An antibacterial hemostatic composite material and a preparation method thereof

Abstract

The application discloses an antibacterial hemostatic composite material and a preparation method thereof. The application adopts polylactic acid glycolic acid which is degradable and has good compatibility, and grafts hydrophilic N-vinyl-2-pyrrolidone, simultaneously utilizes radiation polymerization to form polypyrrolidone, and reacts with iodine to form high polyiodine with strong antibacterial property, and uses the same as an electrospinning base material and combines with notoginseng element, so that the positively charged amino groups on the notoginseng element and the negatively charged high polyiodine ions form a stable structure, the electrospinning solvent phase separation of the base material is regulated, a porous rapid hemostatic fiber material with uniform notoginseng element distribution and high loading capacity is formed, and the material is endowed with antibacterial property, good biocompatibility and notoginseng element stable release performance.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medical materials, and particularly relates to an antibacterial hemostatic composite material and a preparation method thereof. BACKGROUND

[0002] In domestic medical treatment, traditional products such as bandages, gelatin sponges and gauze are often used for hemostasis in the case of massive bleeding caused by external trauma, but these products have slow hemostatic speed and are prone to cause secondary infection of the wound; therefore, it is necessary to develop a hemostatic material which can act on the local part of the human or animal body, has rapid hemostatic speed, can be degraded in the body, is safe and effective.

[0003] In combination with the feature that the porous physical structure can accelerate the blood absorption speed, the prior art proposes to load notoginsenoside with hemostatic function on mesoporous silicon nanomaterial, which has better hemostatic effect than gelatin sponge, but has problems such as great brittleness and poor biocompatibility with the living body. The prior art also reports that a notoginsenoside-modified gelatin solution is blended with chitosan to perform electrospinning, so as to obtain a gelatin-chitosan notoginsenoside composite hemostatic film material, which has advantages such as good biocompatibility and excellent degradation stability, but the notoginsenoside-modified gelatin has low content of loaded notoginsenoside and unstable combination, and cannot be dissolved in water, resulting in poor hemostatic performance of the material. SUMMARY

[0004] In view of the above prior art, the present application provides an antibacterial hemostatic composite material and a preparation method thereof, so as to solve the technical problems of poor hemostatic effect and easy secondary infection of the existing hemostatic material.

[0005] In order to achieve the above-mentioned purpose, the technical scheme adopted by the present application is to provide a preparation method of an antibacterial hemostatic composite material, comprising the following steps:

[0006] S1: dissolving N-vinyl-2-pyrrolidone in an organic solvent to obtain an NVP solution;

[0007] S2: dissolving polylactic glycolic acid in the NVP solution and adding elemental iodine, uniformly stirring, then irradiating with a gamma ray with an irradiation dose of 2-5 kGy for 5-120 min, and then cleaning and drying to obtain a PLGA-g-PVP / (I n - )-polymer;

[0008] S3: dissolving the PLGA-g-PVP / (I n - )-polymer in a mixed solution of dichloromethane and N,N-dimethylacetamide, stirring at room temperature for 10-15 h; then adding notoginsenoside powder into the obtained solution, and continuing to stir for 10-15 h to obtain an electrospinning solution;

[0009] S4: the electrospinning solution is subjected to spinning treatment by using an electrostatic spinning process, and then the spinning sample is naturally dried, thereby obtaining the antibacterial hemostatic composite material.

[0010] Based on the above technical solutions, the application can be further improved as follows.

[0011] Further, the volume ratio of the organic solvent methanol to N-vinyl-2-pyrrolidone is 3:1.

[0012] Further, the ratio of the poly-lactic-glycolic acid to N-vinyl-2-pyrrolidone in S2 is 2-3g:15mL.

[0013] Further, the ratio of the elemental iodine to the poly-lactic-glycolic acid in S2 is 4-5:20-30.

[0014] Further, the irradiation dose of the gamma rays in S2 is 3kGy, and the irradiation time is 30min.

[0015] Further, the washing in S2 is to wash the polymer with methanol for 3-5 times; and the drying is to dry the washed polymer at 40℃ until the weight is constant.

[0016] Further, the volume ratio of dichloromethane to N,N-dimethylacetamide in the mixed solution of dichloromethane and N,N-dimethylacetamide is 20:1.

[0017] Further, the ratio of the panaxin powder to the elemental iodine in S3 is 50:400-500.

[0018] Further, the process parameters of the electrostatic spinning process in S4 are as follows: the distance between the nozzle and the receiver is 15cm, the flow rate of the electrospinning solution is 10mL / h, and the electrospinning voltage is negative pressure 5kv and positive pressure 25kv.

[0019] The application further discloses the antibacterial hemostatic composite material prepared by the preparation method.

[0020] The application has the following beneficial effects:

[0021] 1、PLGA-g-PVP / (I n - The high polyiodine structure in the polymer makes the base material have long-term stable antibacterial property; after the base material composite liquid is added with panaxin, the dichloromethane in the solvent induces the charge shift of the amino group in the panaxin by absorbing the electron group, the amino group is positively charged, and is combined with the negatively charged PLGA-g-PVP / (I n - ) by electrostatic action to form a stable structure, so that the load stability of the panaxin can be effectively ensured.

[0022] 2、The application can graft PVP to the surface of PLGA and form stable chemical bonds by means of gamma ray irradiation, and insoluble PLGA-g-PVP / (I n - Because the high polymer iodine slowly releases free iodine, and the electrostatically combined panaxin slowly releases when acting on the wound, the amino group in the panaxin carries positive charge, and the surface of the red blood cell platelet carries negative charge, the electrostatic action of the two promotes the platelet to gather at the wound, and then the hemostasis effect of the obtained material is guaranteed.

[0023] 3、The mixed solution of dichloromethane and N,N-dimethylacetamide is used to dissolve the PLGA-g-PVP / (I n - The dichloromethane and N,N-dimethylacetamide have different boiling points, wherein the dichloromethane is easy to volatilize, which is beneficial to pore formation, the DMF increases the spinnability, and the two kinds of solvents can form porous electrospun fibers, and the loading amount and loading stability of the panaxin powder on the fibers can be characterized.

[0024] 4、The application adopts the biodegradable and good compatibility polylactic acid glycolic acid (PLGA) grafted with hydrophilic N-vinyl-2-pyrrolidone (NVP), and the polyvinylpyrrolidone (PVP) formed by radiation polymerization is reacted with iodine to form high polymer iodine (I n - ), and the high polymer iodine is combined with panaxin as an electrospun base material, the positively charged amino group on the panaxin and the negatively charged high polymer iodine ion form a stable structure, the electrospun solvent phase separation of the base material is regulated to form a porous rapid hemostatic fiber material with uniform panaxin distribution and high loading amount, and the material is endowed with antibacterial property, good biocompatibility and stable release performance of panaxin. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 It is a liquid chromatogram standard curve of panaxin;

[0026] Figure 2 It is a liquid chromatogram analysis result of the composite material obtained in Example 1;

[0027] Figure 3 It is a coagulation four-item test result;

[0028] Figure 4 It is a rat liver hemostasis situation. DETAILED DESCRIPTION

[0029] The specific embodiments of the application will be described in detail below with reference to the examples.

[0030] Example 1

[0031] An antibacterial hemostatic composite material is prepared by the following steps:

[0032] S1: N-vinyl-2-pyrrolidone and methanol are mixed at a volume ratio of 3:1 to obtain an NVP solution; then 2.1 g of PLGA is added to 20 mL of the freshly prepared NVP solution, and 0.45 g of iodine is added, and after stirring, the sample is sealed, and irradiated in a cobalt 60 gamma ray with an irradiation dose of 3 kGy for 30 min at room temperature to obtain a PLGA-g-PVP / (I n - )-polymer; the polymer is rinsed with methanol for 4 times to remove unreacted monomers and homopolymers, and then dried to constant weight in a vacuum oven at a drying temperature of 40°C;

[0033] S2: a dichloromethane solution and an N,N-dimethylacetamide solution with a volume concentration of 15% are mixed at a volume ratio of 20:1 to obtain a mixed solution; then the PLGA-g-PVP / (I n - )-polymer is dissolved in the mixed solution at a solid-liquid ratio of 1 g:2 mL, and stirred at room temperature for 12 h; then 50 mg of panaxin powder is added to the obtained solution, and continues to be stirred for 12 h to obtain an electrospinning solution;

[0034] S3: the electrospinning solution is transferred to a 10 mL plastic syringe, the distance between the nozzle and the receiver is fixed at 15 cm, the solution flow rate is 10 mL / h, the electrospinning voltage is negative pressure 5 kv and positive pressure 25 kv, and tin foil paper is fixed on the receiver for receiving nanofilaments. After the electrospinning is completed, the obtained sample film is naturally dried at room temperature to prepare a PLGA-g-PVP / (I n - )-panaxin electrospun film, i.e., an antibacterial hemostatic composite material.

[0035] Example 2

[0036] An antibacterial hemostatic composite material is prepared by the following steps:

[0037] S1: N-vinyl-2-pyrrolidone and methanol are mixed at a volume ratio of 3:1 to obtain an NVP solution; then 2.1 g of PLGA is added to 20 mL of the freshly prepared NVP solution, and 0.45 g of iodine is added, and after stirring, the sample is sealed, and irradiated in a cobalt 60 gamma ray with an irradiation dose of 3 kGy for 30 min at room temperature to obtain a PLGA-g-PVP / (I n - )-polymer; the polymer is rinsed with methanol for 4 times to remove unreacted monomers and homopolymers, and then dried to constant weight in a vacuum oven at a drying temperature of 40°C;

[0038] S2: 15% dichloromethane solution and 15% N,N-dimethylacetamide solution were mixed at a volume ratio of 20:1 to obtain a mixed solution; then the PLGA-g-PVP / (I n - )-polymer was dissolved in the mixed solution at a solid-liquid ratio of 1 g:2 mL, and stirred at room temperature for 15 h; 50 mg of notoginseng powder was then added to the obtained solution, and stirring was continued for 10 h to obtain an electrospinning solution;

[0039] S3: The electrospinning solution was transferred to a 10 mL plastic syringe, the distance between the nozzle and the receiver was fixed at 15 cm, the solution flow rate was 10 mL / h, the electrospinning voltage was negative pressure 5 kv and positive pressure 25 kv, and tin foil paper was fixed on the receiver for receiving nanofilaments. After electrospinning, the obtained sample film was naturally dried at room temperature, and a PLGA-g-PVP / (I n - )-notoginseng electrospun film, i.e., an antibacterial hemostatic composite material, was prepared.

[0040] Example 3

[0041] An antibacterial hemostatic composite material was prepared by the following steps:

[0042] S1: N-vinyl-2-pyrrolidone and methanol were mixed at a volume ratio of 3:1 to obtain an NVP solution; then 2 g of PLGA was added to 20 mL of freshly prepared NVP solution, and 0.4 g of iodine was added, and after stirring, the sample was sealed and irradiated in a cobalt 60 gamma ray with an irradiation dose of 2 kGy for 90 min to obtain a PLGA-g-PVP / (I n - )-polymer; the polymer was washed with methanol for 3 times to remove unreacted monomers and homopolymers, and then dried in a vacuum oven at a drying temperature of 40°C to constant weight;

[0043] S2: 15% dichloromethane solution and 15% N,N-dimethylacetamide solution were mixed at a volume ratio of 20:1 to obtain a mixed solution; then the PLGA-g-PVP / (I n - )-polymer was dissolved in the mixed solution at a solid-liquid ratio of 1 g:2 mL, and stirred at room temperature for 10 h; 50 mg of notoginseng powder was then added to the obtained solution, and stirring was continued for 15 h to obtain an electrospinning solution;

[0044] S3: The electrospinning solution was transferred to a 10 mL plastic syringe, the distance between the nozzle and the receiver was fixed at 15 cm, the solution flow rate was 10 mL / h, the electrospinning voltage was negative pressure 5 kv and positive pressure 25 kv, and tin foil was fixed on the receiver for receiving nanofilaments. After electrospinning, the obtained sample film was naturally dried at room temperature, and the PLGA-g-PVP / (I n - ) - Dencichine electrospinning film, that is, an antibacterial hemostatic composite material.

[0045] Experimental examples

[0046] The antibacterial hemostatic composite materials prepared in the above three examples have similar properties, and therefore the antibacterial hemostatic composite material prepared in Example 1 is taken as an example to illustrate its properties.

[0047] 1. Liquid chromatography analysis

[0048] The composite material prepared in Example 1 was taken for liquid analysis, and the results are shown in Figure 1 and 2 It can be seen that the loading amount of dencichine is 30.95 mg, and the utilization rate of dencichine is 61.9%, indicating that dencichine is loaded in large amounts on the composite material.

[0049] 2. Four tests of blood coagulation

[0050] Centrifugal whole blood (3000 rpm, 15 min) was obtained to obtain platelet-poor plasma (PPP). The composite material prepared in Example 1 was immersed in the PPP according to 3 cm 2 / mL, slightly shaken, and incubated for 30 minutes, and then the suspension was detected in an automatic blood coagulation analyzer. The prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen (FIB) were tested, and the results are shown in Figure 3 , wherein the Blank group is a blank control group, the PLGA-g-PVP / (I n - ) group is a material prepared without adding dencichine powder, and the PLGA-g-PVP / (I n - ) - Dencichine is the composite material prepared in Example 1 of the present application.

[0051] As can be seen from the figure, compared with the blank control group, the PT, APTT, TT and FIB of the PLGA-g-PVP / (I n - ) group all decreased, indicating that the introduction of high polyiodine (I n -), can promote blood coagulation to a certain extent. While using the composite material prepared in the present application, the PT, APTT, TT and FIB of the PLGA-g-PVP / (I n - ) group, the PT, APTT, TT and FIB further decrease, indicating that the antibacterial hemostatic composite material prepared in the present application has excellent function of promoting blood coagulation.

[0052] 3. Animal hemostasis experiment

[0053] In order to study the hemostatic ability of the composite material in vivo, the following method was used for experiment: the SD rats were anesthetized in turn, and the liver was placed in a sterile environment; the liquid around the liver was carefully removed in advance with filter paper; a full-thickness wound (unstitchable bleeding wound) with a diameter of 4 mm was created by biopsy punching. In the experimental group, the hemostatic material was applied to the wound site 10 s after injury, and the bleeding condition of the wound was observed after 1 min; the wound of the control group was not treated. Photographs were taken to evaluate the hemostatic effect of the hemostatic material.

[0054] The results are shown in Figure 4 From the figure, it can be seen that, compared with the gauze group (Gauze) and the material group without notigensin (PLGA-g-PVP / (I n - )), after 1 min of coverage, the liver wound no longer bled after being covered with the antibacterial hemostatic composite material in the present application, and the blood appeared to be coagulated, while the wound of the control group was still in a bleeding state. It is indicated that the composite material in the present application can promote blood coagulation and has good hemostatic effect.

[0055] Although the specific embodiments of the present application are described in detail in combination with the embodiments, it should not be understood as limiting the protection scope of the present patent. Various modifications and variations made by those skilled in the art within the scope described in the claims are still within the protection scope of the present patent.

Claims

1. A method of preparing an antibacterial hemostatic composite material, characterized by, The method comprises the following steps: S1: dissolving N-vinyl-2-pyrrolidone in an organic solvent to obtain an NVP solution; S2: dissolve the polylactic acid glycolic acid in the NVP solution, and add elemental iodine, mix well, then irradiate with a gamma ray of 2~5kGy for 5~120min, then clean and dry, to obtain PLGA-g-PVP / (I n - )-polymer; S3: dissolving the PLGA-g-PVP / (I n - polymer in a mixed solution of dichloromethane and N,N-dimethylacetamide, stirring at room temperature for 10-15 h; then adding notoginsenoside powder into the obtained solution, continuing to stir for 10-15 h to obtain an electrospinning solution; the volume ratio of dichloromethane to N,N-dimethylacetamide in the mixed solution is 20:1; S4: performing spinning treatment on the electrospinning solution by using an electrostatic spinning process, and then naturally drying the spinning sample to obtain the product.

2. The method of preparing an antimicrobial hemostatic composite material according to claim 1, characterized in that: The organic solvent is methanol, and the volume ratio of the methanol to N-vinyl-2-pyrrolidone is 3:

1.

3. The method for preparing the antibacterial and hemostatic composite material according to claim 1, characterized in that: The feed liquid ratio of the polylactic acid-glycolic acid to N-vinyl-2-pyrrolidone in S2 is 2-3 g:15 mL.

4. The method of producing an antibacterial hemostatic composite material according to claim 3, characterized in that: The mass ratio of the elemental iodine to the polylactic acid-glycolic acid in S2 is 4-5:20-30.

5. The method of preparing an antimicrobial hemostatic composite material according to claim 1, characterized in that: The irradiation dose of the gamma rays in S2 is 3 kGy, and the irradiation time is 30 min.

6. The method of preparing an antimicrobial hemostatic composite material according to claim 1, characterized in that: The washing in S2 is to rinse the polymer with methanol for 3-5 times, and the drying is to dry the rinsed polymer at 40 ℃ until the weight is constant.

7. The method of claim 4, wherein the antimicrobial hemostatic composite material is prepared by the steps of: The mass ratio of the panaxin powder to the elemental iodine in S3 is 50:400-500.

8. The method of claim 1, wherein the antimicrobial hemostatic composite material is prepared by the steps of: The process parameters of the electrostatic spinning process in S4 are as follows: the distance between the nozzle and the receiver is 15 cm, the electrospinning solution flow rate is 10 mL / h, and the electrospinning voltage is negative pressure 5 kv and positive pressure 25 kv.

9. The antibacterial hemostatic composite material prepared by the preparation method in any one of claims 1-8.

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