A bio-based antibacterial hemostatic sponge, its preparation method and application

Abstract

The present invention relates to the technical field of biomedical materials, and discloses a bio-based antibacterial hemostatic sponge, its preparation method and application. The hemostatic sponge uses chitosan as the main raw material, is crosslinked and modified by using a main crosslinking agent and an auxiliary crosslinking agent, then freeze-dried, and loaded with ginseng aqueous extract. The bio-based antibacterial hemostatic sponge prepared by the present invention is a chitosan-based aerogel sponge loaded with ginseng aqueous extract. Among them, the ginseng aqueous extract can accelerate the hemostasis process by increasing the number of platelets. The presence of guanidine antibacterial groups in the structure of the main crosslinking agent can endow the sponge with good broad-spectrum antibacterial properties. The structure of the auxiliary crosslinking agent contains a large number of hydrophilic ether bonds and hydroxyl groups, etc., which can improve the water absorption rate of the sponge, enable the wound to maintain a dry healing environment continuously, and is beneficial to accelerating the healing of the wound.

Description

Technical Field

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

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

[0003] In surgical operations, the hemostatic sponge is commonly used for small bleeding points, such as skin incisions, mucosal incisions, etc., and for controlling bleeding during tissue or organ separation. In addition, the hemostatic sponge can also be applied to hemostatic powders to further improve the hemostatic effect. In terms of emergency hemostasis, the hemostatic sponge can quickly absorb blood and tissue exudate, accelerate wound healing, and is suitable for occasions such as superficial burns, scalds, and accidental traumas.

[0004] Chitosan sponge is a new type of hemostatic material developed in recent years and has many unique advantages. Chitosan is a natural polymer material with good biocompatibility and biodegradability, and 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 to protect the wound surface and stop bleeding, chitosan has currently been widely used in the field of biomedicine.

[0005] However, the antibacterial effect of chitosan itself is not actually very excellent, 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 the wound. This makes it difficult for chitosan to be directly used as a hemostatic sponge. Therefore, preparing a 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 Problems to be Solved

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

[0008] (2) Technical Solutions

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

[0010]

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

[0012] The co-crosslinking agent is a ring-opening polymer of a double-terminal amide compound and a double-terminal glycidyl ether monomer.

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

[0014] As a further aspect of the present invention, the preparation method of the main crosslinking agent is as follows:

[0015] Add bromosuccinic acid, 1,1,3,3-tetramethylguanidine and toluene solvent into a reactor filled with nitrogen. After adding, start stirring. After forming a uniform mixture, dropwise add an aqueous solution of alkaline hydroxide to the reactor. After the addition is complete, start heating and gradually raise the temperature to 60-70°C. After heat preservation treatment for 12-18 h, evaporate the solvent and separate the product to obtain the main crosslinking agent.

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

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

[0018] Specifically, under the catalytic action of the alkaline hydroxide, the halogen substituent in the structure of bromosuccinic acid can undergo substitution with 1,1,3,3-tetramethylguanidine to obtain a guanidine derivative containing two equivalents of active carboxyl substituents in the structure, that is, the main crosslinking agent.

[0019] As a further aspect of the present invention, the preparation method of the co-crosslinking agent is as follows:

[0020] After uniformly mixing the double-terminal amide compound, the double-terminal glycidyl ether monomer and 1,4-dioxane, pour them into a polymerization kettle, purge with nitrogen to remove oxygen, then control the heating rate at 2-3°C / min, slowly raise the temperature to 70-80°C, after heat preservation polymerization for 8-12 h, reduce the pressure to evaporate and remove the solvent, separate and collect the product to obtain the co-crosslinking agent.

[0021] As a further aspect of the present invention, the double-terminal amide 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.

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

[0023] In the above technical solution, under high-temperature conditions, the terminal amino groups in the structure of the bis(amine) compound can continuously undergo ring-opening addition reactions with the terminal glycidyl ether groups in the structure of the bis(epoxy) monomer, to obtain a polyether macromolecular substance with an alternating block structure, that is, the co-crosslinking agent.

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

[0025] First step: Dissolve chitosan in an aqueous acetic acid solution with a mass fraction of 1-3%, and prepare a chitosan solution with a mass fraction of 15-25%.

[0026] Second step: Add the main crosslinking agent, co-crosslinking agent and catalyst to the chitosan solution, start stirring, and after forming a homogeneous mixture, raise the temperature to 90-100 °C, keep it warm for 12-24 h under continuous stirring, then stop heating, wait for it to cool naturally, pour the material into a mold and place it for 12-18 h, then transfer it to a freeze dryer and perform freeze-drying treatment at a temperature of -40 °C for 24-48 h, take it out to form a sponge base material.

[0027] Third step: Mix the aqueous extract of ginseng with purified water, stir evenly to form an aqueous solution of ginseng extract, then add the sponge base material, perform ultrasonic treatment at an ultrasonic frequency of 80-100 kHz for 4-6 h, take it out, and wait for it to dry naturally to obtain the bio-based antibacterial hemostatic sponge.

[0028] Specifically, the active amino groups in the chitosan structure and the active hydroxyl functional groups generated by the ring-opening reaction in the co-crosslinking agent structure can undergo condensation reactions with the active carboxyl substituents in the main crosslinking agent structure under the catalysis of the catalyst, so as to achieve crosslinking among the three, let the product stand to form a gel, and then through the freeze-drying process, a sponge base material with an aerogel structure can be formed. Finally, under ultrasonic conditions, the aqueous extract of ginseng can enter the sponge base material through the port barrier of the pore structure of the sponge base material to generate a loaded structure, and the bio-based antibacterial hemostatic sponge is obtained.

[0029] An application of a bio-based antibacterial hemostatic sponge, applying the bio-based antibacterial hemostatic sponge to 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 the aqueous extract of ginseng. During use, by slightly squeezing, the aqueous extract of ginseng can flow out from the pores of the sponge. The aqueous extract of ginseng 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 crosslinking agent 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 endow the sponge with good broad-spectrum antibacterial properties, so that the sponge has good antibacterial and anti-inflammatory effects, can inhibit the growth of bacteria, and reduce the risk of postoperative infection.

[0033] (3) The auxiliary crosslinking agent prepared by the present invention contains a large number of hydrophilic ether bonds and hydroxyl groups, etc., which can improve the water absorption rate of the sponge. When exudate appears at the wound, it can quickly absorb the exudate, so that the wound can continuously maintain a dry healing environment, which is beneficial to accelerating the healing of the wound. Specific embodiments

[0034] To facilitate the understanding of the present invention, the present invention will be described more comprehensively 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 content of the present invention more thorough and comprehensive.

[0035] Preparation Example 1

[0036] Preparation of the main crosslinking 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 adding, start stirring. After forming a homogeneous mixture, add 5 mL of a 25% by mass aqueous sodium hydroxide solution dropwise to the reactor. After the dropwise addition is completed, start heating and gradually raise the temperature to 65 °C. After heat preservation treatment for 16 h, evaporate the solvent and separate the product to obtain the main crosslinking agent.

[0038] Preparation Example 2

[0039] The preparation method of the auxiliary crosslinking agent is as follows:

[0040] After mixing 0.5 g of 1,4-butanediol diglycidyl ether, 0.35 g of 1,8-diamino-3,6-dioxaoctane and 1,4-dioxane evenly, pour them into a polymerization kettle, remove oxygen by passing nitrogen, and then control the heating rate at 2 °C / min. Slowly raise the temperature to 70 °C. After heat preservation polymerization for 12 h, evaporate the solvent under reduced pressure and separate and collect the product to obtain the auxiliary crosslinking agent.

[0041] Example 1

[0042] A bio-based antibacterial hemostatic sponge is made from 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] First step: Dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 2%, and prepare a chitosan solution with a mass fraction of 15%.

[0046] Second step: Add the main crosslinking agent, auxiliary crosslinking agent and catalyst into the chitosan solution, start stirring. After forming a uniform mixture, raise the temperature to 90 °C, keep it warm for 24 h under continuous stirring, then stop heating, wait for it to cool naturally, pour the material into a mold and let it stand for 12 h, then transfer it to a freeze dryer and conduct freeze-drying treatment at a temperature of -40 °C for 48 h, take it out to form a sponge base material.

[0047] Third step: Mix the aqueous extract of ginseng with purified water, stir evenly to form an aqueous solution of ginseng extract, then add the sponge base material, conduct ultrasonic treatment at an ultrasonic frequency of 100 kHz for 4 h, take it out, and wait for it to dry naturally to obtain the bio-based antibacterial hemostatic sponge.

[0048] Among them, the deacetylation degree of chitosan is 95%; for the preparation method of the main crosslinking agent, see Preparation Example 1; for the preparation method of the auxiliary crosslinking agent, see Preparation Example 2, and the same applies hereinafter.

[0049] Example 2

[0050] A bio-based antibacterial hemostatic sponge is made from 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] First step: Dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 2%, and prepare a chitosan solution with a mass fraction of 20%.

[0054] Second step: Add the main crosslinking agent, auxiliary crosslinking agent and catalyst into the chitosan solution, start stirring. After forming a uniform mixture, raise the temperature to 95 °C, keep it warm for 18 h under continuous stirring, then stop heating, wait for it to cool naturally, pour the material into a mold and let it stand for 16 h, then transfer it to a freeze dryer and conduct freeze-drying treatment at a temperature of -40 °C for 48 h, take it out to form a sponge base material.

[0055] Third step: Mix the aqueous extract of ginseng with purified water, stir evenly to form an aqueous solution of ginseng extract, then add the sponge base material, conduct ultrasonic treatment at an ultrasonic frequency of 80 kHz for 5 h, take it out, and wait for it to dry naturally to obtain the bio-based antibacterial hemostatic sponge.

[0056] Example 3

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

[0058]

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

[0060] First step: Dissolve chitosan in an aqueous acetic acid solution with a mass fraction of 2%, and prepare a chitosan solution with a mass fraction of 25%.

[0061] Second step: Add the main crosslinking agent, auxiliary crosslinking agent and catalyst to the chitosan solution, start stirring, and after forming a uniform mixture, raise the temperature to 100 °C, keep it warm for 12 h under continuous stirring, then stop heating, wait for it to cool naturally, pour the material into a mold and let it stand for 18 h, then transfer it to a freeze dryer and perform freeze-drying treatment at -40 °C for 48 h, take it out to form a sponge base material.

[0062] Third step: Mix the aqueous extract of ginseng with purified water, stir evenly to form an aqueous solution of ginseng extract, then add the sponge base material, perform ultrasonic treatment at an ultrasonic frequency of 100 kHz for 4 h, take it out, and wait for it to dry naturally to obtain the bio-based antibacterial hemostatic sponge.

[0063] Comparative Example 1

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

[0065]

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

[0067] First step: Dissolve chitosan in an aqueous acetic acid solution with a mass fraction of 2%, and prepare a chitosan solution with a mass fraction of 20%.

[0068] Second step: Add the main crosslinking agent and catalyst to the chitosan solution, start stirring, and after forming a uniform mixture, raise the temperature to 95 °C, keep it warm for 18 h under continuous stirring, then stop heating, wait for it to cool naturally, pour the material into a mold and let it stand for 16 h, then transfer it to a freeze dryer and perform freeze-drying treatment at -40 °C for 48 h, take it out to form a sponge base material.

[0069] Third step: Mix the aqueous extract of ginseng with purified water, stir evenly to form an aqueous solution of ginseng extract, then add the sponge base material, perform ultrasonic treatment at an ultrasonic frequency of 80 kHz for 5 h, take it out, and wait for it to dry naturally to obtain the bio-based antibacterial hemostatic sponge.

[0070] Comparative Example 2

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

[0072]

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

[0074] First step: Dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 2% to prepare a chitosan solution with a mass fraction of 20%.

[0075] Second step: Add a co-crosslinking agent and a catalyst to the chitosan solution, start stirring, after forming a uniform mixture, raise the temperature to 95°C, keep it warm for 18 h under continuous stirring, stop heating, wait for it to cool naturally, pour the material into a mold and place it for 16 h, then transfer it to a freeze dryer, perform freeze-drying treatment at a temperature of -40°C for 48 h, take it out to form a sponge base material.

[0076] Third step: Mix the aqueous extract of ginseng with purified water, stir evenly to form an aqueous solution of ginseng extract, then add the sponge base material, perform ultrasonic treatment at an ultrasonic frequency of 80 kHz for 5 h, take it out, and wait for it to dry naturally to obtain the bio-based antibacterial hemostatic sponge.

[0077] Comparative Example 3

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

[0079]

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

[0081] First step: Dissolve chitosan in an acetic acid aqueous solution with a mass fraction of 2% to prepare a chitosan solution with a mass fraction of 20%.

[0082] Second step: Add a main crosslinking agent, a co-crosslinking agent and a catalyst to the chitosan solution, start stirring, after forming a uniform mixture, raise the temperature to 95°C, keep it warm for 18 h under continuous stirring, stop heating, wait for it to cool naturally, pour the material into a mold and place it for 16 h, then transfer it to a freeze dryer, perform freeze-drying treatment at a temperature of -40°C for 48 h, take it out to obtain the 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 strain, and the antibacterial rate of the bio-based antibacterial hemostatic sponge in the test examples and comparative examples was tested;

[0085] The test method for the water absorption ratio is as follows: The bio-based antibacterial hemostatic sponge was made into a test sample with a specification of 1 cm × 1 cm, weighed, denoted as T1, immersed it in water, took it out after 8 h, waited until no more water droplets dripped, weighed it, and recorded the weight as T2. The water absorption ratio was calculated using the formula (T2 - T1) / T1;

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

[0087] Table 1 - Test results of antibacterial rate

[0088]

[0089]

[0090] Analyzing the test results in Table 1, it can be seen that the sponge prepared by crosslinking chitosan with both the main crosslinking agent and the auxiliary crosslinking agent has good antibacterial and anti-inflammatory effects, and excellent liquid absorption effect.

[0091] After removing the main crosslinking agent, the crosslinking density of the sponge is small and the porosity is low. Therefore, it has an obvious negative impact on the liquid absorption effect of the sponge, and loses the broad-spectrum antibacterial effect brought by the guanidine antibacterial agent, resulting in a significant decline in the antibacterial performance of the sponge.

[0092] After removing the auxiliary crosslinking agent, a large number of hydrophilic ether bonds and the like are missing in the sponge structure. Therefore, the liquid absorption effect of the sponge decreases.

[0093] Test Example 2

[0094] A total of 140 mice with a body weight of 25 ± 2 g were randomly selected as experimental subjects and randomly divided into 7 groups, namely the groups of Example 1 - Example 3, the groups of Comparative Example 1 - Comparative Example 3, and the blank control group. After being fed in the same environment for one week, the back hair of the mice was shaved off, and a wound of 1 cm × 1 cm was artificially created. Immediately, the corresponding group's hemostatic sponge was applied. 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 added with the aqueous extract of ginseng has a significantly faster hemostasis time, indicating that loading the aqueous extract of ginseng has a good positive effect on improving the hemostasis effect of the sponge.

[0098] Based on the inspiration of the ideal embodiments of the present invention, through the above description, relevant personnel can completely make various changes and modifications without departing from the technical idea of this invention. The technical scope of this 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, It is made from raw materials including the following parts by weight: Chitosan: 20 - 30 parts; Main cross-linking agent: 1 - 2.5 parts; Auxiliary cross-linking agent: 2 - 4 parts; Catalyst: 0.1 - 0.2 parts; Ginseng aqueous extract: 0.5 - 1.5 parts; The main cross-linking agent is a tetramethylguanidine derivative; The preparation method of the main cross-linking agent is as follows: Add bromosuccinic acid, 1,1,3,3-tetramethylguanidine and toluene solvent into a reactor filled with nitrogen. After adding, start stirring. After forming a uniform mixture, dropwise add an aqueous solution of alkaline hydroxide into the reactor. After dropping, start heating and gradually raise the temperature to 60 - 70 °C. After heat preservation treatment for 12 - 18 h, evaporate the solvent and separate the product to obtain the main cross-linking agent; The auxiliary cross-linking agent is a ring-opening polymer of a bis-terminal amide and a bis-terminal glycidyl ether monomer; The preparation method of the auxiliary cross-linking agent is as follows: Mix the bis-terminal amide, bis-terminal glycidyl ether monomer and 1,4-dioxane evenly, pour them into a polymerization kettle, remove oxygen by passing nitrogen, then control the heating rate at 2 - 3 °C / min, slowly raise the temperature to 70 - 80 °C, keep the temperature for polymerization for 8 - 12 h, then reduce the pressure to evaporate and remove the solvent, separate and collect the product to obtain the auxiliary cross-linking agent.

2. The bio-based antibacterial hemostatic sponge according to claim 1, characterized in that, The deacetylation degree of the chitosan ≥ 95%; 3. The bio-based antibacterial hemostatic sponge according to claim 1, characterized in that, The alkaline hydroxide is potassium hydroxide or sodium hydroxide; 4. The bio-based antibacterial hemostatic sponge according to claim 1, characterized in that, The mass fraction of the aqueous solution of the alkaline hydroxide is 20 - 30%; 5. The bio-based antibacterial hemostatic sponge according to claim 1, wherein The bis-terminal amide is 1,8-diamino-3,6-dioxaoctane or 2,2'-oxybis(ethylamine); the bis-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; 6. The bio-based antibacterial hemostatic sponge according to claim 1, characterized in that, The catalyst is p-toluenesulfonic acid or trifluoromethanesulfonic acid; 7. A preparation method of the bio-based antibacterial hemostatic sponge as described in claim 1, characterized in that, It includes the following steps: First step: Dissolve chitosan in an aqueous acetic acid solution with a mass fraction of 1 - 3% to prepare a chitosan solution with a mass fraction of 15 - 25%; Second step: Add the main cross-linking agent, auxiliary cross-linking agent and catalyst into the chitosan solution, start stirring. After forming a uniform mixture, raise the temperature to 90 - 100 °C, keep the temperature for heat preservation treatment for 12 - 24 h under continuous stirring, then stop heating and wait for it to cool naturally. Pour the material into a mold and place it for 12 - 18 h, then transfer it to a freeze dryer and perform freeze-drying treatment at a temperature of -40 °C for 24 - 48 h, take it out to form a sponge base material; Third step: Mix the ginseng aqueous extract and purified water, stir evenly to form an aqueous solution of ginseng extract, then add the sponge base material, perform ultrasonic treatment at an ultrasonic frequency of 80 - 100 kHz for 4 - 6 h, take it out and wait for it to dry naturally to obtain the bio-based antibacterial hemostatic sponge.

8. Use of a bio-based antibacterial hemostatic sponge as described in claim 1, characterized in that, Apply the bio-based antibacterial hemostatic sponge to the medical device field.

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