Thin-layer composite membrane hemostatic material with hemostatic and antibacterial capabilities and application of thin-layer composite membrane hemostatic material

Abstract

The invention belongs to the technical field of biomedical materials, and particularly relates to a thin-layer composite membrane hemostatic material with hemostatic and antibacterial capabilities. According to the material, a polysulfone membrane serves as a supporting layer, a thin-layer composite membrane is formed through an interfacial polymerization method, a functional layer is a vancomycin-loaded mesoporous nano-material, and the content of the mesoporous nano-material in the composite membrane is 0.5%. The thin-layer composite membrane has stable mechanical strength, can provide a lasting hemostatic effect, and enhances the blood coagulation process by promoting platelet aggregation and activation. The thickness of the functional layer is about 200nm, and the functional layer is tightly combined with the supporting layer to form a uniform and stable composite membrane structure. In addition, the surface of the mesoporous nano material is provided with negative charges and ammonia hydroxyl groups, and vancomycin can be loaded and slowly released, so that the antibacterial ability is improved. The invention also provides a preparation method of the composite nano hemostatic material, which comprises the following steps: firstly, preparing a mesoporous nano material, chemically modifying the surface of the mesoporous nano material, loading vancomycin, and embedding the mesoporous nano material into a polysulfone membrane support layer formed by an interfacial polymerization method to finally obtain the thin-layer composite membrane hemostatic material with hemostatic and antibacterial capabilities. The material is good in biocompatibility and free of toxic and side effects, and has wide application potential in the fields of hemostasis and antibiosis.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedical materials, and particularly relates to a thin-layer composite membrane hemostatic material with hemostatic and antibacterial capabilities, a preparation method thereof, and applications in traumatic hemostasis and antibacterial treatment. Background Art

[0002] Traditional hemostatic materials, such as hemostatic sponges and hemostatic gauzes, although able to provide hemostatic effects, have poor antibacterial properties and problems such as uneven drug release and short-term hemostatic effects. To overcome these deficiencies, there is an urgent need to develop composite materials that can provide a long-lasting hemostatic effect at the trauma site and simultaneously inhibit infection.

[0003] Mesoporous nanomaterials, with their high specific surface area and uniform pore structure, can promote platelet aggregation, improve the hemostatic effect, and inhibit wound infection while hemostasis by loading antibacterial drugs.

[0004] Combining polysulfone membranes with mesoporous nanomaterials to form a composite material with stable mechanical strength and antibacterial effects has become a potential solution to improve the effect of trauma treatment and accelerate healing.

[0005] When a trauma occurs, the aggregation and activation of platelets are key steps in the hemostasis process. By using a composite membrane material, platelet aggregation and activation can be promoted, the blood coagulation process can be enhanced, thereby improving the hemostatic effect of the material. In addition, the composite membrane can also promote water penetration, further helping to increase the platelet concentration, thus enhancing the hemostatic effect. Summary of the Invention

[0006] The first object of the present invention is to design a thin-layer composite membrane hemostatic material with both hemostatic and antibacterial capabilities; the second object of the present invention is to provide a preparation method of the above material; the third object of the present invention is to provide the application of the above material. The present invention is used for the treatment of traumatic hemostasis and prevention of wound infection.

[0007] To achieve the above first object, the present invention makes the following design: using a polysulfone membrane as the support layer, forming a thin-layer composite membrane by the interfacial polymerization method, and embedding mesoporous nanomaterials loaded with vancomycin as the functional layer.

[0008] The further description of the above design is as follows:

[0009] The thin-layer composite membrane makes the support layer and the functional layer closely combined through the interfacial polymerization technology, ensuring that the composite membrane has excellent mechanical strength and stability.

[0010] The mesoporous nanomaterials are a kind of mesoporous material with a size between 100 - 200 nm.

[0011] The special pore structure of the mesoporous nanomaterial not only improves the antibacterial drug loading capacity, but also promotes the aggregation and activation of platelets, thereby accelerating blood coagulation and enhancing the hemostatic effect.

[0012] To achieve the second above-mentioned purpose, the present invention adopts the following technical solutions: First, prepare the mesoporous nanomaterial, chemically modify its surface, and after loading vancomycin, embed it on the polysulfone membrane support layer formed by the interfacial polymerization method to finally obtain a thin-film composite membrane hemostatic material with hemostatic and antibacterial capabilities.

[0013] The further description of the above technical solutions is as follows:

[0014] The method for preparing the thin-film composite membrane hemostatic material is the interfacial polymerization method.

[0015] The mesoporous nanomaterial itself has mesoporous channels, and the mesoporous channels are between 2 and 10 nm.

[0016] The chemical modification of the surface of the mesoporous nanomaterial is to amino-functionalize its surface to ensure its ability to load vancomycin.

[0017] To achieve the third above-mentioned purpose, the present invention adopts the following technical solutions: Apply the thin-film composite membrane hemostatic material to the wound site, effectively control bleeding through its hemostatic function, and at the same time inhibit the growth of bacteria at the wound site and reduce the risk of infection by the slow release of the loaded vancomycin.

[0018] Use the mouse liver injury coagulation test to examine the hemostatic effect of the material: Expose the liver lobe of the mouse, make a wound 0.5 cm long and 0.1 cm deep on the liver with a scalpel, cover it with the material until the bleeding stops, and record the hemostasis time and the amount of bleeding.

[0019] Use the plate coating antibacterial experiment to examine the antibacterial effect of the material: Coat and culture the LB solid medium, and use the mesoporous nanomaterial suspension for antibacterial testing, and observe and record the changes in the antibacterial zone at different time periods. Description of the Drawings

[0020] Figure 1 Scanning electron microscope image of the mesoporous nanomaterial,

[0021] Figure 2 Scanning electron microscope image of the thin-film composite membrane hemostatic material. Detailed Embodiments

[0022] Example 1 of Material Preparation 1. Preparation of the mesoporous nanomaterial: TEOS was dissolved in cyclohexane as the upper oil phase, CTAC was used as the template agent, and a certain amount of TEA was added and dissolved in water as the lower water phase to construct an oil-water two-phase stratified system. Take 36 mL of distilled water, 24 mL of a pre-prepared 25 wt.% CTAB solution, and 0.18 g of TEA, stir for 1 h in an oil bath at 60 °C, slowly add the pre-prepared cyclohexane solution of 20 vv.% TEOS to the upper layer of the water phase, and continue to stir slowly at 60 °C for 12 h. After the reaction, take the water phase and centrifuge it at 15000 rpm for 30 min, collect the product, disperse it in a dilute hydrochloric acid ethanol solution, and extract it twice at 60 °C for 6 h each time. Finally, the obtained precipitate was washed three times with ethanol and deionized water respectively, centrifuged and dried in vacuum at room temperature to obtain a three-dimensional dendritic mesoporous silica nanomaterial. 2. Modification of mesoporous nanomaterials: Weigh 200 mg of the nanomaterial and place it in a round-bottom flask. Take 20 mL of toluene as the solvent, add 20 μL of APTES, reflux and cool at 80 °C, stir continuously under argon protection, after reacting for 6 h, pour off the supernatant, repeat the above experimental steps once, react for a total of 12 h, centrifuge and wash 2 - 3 times with an appropriate amount of toluene and absolute ethanol, and then dry it in an 80 °C drying oven to obtain the amino-functionalized modified nanomaterial. 3. Loading vancomycin: In this experiment, using the principle of concentration difference of drugs in the solution system, Van was loaded into the pores of mesoporous nanoparticles. Dispersed mesoporous silica nanoparticles into ultrapure water, added vancomycin to the MSNs dispersion system, controlled the mass ratio of MSNs to vancomycin to be 1:1, stirred at room temperature for 20 - 28 h to load it into the pores of MSNs, washed three times with deionized water, and freeze-dried to obtain a mesoporous silica nanocarrier loaded with vancomycin. 4. Preparation of thin-layer composite membrane hemostatic material: In this experiment, a composite polyamide membrane was prepared by the interfacial polymerization reaction of two active monomers, TMC and MPD, on the surface of a polysulfone porous support. Drug preparation: 150 - 200 mL of MPD; Place TMC in 300 mL of Isopar-G, add 0.4% of the mesoporous nanomaterial, and ultrasonicate for 1 h until completely dissolved. Sealing the membrane: Lay the polysulfone membrane flat on a glass plate and seal the four sides with waterproof tape. Here, in addition to the polymerization reaction between the amino group and the acyl chloride group, there is also an acyl hydrolysis reaction (generating carboxylic acid groups), which improves the hydrophilicity of the composite membrane and makes the membrane surface negatively charged. The preparation of the TFC membrane doped with mesoporous nanoparticles is to mix the mesoporous nanomaterials with a fixed concentration with the TMC / Isopar-G solution. The mixture is ultrasonically treated for 0.5 h to make it completely dispersed, and then interfacial polymerization is carried out immediately.

[0025] Example 1 of material application Expose the liver lobe of the mouse, make a wound with a length of 0.5 cm and a depth of 0.1 cm on the liver with a scalpel, and cover it with the material until the bleeding stops. In this experiment, a control group, a positive control group, and each nano hemostatic material group are used. Record the hemostasis time and the amount of bleeding. The amount of bleeding is the difference in the mass of the weighed gauze at the beginning and end of the experiment, and the same experiment is repeated three times.

[0026] Example 2 of material application Sterilize the LB solid medium at 120 °C for 2 h, take it out while it is hot, quickly add 20 mL of LB solid medium to the petri dish on the ultra-clean bench, dry it at room temperature for 30 min under ultraviolet irradiation, and wait for the inoculation of the bacterial solution. Determination of the bacterial solution concentration: The OD value is measured by an ultraviolet spectrophotometer to roughly estimate that the concentration of the original bacterial solution is between 10 7 ~10 8 CFU / mL. Dilute the original bacterial solution with the medium to a concentration of 1×10 7 CFU / mL, spread the bacterial solution evenly with a glass triangular spreader, let it dry, divide the petri dish lid into six large areas, drop 20 μL of the MSN suspension with a fixed concentration (10 mg / mL) at the center of each area, use sterile water as a negative control, and place it upside down in an incubator at 37 °C for 16 - 80 h of incubation. During the incubation process, observe the antibacterial zone situation of the samples and take pictures to record the antibacterial zone phenomenon.

Claims

1. A thin layer composite film hemostatic material having both hemostatic and antibacterial capabilities, characterized in that: The polysulfone membrane is used as a supporting layer, an interfacial polymerization method is adopted to form a thin composite membrane, and a mesoporous nanomaterial loaded with vancomycin is embedded as a functional layer. The content of the mesoporous nanomaterial in the composite hemostatic membrane is 0.5%.

2. A thin layer composite film hemostatic material having both hemostatic and antibacterial capabilities according to claim 1, characterized in that: The thin layer composite film hemostatic material mainly comprises a supporting layer and a functional layer.

3. A thin layer composite film hemostatic material having both hemostatic and antibacterial capabilities according to claim 1, characterized in that: The polysulfone membrane as a supporting layer can provide stable mechanical strength, ensure the lasting effect of the composite membrane on the wound site, and ensure the hemostatic effect.

4. A thin layer composite film hemostatic material having both hemostatic and antibacterial capabilities according to claim 1, characterized in that: The thickness of the functional layer is about 200 nm, and the functional layer is tightly combined with the support layer through an interfacial polymerization method to form a uniform and stable composite membrane structure.

5. A thin layer composite film hemostatic material having both hemostatic and antibacterial capabilities according to claim 1, characterized in that: The mesoporous nanomaterial is a mesoporous material with a size between 100 and 200 nm, and has a large number of uniform and regular mesoporous channels, and the channels are between 2 and 10 nm.

6. A thin layer composite film hemostatic material having both hemostatic and antibacterial capabilities according to claim 1, characterized in that: The surface of the mesoporous nanomaterial has negative charges and aminohydroxyl groups, and can load and slowly release vancomycin antibiotics, thereby improving the antibacterial ability of the material.

7. A thin layer composite film hemostatic material having both hemostatic and antibacterial capabilities according to claim 1, characterized in that: The thin layer composite membrane hemostatic material can promote water to pass through the functional layer into the supporting layer, intercept other substances, increase the platelet concentration, and thus improve the hemostatic effect of the material.

8. A thin layer composite film hemostatic material having both hemostatic and antibacterial capabilities according to claim 1, characterized in that: The thin layer composite film hemostatic material can promote platelet aggregation and activation, enhance the blood coagulation process, and thus improve the hemostatic effect of the material.

9. A thin layer composite film hemostatic material having both hemostatic and antibacterial capabilities according to claim 1, wherein the preparation method comprises the following steps: Firstly, mesoporous nanomaterials are prepared, and the surfaces of the mesoporous nanomaterials are chemically modified to introduce amino groups; Vancomycin is loaded on the surface of the modified mesoporous nanomaterial; the mesoporous nanomaterial loaded with vancomycin is embedded into a thin layer composite membrane formed on the surface of a polysulfone membrane by interfacial polymerization, and finally a thin layer composite membrane hemostatic material having both hemostatic and antibacterial capabilities is formed.

10. Use of the thin layer composite film hemostatic material prepared by the preparation method according to any one of claims 1 to 9 in hemostasis and antibacterial.

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