Bio-based antibacterial hemostatic sponge as well as preparation method and application thereof

Abstract

The invention relates to the technical field of biomedical materials, and discloses a bio-based antibacterial hemostatic sponge and a preparation method and application thereof.The hemostatic sponge is prepared by taking chitosan as a main raw material, conducting cross-linking modification through a main cross-linking agent and an auxiliary cross-linking agent, then conducting freeze drying and loading a ginseng water extract, the bio-based antibacterial hemostatic sponge prepared by the method is chitosan-based aerogel sponge loaded with the ginseng aqueous extract, the ginseng aqueous extract can accelerate the hemostatic process by increasing the number of platelets, and the sponge can be endowed with good broad-spectrum antibacterial performance due to existence of guanidine antibacterial groups in a main cross-linking agent structure; the structure of the auxiliary cross-linking agent contains a large number of hydrophilic ether bonds, hydroxyl groups and the like, so that the water absorption rate of the sponge can be improved, a dry healing environment can be continuously kept at the wound, and the healing of the wound can be accelerated.

Description

Technical Field

[0001] The invention relates to the technical field of biomedical materials, and in particular to a bio-based antibacterial hemostatic sponge and a preparation method and application thereof. Background Art

[0002] Hemostatic sponge is a tiny sponge with strong water absorption that can directly act on the human body. In the field of modern medicine, hemostatic sponge is an important auxiliary material and is widely used in surgical operations, emergency hemostasis and trauma treatment. It can not only effectively control bleeding, but also promote wound healing and reduce the risk of postoperative infection.

[0003] In surgical operations, hemostatic sponges are often used for smaller bleeding points, such as skin incisions, mucosal incisions, etc., as well as for bleeding control during tissue or organ separation. In addition, hemostatic sponges can also be used in hemostatic powders to further improve the hemostatic effect. In terms of emergency hemostasis, hemostatic sponges can quickly absorb blood and tissue exudate, accelerate wound healing, and are suitable for occasions such as shallow burns, scalds, and accidental trauma.

[0004] Chitosan sponge is a new type of hemostatic material developed in recent years, which has many unique advantages. Chitosan is a natural polymer material with good biocompatibility and degradability. It can be degraded into safe and non-toxic products in the human body. Since chitosan can form a uniform chitosan gel layer on the wound surface, it plays a role in protecting the wound surface and stopping bleeding. Therefore, chitosan has been widely used in the biomedical field.

[0005] However, the antibacterial effect of chitosan itself is not very good, and it cannot completely prevent wound infection problems caused by possible bacterial growth. Moreover, the hemostatic effect of chitosan is not good, and it is difficult to achieve rapid hemostasis of wounds. This makes it difficult to use chitosan directly as a hemostatic sponge. Therefore, the preparation of chitosan sponge with rapid hemostasis and excellent antibacterial properties is of great significance for its application in the field of medical devices. Summary of the invention

[0006] 1. Technical issues to be resolved

[0007] In view of the deficiencies in the prior art, the present invention provides a bio-based antibacterial hemostatic sponge and a preparation method and application thereof.

[0008] (II) Technical solution

[0009] A bio-based antibacterial hemostatic sponge is made of the following raw materials in parts by weight:

[0010]

[0011] The main cross-linking agent is a tetramethylguanidine derivative;

[0012] The auxiliary cross-linking agent is a ring-opening polymer of a double-terminal amine compound and a double-terminal glycidyl ether monomer.

[0013] As a further embodiment of the present invention, the deacetylation degree of the chitosan is ≥95%.

[0014] As a further embodiment of the present invention, the preparation method of the main cross-linking agent is as follows:

[0015] Add bromosuccinic acid, 1,1,3,3-tetramethylguanidine and toluene solvent to a reactor filled with nitrogen. After the addition, start stirring. After a uniform mixed liquid is formed, add an alkaline hydroxide aqueous solution dropwise to the reactor. After the addition is completed, start heating and gradually increase the temperature to 60-70° C. After heat preservation for 12-18 hours, evaporate the solvent, separate the product, and obtain the main cross-linking agent.

[0016] As a further embodiment of the present invention, the alkaline hydroxide is potassium hydroxide or sodium hydroxide.

[0017] As a further embodiment of the present invention, the mass fraction of the alkaline hydroxide aqueous solution is 20-30%.

[0018] Specifically, under the catalytic action of alkaline hydroxide, the halogen substituents in the bromosuccinic acid structure can be replaced by 1,1,3,3-tetramethylguanidine to obtain a guanidine derivative containing two equivalents of active carboxyl substituents in the structure, namely the main cross-linking agent.

[0019] As a further embodiment of the present invention, the preparation method of the auxiliary cross-linking agent is as follows:

[0020] After the double-terminal amine, double-terminal glycidyl ether monomer and 1,4-dioxane are evenly mixed, they are poured into a polymerization kettle, nitrogen is passed to deoxygenate, and then the heating rate is controlled to be 2-3°C / min, and the temperature is slowly increased to 70-80°C. After heat preservation and polymerization for 8-12 hours, the solvent is evaporated under reduced pressure, and the product is separated and collected to obtain the auxiliary cross-linking agent.

[0021] As a further embodiment of the present invention, the double-terminated aminated compound is 1,8-diamino-3,6-dioxaoctane or 2,2'-oxybis(ethylamine); the double-terminated glycidyl ether monomer is any one of ethylene glycol diglycidyl ether, dipropylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether or 1,2-cyclohexanediol diglycidyl ether.

[0022] As a further embodiment of the present invention, the catalyst is p-toluenesulfonic acid or trifluoromethanesulfonic acid.

[0023] In the above technical scheme, under high temperature conditions, the terminal amine group in the double-terminal amine structure can undergo continuous ring-opening addition reaction with the terminal glycidyl ether group in the double-terminal glycidyl ether monomer structure to produce a polyether macromolecular substance with an alternating block structure, namely, an auxiliary cross-linking agent.

[0024] A method for preparing a bio-based antibacterial hemostatic sponge comprises the following steps:

[0025] The first step is to dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 1-3% to form a chitosan solution with a mass fraction of 15-25%;

[0026] The second step is to add the main cross-linking agent, the auxiliary cross-linking agent and the catalyst to the chitosan solution, start stirring, and after a uniform mixed solution is formed, the temperature is raised to 90-100°C, and the mixture is kept warm for 12-24 hours under continuous stirring, and then the heating is stopped and allowed to cool naturally. The material is poured into a mold and placed for 12-18 hours, and then transferred to a freeze dryer, freeze-dried at -40°C for 24-48 hours, and taken out to form a sponge base material;

[0027] The third step is to mix the ginseng water extract with purified water, stir evenly to form a ginseng extract aqueous solution, then add the sponge base material, ultrasonically treat it at an ultrasonic frequency of 80-100kHz for 4-6 hours, take it out, and let it dry naturally to obtain a bio-based antibacterial hemostatic sponge.

[0028] Specifically, the active amino groups in the chitosan structure and the active hydroxyl functional groups in the auxiliary cross-linking agent structure produced by the ring-opening reaction can undergo condensation reactions with the active carboxyl substituents in the main cross-linking agent structure under the catalytic action of the catalyst, thereby achieving cross-linking among the three. The product is allowed to stand to form a gel, and then undergoes a freeze-drying process to form a sponge base material with an aerogel structure. Finally, under ultrasonic conditions, the ginseng water extract can pass through the port potential barrier of the sponge base material's pore structure and enter the sponge base material, producing a loaded structure to produce a bio-based antibacterial hemostatic sponge.

[0029] The invention discloses an application of a bio-based antibacterial hemostatic sponge, and the bio-based antibacterial hemostatic sponge is applied in the field of medical devices.

[0030] (III) Beneficial technical effects

[0031] (1) The bio-based antibacterial hemostatic sponge prepared by the present invention is a chitosan-based aerogel sponge loaded with ginseng water extract. During use, the ginseng water extract can flow out of the pores of the sponge by slight squeezing. The ginseng water extract can increase the number of platelets, form a platelet thrombus at the wound, and then accelerate the hemostasis process to achieve the purpose of rapid hemostasis.

[0032] (2) The main cross-linking agent structure prepared by the present invention contains a guanidine antibacterial agent with broad-spectrum antibacterial properties. After introducing it into the sponge structure, the excellent antibacterial effect of the guanidine antibacterial agent can be utilized to give the sponge good broad-spectrum antibacterial properties, thereby making the sponge have good antibacterial and anti-inflammatory effects, inhibiting bacterial growth and reducing the risk of postoperative infection.

[0033] (3) The auxiliary cross-linking agent prepared by the present invention contains a large number of hydrophilic ether bonds and hydroxyl groups in its structure, which can improve the water absorption rate of the sponge. When exudate appears at the wound, the exudate can be quickly absorbed, so that the wound can continue to maintain a dry healing environment, which is conducive to accelerating the healing of the wound. DETAILED DESCRIPTION

[0034] In order to facilitate the understanding of the present invention, the present invention will be described more fully below. Preferred embodiments of the present invention are given below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0035] Preparation Example 1

[0036] Preparation of main cross-linking agent:

[0037] Add 0.3 g of bromosuccinic acid, 0.18 g of 1,1,3,3-tetramethylguanidine and toluene solvent to a reactor filled with nitrogen. After the addition, start stirring. After a uniform mixed solution is formed, add 5 mL of a 25% by mass sodium hydroxide aqueous solution dropwise to the reactor. After the addition is complete, start heating and gradually increase the temperature to 65°C. After heat preservation for 16 hours, evaporate the solvent and separate the product to obtain the main cross-linking agent.

[0038] Preparation Example 2

[0039] The preparation method of the auxiliary cross-linking agent is as follows:

[0040] 0.5 g of 1,4-butanediol diglycidyl ether, 0.35 g of 1,8-diamino-3,6-dioxaoctane and 1,4-dioxane were mixed evenly, poured into a polymerization kettle, and nitrogen was passed to deoxygenate. Then, the heating rate was controlled to be 2°C / min, and the temperature was slowly increased to 70°C. After heat preservation and polymerization for 12 hours, the solvent was evaporated under reduced pressure, and the product was separated and collected to obtain an auxiliary cross-linking agent.

[0041] Example 1

[0042] A bio-based antibacterial hemostatic sponge is made of the following raw materials in parts by weight:

[0043]

[0044] The preparation method of the bio-based antibacterial hemostatic sponge comprises the following steps:

[0045] The first step is to dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 2% to form a chitosan solution with a mass fraction of 15%;

[0046] The second step is to add the main cross-linking agent, the auxiliary cross-linking agent and the catalyst to the chitosan solution, start stirring, and after a uniform mixed solution is formed, the temperature is raised to 90°C, and the mixture is kept warm for 24 hours under continuous stirring, and then the heating is stopped and allowed to cool naturally. The material is poured into a mold and placed for 12 hours, and then transferred to a freeze dryer, freeze-dried at -40°C for 48 hours, and taken out to form a sponge base material;

[0047] The third step is to mix the ginseng water extract with purified water, stir evenly to form a ginseng extract aqueous solution, then add the sponge base material, ultrasonically treat it at an ultrasonic frequency of 100 kHz for 4 hours, take it out, and let it dry naturally to obtain a bio-based antibacterial hemostatic sponge.

[0048] The deacetylation degree of chitosan is 95%. The preparation method of the main cross-linking agent is shown in Preparation Example 1; the preparation method of the auxiliary cross-linking agent is shown in Preparation Example 2, and the same applies to the following.

[0049] Example 2

[0050] A bio-based antibacterial hemostatic sponge is made of the following raw materials in parts by weight:

[0051]

[0052] The preparation method of the bio-based antibacterial hemostatic sponge comprises the following steps:

[0053] The first step is to dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 2% to form a chitosan solution with a mass fraction of 20%;

[0054] The second step is to add the main cross-linking agent, the auxiliary cross-linking agent and the catalyst to the chitosan solution, start stirring, and after a uniform mixture is formed, the temperature is raised to 95°C, and the mixture is kept warm for 18 hours under continuous stirring, and then the heating is stopped and allowed to cool naturally. The mixture is poured into a mold and placed for 16 hours, and then transferred to a freeze dryer, freeze-dried at -40°C for 48 hours, and taken out to form a sponge base material;

[0055] The third step is to mix the ginseng water extract with purified water, stir evenly to form a ginseng extract aqueous solution, then add the sponge base material, ultrasonically treat it at an ultrasonic frequency of 80 kHz for 5 hours, take it out, and let it dry naturally to obtain a bio-based antibacterial hemostatic sponge.

[0056] Example 3

[0057] A bio-based antibacterial hemostatic sponge is made of the following raw materials in parts by weight:

[0058]

[0059] The preparation method of the bio-based antibacterial hemostatic sponge comprises the following steps:

[0060] The first step is to dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 2% to form a chitosan solution with a mass fraction of 25%;

[0061] The second step is to add the main cross-linking agent, the auxiliary cross-linking agent and the catalyst to the chitosan solution, start stirring, and after a uniform mixed solution is formed, the temperature is raised to 100°C, and the mixture is kept warm for 12 hours under continuous stirring, and then the heating is stopped. After it is cooled naturally, the material is poured into a mold and placed for 18 hours, and then transferred to a freeze dryer, freeze-dried at -40°C for 48 hours, and taken out to form a sponge base material;

[0062] The third step is to mix the ginseng water extract with purified water, stir evenly to form a ginseng extract aqueous solution, then add the sponge base material, ultrasonically treat it at an ultrasonic frequency of 100 kHz for 4 hours, take it out, and let it dry naturally to obtain a bio-based antibacterial hemostatic sponge.

[0063] Comparative Example 1

[0064] A bio-based antibacterial hemostatic sponge is made of the following raw materials in parts by weight:

[0065]

[0066] The preparation method of the bio-based antibacterial hemostatic sponge comprises the following steps:

[0067] The first step is to dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 2% to form a chitosan solution with a mass fraction of 20%;

[0068] The second step is to add the main cross-linking agent and the catalyst to the chitosan solution, start stirring, and after a uniform mixture is formed, the temperature is raised to 95°C, and the mixture is kept warm for 18 hours under continuous stirring, and then the heating is stopped and allowed to cool naturally. The mixture is poured into a mold and placed for 16 hours, and then transferred to a freeze dryer, and freeze-dried at -40°C for 48 hours, and then taken out to form a sponge base material;

[0069] The third step is to mix the ginseng water extract with purified water, stir evenly to form a ginseng extract aqueous solution, then add the sponge base material, ultrasonically treat it at an ultrasonic frequency of 80 kHz for 5 hours, take it out, and let it dry naturally to obtain a bio-based antibacterial hemostatic sponge.

[0070] Comparative Example 2

[0071] A bio-based antibacterial hemostatic sponge is made of the following raw materials in parts by weight:

[0072]

[0073] The preparation method of the bio-based antibacterial hemostatic sponge comprises the following steps:

[0074] The first step is to dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 2% to form a chitosan solution with a mass fraction of 20%;

[0075] The second step is to add the auxiliary cross-linking agent and the catalyst to the chitosan solution, start stirring, and after a uniform mixed solution is formed, the temperature is raised to 95°C, and the mixture is kept warm for 18 hours under continuous stirring, and then the heating is stopped and allowed to cool naturally. The mixture is poured into a mold and placed for 16 hours, and then transferred to a freeze dryer, and freeze-dried at -40°C for 48 hours, and then taken out to form a sponge base material;

[0076] The third step is to mix the ginseng water extract with purified water, stir evenly to form a ginseng extract aqueous solution, then add the sponge base material, ultrasonically treat it at an ultrasonic frequency of 80 kHz for 5 hours, take it out, and let it dry naturally to obtain a bio-based antibacterial hemostatic sponge.

[0077] Comparative Example 3

[0078] A bio-based antibacterial hemostatic sponge is made of the following raw materials in parts by weight:

[0079]

[0080] The preparation method of the bio-based antibacterial hemostatic sponge comprises the following steps:

[0081] The first step is to dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 2% to form a chitosan solution with a mass fraction of 20%;

[0082] In the second step, the main cross-linking agent, the auxiliary cross-linking agent and the catalyst are added to the chitosan solution, and stirring is started until a uniform mixture is formed. The temperature is then raised to 95°C, and the mixture is kept warm for 18 hours under continuous stirring. Then, the heating is stopped and the mixture is allowed to cool naturally. The mixture is poured into a mold and placed for 16 hours. The mixture is then transferred to a freeze dryer and freeze-dried at -40°C for 48 hours. The mixture is then taken out to obtain a bio-based antibacterial hemostatic sponge.

[0083] Test Example 1

[0084] According to the standard GB / T 20944.3-2008, Staphylococcus aureus was selected as the test bacteria, and the antibacterial rate of the bio-based antibacterial hemostatic sponge in the test examples and comparative examples was tested;

[0085] The test method of water absorption rate is as follows: a bio-based antibacterial hemostatic sponge is made into a test sample with a specification of 1 cm×1 cm, weighed, recorded as T1, immersed in water, taken out after 8 hours, and weighed when no water drops fall, and the weight is recorded as T2. The water absorption rate is calculated using the formula (T2-T1) / T1;

[0086] The test results are shown in Table 1:

[0087] Table 1 - Antibacterial rate test results

[0088]

[0089]

[0090] Analysis of the test results in Table 1 shows that the sponge prepared by simultaneously cross-linking chitosan with a primary cross-linking agent and an auxiliary cross-linking agent has good antibacterial and anti-inflammatory effects and excellent liquid absorption effect.

[0091] After the main cross-linking agent is removed, the cross-linking density and porosity of the sponge are lower, which has a significant negative impact on the liquid absorption effect of the sponge. In addition, the broad-spectrum antibacterial effect brought by the guanidine antibacterial agent is lost, resulting in a significant decrease in the antibacterial properties of the sponge.

[0092] After the auxiliary cross-linking agent is removed, a large number of hydrophilic ether bonds are missing from the sponge structure, and thus the liquid absorption effect of the sponge is reduced.

[0093] Test Example 2

[0094] 140 mice weighing 25±2g were randomly selected as experimental subjects and randomly divided into 7 groups, namely Example 1-Example 3 groups, Comparative Example 1-Comparative Example 3 groups and a blank control group. After being fed in the same environment for one week, the back hair of the mice was shaved, and a 1 cm×1 cm wound was artificially created. The corresponding group of hemostatic sponges were immediately used for application. The blank control group was not treated. The complete hemostasis time was recorded, and the average hemostasis time of each group of mice was calculated and recorded in Table 2:

[0095] Table 2

[0096]

[0097] Analysis shows that the hemostatic sponge with ginseng water extract added has significantly faster hemostasis time, indicating that loading ginseng water extract has a good positive effect on improving the hemostatic effect of the sponge.

[0098] Based on the ideal embodiment of the present invention, through the above description, relevant personnel can make various changes and modifications without departing from the technical concept of the present invention. The technical scope of the present invention is not limited to the content in the specification, and the technical scope must be determined according to the scope of the claims.

Claims

1. A bio-based antibacterial hemostatic sponge, characterized in that: Made from the following raw materials in parts by weight: The main cross-linking agent is a tetramethylguanidine derivative; The auxiliary cross-linking agent is a ring-opening polymer of a double-terminal amine compound and a double-terminal glycidyl ether monomer.

2. The bio-based antibacterial hemostatic sponge according to claim 1, characterized in that: The deacetylation degree of the chitosan is ≥95%.

3. The bio-based antibacterial hemostatic sponge according to claim 1, characterized in that: The preparation method of the main cross-linking agent is as follows: Add bromosuccinic acid, 1,1,3,3-tetramethylguanidine and toluene solvent to a reactor filled with nitrogen. After the addition, start stirring. After a uniform mixed liquid is formed, add an alkaline hydroxide aqueous solution dropwise to the reactor. After the addition is completed, start heating and gradually increase the temperature to 60-70° C. After heat preservation for 12-18 hours, evaporate the solvent, separate the product, and obtain the main cross-linking agent.

4. The bio-based antibacterial hemostatic sponge according to claim 3, characterized in that: The alkaline hydroxide is potassium hydroxide or sodium hydroxide.

5. The bio-based antibacterial hemostatic sponge according to claim 3, characterized in that: The mass fraction of the alkaline hydroxide aqueous solution is 20-30%.

6. The bio-based antibacterial hemostatic sponge according to claim 1, characterized in that: The preparation method of the auxiliary cross-linking agent is as follows: After the double-terminal amine, double-terminal glycidyl ether monomer and 1,4-dioxane are evenly mixed, they are poured into a polymerization kettle, nitrogen is passed to deoxygenate, and then the heating rate is controlled to be 2-3°C / min, and the temperature is slowly increased to 70-80°C. After heat preservation and polymerization for 8-12 hours, the solvent is evaporated under reduced pressure, and the product is separated and collected to obtain the auxiliary cross-linking agent.

7. The bio-based antibacterial hemostatic sponge according to claim 6, characterized in that: The double-terminal aminated compound is 1,8-diamino-3,6-dioxaoctane or 2,2'-oxybis(ethylamine); the double-terminal glycidyl ether monomer is any one of ethylene glycol diglycidyl ether, dipropylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether or 1,2-cyclohexanediol diglycidyl ether.

8. The bio-based antibacterial hemostatic sponge according to claim 1, characterized in that: The catalyst is p-toluenesulfonic acid or trifluoromethanesulfonic acid.

9. A method for preparing the bio-based antibacterial hemostatic sponge as claimed in claim 1, characterized in that: The following steps are involved: The first step is to dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 1-3% to form a chitosan solution with a mass fraction of 15-25%; The second step is to add the main cross-linking agent, the auxiliary cross-linking agent and the catalyst to the chitosan solution, start stirring, and after a uniform mixed solution is formed, the temperature is raised to 90-100°C, and the mixture is kept warm for 12-24 hours under continuous stirring, and then the heating is stopped and allowed to cool naturally. The material is poured into a mold and placed for 12-18 hours, and then transferred to a freeze dryer, freeze-dried at -40°C for 24-48 hours, and taken out to form a sponge base material; The third step is to mix the ginseng water extract with purified water, stir evenly to form a ginseng extract aqueous solution, then add the sponge base material, ultrasonically treat it at an ultrasonic frequency of 80-100kHz for 4-6 hours, take it out, and let it dry naturally to obtain a bio-based antibacterial hemostatic sponge.

10. A use of the bio-based antibacterial hemostatic sponge as claimed in claim 1, characterized in that: The bio-based antibacterial hemostatic sponge is applied in the field of medical devices.

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