CN113842494A

CN113842494A - Injectable hemostatic crystal gel for promoting tissue regeneration and preparation method and application thereof

Info

Publication number: CN113842494A
Application number: CN202111062691.4A
Authority: CN
Inventors: 范代娣; 米钰; 马晨辉; 蒋弘波; 朱晨辉; 马沛; 段志广
Original assignee: Northwest University
Current assignee: Northwest University
Priority date: 2021-09-10
Filing date: 2021-09-10
Publication date: 2021-12-28
Anticipated expiration: 2041-09-10
Also published as: CN113842494B

Abstract

The invention discloses an injectable hemostatic crystal gel for promoting tissue regeneration, a preparation method and application thereof, wherein the preparation method comprises the following steps: ultrasonically dispersing polydopamine, chitosan and collagen in deionized water, adding glacial acetic acid, and stirring until the polydopamine, the chitosan and the collagen are dissolved; dissolving oxidized dextran in deionized water, adding the oxidized dextran solution into the mixed solution under vigorous stirring, transferring to a refrigerator for reaction, and melting at room temperature to obtain the injectable hemostatic crystal gel for promoting tissue regeneration. The crystal gel has the aperture of 50-200 mu m, has a mutually connected macroporous structure and stable mechanical strength, can be conveyed to a narrow and deep wound by an injector after being compressed and fixed, quickly restores the shape after blood suction to block the wound and form a physical barrier, and activates the blood coagulation process by concentrated blood to achieve the effect of stopping bleeding. The crystal gel prepared by the invention has the advantages of rapid shape recovery capability, injectability, good hemostatic capability and the like.

Description

Injectable hemostatic crystal gel for promoting tissue regeneration and preparation method and application thereof

Technical Field

The invention belongs to the technical field of biomedical materials, and particularly relates to an injectable hemostatic crystal gel for promoting tissue regeneration, and a preparation method and application thereof.

Background

Bleeding complications from trauma, surgery, congenital diseases or drug-induced hematological disorders result in significant morbidity and mortality. Bleeding has been reported to result in over 30% traumatic mortality. Therefore, the hemostatic bag has extremely important significance in timely hemostasis in emergency situations such as battlefields, operations, car accidents and the like. In these emergency situations, rapid hemostasis cannot be achieved by the body's hemostatic mechanisms alone. Thus, there is a need for suitable hemostatic agents to promote, enhance, compensate for, or mimic the natural mechanisms of hemostasis. Among the deaths caused by bleeding, non-compression bleeding accounted for 50% of them. Wounds caused by bullets, bombs and explosives in a battlefield are irregular in shape and cannot be pressed, and the conventional hemostasis method only fills gauze in the wounds, but needs secondary operations to remove the gauze, so that secondary damage is caused to patients. The hemostatic applied to the surface of a wound can achieve a good hemostatic effect, but cannot be applied to non-compressible, deep and narrow bleeding wounds, and in addition, the traditional pressurization-assisted hemostatic method is not suitable for puncture wounds and wounds in the axillary or inguinal regions. In addition, large-area wounds are often difficult to repair, and health is seriously affected. Therefore, the prepared hemostatic material which can rapidly treat deep and narrow non-compressive hemorrhage, can be degraded in vivo and has the capability of promoting wound repair has extremely important clinical significance.

The crystalloid glue with interconnected macroporous structure and stable mechanical strength can be compressed and fixed and then delivered to narrow and deep wounds through a syringe, and the fixed crystalloid glue can quickly recover to the shape after blood suction to block bleeding wounds and form a physical barrier to stop bleeding. The conventional crosslinking mode for preparing the crystal gel is free radical polymerization, EDC/NHS crosslinked protein, or Michael addition and the like, so that not only are complicated vinyl functionalization carried out on natural polymers such as gelatin, sodium alginate, chitosan and the like required, but also residual crosslinking agents such as APS/TEMED and the like have obvious cytotoxicity. Chitosan cryogels may be formed from dialdehyde crosslinking agents such as glutaraldehyde and glyoxal. However, their high reactivity and toxicity limit their use as cross-linking agents for biomedical systems. Therefore, a safe crosslinking agent is needed to prepare the desired crystal gel.

Disclosure of Invention

The invention aims to overcome the problems in the prior art and provides an injectable hemostatic crystal gel for promoting tissue regeneration as well as a preparation method and application thereof. The crystal glue can rapidly treat non-compression bleeding with narrow depth and large deep lethal bleeding amount. Can be degraded in vivo, and can promote tissue regeneration. The raw materials are low in price, the process is simple, the biocompatibility is good, the shape memory capacity is good, and a physical barrier can be formed when the raw materials contact blood to block bleeding wounds.

In order to solve the technical problems, the invention adopts the technical scheme that: a preparation method of an injectable hemostatic crystal gel for promoting tissue regeneration is characterized by comprising the following steps:

step one, adding collagen and chitosan into deionized water, and uniformly dispersing to obtain a mixed solution; or adding collagen, chitosan and polydopamine into deionized water, and uniformly dispersing to obtain a mixed solution;

step two, adding glacial acetic acid into the mixed solution obtained in the step one, and stirring to completely dissolve the chitosan; then adding oxidized dextran solution and mixing uniformly; and placing the uniformly mixed solution in a refrigerator for reaction, and thawing at normal temperature after the reaction is finished to obtain the injectable hemostatic crystal gel for promoting tissue regeneration.

The preparation method of the injectable hemostatic crystal gel for promoting tissue regeneration is characterized in that in the first step, the mass percentage content of chitosan in the mixed solution is 0.5-1%, and the content of polydopamine in the mixed solution is not more than 2 mg/mL; the mass ratio of the collagen to the chitosan is 1 (0.5-2); the mass ratio of the oxidized glucan in the second step to the chitosan in the first step is (0.1-1): 1.

The preparation method of the injectable hemostatic crystal gel for promoting tissue regeneration is characterized in that the reaction temperature in the step two is-8 ℃ to-20 ℃, and the reaction time is 8h to 36 h.

The preparation method of the injectable hemostatic gel for promoting tissue regeneration is characterized in that the preparation method of polydopamine in the first step comprises the following steps: uniformly mixing ammonia water, absolute ethyl alcohol and deionized water, reacting at room temperature for 10-30 min, then adding a dopamine hydrochloride solution, reacting at room temperature for 12-24 h, and centrifuging to obtain the polydopamine.

The preparation method of the injectable hemostatic crystal gel for promoting tissue regeneration is characterized in that the form of the polydopamine is spherical with the diameter of 200-300 nm.

The preparation method of the injectable hemostatic crystal gel for promoting tissue regeneration is characterized in that the volume ratio of ammonia water, absolute ethyl alcohol and deionized water is (0.5-1.5) to 20 (30-45), the concentration of a dopamine hydrochloride solution is 20 mg/mL-50 mg/mL, and the volume ratio of the dopamine hydrochloride solution to water is 1 (5-10).

The preparation method of the injectable hemostatic gel for promoting tissue regeneration is characterized in that the preparation method of oxidized dextran in the second step comprises the following steps: dissolving glucan in deionized water, adding sodium periodate, and reacting for 18-24 h at room temperature in a dark place; then adding ethylene glycol to stop further oxidation of glucan, continuously stirring for 1-2 h, and then dialyzing the solution in a dialysis bag by using water until the solution is colorless, and then carrying out freeze drying to obtain the oxidized glucan.

The preparation method of the injectable hemostatic crystal gel for promoting tissue regeneration is characterized in that the molar ratio of dextran to sodium periodate is 1 (0.5-1), and the molar ratio of sodium periodate to glycol is 1 (0.5-1); the molecular weight of the dialysis bag is 8000-12000.

Further, the invention also provides an injectable hemostatic crystal gel for promoting tissue regeneration, which is prepared according to the method.

Further, the invention also provides application of the injectable hemostatic crystal gel for promoting tissue regeneration prepared by the method in hemostatic materials.

Compared with the prior art, the invention has the following advantages:

1. the invention adopts collagen, chitosan, polydopamine and oxidized dextran, the hemostatic activity of the chitosan is mainly derived from positive charges on the surface of the chitosan, and the characteristic does not depend on the coagulation mechanism of the chitosan, so the chitosan hemostatic agent can be suitable for patients suffering from blood coagulation disorder. Collagen, chitosan and oxidized dextran can mimic the extracellular matrix, promoting wound healing. The catechol group carried by polydopamine provides tissue adhesion and may increase the efficiency of coagulation in vivo.

2. The raw materials in the invention have good biocompatibility and can be degraded in vivo, and no new cross-linking agent is added in the gel, thereby further ensuring the non-toxicity and non-side effects of the material.

3. The preparation method has the advantages of cheap and easily-obtained raw materials, low cost and simple synthesis steps, can quickly expand to form a physical barrier to prevent bleeding when contacting blood, and has high blood absorption capacity and expansion coefficient.

4. Compared with other shape memory hemostatic materials, the crystal gel prepared by the invention has higher recovery speed and higher swelling rate, the crystal gel prepared by the invention is compressed to 80% of the volume after being swelled and balanced, the recovery time in deionized water is 1.8 s-9.7 s, and the swelling rate is 4500% -6000% after being swelled and balanced in deionized water. Rapid shape recovery can more rapidly occlude bleeding wounds, apply pressure around the wound and physically occlude hemostasis. The high swelling ratio can better concentrate the blood coagulation factors, thereby rapidly stopping bleeding.

5. The aperture of the crystal glue is 50 mu m _～200 μm, has interconnected macroporous structure and stable mechanical strength, can be delivered to narrow and deep wounds by a syringe after compression fixation, rapidly recovers shape after blood suction to block the wound and form a physical barrier, and achieves the effect of stopping bleeding by concentrating blood and activating a blood coagulation process, and can be used as a hemostatic material.

The technical solution of the present invention will be described in further detail with reference to the following detailed description and accompanying drawings.

Drawings

Fig. 1 is an SEM image of polydopamine prepared in example 1 of the present invention.

FIG. 2 is a FTIR plot of oxidized dextran prepared in example 4 of the present invention.

FIG. 3 is a graph of oxidized dextran prepared in example 4 of the present invention¹HNMR map.

FIG. 4 shows the swelling ratios of the crystal gels prepared in examples 10 to 14 of the present invention.

FIG. 5 is a graph of compressive strain for the gels prepared in examples 10-14 of the present invention.

FIG. 6 is a graph of whole blood clots of cryogels prepared in examples 10-14 of the present invention.

FIG. 7 shows the hemolysis ratio of the gels prepared in examples 10-14 of the present invention.

FIG. 8 is a diagram showing the results of MTT assay for cytotoxicity of the gels prepared in examples 10 to 14 of the present invention.

Fig. 9 is a photograph of a wound bed when hemostasis is performed in vivo with different hemostatic materials.

FIG. 10 is a graph of the amount of bleeding from hemostasis in vivo using different hemostatic materials.

Fig. 11 is a graph showing the repairing effect of the crystal glue prepared in example 11 and example 13 of the present invention on the rat full cortex defect wound.

FIG. 12 is a graph showing the degradation of the crystal gels prepared in examples 11 and 13 of the present invention in lysozyme.

Detailed Description

Preparation of polydopamine

Example 1

1mL of an aqueous ammonia solution, 20mL of absolute ethanol, and 45mL of deionized water were stirred at room temperature for 20 minutes. Then, 5mL of a 50mg/mL dopamine hydrochloride solution was rapidly added to the above mixed solution. After 1 minute, the solution became light yellow in color. After vigorous stirring at room temperature for 20 hours, the resulting solution turned dark brown. And finally, centrifuging to obtain the poly-dopamine nano-particles, and washing for 3 times by using deionized water. The product is detected by scanning electron microscope to be nanosphere with the particle size of about 200 nm-300 nm, as shown in figure 1.

Example 2

1.5mL of an aqueous ammonia solution, 20mL of absolute ethanol, and 40mL of deionized water were stirred at room temperature for 30 minutes. Then, 4mL of a dopamine hydrochloride solution having a concentration of 40mg/mL was rapidly added to the above mixed solution. After 1 minute, the solution became light yellow in color. After 12 hours of vigorous stirring at room temperature, the resulting solution turned dark brown. And finally, centrifuging to obtain the poly-dopamine nano-particles, and washing for 3 times by using deionized water. Its physicochemical properties were similar to those of example 1.

Example 3

0.5mL of an aqueous ammonia solution, 20mL of absolute ethanol, and 30mL of deionized water were stirred at room temperature for 10 minutes. Then, 6mL of a dopamine hydrochloride solution having a concentration of 20mg/mL was rapidly added to the above mixed solution. After 1 minute, the solution became light yellow in color. After vigorous stirring at room temperature for 24 hours, the resulting solution turned dark brown. And finally, centrifuging to obtain the poly-dopamine nano-particles, and washing for 3 times by using deionized water. Its physicochemical properties were similar to those of example 1.

Preparation of oxidized dextran

Example 4

4g of dextran of 70000 molecular weight and 3.4g of sodium periodate were dissolved in 50mL of deionized water with magnetic stirring. The mixture was left to react for 24h at room temperature in the absence of light, and the solution was pale yellow. 1g of ethylene glycol was then added to react with the mixture for 2h to terminate further oxidation of the glucan. And (3) continuously dialyzing the product with deionized water for 3 days (the cut-off molecular weight of a dialysis bag is 8000-12000), replacing the dialysis water for 5 times every day, and then freeze-drying the solution in the dialysis bag to obtain the oxidized glucan.

Fourier transform infrared absorption Spectroscopy analysis of oxidized dextran prepared in this exampleOxidized dextran at 1732cm^-1There was a carbonyl stretch peak (confirming successful oxidative modification of dextran) as shown in fig. 2 and 3. By passing¹The oxidation of dextran, a weak resonance generated by oxidized dextran at 9.6ppm, which is attributed to the aldehyde proton, was confirmed qualitatively by H-NMR analysis, and the absence of these peaks on the dextran spectrum, which confirms the oxidation of dextran.

Example 5

4g of dextran of 70000 molecular weight and 2.6g of sodium periodate were dissolved in 50mL of deionized water with magnetic stirring. The mixture was left to react for 18h at room temperature in the absence of light, and the solution was pale yellow. 0.6g of ethylene glycol was then added to react with the mixture for 1h to terminate further oxidation of the glucan. And (3) continuously dialyzing the product with deionized water for 3 days (the cut-off molecular weight of a dialysis bag is 8000-12000), replacing the dialysis water for 5 times every day, and then freeze-drying the solution in the dialysis bag to obtain the oxidized glucan.

The oxidized dextran prepared in this example was analyzed by Fourier transform infrared absorption Spectroscopy at 1732cm^-1There was a carbonyl stretch peak (confirming successful oxidative modification of dextran). By passing¹The oxidation of dextran, a weak resonance generated by oxidized dextran at 9.6ppm, which is attributed to the aldehyde proton, was confirmed qualitatively by H-NMR analysis, and the absence of these peaks on the dextran spectrum, which confirms the oxidation of dextran.

Example 6

4g of dextran of 70000 molecular weight and 5.3g of sodium periodate were dissolved in 50mL of deionized water with magnetic stirring. The mixture was left to react for 20h at room temperature in the absence of light, and the solution was pale yellow. 0.8g of ethylene glycol was then added to react with the mixture for 1.5h to stop further oxidation of the glucan. And (3) continuously dialyzing the product with deionized water for 3 days (the cut-off molecular weight of a dialysis bag is 8000-12000), replacing the dialysis water for 5 times every day, and then freeze-drying the solution in the dialysis bag to obtain the oxidized glucan.

Preparation of crystal glue

Example 7

Step one, adding 0.05g of collagen and 0.1g of chitosan into deionized water, and stirring until the collagen is completely dissolved to obtain 10mL of mixed solution;

and step two, adding 50 mu L of glacial acetic acid into the mixed solution until chitosan is uniformly dissolved, dissolving 1g of oxidized dextran prepared in the embodiment 4 into 10mL of deionized water, adding 100 mu L of oxidized dextran solution into the chitosan mixed solution under vigorous stirring, then reacting for 8h at-8 ℃, and unfreezing at normal temperature to obtain the injectable hemostatic gel for promoting tissue regeneration.

Example 8

Step one, adding 0.1g of collagen and 0.1g of chitosan into deionized water, and stirring until the collagen is completely dissolved to obtain 10mL of mixed solution;

and step two, adding 50 mu L of glacial acetic acid into the mixed solution until chitosan is uniformly dissolved, dissolving 1g of oxidized dextran prepared in the embodiment 4 into 10mL of deionized water, adding 200 mu L of oxidized dextran solution into the chitosan mixed solution under vigorous stirring, then reacting for 16h at-12 ℃, and unfreezing at normal temperature to obtain the injectable hemostatic gel for promoting tissue regeneration.

Example 9

Step one, adding 0.1g of collagen and 0.05g of chitosan into deionized water, and stirring until the collagen is completely dissolved to obtain 10mL of mixed solution;

and step two, adding 50 mu L of glacial acetic acid into the mixed solution until chitosan is uniformly dissolved, dissolving 1g of oxidized dextran prepared in the embodiment 4 into 10mL of deionized water, adding 500 mu L of oxidized dextran solution into the chitosan mixed solution under vigorous stirring, then reacting the mixture at-18 ℃ for 24h, and unfreezing the mixture at normal temperature to obtain the injectable hemostatic gel for promoting tissue regeneration.

Example 10

Step one, adding 0.05g of collagen, 0.1g of chitosan and 2.5mg of polydopamine into deionized water, performing ultrasonic treatment and stirring until the collagen and the polydopamine are dissolved to obtain 10mL of mixed solution;

and step two, adding 50 mu L of glacial acetic acid into the mixed solution until chitosan is uniformly dissolved, dissolving 1g of oxidized dextran prepared in the embodiment 4 into 10mL of deionized water, adding 100 mu L of oxidized dextran solution into the chitosan mixed solution under vigorous stirring, then reacting for 36h at-20 ℃, and unfreezing at normal temperature to obtain the injectable hemostatic gel for promoting tissue regeneration. Named CS/HLC/PDA 0.25.

The polydopamine used in this example may be polydopamine prepared in any of examples 1 to 3, or polydopamine having a commercially available particle size of about 200nm to 300 nm.

Example 11

And (3) controlling the final concentration of PDA (polydopamine) in the mixed solution in the first step to be 0mg/mL, and obtaining the CS/HLC crystal gel under the other conditions in the same way as in the example 10.

Example 12

And (3) controlling the final concentration of the PDA in the mixed solution in the first step to be 0.5mg/mL, and obtaining the CS/HLC/PDA0.5 crystal gel under the other conditions in the same way as the example 10.

Example 13

And (3) controlling the final concentration of the PDA in the mixed solution in the first step to be 1.0mg/mL, and obtaining the CS/HLC/PDA1.0 crystal gel under the same other conditions as the example 10.

Example 14

And (3) controlling the final concentration of the PDA in the mixed solution in the first step to be 2.0mg/mL, and obtaining the CS/HLC/PDA2.0 crystal gel under the same other conditions as the example 10.

The gel prepared in examples 7-10 of the present invention was measured for its swelling rate in deionized water at equilibrium, as shown in fig. 4, a high swelling rate can rapidly absorb wound exudate and blood, reduce bacterial infection, and also contribute to the concentration of blood coagulation factors, thereby increasing the hemostasis rate. As shown in table 1, as the content of polydopamine increases, the time for the gel to recover in deionized water increases, and the gel can expand rapidly in an emergency situation in response to bleeding, completely fill the wound, form a physical barrier, and prevent blood from flowing out.

TABLE 1 recovery time in Water for the crystal gels prepared in examples 7-10 of the present invention

The compression strengths of the crystal gels prepared in examples 10 to 14 were measured by a universal tester, and the results are shown in FIG. 5. Because reactive quinone groups carried by polydopamine can form Schiff base bonds with amino groups on chitosan and collagen, the cross-linking density of the polydopamine is increased by adding the polydopamine, and the mechanical properties of the crystal gel are gradually increased along with the continuous increase of the polydopamine content.

Fig. 6 is a cryogel in vitro coagulation performance test. The coagulation performance is measured by testing the dynamic whole blood coagulation index of the crystal gel, and the higher the coagulation index is, the worse the coagulation performance is. As can be seen from the results of fig. 6, the blank group, the gelatin sponge group and the gauze group still exhibited higher blood coagulation indexes after 150 seconds, whereas all the gel groups exhibited lower blood coagulation indexes at the same time point than the gelatin sponge and the gauze groups.

After the erythrocytes are treated by the crystal gel suspension with the concentration of 0.5mg/mL to 4mg/mL, the hemolysis rate is lower than 5 percent, which indicates that the prepared crystal gel has good blood compatibility. As shown in fig. 7.

Fig. 8 is a cell compatibility experiment of the gels prepared in examples 10-14, and the cell survival rates were all above 95% after 24, 48, and 72 hours of culture, demonstrating that the gels prepared have good biocompatibility.

Rats were anesthetized by injection of 1mL 10% chloral hydrate. Prepared on rat liver with punch (5X 3 mm)²) As a wound model for non-pressure bleeding. The gel prepared in example 11 was immediately inserted into a cylindrical wound and the amount of blood loss was recorded. Commercial gauze, gelatin sponge and chitosan hemostatic powder were used as controls. Fig. 9 is a photograph of a wound surface during hemostasis, and fig. 10 is a photograph of bleeding amount of a rat, which shows that the crystal gel prepared by the invention has good hemostasis capability.

Fig. 11 is a study on the repairing effect of the crystal glue on the rat full cortex defect wound. As can be seen from the results in FIG. 11, the CS/HLC and CS/HLC/PDA1.0 crystal gels showed significant differences in wound healing after 3 days of treatment compared to the blank. After 7 days of treatment, the CS/HLC and CS/HLC/PDA1.0 crystal gels show higher wound healing rate than the blank, and the wound healing rate of the CS/HLC/PDA1.0 crystal gel group is obviously higher than that of the CS/HLC group. After 13 days of treatment, the CS/HLC and CS/HLC/PDA1.0 crystal gels showed 100% healing rate, while the blank did not heal completely. The crystal gel has interconnected macroporous structures, which is helpful for accelerating wound repair by promoting vascularization, and the crystal gel added with polydopamine has antioxidant activity, can eliminate active oxygen excessively generated in the inflammatory reaction process, and accelerates wound healing.

FIG. 12 is a graph showing the degradation of the gel in 5mg/mL lysozyme, indicating that the gel can be degraded by lysozyme, and the addition of polydopamine increases the crosslinking density, resulting in a slower degradation rate. The crystal gel can be degraded by lysozyme in body fluid.

The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, changes and equivalent structural changes made to the above embodiment according to the technical spirit of the present invention still fall within the protection scope of the technical solution of the present invention.

Claims

1. A preparation method of an injectable hemostatic crystal gel for promoting tissue regeneration is characterized by comprising the following steps:

2. The preparation method of the injectable hemostatic crystal gel for promoting tissue regeneration according to claim 1, wherein in the first step, the mass percentage of chitosan in the mixed solution is 0.5% -1%, and the content of polydopamine in the mixed solution is not more than 2 mg/mL; the mass ratio of the collagen to the chitosan is 1 (0.5-2); the mass ratio of the oxidized glucan in the second step to the chitosan in the first step is (0.1-1): 1.

3. The method for preparing an injectable hemostatic crystal gel for promoting tissue regeneration according to claim 1, wherein the reaction temperature in step two is-8 ℃ to-20 ℃ and the reaction time is 8h to 36 h.

4. The method for preparing an injectable hemostatic gel for promoting tissue regeneration according to claim 1, wherein the method for preparing polydopamine in step one comprises: uniformly mixing ammonia water, absolute ethyl alcohol and deionized water, reacting at room temperature for 10-30 min, then adding a dopamine hydrochloride solution, reacting at room temperature for 12-24 h, and centrifuging to obtain the polydopamine.

5. The method for preparing an injectable hemostatic crystal gel for promoting tissue regeneration according to claim 1, wherein the form of polydopamine is spherical with a diameter of 200nm to 300 nm.

6. The preparation method of the injectable hemostatic crystal gel for promoting tissue regeneration according to claim 4, wherein the volume ratio of ammonia water, absolute ethyl alcohol and deionized water is (0.5-1.5) to 20 (30-45), the concentration of the dopamine hydrochloride solution is 20 mg/mL-50 mg/mL, and the volume ratio of the dopamine hydrochloride solution to water is 1 (5-10).

7. The method for preparing an injectable hemostatic gel for promoting tissue regeneration according to claim 1, wherein the method for preparing oxidized dextran in step two comprises: dissolving glucan in deionized water, adding sodium periodate, and reacting for 18-24 h at room temperature in a dark place; then adding ethylene glycol to stop further oxidation of glucan, continuously stirring for 1-2 h, and then dialyzing the solution in a dialysis bag by using water until the solution is colorless, and then carrying out freeze drying to obtain the oxidized glucan.

8. The method for preparing an injectable hemostatic crystal gel for promoting tissue regeneration according to claim 7, wherein the molar ratio of dextran to sodium periodate is 1 (0.5-1), and the molar ratio of sodium periodate to ethylene glycol is 1 (0.5-1); the molecular weight of the dialysis bag is 8000-12000.

9. An injectable haemostatic gel prepared according to the method of any of claims 1 to 8 for promoting tissue regeneration.

10. Use of an injectable haemostatic gel prepared according to the method of any of claims 1 to 8 to promote tissue regeneration in a haemostatic material.