A method for preparing hemostatic hydrogel containing natural polysaccharides

Abstract

The invention discloses a method for preparing a hemostatic hydrogel containing natural polysaccharides, comprising the following steps: (1) preparing a natural polysaccharide-clay aqueous solution: adding polyphenols and natural polysaccharides to a clay aqueous solution under stirring, adjusting the pH value of the system to 8-9, and forming a natural polysaccharide-clay aqueous solution; (2) preparing a hydrogel: adding a monomer, an initiator and a cross-linking agent to the natural polysaccharide-clay aqueous solution to form a mixed solution, stirring evenly, and reacting at 10-40° C. for 10-60 min to obtain the hemostatic hydrogel containing natural polysaccharides; wherein the natural polysaccharide is at least one of sodium alginate, pectin, and cellulose and chitosan. The prepared hydrogel has excellent adhesion, self-repairing, biocompatibility, antibacterial and hemostatic properties.

Description

Technical Field

[0001] The invention belongs to the technical field of hydrogels, and in particular relates to a method for preparing a hemostatic hydrogel containing natural polysaccharides. Background Art

[0002] Hydrogel is a kind of material with a three-dimensional network structure formed by hydrophilic monomers through physical or chemical crosslinking. Because hydrogel has good biocompatibility and flexibility, it can be used in fields such as tissue engineering, drug delivery and wound dressing. Hydrogels with adhesion properties can achieve wound closure on the tissue surface, thereby achieving the purpose of hemostasis, but most adhesion hydrogels have the disadvantages of poor mechanical properties and easy damage. In addition, hydrogels used for wound dressings should also have good antibacterial properties to prevent wound infection. Therefore, it is urgent to develop hydrogel materials with excellent adhesion properties, self-repairing, good biocompatibility, antibacterial, and hemostatic properties.

[0003] Natural polysaccharides are widely used to make biomedical hydrogels because of their good hydrophilicity, excellent biodegradability, good bioactivity and high biocompatibility. However, hydrogels based on natural polysaccharides cannot achieve repeated adhesion, self-repairing, antibacterial and hemostatic properties at the same time. The cumbersome preparation and chemical modification process also greatly affects the biocompatibility of hydrogels, which greatly limits their application in the biomedical field. Therefore, it is still challenging to prepare natural polysaccharide hydrogels that have excellent adhesion, self-repairing, antibacterial and hemostatic properties as well as excellent biocompatibility. Summary of the invention

[0004] In view of the defects of the prior art, the present invention provides a method for preparing a hemostatic hydrogel containing natural polysaccharides. The hydrogel prepared by compounding natural polysaccharides, polyphenols and clay under alkaline conditions can overcome the defects of the hydrogel in the prior art that it cannot have excellent adhesion, self-repairing, biocompatibility, antibacterial and hemostatic properties.

[0005] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0006] (1) Preparing a natural polysaccharide-clay aqueous solution: adding polyphenols and natural polysaccharides to a clay aqueous solution under stirring, and adjusting the pH value of the system to 8-9 to form a natural polysaccharide-clay aqueous solution;

[0007] (2) Preparing a hydrogel: adding a monomer, an initiator and a cross-linking agent to the natural polysaccharide-clay aqueous solution to form a mixed solution, stirring evenly, and reacting at 10-40° C. for 10-60 minutes to obtain the hemostatic hydrogel containing the natural polysaccharide;

[0008] Wherein, the natural polysaccharide is at least one of sodium alginate, pectin, cellulose and chitosan.

[0009] Preferably, the polyphenolic substance is at least one of lignin and tannic acid.

[0010] Preferably, the clay is any one of lithium magnesium silicate, aluminum magnesium silicate, kaolin, montmorillonite or hydrotalcite.

[0011] Preferably, the monomer is at least one of acrylamide, methacrylamide and hydroxyethyl methacrylate.

[0012] Preferably, the natural polysaccharide accounts for 10-200% of the mass of the clay, and the polyphenolic substance accounts for 20-200% of the mass of the clay.

[0013] Preferably, the concentration of the clay aqueous solution is 1-3 wt %.

[0014] Preferably, the clay is 1-10% by weight of the monomer.

[0015] Preferably, the initiator is 1-3% of the mass of the monomer, and the cross-linking agent is 0.1-0.3% of the mass of the monomer.

[0016] Preferably, the mass ratio of at least one of sodium alginate, pectin and cellulose in the natural polysaccharide to chitosan is 5:(2.5-20).

[0017] Preferably, the initiator is a mixture of ammonium persulfate and tetramethylethylenediamine; and the cross-linking agent is N,N-methylenebisacrylamide.

[0018] Preferably, in step (1), NaOH solution is used to adjust the pH value of the system to 8-9.

[0019] Advantages of the present invention:

[0020] (1) In the present invention, natural polysaccharides, polyphenols and clay are compounded under alkaline conditions, and the mechanical properties of the hydrogel are enhanced through Michael addition between polyphenols and natural polysaccharides and multiple non-covalent interactions between natural polysaccharides, polyphenols and clay, including hydrogen bonds, electrostatic interactions, etc.

[0021] (2) The present invention adopts a one-step method to prepare the hydrogel, which is simple and easy to operate. The prepared hemostatic hydrogel containing natural polysaccharides has excellent self-repairing and adhesion properties.

[0022] (3) In addition, the hydrogel has excellent biocompatibility and good antibacterial properties. The cell survival rate is high after 24 h of co-culture of the gel extract and cells, and the hydrogel has a high bactericidal efficiency against Escherichia coli.

[0023] (4) The hydrogel also has excellent hemostatic properties. Compared with the blank control group, the amount of bleeding and bleeding time of the hydrogel were significantly reduced. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a test diagram of the mechanical properties of the hydrogel prepared in Example 1;

[0025] Figure 2 is a test diagram of the adhesion performance of the hydrogel prepared in Example 1;

[0026] Figure 3 This is a self-healing performance test diagram of the hydrogel prepared in Example 1;

[0027] Figure 4 is a cytotoxicity graph of the hydrogel prepared in Example 1;

[0028] Figure 5 is a graph showing the antibacterial performance of the hydrogel prepared in Example 1;

[0029] Figure 6 This is a graph showing the hemostatic performance of the hydrogel prepared in Example 1. DETAILED DESCRIPTION

[0030] Example 1

[0031] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0032] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 62.5 mg of lithium magnesium silicate in 5 mL of deionized water to obtain a lithium magnesium silicate aqueous solution; adding 120 mg of lignin, 80 mg of chitosan, and 45 mg of sodium alginate to the lithium magnesium silicate aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a black mixed solution;

[0033] (2) Preparation of hydrogel: 1.25 g of acrylamide monomer, 25 mg of ammonium persulfate and 26 μL (20.02 μg) of tetramethylethylenediamine as an initiator and 1.9 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the mixture was reacted at 25° C. for 30 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a black hydrogel.

[0034] Example 2

[0035] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0036] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 125 mg of magnesium aluminum silicate in 5 mL of deionized water to obtain a magnesium aluminum silicate aqueous solution; adding 110 mg of lignin, 125 mg of chitosan, and 100 mg of cellulose to the magnesium aluminum silicate aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a black mixed solution;

[0037] (2) Preparation of hydrogel: 1.25 g of methacrylamide monomer, 12.5 mg of ammonium persulfate and 16.1 μL (12.86 μg) of tetramethylethylenediamine as an initiator and 1.25 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the solution was reacted at 30° C. for 40 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a black hydrogel.

[0038] Example 3

[0039] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0040] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 125 mg of kaolin in 5 mL of deionized water to obtain a kaolin aqueous solution; adding 120 mg of lignin, 125 mg of chitosan, and 100 mg of pectin to the kaolin aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a black mixed solution;

[0041] (2) Preparation of hydrogel: 1.25 g of hydroxyethyl methacrylate monomer, 37.5 mg of ammonium persulfate and 48 μL (36.96 μg) of tetramethylethylenediamine as an initiator, and 3.75 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the solution was reacted at 20° C. for 20 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a black hydrogel.

[0042] Example 4

[0043] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0044] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 50 mg of montmorillonite in 5 mL of deionized water to obtain a montmorillonite aqueous solution; adding 100 mg of tannic acid, 25 mg of chitosan, 25 mg of sodium alginate, and 25 mg of cellulose to the montmorillonite aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a light yellow mixed solution;

[0045] (2) Preparation of hydrogel: 0.5 g of hydroxyethyl methacrylate monomer, 5 mg of ammonium persulfate and 6.4 μL (4.93 μg) of tetramethylethylenediamine as an initiator and 0.5 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the solution was reacted at 40° C. for 50 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a light yellow hydrogel.

[0046] Example 5

[0047] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0048] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 50 mg of hydrotalcite in 5 mL of deionized water to obtain a hydrotalcite aqueous solution; adding 50 mg of tannic acid, 50 mg of chitosan, and 50 mg of sodium alginate to the hydrotalcite aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a light yellow mixed solution;

[0049] (2) Preparation of hydrogel: 0.5 g acrylamide monomer, 10 mg ammonium persulfate and 12.8 μL (9.86 μg) tetramethylethylenediamine as an initiator and 1 mg N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the solution was reacted at 40° C. for 60 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a light yellow hydrogel.

[0050] Example 6

[0051] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0052] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 50 mg of montmorillonite in 5 mL of deionized water to obtain a montmorillonite aqueous solution; adding 20 mg of tannic acid, 30 mg of chitosan, and 10 mg of sodium alginate to the montmorillonite aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a light yellow mixed solution;

[0053] (2) Preparation of hydrogel: 0.5 g of methacrylamide monomer, 15 mg of ammonium persulfate and 19.2 μL (14.78 μg) of tetramethylethylenediamine as an initiator and 1.5 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the solution was reacted at 10° C. for 20 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a light yellow hydrogel.

[0054] Example 7

[0055] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0056] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 50 mg of hydrotalcite in 5 mL of deionized water to obtain a hydrotalcite aqueous solution; adding 10 mg of tannic acid, 4 mg of chitosan, and 1 mg of pectin to the hydrotalcite aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a light yellow mixed solution;

[0057] (2) Preparation of hydrogel: 0.5 g of hydroxyethyl methacrylate monomer, 5 mg of ammonium persulfate and 6.4 μL (4.93 μg) of tetramethylethylenediamine as an initiator and 0.5 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the solution was reacted at 20° C. for 10 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a light yellow hydrogel.

[0058] Example 8

[0059] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0060] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 150 mg of kaolin in 5 mL of deionized water to obtain a kaolin aqueous solution; adding 175 mg of tannic acid, 150 mg of chitosan, and 100 mg of cellulose to the kaolin aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a light yellow mixed solution;

[0061] (2) Preparation of hydrogel: 1.75 g of methacrylamide monomer, 52.5 mg of ammonium persulfate and 67.7 μL (52.13 μg) of tetramethylethylenediamine as an initiator and 5.25 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the solution was reacted at 30° C. for 60 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a light yellow hydrogel.

[0062] Example 9

[0063] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0064] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 150 mg of magnesium aluminum silicate in 5 mL of deionized water to obtain a magnesium aluminum silicate aqueous solution; adding 50 mg of tannic acid, 75 mg of chitosan, and 125 mg of pectin to the magnesium aluminum silicate aqueous solution in sequence under stirring, and adjusting the pH of the system to 8 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a light yellow mixed solution;

[0065] (2) Preparation of hydrogel: 1.75 g of methacrylamide monomer, 35 mg of ammonium persulfate and 45.1 μL (34.73 μg) of tetramethylethylenediamine as an initiator and 3.5 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the solution was reacted at 40° C. for 30 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a light yellow hydrogel.

[0066] Example 10

[0067] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0068] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 150 mg of lithium magnesium silicate in 5 mL of deionized water to obtain a lithium magnesium silicate aqueous solution; adding 75 mg of lignin, 150 mg of tannic acid, 75 mg of chitosan, and 150 mg of sodium alginate to the lithium magnesium silicate aqueous solution in sequence under stirring, and adjusting the pH of the system to 9 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a light yellow mixed solution;

[0069] (2) Preparation of hydrogel: 1.75 g of acrylamide monomer, 17.5 mg of ammonium persulfate and 22.6 μL (17.4 μg) of tetramethylethylenediamine as an initiator and 1.75 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the solution was reacted at 10° C. for 50 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a light yellow hydrogel.

[0070] Comparative Example 1

[0071] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0072] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 62.5 mg of lithium magnesium silicate in 5 mL of deionized water to obtain a lithium magnesium silicate aqueous solution; adding 120 mg of lignin, 80 mg of chitosan, and 45 mg of sodium alginate to the lithium magnesium silicate aqueous solution in sequence under stirring, and stirring to obtain a natural polysaccharide-clay aqueous solution, which is a black mixed solution;

[0073] (2) Preparation of hydrogel: 1.25 g of acrylamide monomer, 25 mg of ammonium persulfate and 26 μL (20.02 μg) of tetramethylethylenediamine as an initiator and 1.9 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the mixture was reacted at 25° C. for 30 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a black hydrogel.

[0074] Comparative Example 2

[0075] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0076] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 62.5 mg of lithium magnesium silicate in 5 mL of deionized water to obtain a lithium magnesium silicate aqueous solution; adding 80 mg of chitosan and 45 mg of sodium alginate to the lithium magnesium silicate aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a light yellow mixed solution;

[0077] (2) Preparation of hydrogel: 1.25 g of acrylamide monomer, 25 mg of ammonium persulfate and 26 μL (20.02 μg) of tetramethylethylenediamine as an initiator and 1.9 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the mixture was reacted at 25° C. for 30 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a light yellow hydrogel.

[0078] Comparative Example 3

[0079] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0080] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 62.5 mg of lithium magnesium silicate in 5 mL of deionized water to obtain a lithium magnesium silicate aqueous solution; adding 120 mg of lignin and 80 mg of chitosan to the lithium magnesium silicate aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a black mixed solution;

[0081] (2) Preparation of hydrogel: 1.25 g of acrylamide monomer, 25 mg of ammonium persulfate and 26 μL (20.02 μg) of tetramethylethylenediamine as an initiator and 1.9 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to the natural polysaccharide-clay aqueous solution obtained in step (1) to form a mixed solution. After stirring evenly, the mixture was reacted at 25° C. for 30 min to obtain the hemostatic hydrogel containing natural polysaccharides, which was a black hydrogel.

[0082] Comparative Example 4

[0083] A method for preparing a hemostatic hydrogel containing natural polysaccharides comprises the following steps:

[0084] (1) Preparing a natural polysaccharide-clay aqueous solution: dissolving 62.5 mg of lithium magnesium silicate in 5 mL of deionized water to obtain a lithium magnesium silicate aqueous solution; adding 120 mg of lignin and 45 mg of sodium alginate to the lithium magnesium silicate aqueous solution in sequence under stirring, and adjusting the pH of the system to 8.5 with a NaOH solution to obtain a natural polysaccharide-clay aqueous solution, which is a black mixed solution;

[0085] (2) To the natural polysaccharide-clay aqueous solution obtained in step (1), 1.25 g of acrylamide monomer, 25 mg of ammonium persulfate and 26 μL (20.02 μg) of tetramethylethylenediamine as an initiator and 1.9 mg of N,N-methylenebisacrylamide as a cross-linking agent were added to form a mixed solution. After stirring evenly, the solution was reacted at 25°C for 30 min, but no gel was formed.

[0086] Performance Testing

[0087] 1. Mechanical properties testing

[0088] The tensile and compressive properties of the hydrogel in Example 1 were tested. Figure 1 .

[0089] For the tensile test: The tensile properties of the hydrogels were measured by uniaxial tensile testing at 25 °C at a tensile speed of 50 mm / min. Figure 1 (a). Figure 1 As shown in (a), the hydrogel sample has excellent tensile properties, and its fracture strain reaches 1382%. The calculated values ​​are: Young's modulus is 2.94 kPa, and toughness is 103.6 kJ / m 3 At the same time, the tensile properties of the hydrogels of Comparative Examples 1-3 were detected, and their fracture strains were calculated. The results are shown in Table 1. Among them, Comparative Example 4 did not form a gel, so its mechanical properties are not shown in the table.

[0090] For compression experiments, the hydrogel was subjected to cyclic compression measurement at room temperature at a compression speed of 5 mm / min through a tensile tester to detect its compression properties. Figure 1 (b). Figure 1 As shown in (b), the hydrogel is compressed to 80% of the initial strain and can recover to its original state after the pressure is removed. This shows that it can withstand a large compressive force and quickly recover to its initial state.

[0091] 2. Adhesion performance test

[0092] The hydrogel obtained in Example 1 was subjected to an adhesion test. The shear adhesion strength test was performed by peeling the gel from between two substrates at a tensile speed of 30 mm / min, and recording the maximum force during the peeling process. The shear adhesion strength was calculated by dividing the maximum force by the overlapping area between the two substrates. Figure 2 As shown in the figure, with the extension of adhesion time, the adhesion strength of the hydrogel is significantly enhanced. When the adhesion time increases from 1 h to 7 h, the adhesion strength of the hydrogel on the stainless steel surface increases from 16 kPa to 78 kPa, indicating that the hydrogel has good adhesion properties on stainless steel.

[0093] At the same time, the adhesion strength of the hydrogels of Comparative Examples 1-3 was tested, and the results are shown in Table 1, wherein the adhesion strength is based on the adhesion time of 7 hours; wherein, Comparative Example 4 did not form a gel, so its adhesion strength is not shown in the table.

[0094] Table 1 Fracture strain and adhesion strength

[0095]

[0096] 3. Tensile self-repair performance test

[0097] The self-healing performance test of the hydrogel obtained in Example 1 was performed. The hydrogel was prepared into a cylindrical sample for self-healing performance test. The stretching speed was 50 mm / min, and samples with different self-healing times were taken for experiments. The self-healing efficiency is the ratio of the fracture stress of the tensile stress-strain curve of the hydrogel sample after self-healing to the fracture stress of the tensile stress-strain curve of the original sample, see Figure 3 .Depend on Figure 3 It can be seen that when the self-healing time increases from 1 h to 24 h, the self-healing efficiency increases from 35.7% to 102.5%, indicating that the hydrogel prepared in the present invention has excellent self-healing properties.

[0098] IV. Cytotoxicity Test

[0099] The hydrogel obtained in Example 1 was subjected to a cytotoxicity test. In the cytotoxicity test, the hydrogel oC for 24 h to obtain hydrogel extracts; co-culture the cells with hydrogel extracts of different concentrations for 24 h; use the MTT method to quantitatively test the optical density (OD) of the solution with a Spectra Max 190 microplate reader to calculate cell viability, cell viability (%) = [OD] tested / [OD] control × 100, the result is Figure 4 As shown. Figure 4 It can be seen that when the concentration of the extract of the hydrogel obtained in Example 1 is as high as 75%, the cell survival rate of the hydrogel is also maintained at about 85%, indicating that the hydrogel has excellent biocompatibility.

[0100] 5. Antibacterial performance test

[0101] The hydrogel obtained in Example 1 was tested for its antibacterial properties. In the antibacterial performance test study, Escherichia coli was used to characterize the antibacterial properties of the gel. First, the bacterial solution and the sterilized hydrogel were cultured in a mold incubator for 4 hours. Then, 8 μL of the gel and bacterial co-culture solution were evenly spread on the LB agar culture plate, and cultured in a mold incubator at 37°C for 18 hours to calculate the number of colonies. The calculation formula for the sterilization rate is as follows: Kill% = (average number of colonies in the control sample - average number of colonies in the sample) / average number of colonies in the control sample x 100%. Among them, the sample without hydrogel was used as the control group. Figure 5 As shown, the bactericidal rate of the hydrogel can reach 86%, indicating that the hydrogel has excellent antibacterial properties.

[0102] 6. Hemostasis performance test

[0103] The hydrogel obtained in Example 1 was tested for hemostatic performance. In the hemostatic performance test study, a rat (normal standard rat, 160-180 g) hemorrhage liver model was used. First, the abdomen of the anesthetized mouse was incised and the liver was exposed. Then, liver bleeding was induced with a needle, and the gel was immediately applied to the bleeding site, and the bleeding time and amount were recorded. The mouse without any hemostatic treatment was used as a blank control, and the mouse with gauze hemostasis was used as a comparative example. The results are shown in Figure 2. Figure 6 As shown. It can be seen that the bleeding volume and hemostasis time of the hydrogel in Example 1 are 43 mg and 24 s, respectively. Compared with the blank control group, the bleeding volume and bleeding time of the hydrogel in Example 1 are reduced by 74.3% and 84.7%, respectively; compared with the mice using gauze to stop bleeding, it also has a significantly faster hemostasis effect. The above results show that the hydrogel in Example 1 has good in vivo hemostasis ability.

Claims

1. A method for preparing a hemostatic hydrogel containing natural polysaccharides, Features: The following steps are involved: (1) Preparing a natural polysaccharide-clay aqueous solution: adding polyphenols and natural polysaccharides to a clay aqueous solution under stirring, and adjusting the pH value of the system to 8-9 to form a natural polysaccharide-clay aqueous solution; (2) Preparing a hydrogel: adding a monomer, an initiator and a cross-linking agent to the natural polysaccharide-clay aqueous solution to form a mixed solution, stirring evenly, and reacting at 10-40° C. for 10-60 minutes to obtain the hemostatic hydrogel containing the natural polysaccharide; Wherein, the natural polysaccharide is at least one of sodium alginate, pectin, cellulose and chitosan.

2. The method for preparing the hemostatic hydrogel containing natural polysaccharides according to claim 1, Features: The polyphenolic substance is at least one of lignin and tannic acid.

3. The method for preparing the hemostatic hydrogel containing natural polysaccharides according to claim 2, Features: The clay is any one of lithium magnesium silicate, aluminum magnesium silicate, kaolin, montmorillonite or hydrotalcite.

4. The method for preparing the hemostatic hydrogel containing natural polysaccharides according to claim 3, Features: The monomer is at least one of acrylamide, methacrylamide and hydroxyethyl methacrylate.

5. The method for preparing the hemostatic hydrogel containing natural polysaccharides according to claim 4, Features: The natural polysaccharide accounts for 10-200% of the mass of the clay, and the polyphenolic substances account for 20-200% of the mass of the clay.

6. The method for preparing the hemostatic hydrogel containing natural polysaccharides according to claim 5, Features: The concentration of the clay aqueous solution is 1-3 wt %.

7. The method for preparing the hemostatic hydrogel containing natural polysaccharides according to claim 6, Features: The clay accounts for 1-10% of the monomer mass.

8. The method for preparing the hemostatic hydrogel containing natural polysaccharides according to claim 7, Features: The initiator accounts for 1-3% of the monomer mass, and the cross-linking agent accounts for 0.1-0.3% of the monomer mass.

9. The method for preparing the hemostatic hydrogel containing natural polysaccharides according to claim 8, Features: The mass ratio of at least one of sodium alginate, pectin and cellulose in the natural polysaccharide to chitosan is 5:(2.5-20).

10. The method for preparing the hemostatic hydrogel containing natural polysaccharides according to claim 8, Features: The initiator is a mixture of ammonium persulfate and tetramethylethylenediamine; and the crosslinking agent is N,N-methylenebisacrylamide.

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