the silk fibroin/chitosan/tannin adhesive, so that the effect of quickly stopping bleeding is realized. The main components of the material of the invention, namely chitosan, fibroin, tannic acid and silver alginate, are natural extracts, and the material has good biocompatibility of tissues and blood through in vivo and in vitro experiments. Meanwhile, the efficacy and the biological safety of the novel hemostatic material in the aspects of hemostasis, wound closure, bacteriostasis, healing promotion, scar hyperplasia inhibition and the like are researched through the construction of various animal wound models, and the military and clinical application values of the novel hemostatic material are systematically evaluated.

Experiment three: the adjuvants prepared in examples 7 to 14 of the present invention were subjected to medical performance tests to obtain the results shown in table 2.

TABLE 2 Performance testing of adjuvants prepared in inventive examples 7-14

As can be seen from Table 2, the adjuvants prepared by the present invention have good medical properties.

Experiment four, in vitro hemostasis test

The large-area wound emergency dressing prepared in example 7 was subjected to a mouse femoral artery puncture hemostasis test, and the test results are shown in fig. 2, and a blank test was used as a control, and the results are shown in fig. 3. As shown in FIG. 2, the femoral vein puncture hemostasis test was performed on mice. After the first-aid dressing is added, blood does not flow out, and the dressing is proved to have the rapid hemostasis capability, mainly because of the cationic effect of the dodecyl chitosan and the characteristic that natural alginic acid fiber is in a stereo molecular net structure in blood, red blood cells and platelets are rapidly captured and activated, and the release of blood coagulation factors is accelerated; by means of self-assembly between polyphenol compounds and silk fibroin biomacromolecules and chelation between the dodecyl chitosan and the polyphenol polymers, a double network is formed in the adhesive. Meanwhile, after silver ions released by the silver alginate are effectively combined with the bacterial targeting function of the dodecyl chitosan, the wound healing function of the dressing is further improved based on the sterilization and anti-inflammation effects. Therefore, the multifunctional novel large-area wound first-aid dressing has very considerable clinical prospect.

Experiment five, comparative experiment

The comparative experiment shows that: the hemostatic effect and the bactericidal effect of the large-area wound emergency dressing of the embodiment 7 of the invention are both obviously superior to those of the comparative examples 1-7,

in the hemostasis experiment, experimental animals with the same growth condition are respectively set as an example 7 group, a comparative example 1 group and a comparative example 2 group, the three groups of experimental animals are respectively caused to cause the same wound at the same position, after the wound position is respectively correspondingly processed by using the auxiliary materials of the example 7 and the comparative examples 1 and 2,

respectively measuring the covering areas of the dressings and the blood flowing out after hemostasis, wherein the larger the covering area is, the poorer the hemostasis effect is;

the total number of bacteria at the wound site was determined separately: the total number of bacterial colonies grown around the hydrogel was detected by placing 1mL of hydrogel in a bacterial incubation environment at 37 ℃.

The above results are shown in table 1, and the data in table 3 are all average values within the error tolerance range of a plurality of effective experimental data:

TABLE 3 evaluation of hemostatic and antibacterial Properties of Large-area wound Emergency dressings

	Area of bleeding (cm)²)	Total colony count CFU/mL
			Comparative example 1	2.45±0.80	4500±550
Comparative example 2	1.20±0.45	2700±420
			Example 7	0.18±0.05	350±50

As can be seen from the data in Table 3, the bleeding area and colony count of the adjuvant of example 7 are different by an order of magnitude from those of comparative examples 1 and 2, indicating that: the silk fibroin solution and the catechol solution have great synergistic effect on the hemostasis and antibiosis of the auxiliary material, and have unexpected technical effect.

Compared with the auxiliary materials of comparative examples 1 and 2, the auxiliary material of the embodiment 7 of the invention has better and obvious bacteriostatic effect and meets the application requirement of products lower than 500 CFU/mL.

In addition, experiments show that specific numerical values are not exhaustive, the bleeding area and the total number of bacterial colonies of the auxiliary material in the embodiment 7 are different by more than one order of magnitude compared with those of the comparative examples 3-7, the hemostatic effect and the antibacterial effect of the auxiliary material in the embodiment 7 are obviously better than those of the comparative examples 3-7, and the twelve-position alkylated chitosan, the silk fibroin, the catechol and the silver alginate have great synergistic effect on the hemostasis and the antibacterial aspects of the auxiliary material in the invention, and have unexpected technical effect.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A large-area wound first-aid dressing is characterized by comprising the following components: modified chitosan, biomass protein fiber and polyphenol compounds.

2. The large-area wound emergency dressing as claimed in claim 1, wherein the mass ratio of the modified chitosan, the biomass protein fiber and the polyphenol compound is 1-1000: 1-1000; preferably: the mass ratio of the modified chitosan to the biomass protein fiber to the polyphenol compound is 1-15: 1-15; more preferably: the mass ratio of the modified chitosan to the biomass protein fiber to the polyphenol compound is 3-6: 1-10: 1-15; further preferably: the mass ratio of the modified chitosan to the biomass protein fiber to the polyphenol compound is 15: 1: 15.

3. The large area wound first aid dressing of claim 1 or 2, wherein: the biological protein fiber is one or the combination of more than two of silk fibroin, keratin, collagen and marine mussel protein.

4. The large area wound first aid dressing of claim 1 or 2, wherein: the modified chitosan is one or the combination of more than two of dodecyl chitosan, quaternary ammonium salt chitosan and carboxyl chitosan.

5. The large area wound first aid dressing of claim 1 or 2, wherein: the polyphenol compound is one or the combination of more than two of phenolic hydroxyl compound, tannin compound, phenolic acid compound and anthocyanin; preferably one or a combination of more than two of flavone, tannic acid, gallic acid, dopamine and catechol.

6. The large area wound first aid dressing of claim 1, further comprising one or more of the following components: silver-containing compounds, antimicrobial peptides, aspirin, curcumin, celecoxib, or analgesics.

7. The large area wound first aid dressing of claim 1, comprising a first ingredient that is a modified chitosan and a second ingredient that is a mixture of a bioprotein fiber and a polyphenolic compound.

8. The method of preparing the large area wound first aid dressing of claim 1, comprising the steps of: and adding a biomass protein fiber solution and a polyphenol compound solution into the modified chitosan solution, and fully and uniformly mixing to obtain the dressing.

9. The method of preparing the large area wound first aid dressing of claim 8, wherein: the mixing speed is 0.01-1L/min.

10. The large area wound first aid dressing of claim 8, wherein: the modified chitosan solution is mainly prepared by dissolving modified chitosan in aluminum chloride hexahydrate;

or/and