Aerogel material as well as preparation method and application thereof

Abstract

The invention provides an aerogel material as well as a preparation method and application thereof, and belongs to the technical field of biological materials and hemostatic agents. The aerogel material is prepared by carrying out chemical crosslinking on aldehyde hyaluronic acid and epsilon-polylysine in a solution to form self-healing hydrogel, and freeze-drying the self-healing hydrogel. The aerogel material has excellent biocompatibility and biodegradability, shows rapid closed hemostasis and expandable hemostasis in a rat femoral artery cross section and liver punching model, can be easily removed through normal saline after hemostasis, and is free of adhesion and tissue damage. The invention provides an innovative scheme for emergency accident first aid, surgical wound hemostasis and rescue of injured people in war, and is expected to be further applied to precise treatment of complex wounds.

Description

Technical Field

[0001] The present invention relates to the technical field of biomaterials and hemostatic agents, and particularly to an aerogel material, a preparation method thereof, and an application thereof. Background Art

[0002] In recent years, with the continuous progress of medical technology, traumatic hemostatic materials have been more and more widely used in first aid, surgery, and battlefield rescue. Effective hemostatic materials can not only stop bleeding quickly, but also reduce postoperative complications and promote wound healing. Although traditional hemostatic materials meet the clinical needs to a certain extent, there are still some deficiencies, which prompt researchers to continuously explore and develop new hemostatic materials.

[0003] Traditional hemostatic materials mainly include gauze, gelatin sponge, oxidized cellulose, microporous expanded starch, etc. The hemostatic mechanisms of these materials mainly rely on physical barrier effects and the ability to adsorb blood components. However, these materials have the following deficiencies in practical applications: unsatisfactory hemostatic effects, difficult to degrade, not easy to peel off, and physiological toxicity. Summary of the Invention

[0004] The purpose of the present invention is to provide an aerogel material, a preparation method thereof, and an application thereof, so as to solve the technical problems of unsatisfactory hemostatic effects, difficult to degrade, not easy to peel off, and having physiological toxicity of the existing hemostatic materials.

[0005] In order to achieve the above-mentioned invention purpose, the present invention provides the following technical solutions:

[0006] The present invention provides an aerogel material obtained by freeze-drying a hydrogel cross-linked from an aldehyde-modified hyaluronic acid substance and an ε-polylysine substance.

[0007] Further, the aldehyde-modified hyaluronic acid substance includes aldehyde-modified hyaluronic acid and / or sodium aldehyde-modified hyaluronate, and the ε-polylysine substance includes ε-polylysine and / or ε-polylysine hydrochloride.

[0008] Further, the degree of aldehyde modification of the aldehyde-modified hyaluronic acid substance is 10-50%, and the weight-average molecular weight is 5-500 kDa;

[0009] The weight-average molecular weight of the ε-polylysine substance is 1-200 kDa.

[0010] The present invention provides a preparation method of an aerogel material, which is prepared by including the following steps:

[0011] Step 1): Mix a hyaluronic acid substance solution with an oxidant solution and carry out an oxidation reaction, and then sequentially dialyze and freeze-dry the reaction solution to obtain an aldehyde-modified hyaluronic acid substance;

[0012] Step 2): Prepare an aldehyde-functionalized hyaluronic acid-based material solution. Mix the aldehyde-functionalized hyaluronic acid-based material solution and an ε-polylysine-based material solution, and then carry out a cross-linking reaction to obtain a hydrogel. After subjecting the obtained hydrogel to freeze-drying treatment, an aerogel material is obtained.

[0013] Further, in Step 1), the oxidant in the oxidant solution includes one or more of sodium periodate, manganese dioxide, and sodium hypochlorite;

[0014] The concentration of the oxidant solution is 20 - 100 mg / mL, and the dosage ratio of the hyaluronic acid-based material solution to the oxidant solution is 10 - 50:1;

[0015] The temperature of the oxidation is 20 - 60 °C, and the time of the oxidation is 1 - 24 h.

[0016] Further, in Step 1), the time of the freeze-drying is 48 - 96 h.

[0017] Further, in Step 2), the volume ratio of the aldehyde-functionalized hyaluronic acid-based material solution to the ε-polylysine-based material solution is 1 - 8:1;

[0018] The concentration of the aldehyde-functionalized hyaluronic acid-based material solution is 1 - 200 mg / mL, and the concentration of the ε-polylysine-based material solution is 1 - 200 mg / mL.

[0019] Further, the pH of the ε-polylysine-based material solution is 7 - 12.

[0020] Further, in Step 2), the time of the cross-linking reaction is 10 - 600 s, and the temperature of the cross-linking reaction is 23 - 35 °C.

[0021] Further, in Step 2), the time of the freeze-drying is 24 - 72 h.

[0022] The present invention provides a preparation method of the above-mentioned aerogel material.

[0023] The present invention also provides an application of the above-mentioned aerogel material as a hemostatic dressing.

[0024] Advantages of the present invention:

[0025] The aerogel hemostatic material of the present invention has remarkable beneficial effects, including rapid hemostasis ability, excellent biocompatibility, self-healing property, easy removal, good mechanical properties, high water absorbency, applicability to various bleeding situations, simple operation, promotion of wound healing, and biodegradability.

[0026] The aerogel hemostatic material of the present invention can rapidly absorb the moisture in the blood, quickly expand to form a physical barrier, effectively seal the wound, and achieve rapid hemostasis. Since both hyaluronic acid and ε-polylysine are naturally occurring substances in the human body, the material of the present invention exhibits excellent biocompatibility, reducing the possibility of immune response and tissue rejection when used by patients.

[0027] For the aerogel hemostatic material of the present invention, the hydrogel before freeze-drying has self-healing properties. This property enables the material to maintain its hemostatic effect even when slightly damaged. After successful hemostasis, by wetting the material with physiological saline, the viscosity of the aerogel material can be reduced, making it easy to remove and reducing secondary damage to the wound. In addition, the biodegradability of the material means that patients do not need to undergo a second operation to remove it after hemostasis, reducing the physical burden and economic cost of the patients.

[0028] The aerogel hemostatic material of the present invention can not only stop bleeding, but also provide a moist and antibacterial environment for the wound, helping to promote the wound healing process, showing great potential and advantages in clinical applications, and is expected to become a new generation of highly efficient hemostatic materials. Brief Description of the Drawings

[0029] Figure 1 It is a picture of the aerogel material obtained in Example 1.

[0030] Figure 2 It is an application diagram of the aerogel material obtained in Example 1 in the hemostasis experiment of transection of the femoral artery in rats;

[0031] Figure 3 It is a schematic diagram of the non-destructive removal effect of adding water in the hemostasis experiment of transection of the femoral artery in rats using the aerogel material obtained in Example 1;

[0032] Figure 4 It is an application diagram of the aerogel material obtained in Example 1 in the hemostasis experiment of liver perforation in rats. Detailed Description of the Invention

[0033] The present invention provides an aerogel material obtained by freeze-drying a hydrogel crosslinked from an aldehyde-modified hyaluronic acid substance and an ε-polylysine substance.

[0034] In the present invention, the aldehyde-modified hyaluronic acid substance includes aldehyde-modified hyaluronic acid and / or sodium aldehyde-modified hyaluronate, preferably aldehyde-modified hyaluronic acid; the ε-polylysine substance includes ε-polylysine and / or ε-polylysine hydrochloride, preferably ε-polylysine.

[0035] In the present invention, the degree of aldehyde group modification of the aldehyde group-modified hyaluronic acid substances is 10-50%, preferably 20-40%; the weight-average molecular weight is 5-500 kDa, preferably 50-150 kDa;

[0036] The weight-average molecular weight of the ε-polylysine substances is 1-200 kDa, preferably 1-50 kDa.

[0037] In the present invention, the aldehyde group-modified hyaluronic acid substances are white flocculent.

[0038] In the present invention, hyaluronic acid, as a polysaccharide widely present in human tissues, plays an important role in the extracellular matrix. It not only provides necessary physical support for cells but also participates in physiological processes such as cell signal transduction and migration. Aldehyde group-modified hyaluronic acid enhances the chemical cross-linking ability of hyaluronic acid by introducing aldehyde groups, enabling hyaluronic acid molecules to form a three-dimensional cross-linked network structure with amino-containing biological macromolecules, such as ε-polylysine, through the formation of dynamic Schiff base bonds. This unique chemical reaction not only enhances the cross-linking ability of hyaluronic acid but also endows the material with more excellent mechanical properties and plasticity.

[0039] In the present invention, on the one hand, ε-polylysine achieves rapid and effective tissue adhesion through the electrostatic interaction between its unique cationic amino group and the negative charges on the cell membrane. On the other hand, due to the cationic nature of ε-polylysine, it can interact with negatively charged molecules on the bacterial cell membrane, causing changes in the membrane structure and thus affecting the normal physiological functions of bacteria. In the hyaluronic acid-based aerogel hemostatic material prepared in the present invention, in addition to the above two advantages, ε-polylysine, as a natural antibacterial polypeptide, is positively charged and can attract and concentrate blood coagulation factors. Blood coagulation factors are indispensable proteins in the blood coagulation process, and they play a crucial role in the initial stage of wound healing. By concentrating blood coagulation factors, the material can accelerate blood coagulation, thereby achieving rapid hemostasis, which is of great significance for controlling bleeding and preventing infection.

[0040] The present invention provides a preparation method of an aerogel material, which is prepared by including the following steps:

[0041] Step 1): Mix the solution of the aldehyde group-modified hyaluronic acid substances with the oxidant solution and then carry out an oxidation reaction. After that, dialyze and freeze-dry the reaction solution in sequence to obtain the aldehyde group-modified hyaluronic acid substances;

[0042] Step 2): Prepare a solution of the aldehyde group-modified hyaluronic acid substances, mix the solution of the aldehyde group-modified hyaluronic acid substances with the solution of the ε-polylysine substances and then carry out a cross-linking reaction to obtain a hydrogel. After subjecting the obtained hydrogel to freeze-drying treatment, the aerogel material is obtained.

[0043] In the present invention, the solvent in the hyaluronic acid-based substance solution, ε-polylysine-based substance solution, and aldehyde-modified hyaluronic acid-based substance solution is independently one or more of deionized water, sterile water, physiological saline, and phosphate buffer solution, preferably physiological saline.

[0044] In the present invention, in step 1), the oxidation reaction is carried out under light-shielded conditions.

[0045] In the present invention, in step 1), the oxidant in the oxidant solution includes one or more of sodium periodate, manganese dioxide, and sodium hypochlorite, preferably sodium periodate and / or manganese dioxide, and further preferably sodium periodate;

[0046] The concentration of the oxidant solution is 20 - 100 mg / mL, preferably 75 mg / mL, and the dosage ratio of the hyaluronic acid-based substance solution to the oxidant solution is 10 - 50:1, preferably 30:1;

[0047] The temperature of the oxidation is 20 - 60 °C, preferably 30 °C, and the oxidation time is 1 - 24 h, preferably 6 h.

[0048] In the present invention, after the oxidation reaction is completed, polyethylene glycol is added to the reaction solution to neutralize the remaining oxidant, and the reaction solution is continuously stirred to ensure complete neutralization reaction.

[0049] In the present invention, in step 1), the freeze-drying time is 48 - 96 h, preferably 72 h.

[0050] In the present invention, in step 1), the dialysis is to load the reaction solution into a dialysis bag and place it in deionized water for dialysis for 3 days, during which the dialysis water is replaced regularly to ensure that small molecule impurities in the solution are completely removed.

[0051] In the present invention, in step 2), the volume ratio of the aldehyde-modified hyaluronic acid-based substance solution to the ε-polylysine-based substance solution is preferably 1 - 8:1;

[0052] The concentration of the aldehyde-modified hyaluronic acid-based substance solution is preferably 1 - 200 mg / mL, and the concentration of the ε-polylysine-based substance solution is preferably 1 - 200 mg / mL.

[0053] In the present invention, the pH of the ε-polylysine-based substance solution is 7 - 12, preferably 8.5.

[0054] In the present invention, the pH of the ε-polylysine-based substance solution is adjusted using a sodium hydroxide solution, and after the adjustment is completed, it is left standing for a period of time to ensure that small molecule impurities in the solution are completely removed.

[0055] In the present invention, in step 2), the time of the cross-linking reaction is 10 to 600 s, preferably 100 to 400 s, and more preferably 200 s; the temperature of the cross-linking reaction is 23 to 35 °C, preferably 28 to 32 °C, and more preferably 30 °C.

[0056] In the present invention, in step 2), the time of the freeze-drying is 24 to 72 h, preferably 40 to 60 h, and more preferably 48 h.

[0057] The present invention provides an aerogel material prepared by the above preparation method.

[0058] The present invention also provides an application of the above aerogel material as a hemostatic dressing.

[0059] In the present invention, when the aerogel material is used as a hemostatic dressing, the aerogel material is pressed onto the bleeding point. After hemostasis is completed, the aerogel can be wetted with an aqueous solution to reduce the viscosity of the aerogel material, so that it can be easily removed without damage.

[0060] In the present invention, the aqueous solution is preferably one or more of sterile water, physiological saline and phosphate buffer solution, and more preferably physiological saline.

[0061] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0062] Example 1

[0063] Under light-shielded conditions, a sodium periodate solution was slowly dropped into a hyaluronic acid solution with a concentration of 5 mg / mL and continuously stirred to complete the oxidation reaction. After the reaction was completed, ethylene glycol was dropped into the system to neutralize the remaining sodium periodate and stirring was continued. Then the reaction solution was put into a dialysis bag and dialyzed in deionized water for 3 days. The reaction solution was taken out and freeze-dried for 72 h to obtain aldehyde-group modified hyaluronic acid;

[0064] ε-Polylysine hydrochloride was dissolved in deionized water to obtain an ε-polylysine solution with a concentration of 30 mg / mL. Then, a sodium hydroxide solution was added dropwise to the ε-polylysine solution to adjust the pH of the solution to 8.5;

[0065] An aldehyde-group modified hyaluronic acid solution with a concentration of 30 mg / mL was prepared and mixed with an ε-polylysine solution with a pH of 8.5 and a concentration of 30 mg / mL in a volume ratio of 3:1, and a cross-linking reaction was carried out at 25 °C. The time for completing the cross-linking was 22 s. The obtained hydrogel was freeze-dried for 48 h to obtain an aerogel material, denoted as OPA 1 。

[0066] Example 2

[0067] In this embodiment, different from Embodiment 1, the volume ratio of the aldehyde group-modified hyaluronic acid solution to the ε-polylysine solution is 4:1, the time to complete crosslinking is 17 s, and the obtained aerogel material is denoted as OPA 2 。

[0068] Embodiment 3

[0069] In this embodiment, different from Embodiment 1, the volume ratio of the aldehyde group-modified hyaluronic acid solution to the ε-polylysine solution is 5:1, the time to complete crosslinking is 19 s, and the obtained aerogel material is denoted as OPA 3 。

[0070] Embodiment 4

[0071] In this embodiment, different from Embodiment 1, the concentration of the aldehyde group-modified hyaluronic acid solution is 20 mg / mL, and the concentration of the ε-polylysine solution is 20 mg / mL, obtaining a dense hyaluronic acid-based aerogel material

[0072] Embodiment 5

[0073] In this embodiment, different from Embodiment 4, the volume ratio of the aldehyde group-modified hyaluronic acid solution to the ε-polylysine solution is 4:1, obtaining a dense hyaluronic acid-based aerogel material

[0074] Embodiment 6

[0075] In this embodiment, different from Embodiment 4, the volume ratio of the aldehyde group-modified hyaluronic acid solution to the ε-polylysine solution is 5:1, obtaining a dense hyaluronic acid-based aerogel material

[0076] Appropriately amount of water was dropped onto the aerogel materials obtained in Embodiments 1 to 3 respectively, and the absorption rate and absorption amount of the aerogel were observed and recorded. Table 1 shows the absorption rates of the aerogel materials prepared in Embodiments 1 to 3 respectively

[0077] Table 1 Absorption rates of the aerogel materials prepared in Embodiments 1 to 3 respectively

[0078] Example Group Absorption rate / ms 1 <![CDATA[OPA 1 > 1033 2 <![CDATA[OPA 2 > 267 3 <![CDATA[OPA 3 > 667

[0079] Application Example 1

[0080] Transverse Hemostasis Experiment of Rat Inguinal Artery: Prepare 5 male SD rats weighing about 250 g. After anesthetizing the rats, perform transverse section treatment on the inguinal artery. Quickly press the aerogel onto the bleeding point, record the hemostasis time (from the start of pressing to the stop of bleeding), and record the blood loss (using the weighing method). After hemostasis is completed, rinse the aerogel with physiological saline and observe its effect of losing adhesiveness and being easily peeled off. Table 2 shows the hemostasis time and blood loss of rats in the transverse hemostasis of the inguinal artery using the aerogel materials obtained from the blank group (Blank) and Examples 1-3 respectively.

[0081] Table 2 Hemostasis Time and Blood Loss of Rats in the Transverse Hemostasis of the Inguinal Artery Using the Aerogel Materials Obtained from the Blank Group (Blank) and Examples 1-3 Respectively

[0082] Group Hemostasis time / s Blood loss / g Blank 130 4.85 <![CDATA[OPA 1 > 67 0.31 <![CDATA[OPA 2 > 41 0.19 <![CDATA[OPA 3 > 60 0.31

[0083] Application Example 2

[0084] Liver Perforation Hemostasis Experiment of Rats: Prepare 5 male SD rats weighing about 250 g. After anesthetizing the rats, perform liver perforation treatment. Quickly fill the aerogel into the bleeding hole, record the blood loss (using the weighing method). After hemostasis is completed, observe the removal situation of the aerogel and record its effect of being easily removed. Table 3 shows the hemostasis time and blood loss of rats in the liver perforation hemostasis using the aerogel materials prepared from the blank group (Blank) and Examples 1-3 respectively.

[0085] Table 3 Hemostasis Time and Blood Loss of Rats in the Liver Perforation Hemostasis Using the Aerogel Materials Prepared from the Blank Group (Blank) and Examples 1-3 Respectively

[0086]

[0087]

[0088] As can be seen from the above examples, the present invention provides an aerogel material, its preparation method and application. Figure 1 It is a picture of the aerogel material obtained in Example 1. Figure 2 It is an application diagram of the aerogel material obtained in Example 1 in the hemostasis experiment of transverse section treatment of rat inguinal artery. Figure 3 It is a schematic diagram of the non-destructive removal effect of adding water in the hemostasis experiment of transverse section treatment of rat inguinal artery using the aerogel material obtained in Example 1. Figure 4 It is an application diagram of the aerogel material obtained in Example 1 in the liver perforation hemostasis experiment of rats. From Table 1, Table 2 and Table 3 and Figures 1 - 4It can be seen that the aerogel material prepared by the present invention has high water absorbency and rapid hemostatic ability, can rapidly expand to form a physical barrier, effectively seal the wound, and achieve rapid hemostasis. The present invention uses naturally occurring hyaluronic acid and ε-polylysine in the human body to prepare an aerogel hemostatic material, which not only has excellent biocompatibility and self-healing properties, but also can control the mechanical properties and degradation rate of the hydrogel by adjusting the ratio of hyaluronic acid and cross-linking agent. The aerogel material prepared thereby has different mechanical properties to meet different clinical needs. It can be seen that the present invention solves the technical problems of the existing hemostatic materials, such as unsatisfactory hemostatic effect, difficult degradation, difficult peeling, and physiological toxicity, and has important significance for applications in first aid, surgery, and battlefield rescue.

[0089] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. An aerogel material, characterized in that: The hydrogel is prepared by cross-linking aldehyde-modified hyaluronic acid and ε-polylysine and then freeze-drying.

2. An aerogel material according to claim 1, characterized in that: The aldehyde-hydrated hyaluronic acid substances include aldehyde-hydrated hyaluronic acid and / or aldehyde-hydrated sodium hyaluronate, and the ε-polylysine substances include ε-polylysine and / or ε-polylysine hydrochloride.

3. An aerogel material according to claim 1 or 2, characterized in that: The degree of hydroformylation of the hydroformyl hyaluronic acid substance is 10-50%, and the weight average molecular weight is 5-500 kDa; The weight average molecular weight of the ε-polylysine substance is 1-200 kDa.

4. The method for preparing an aerogel material according to any one of claims 1 to 3, characterized in that: The method is prepared by comprising the following steps: Step 1), mixing the hyaluronic acid substance solution and the oxidant solution and performing an oxidation reaction, and then dialyzing and freeze-drying the reaction solution in sequence to obtain the aldehyde-modified hyaluronic acid substance; Step 2), prepare a aldehyde-modified hyaluronic acid substance solution, mix the aldehyde-modified hyaluronic acid substance solution and the ε-polylysine substance solution, and then perform a cross-linking reaction to obtain a hydrogel, and freeze-dry the obtained hydrogel to obtain an aerogel material.

5. The method for preparing an aerogel material according to claim 4, characterized in that: In step 1), the oxidant in the oxidant solution includes one or more of sodium periodate, manganese dioxide and sodium hypochlorite; The concentration of the oxidant solution is 20 to 100 mg / mL, and the dosage ratio of the hyaluronic acid solution to the oxidant solution is 10 to 50:1; The oxidation temperature is 20-60° C., and the oxidation time is 1-24 hours. The freeze-drying time is 48 to 96 hours.

6. The method for preparing an aerogel material according to claim 1 or 5, characterized in that: In step 2), the volume ratio of the aldehyded hyaluronic acid substance solution to the ε-polylysine substance solution is 1 to 8:1; The concentration of the aldehyde-modified hyaluronic acid substance solution is 1-200 mg / mL, and the concentration of the ε-polylysine substance solution is 1-200 mg / mL.

7. The method for preparing an aerogel material according to claim 4 or 5, characterized in that: The pH of the ε-polylysine substance solution is 7-12.

8. The method for preparing an aerogel material according to claim 6, characterized in that: In step 2), the cross-linking reaction time is 10 to 600 seconds, and the cross-linking reaction temperature is 23 to 35°C.

9. The method for preparing an aerogel material according to claim 7, characterized in that: In step 2), the freeze-drying time is 24 to 72 hours.

10. Use of the aerogel material according to any one of claims 1 to 3 as a hemostatic dressing.

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