A chitosan-based polyelectrolyte / surfactant composite sponge and a preparation method and application thereof

Abstract

The application discloses a chitosan-based polyelectrolyte / surfactant composite sponge and a preparation method and application thereof. Basic quaternary amine chitosan is used as a cationic polyelectrolyte, and L-hydroxyproline alkylated derivatives are used as an anionic surfactant. The polyelectrolyte / surfactant composite is formed through hydrogen bonding and electrostatic interaction. The chitosan-based sponge is prepared by a freezing method and is used as a support framework. The composite is chemically crosslinked with the sponge framework by using epichlorohydrin, and the chitosan-based polyelectrolyte / surfactant composite sponge with a porous structure is prepared. The application of the chitosan-based polyelectrolyte / surfactant composite sponge effectively expands the application range of the polyelectrolyte / surfactant composite. The chitosan-based polyelectrolyte / surfactant composite sponge has excellent properties such as light weight, degradability, antibacterial property, hemostasis and good biocompatibility. The preparation method is simple, the cost is low, and the chitosan-based polyelectrolyte / surfactant composite sponge is convenient for practical promotion.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of high polymer materials and biomedical materials, and particularly relates to a chitosan-based polyelectrolyte / surfactant composite sponge, a preparation method and application thereof. BACKGROUND

[0002] Skin is the largest tissue organ of the human body, which can maintain normal metabolism of the body and effectively resist the invasion of external environment and substances. However, when the skin is damaged due to surgery, accidental injury or chronic disease, etc., different degrees of skin trauma will be caused, and uncontrolled bleeding may lead to severe shock and multiple organ failure, resulting in a mortality rate of more than 30%. Therefore, it is very important to develop a functional wound dressing capable of quickly and efficiently stopping bleeding for treating emergency trauma. At present, commercially available hemostatic materials include bandages, gauze, gelatin sponge, zeolite powder and hydrogel, etc. However, these materials have relatively single function, generally lack antibacterial properties and are not easy to degrade. An ideal hemostatic dressing should have the advantages of low cost, high hemostatic efficiency, good clinical safety, antibacterial property, degradability and light quality, etc.

[0003] Chitosan is a renewable natural high molecular polysaccharide derived from crustaceans, which is widely used in medical materials and has antibacterial properties, good adsorption and chelation, biocompatibility, biological safety and biodegradability. As a derivative of chitosan, quaternized chitosan is a typical cationic polymer containing a large number of quaternary ammonium groups, which exhibits more excellent water solubility and antibacterial activity. In addition, studies have shown that both chitosan and quaternized chitosan are polyelectrolytes with dissociable groups, which can form complexes with polyelectrolytes or surfactants with opposite charges in solution, i.e. polyelectrolyte complexes. Generally, polyelectrolyte complexes, as a kind of electrostatic crosslinking material, can play a role in the synthesis and preparation of many materials. Many antibacterial materials are quaternary ammonium salt compounds modified with long alkyl chains, such as benzalkonium chloride, cetyltrimethylammonium bromide and cetylpyridinium chloride, etc. Due to the hydrophobic alkyl chain, these materials can damage the cell membrane of bacteria and thus synergistically enhance the antibacterial performance of the material. However, the raw materials of the current antibacterial hemostatic dressing are relatively expensive and the production method is complicated. In view of the above problems, it is necessary to design a new type of chitosan-based polyelectrolyte / surfactant composite sponge and its preparation method and application, which can overcome the problems existing in the current antibacterial hemostatic dressing. SUMMARY

[0004] In order to solve the problems in the prior art, the purpose of the present application is to provide a new type of chitosan-based polyelectrolyte / surfactant composite sponge, which is cheap and easy to obtain, has a simple preparation method, is degradable and environment-friendly, and a preparation method and application thereof.

[0005] The technical scheme adopted by the present application to achieve the above-mentioned purpose is: a chitosan-based polyelectrolyte / surfactant composite sponge, comprising a polyelectrolyte / surfactant composite and a chitosan-based composite sponge skeleton material; the polyelectrolyte / surfactant composite is a composite formed by hydrogen bonding and electrostatic interaction between basic quaternary aminated chitosan and L-hydroxyproline alkylated derivatives; and the polyelectrolyte / surfactant composite is bonded to the chitosan-based composite sponge skeleton material by using epichlorohydrin as a crosslinking agent.

[0006] The chitosan-based polyelectrolyte / surfactant composite sponge according to some embodiments of the present application has a chemical structural formula as shown below,

[0007]

[0008] The polyelectrolyte / surfactant composite comprises two components, namely cationic polyelectrolyte and anionic surfactant, i.e. basic quaternary aminated chitosan as cationic polyelectrolyte and amphiphilic L-hydroxyproline alkylated derivatives as anionic surfactant. The polyelectrolyte / surfactant composite rapidly aggregates and self-assembles to form a composite in an aqueous solution through synergistic electrostatic and hydrophobic interaction between the oppositely charged quaternary ammonium salt groups and carboxylic acid groups.

[0009] The present application also provides a preparation method of the above-mentioned chitosan-based polyelectrolyte / surfactant composite sponge, comprising the following steps:

[0010] S1. Quaternary aminated chitosan is prepared into a quaternary aminated chitosan aqueous solution with a mass fraction of 0.5%-1.5%, and subjected to strong alkaline ion exchange resin activation and ion exchange chromatography treatment, so that 90%-100% of the chloride ions in the quaternary aminated chitosan are exchanged into hydroxide ions. After vacuum freeze-drying of the effluent, basic quaternary aminated chitosan is obtained;

[0011] S2. The basic quaternary aminated chitosan obtained in step S1 is prepared into a basic quaternary aminated chitosan aqueous solution with a mass fraction of 0.2%-0.22%, and L-hydroxyproline alkylated derivatives are prepared into an L-hydroxyproline alkylated derivative ethanol solution with a mass fraction of 4.5%-5.5%. The L-hydroxyproline alkylated derivative ethanol solution is slowly added to the basic quaternary aminated chitosan aqueous solution, and the molar ratio of the basic quaternary aminated chitosan to the L-hydroxyproline alkylated derivatives is 10-1.5:1. When the mixed solution gradually changes from colorless and transparent to white turbidity, centrifugal filtration and freeze-drying are performed to obtain a polyelectrolyte / surfactant composite;

[0012] S3. After adding the epichlorohydrin and the polyelectrolyte / surfactant complex prepared in step S2 into the deionized water, ultrasonic treatment is performed to mix them thoroughly. The chitosan-based composite sponge skeleton material is immersed in the mixture and stirred. The mass ratio of the polyelectrolyte / surfactant complex to the chitosan-based composite sponge skeleton material is 10%-30%, the volume ratio of the epichlorohydrin to the deionized water is 9%-11%, and the stirring temperature is 45-55°C. After the reaction stops, the obtained product is repeatedly immersed in deionized water and ethanol to remove impurities, and then is subjected to freeze-drying and sterilization treatment to obtain the chitosan-based polyelectrolyte / surfactant complex sponge.

[0013] In the preparation method of the chitosan-based polyelectrolyte / surfactant complex sponge according to some embodiments of the present application, in step S1, the quaternary ammonium chitosan is prepared into a quaternary ammonium chitosan aqueous solution with a mass fraction of 1%.

[0014] In the preparation method of the chitosan-based polyelectrolyte / surfactant complex sponge according to some embodiments of the present application, in step S1, the preparation of the quaternary ammonium chitosan includes dissolving chitosan in an acetic acid aqueous solution, adding glycidyltrimethylammonium chloride under stirring at room temperature, and heating to 55°C for 18 hours. After the reaction ends, the reaction solution is centrifuged and filtered to obtain filter residue, which is repeatedly washed with acetone and vacuum dried to obtain the quaternary ammonium chitosan.

[0015] In the preparation method of the chitosan-based polyelectrolyte / surfactant complex sponge according to some embodiments of the present application, in step S2, the feeding ratio of the basic quaternary ammonium chitosan to the L-hydroxyproline alkylated derivative is generally required to be excess of the basic quaternary ammonium chitosan, and the formed complex does not settle to be fed. The feeding molar ratio of the basic quaternary ammonium chitosan to the L-hydroxyproline alkylated derivative is one of 10:1, 8:1, 5:1, or 3:2.

[0016] In the preparation method of the chitosan-based polyelectrolyte / surfactant complex sponge according to some embodiments of the present application, in step S2, the L-hydroxyproline alkylated derivative is one of N-stearoyl-L-hydroxyproline, N-lauroyl-L-hydroxyproline, N-myristoyl-L-hydroxyproline, or N-palmitoyl-L-hydroxyproline.

[0017] In the preparation method of the chitosan-based polyelectrolyte / surfactant complex sponge according to some embodiments of the present application, in step S3, the mass ratio of the polyelectrolyte / surfactant complex to the chitosan-based composite sponge skeleton material is 10%-25%.

[0018] According to the preparation method of the chitosan-based polyelectrolyte / surfactant complex sponge according to some embodiments of the present application, in step S3, the chitosan-based complex sponge skeleton material is a chitosan-based sponge material, a chitosan sodium alginate complex sponge, or a chitosan cellulose complex sponge, and the preparation method of the chitosan-based sponge material comprises dissolving chitosan in an aqueous solution containing urea and sodium hydroxide, adding epichlorohydrin and stirring, the stirring temperature is 50 DEG C, the stirring time is 30 minutes, the mixed solution is subjected to crosslinking reaction to obtain a white hydrogel, the obtained white hydrogel is repeatedly immersed and washed with deionized water and ethanol to remove impurities, and the hydrogel after immersion and washing is subjected to freeze-drying to obtain the chitosan-based sponge skeleton material.

[0019] According to the preparation method of the chitosan-based polyelectrolyte / surfactant complex sponge according to some embodiments of the present application, in step S3, the volume ratio of epichlorohydrin to deionized water is 10%, the stirring temperature is 50 DEG C, and the stirring time is 30 minutes.

[0020] According to the preparation method of the chitosan-based polyelectrolyte / surfactant complex sponge according to some embodiments of the present application, the degree of deacetylation of chitosan is greater than or equal to 95%.

[0021] The present application also provides applications of the above-mentioned chitosan-based polyelectrolyte / surfactant complex sponge, and the chitosan-based polyelectrolyte / surfactant complex sponge can be practically applied to wound dressings for hemostasis and wound healing promotion, non-compressible wound hemostatic agents, and soft tissue repair and regeneration materials.

[0022] The chitosan-based polyelectrolyte / surfactant complex sponge and the preparation method and applications thereof according to the present application use quaternary ammonium chitosan as a cationic polyelectrolyte, which is widely available, low in cost, biocompatible, and non-toxic, and uses alkylated modified amino acids as an anionic surfactant, which is co-assembled into a novel polyelectrolyte complex under the action of hydrogen bonds and electrostatic forces, and the preparation process is simple, fast, and non-toxic reagents are not used, so that large-scale production can be realized. The chitosan-based polyelectrolyte / surfactant complex sponge contains quaternary ammonium chitosan and long-alkyl-chain surfactants in structure, and the cationic groups and hydrophobic alkyl chains on the molecular chains can effectively inhibit bacteria and accelerate thrombus formation, which is conducive to preventing wound infection and promoting wound healing. The chitosan sponge is used as a support skeleton, is light in weight, has a connected macroporous structure, can rapidly and effectively absorb blood, and can concentrate blood and promote blood coagulation during the hemostasis process. The chitosan-based polyelectrolyte / surfactant complex sponge is mainly based on chitosan and its derivative materials, can be naturally degraded by lysozyme in the body, has good biocompatibility and degradability, and is expected to become a multifunctional antibacterial hemostatic and healing-promoting material, and has great clinical application potential. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1The physical picture of the sponge prepared in Example 1 of the present application;

[0024] Figure 2 The scanning electron microscope picture of the microstructure of the sponge prepared in Example 1 of the present application;

[0025] Figure 3 The compression stress-strain curve of the chitosan sponge and the sponge prepared in Example 1 and Example 2 of the present application;

[0026] Figure 4 The liquid absorption capacity of the sponge sample of the present application;

[0027] Figure 5 The control picture of the antibacterial performance test of the sponge sample of the present application;

[0028] Figure 6 The hemostatic effect picture of the sponge sample of the present application in the rat liver hemorrhage model;

[0029] Figure 7 The bar chart of the hemostatic time and the amount of bleeding of the sponge sample of the present application in the rat liver hemorrhage model. DETAILED DESCRIPTION

[0030] The embodiments of the present application will be further described in detail below with reference to the accompanying drawings and examples. The following examples are used to illustrate the present application, but cannot be used to limit the scope of the present application.

[0031] Example 1

[0032] (1) Preparation of polyelectrolyte / surfactant complex

[0033] First, 0.5 g of chitosan with a degree of deacetylation ≥ 95% is dissolved in 18 mL of 0.5% acetic acid aqueous solution, 773 μL of glycidyltrimethylammonium chloride is added under stirring at room temperature, the temperature is raised to 55°C, and the reaction is carried out for 18 hours. Subsequently, the reaction solution is centrifuged and filtered to obtain filter residue, which is repeatedly washed with acetone and vacuum dried to obtain quaternized chitosan. 1.0 g of the quaternized chitosan is dissolved in 100 mL of deionized water, and the chloride ions Cl - in the quaternized chitosan are exchanged into hydroxide ions OH -The effluent phase was subjected to freeze-drying to obtain the basic quaternary aminated chitosan. Then, 1 mL of N-stearoyl-L-hydroxyproline ethanol solution, in which the weight of N-stearoyl-L-hydroxyproline was 39.76 mg, was slowly added dropwise into 50 mL of the aqueous solution of the basic quaternary aminated chitosan, in which the weight of the basic quaternary aminated chitosan was 0.1 g. When the mixed solution changed from colorless and transparent to white turbidity, it was proved that the polyelectrolyte / surfactant complex had been synthesized. After centrifugal filtration and freeze-drying, white crystals were obtained, which were the polyelectrolyte / surfactant complex.

[0034] (2) Preparation of the chitosan-based polyelectrolyte / surfactant complex sponge

[0035] 0.5 g of chitosan was dissolved in 50 mL of the aqueous solution containing urea and sodium hydroxide, in which the weight content of urea was 7% and the weight content of sodium hydroxide was 15%, and then 3 mL of epichlorohydrin was added as a crosslinking agent. The white hydrogel was obtained after reaction at 50°C under stirring for 30 minutes. The obtained white hydrogel was repeatedly immersed in deionized water and ethanol to remove impurities, and then subjected to freeze-drying to obtain the chitosan-based sponge skeleton material. 5 mg of the polyelectrolyte / surfactant complex and 100 μL of epichlorohydrin were added to 1 mL of deionized water, and ultrasonic treatment was performed for 2 minutes to fully mix them. 20 mg of the chitosan-based sponge skeleton was placed in the fully mixed mixture to be completely immersed. The reaction was performed at 50°C under stirring for 30 minutes. The obtained sponge was repeatedly immersed in deionized water and ethanol to remove impurities, and then subjected to freeze-drying and sterilization treatment to obtain the chitosan-based polyelectrolyte / surfactant complex sponge.

[0036] Example 2

[0037] (1) The synthesis of the basic quaternary aminated chitosan and the preparation of the polyelectrolyte / surfactant complex were the same as in Example 1.

[0038] (2) Preparation of the chitosan-based polyelectrolyte / surfactant complex sponge

[0039] Take 0.5 g of chitosan and dissolve it in 50 mL of an aqueous solution containing urea and sodium hydroxide, wherein the weight content of urea is 7% and the weight content of sodium hydroxide is 15%, then add 3 mL of epichlorohydrin as a crosslinking agent, and react under stirring at 50°C for 30 minutes to obtain a white hydrogel. The obtained white hydrogel is repeatedly immersed in deionized water and ethanol to remove impurities, and then freeze-dried to obtain a chitosan-based sponge skeleton material. Add 2 mg of polyelectrolyte / surfactant complex and 100 μL of epichlorohydrin to 1 mL of deionized water, and ultrasonically treat for 2 minutes to fully mix. Take 20 mg of chitosan-based sponge skeleton and place it in the above fully mixed mixture so that it is completely immersed. React under stirring at 50°C for 30 minutes. The obtained sponge is repeatedly immersed in deionized water and ethanol to remove impurities, and then freeze-dried and sterilized to obtain a chitosan-based polyelectrolyte / surfactant complex sponge.

[0040] Example 3

[0041] (1) The synthesis of basic quaternary ammonium chitosan and the preparation of polyelectrolyte / surfactant complex are the same as in Example 1.

[0042] (2) Preparation of chitosan-based polyelectrolyte / surfactant complex sponge

[0043] Take 0.3 g of chitosan and dissolve it in 30 mL of an aqueous solution containing urea and sodium hydroxide, wherein the weight content of urea is 7% and the weight content of sodium hydroxide is 15%. Take 0.2 g of oxidized sodium alginate and dissolve it in 20 mL of deionized water. Then add the oxidized sodium alginate solution to the chitosan solution. After stirring at room temperature for 1 hour, the mixture is pre-frozen at -45°C. Then, repeatedly immerse it in deionized water and ethanol to remove impurities, and then freeze-dry to obtain a chitosan / alginate composite sponge skeleton material. Add 2 mg of polyelectrolyte / surfactant complex and 100 μL of epichlorohydrin to 1 mL of deionized water, and ultrasonically treat for 2 minutes to fully mix. Take about 20 mg of chitosan / alginate composite sponge skeleton and place it in the above mixture so that it is completely immersed. React under stirring at 50°C for 30 minutes. The obtained sponge is repeatedly immersed in deionized water and ethanol to remove impurities, and then freeze-dried and sterilized to obtain a chitosan-based polyelectrolyte / surfactant complex sponge.

[0044] Example 4

[0045] The chitosan-based polyelectrolyte / surfactant complex sponge prepared in Example 1 above is tested for the following relevant performance:

[0046] (1) The actual photo of the chitosan-based polyelectrolyte / surfactant complex sponge prepared in Example 1 is shown in Figure 1.Figure 1 As shown, the chitosan-based polyelectrolyte / surfactant composite sponge is lightweight, compressible, absorbs water and expands to recover its shape, and rapidly absorbs blood.

[0047] (2) The morphology of the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 was characterized by scanning electron microscopy, as shown in the attached figure. Figure 2 As shown, the chitosan-based polyelectrolyte / surfactant composite sponge has a three-dimensional porous structure with continuous layers, and the interpenetrating, interconnected macroporous structure is conducive to rapid water absorption and blood concentration.

[0048] (3) Compression stress-strain tests were performed on the chitosan-based polyelectrolyte / surfactant composite sponges prepared in Examples 1 and 2. A universal testing machine was used to test the compression performance of chitosan-based polyelectrolyte / surfactant composite sponge samples with a diameter of 1.5 cm and a height of 0.5 cm prepared in Examples 1 and 2. The samples were compressed to 80% strain at a displacement rate of 2 mm / min. The measurement results are shown in the attached figure. Figure 3 As shown in the figure, the compressive modulus of the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 was 254.54 ± 12.03 kPa, the compressive modulus of the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 2 was 218.86 ± 14.06 kPa, and the compressive modulus of the chitosan-based sponge was 151.29 ± 18.01 kPa. The compressive modulus shows that as the content of the polyelectrolyte / surfactant composite increases, the chitosan-based polyelectrolyte / surfactant composite sponge exhibits a higher compressive modulus, indicating that the addition of the polyelectrolyte / surfactant composite significantly enhances the compressive strength of the chitosan-based polyelectrolyte / surfactant composite sponge.

[0049] (4) The liquid absorption performance of the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 was tested. Three initial masses (W0) of each of the chitosan-based polyelectrolyte / surfactant composite sponge and the control group (gelatin sponge and chitosan sponge) were weighed beforehand. The three chitosan-based polyelectrolyte / surfactant composite sponges were immersed in 50 mL of deionized water, 50 mL of phosphate buffer, and 50 mL of simulated sweat, respectively, at 37°C. The gelatin sponge and chitosan sponge underwent the same procedure. After the samples absorbed each liquid to saturation, they were removed and weighed again (W0). aq ), calculate the sample's liquid absorption capacity according to the formula: Liquid absorption rate (%) = [(W aq -W0) / W0]×100%. The results are attached. Figure 4As shown, the absorption capacity of the chitosan sponge and the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 to the three kinds of liquids is obviously higher than that of the gelatin sponge, and the absorption capacity of the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 to the three kinds of liquids is better, indicating that the chitosan-based polyelectrolyte / surfactant composite sponge has good liquid absorption capacity, which is beneficial to adsorb wound exudates in actual wound treatment and promote wound healing.

[0050] (5) The chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 was subjected to antibacterial performance test. S. aureus and E. coli were used as model microorganisms. 100 μL of S. aureus liquid was taken by using a pipette gun and inoculated on 0.2 g of the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1, and 100 μL of E. coli liquid was taken by using a pipette gun and inoculated on 0.2 g of the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1, and the two samples were incubated at 37°C for 4 hours. The same operation was performed on a blank sample and a chitosan sponge as a comparative example. After the six samples were incubated for 4 hours, the six samples were transferred into centrifuge tubes containing 1 mL of phosphate buffer solution, and vortexed for 10 minutes to resuspend the adhered bacteria. The bacterial suspension was further diluted to a countable range, and uniformly plated on 1.5% LB agar plates, and incubated at 37°C for 24 hours. The number of colonies of the formed microorganisms was observed and counted. As shown in the following table, the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 can effectively inhibit the growth and reproduction of S. aureus and E. coli, and has excellent antibacterial performance. Figure 5 As shown in the following table, the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 can effectively inhibit the growth and reproduction of S. aureus and E. coli, and has excellent antibacterial performance.

[0051] (6) The chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 was applied to animal in vivo hemostasis experiment.

[0052] The experimental method includes: 9 healthy SD rats with a body weight of 60-75 g were selected, all were male, and randomly divided into three groups for experiment. First, a rat liver bleeding model was established. The rats were anesthetized by intraperitoneal injection of a certain amount of 10% chloral hydrate, and fixed on the experimental plate. The liver was exposed by making an incision on the abdomen of the rat with a surgical knife, and the serous fluid around the liver was carefully removed to prevent interference with the blood measurement in the experiment. Then, a wound about 0.5 cm long and 0.2 cm deep was made on the surface of the liver with a surgical knife, and the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 and the gelatin sponge for control were gently placed on the wound bleeding point, and a filter paper was placed at the bottom to evaluate the amount of bleeding. The sample was quickly moved away every 5 seconds, and the bleeding of the wound was observed until hemostasis was completed.Figure 6 The image shows the hemostatic effect of a rat liver hemorrhage model. Compared with the hemostatic effect of gelatin sponge, the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 has a significant hemostatic effect.

[0053] In addition, the relationship between bleeding volume and hemostasis time was tested using the above experimental methods on medical gauze, gelatin sponge, chitosan sponge, the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 2, and the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1. The experimental results are as follows: Figure 7 As shown, by appendix Figure 7 The bar chart showing the bleeding volume versus hemostasis time indicates that the longest hemostasis time was 235.6 s for the blank control group, followed by 218.4 s for medical gauze and 163.5 s for gelatin sponge. The chitosan sponge had a hemostasis time of 80.6 s, the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 2 had a hemostasis time of 52.4 s, and the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 had a hemostasis time of 27.2 s. The hemostasis times of the chitosan-based polyelectrolyte / surfactant composite sponges prepared in Examples 1 and 2 were shorter than those of the chitosan sponge alone, demonstrating that the higher the content of the polyelectrolyte / surfactant composite in the sponge, the shorter the hemostasis time. Furthermore, regarding blood loss after liver wound treatment, the highest blood loss was observed in the blank control group (505.6 mg), followed by 443.5 mg with medical gauze and 309.1 mg with gelatin sponge. The blood loss with chitosan sponge was 243.9 mg, the blood loss with the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 2 was 176.4 mg, and the blood loss with the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 was 97.1 mg. In summary, using the chitosan-based polyelectrolyte / surfactant composite sponge prepared in Example 1 resulted in the least blood loss and the shortest hemostasis time, demonstrating excellent in vivo hemostatic effect.

[0054] The embodiments of the present invention are given for illustrative and descriptive purposes only, and are not intended to be exhaustive or to limit the invention to the forms disclosed. Many modifications and variations will be apparent to those skilled in the art. The embodiments were chosen and described in order to better illustrate the principles and practical application of the invention, and to enable those skilled in the art to understand the invention and to design various embodiments with various modifications suitable for a particular purpose.

Claims

1. A chitosan-based polyelectrolyte / surfactant composite sponge, characterized in that, The invention comprises a polyelectrolyte / surfactant complex and a chitosan-based composite sponge skeleton material; the polyelectrolyte / surfactant complex is a complex formed by basic quaternary ammonium chitosan and L-hydroxyproline alkylated derivatives through hydrogen bonding and electrostatic interaction; the polyelectrolyte / surfactant complex is bonded to the chitosan-based composite sponge skeleton material using epichlorohydrin as a crosslinking agent; the chitosan-based composite sponge skeleton material is one of chitosan-based sponge material, chitosan sodium alginate composite sponge, or chitosan cellulose composite sponge.

2. The chitosan-based polyelectrolyte / surfactant composite sponge according to claim 1, characterized in that, The chemical structural formula of the polyelectrolyte / surfactant complex is shown below. 。 3. A method for preparing the chitosan-based polyelectrolyte / surfactant composite sponge according to claim 1, characterized in that, Includes the following steps: S1. Quaternized chitosan is prepared into an aqueous solution of quaternized chitosan with a mass fraction of 0.5%-1.5%, and then activated with a strong basic ion exchange resin and subjected to ion exchange chromatography to exchange 90%-100% of the chloride ions in the quaternized chitosan into hydroxide ions. The effluent phase is then freeze-dried under vacuum to obtain basic quaternized chitosan. S2. Prepare an aqueous solution of alkaline quaternized chitosan with a mass fraction of 0.2%-0.22% by preparing an ethanolic solution of L-hydroxyproline alkylated derivative with a mass fraction of 4.5%-5.5% by preparing an ethanolic solution of L-hydroxyproline alkylated derivative. Slowly add the ethanolic solution of L-hydroxyproline alkylated derivative to the aqueous solution of alkaline quaternized chitosan. The molar ratio of alkaline quaternized chitosan to L-hydroxyproline alkylated derivative is 10-1.5:

1. When the mixed solution gradually changes from colorless and transparent to white and turbid, centrifuge, filter and freeze dry to obtain a polyelectrolyte / surfactant complex. S3. After adding epichlorohydrin and the polyelectrolyte / surfactant composite obtained in step S2 to deionized water, ultrasonic treatment is performed to ensure thorough mixing. Chitosan-based composite sponge skeleton material is placed in the mixture, completely submerged, and stirred. The mass ratio of polyelectrolyte / surfactant composite to chitosan-based composite sponge skeleton material is 10%-30%, and the volume ratio of epichlorohydrin to deionized water is 9%-11%. The stirring temperature is 45℃-55℃. After the reaction stops, the obtained product is repeatedly washed with deionized water and ethanol to remove impurities. After washing, it is freeze-dried and sterilized to obtain chitosan-based polyelectrolyte / surfactant composite sponge.

4. The method for preparing a chitosan-based polyelectrolyte / surfactant composite sponge according to claim 3, characterized in that, In step S1, the quaternized chitosan is prepared into a quaternized chitosan aqueous solution with a mass fraction of 1%.

5. The method for preparing a chitosan-based polyelectrolyte / surfactant composite sponge according to claim 3, characterized in that, In step S2, the molar ratio of alkaline quaternized chitosan to L-hydroxyproline alkylated derivative is one of 10:1, 8:1, 5:1 or 3:

2.

6. The method for preparing a chitosan-based polyelectrolyte / surfactant composite sponge according to claim 3, characterized in that, In step S2, the L-hydroxyproline alkylation derivative is one of N-stearoyl-L-hydroxyproline, N-lauroyl-L-hydroxyproline, N-myristoyl-L-hydroxyproline, or N-palmacloyl-L-hydroxyproline.

7. The method for preparing a chitosan-based polyelectrolyte / surfactant composite sponge according to claim 3, characterized in that, In step S3, the mass ratio of the polyelectrolyte / surfactant composite to the chitosan-based composite sponge skeleton material is 10%-25%.

8. The method for preparing a chitosan-based polyelectrolyte / surfactant composite sponge according to claim 3, characterized in that, In step S3, the preparation method of the chitosan-based sponge material includes dissolving chitosan in an aqueous solution containing urea and sodium hydroxide, adding epichlorohydrin and stirring at a temperature of 50°C for 30 minutes, allowing the mixture to undergo a cross-linking reaction to obtain a white hydrogel, repeatedly rinsing the obtained white hydrogel with deionized water and ethanol to remove impurities, and then freeze-drying it to obtain the chitosan-based sponge skeleton material.

9. The method for preparing a chitosan-based polyelectrolyte / surfactant composite sponge according to claim 3, characterized in that, In step S3, the volume ratio of epichlorohydrin to deionized water is 10%, the stirring temperature is 50°C, and the stirring time is 30 minutes.

10. The application of a chitosan-based polyelectrolyte / surfactant composite sponge as described in any one of claims 1-2.

Images