Composite hemostatic material as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medical materials, and particularly relates to a composite hemostatic material and a preparation method and application thereof. Nanometer zeolite with a higher specific surface area and a larger pore volume is used as a basis of the composite hemostatic material, and a polyphenol compound is introduced into a traditional zeolite hemostatic material to serve as a barrier for inhibiting heat production, so that possible heat damage to surrounding tissues in the rapid water absorption process of zeolite is effectively avoided, secondary damage is avoided, and the hemostatic material has a good application prospect. The safety of the hemostatic material is improved; by compounding the nano-zeolite and the polyphenol compound, agglomeration among the nano-zeolite is reduced, more microporous structures are exposed, on the basis of promoting an exogenous blood coagulation mechanism, the blood coagulation performance is further improved from an endogenous blood coagulation mechanism, antibacterial performance is introduced while the blood coagulation performance is improved, heat damage is eliminated, and the blood coagulation activity is improved. The application range and safety of the composite hemostatic material as a hemostatic material are greatly expanded, so that the composite hemostatic material has a wide application prospect.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical materials, and in particular relates to a composite hemostatic material and a preparation method and application thereof. Background Art

[0002] Zeolite is a natural molecular sieve and a natural aluminosilicate, composed of silicon-oxygen tetrahedrons and aluminum-oxygen tetrahedrons. The AlO4 and SiO4 tetrahedrons are interconnected by oxygen atoms, forming a network of cavities with a three-dimensional microporous structure. The aluminum-oxygen tetrahedrons are negatively charged, and to maintain charge balance, alkali and alkaline earth metal ions (such as Na, Ca, and Mg) enter the cavities within the zeolite structure. Zeolite's unique structure and properties offer long-term physical and chemical stability, the ability to absorb large amounts of water, and the ability to exchange ions. Furthermore, zeolite contains no proteins that could cause allergic reactions and is non-biotoxic. These properties make zeolite one of the most common inorganic hemostatic materials, offering enormous application prospects.

[0003] The hemostatic mechanism of zeolite is that its cage-like cavity can absorb water in the blood, causing the aggregation and concentration of platelets, fibrin and coagulation factors in the blood. At the same time, the Ca2+ released from the inside of the zeolite 2+ As a coagulation factor, it activates the endogenous coagulation cascade, thereby promoting hemostasis. For example, patent CN 100571711C discloses a process for preparing a zeolite-based rapid hemostatic agent, using crushed zeolite as a hemostatic agent, which can achieve rapid hemostasis. However, during the hemostatic process, the heat released by the exchange of cations between zeolite and nano-zeolite and water inevitably burns the surrounding tissue, putting the wound at risk of infection. This significantly limits the application of zeolite in hemostatic materials. Summary of the Invention

[0004] The present invention aims to overcome the defects of the existing zeolite materials in the art, such as insufficient hemostatic ability, easy thermal damage to surrounding tissues during the hemostatic process, and the disadvantage of possible secondary damage to the wound due to bacterial infection. A composite hemostatic material and its preparation method and application are provided to overcome the above-mentioned defects.

[0005] To achieve the above-mentioned purpose, the present invention is implemented through the following technical solutions: A method for preparing a nanocomposite hemostatic material comprises the following steps: S1, ball milling the natural zeolite to obtain zeolite powder; S2, stirring the zeolite powder in water to obtain a suspension, and obtaining nano-zeolite colloid after gravity sedimentation; S3, adding polyphenol compounds to the nano-zeolite colloid, heating and stirring to obtain a nano-composite colloid; S4. Freeze-drying the nanocomposite colloid to obtain the nanocomposite hemostatic material.

[0006] Zeolite, a common aluminosilicate in nature, can be used as a hemostatic material for two main reasons: First, the three-dimensional cage structure of zeolite gives it a naturally microporous structure, resulting in a large specific surface area. When zeolite contacts a wound, the cage-like cavity can quickly absorb water from the blood, concentrating coagulation factors and platelets, thereby promoting coagulation; second, when zeolite absorbs water from the blood, it undergoes an ion exchange reaction with the blood, and calcium ions enter the blood, promoting the formation of fibrin by participating in the endogenous coagulation mechanism, thereby accelerating blood coagulation. However, in this process, the alkali metal ions in the zeolite framework undergo hydration reaction, releasing a large amount of heat; the concentration of coagulation factors in the blood increases, accelerating the coagulation reaction, and this process also releases heat. This heat can cause thermal damage to the tissues around the wound, posing a risk of wound infection.

[0007] Therefore, considering the inherent thermal damage problem of zeolite and the necessary antibacterial performance requirements, the present invention grafts polyphenolic compounds with antibacterial properties on the surface of zeolite particles as a barrier to inhibit heat production. Polyphenolic compounds have multiple phenolic hydroxyl groups, which can be combined with zeolite particles through hydrogen bonds and covalent bonds to achieve stable compounding. When zeolite comes into contact with blood, polyphenols reduce the hydration capacity of zeolite by absorbing part of the water and covering the zeolite, thereby reducing heat release. Polyphenolic compounds destroy the cell membrane and cell wall of bacteria by interacting with proteins and lipids on the surface of bacteria, causing the cell contents to leak out; interfere with bacterial metabolism and growth by inhibiting enzyme activity; and inhibit the synthesis of cell walls or directly interact with the surface of cell walls to destroy their integrity, thereby achieving antibacterial effects and reducing the risk of wound infection.

[0008] However, the hemostatic ability of natural zeolite is very limited and often cannot achieve the effect of rapid hemostasis. Therefore, the present invention optimizes the preparation process and, before compounding the polyphenolic compound, first performs nano-processing on the zeolite. Nano-zeolite presents better hemostatic performance because: nano-zeolite has a higher specific surface area, smaller crystal size, higher porosity and more regular shape, and has a faster mass transfer rate and larger storage space than natural zeolite, so it has a stronger water absorption capacity and improves coagulation efficiency. Compounding is carried out on the basis of nano-zeolite, so that the polyphenolic compound is loaded on the surface of the nano-zeolite through hydrogen bonds and covalent bonds, and the repulsive force between the polyphenolic compounds causes the nano-zeolite to be uniformly dispersed, exposing more microporous structures, so that the nano-composite material releases more Ca in the hemostatic process. 2+ , accelerating the intrinsic coagulation pathway and greatly improving hemostatic performance.

[0009] In summary, the present invention optimizes the preparation process of traditional zeolite hemostatic materials, adjusts the particle size of natural zeolite to nanometer level through ball milling and gravity sedimentation, and then adds polyphenol compounds to the zeolite colloid. A composite structure of nano-zeolite particles and polyphenol compounds is formed through heating and stirring steps. The nano-zeolite particles introduce a mesoporous structure while retaining the zeolite's own microporous framework. Compared with natural zeolite, they have a faster mass transfer rate and a larger storage space. They not only retain the stability brought by the zeolite framework, but also greatly improve the water adsorption and diffusion capacity through the mesoporous structure. After the polyphenol compounds are introduced into the nano-zeolite particles, the nano-zeolite particles in the composite colloid are dispersed from each other due to the repulsive force between the polyphenol compounds, which effectively avoids the agglomeration of the zeolite particles. Characterization has proved that the nano-composite material of the present invention is mainly primary particle size and has a smaller particle size than nano-zeolite. At the same time, the polyphenol compounds coated on the surface of the nano-zeolite can effectively reduce the heat generated during the water absorption process of the zeolite, avoid the occurrence of thermal damage, and form a highly safe hemostatic material with excellent hemostatic and antibacterial properties.

[0010] Preferably, in step S3, the polyphenolic compounds include one or more of tannic acid, catechin, proanthocyanidin, and quercetin.

[0011] Preferably, in step S3, the concentration of the polyphenol compound in the zeolite colloid is 0.5-2.0 mg / mL.

[0012] Preferably, the milling parameters in step S1 are: a ball-to-material mass ratio of 4:1, a bead ratio of 2 mm: 4 mm: 6 mm of 1:4:16, a ball mill speed of 600 rpm, a milling time of 10 to 25 hours, and an alternating time of 10 minutes with a 5-minute interval. In practice, the milling rotation mode is alternating between forward and reverse rotation.

[0013] Preferably, in step S3, the heating temperature is 30-75°C.

[0014] Preferably, in step S3, the heating time is 12 to 48 hours.

[0015] The present invention also discloses a nano-composite hemostatic material prepared by the above steps. The nano-composite material comprises nano-zeolite particles and polyphenol compounds grafted on the surface of the nano-zeolite particles through hydrogen bonds and covalent bonds.

[0016] Preferably, the particle size of the nanocomposite hemostatic material is 5-10 nm.

[0017] Preferably, the volume of micropores in the composite hemostatic material accounts for 20-40% of the total pore volume.

[0018] The invention also discloses the application of the composite hemostatic material in the preparation of medical hemostatic products.

[0019] Therefore, the present invention has the following beneficial effects: (1) The present invention introduces polyphenol compounds into the traditional zeolite hemostatic material as a barrier to inhibit heat production, effectively avoiding the thermal damage to the surrounding tissues during the rapid water absorption of the zeolite, avoiding secondary damage, and improving the safety of the hemostatic material.

[0020] (2) The present invention improves the water absorption performance of the hemostatic material through the higher specific surface area and larger pore volume of nano-zeolite, thereby promoting the coagulation efficiency from the exogenous coagulation mechanism.

[0021] (3) The present invention reduces the aggregation of nano-zeolites and exposes more microporous structures by compounding nano-zeolites with polyphenol compounds. On the basis of promoting the exogenous coagulation mechanism, the coagulation performance is further improved from the endogenous coagulation mechanism. While improving the coagulation performance, antibacterial properties are introduced and thermal damage is eliminated, which greatly expands the scope of application and safety of the present invention as a hemostatic material. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a scanning electron microscope image of the natural zeolite (Z) of Comparative Example 3.

[0023] Figure 2 This is a scanning electron microscope image of the nano zeolite (NZ) of Comparative Example 1.

[0024] Figure 3 This is a scanning electron microscope image of the composite hemostatic material (TA / NZ-1) of Example 1 of the present invention.

[0025] Figure 4 This is a scanning electron microscope image of the composite hemostatic material (TA / NZ-2) of Example 2 of the present invention.

[0026] Figure 5 This is a scanning electron microscope image of the composite hemostatic material (TA / NZ-3) of Example 3 of the present invention.

[0027] Figure 6 This is a scanning electron microscope image of the composite hemostatic material (TA / NZ-4) of Example 4 of the present invention.

[0028] Figure 7 This is a scanning electron microscope image of the composite hemostatic material (TA / NZ-5) of Example 5 of the present invention.

[0029] Figure 8 This is a further magnified scanning electron microscope image of the composite hemostatic material (TA / NZ-5) of Example 5 of the present invention; Figure 9 This is a scanning electron microscope image of the composite hemostatic material (TA / NZ-6) of Example 6 of the present invention.

[0030] Figure 10 This is a scanning electron microscope image of the composite hemostatic material (TA / NZ-7) of Example 7 of the present invention.

[0031] Figure 11 This is a scanning electron microscope image of the composite hemostatic material (TA / NZ-8) of Example 8 of the present invention.

[0032] Figure 12 This is a scanning electron microscope image of the composite hemostatic material (TA / NZ-9) of Example 9 of the present invention.

[0033] Figure 13 This is a scanning electron microscope image of the composite hemostatic material (TA / NZ-10) according to Example 10 of the present invention.

[0034] Figure 14 This is a comparison chart of Fourier transform infrared spectra (FT-IR) of the composite hemostatic material (TA / NZ-2) of Example 2 of the present invention, NZ and TA of Comparative Example 1.

[0035] Figure 15 This is the EDS energy spectrum analysis diagram of the nano zeolite (NZ) of Comparative Example 1.

[0036] Figure 16 This is an EDS energy spectrum analysis diagram of the composite hemostatic material (TA / NZ-2) of Example 2 of the present invention.

[0037] Figure 17 The adsorption-desorption curve and pore size distribution diagram of the natural zeolite (Z) of Comparative Example 3.

[0038] Figure 18 The adsorption-desorption curve and pore size distribution diagram of the nano zeolite (NZ) of Comparative Example 1.

[0039] Figure 19 1 and 2 show the adsorption-desorption curve and pore size distribution diagram of the composite hemostatic material (TA / NZ-2) of Example 2 of the present invention.

[0040] Figure 20 This is a comparison chart of the in vitro coagulation time and coagulation index of the composite hemostatic material (TA / NZ-2) of Example 2 of the present invention, the nano zeolite (NZ) of Comparative Example 1, and the natural zeolite (Z) of Comparative Example 3.

[0041] Figure 21 Graphs showing temperature changes during coagulation tests of the composite hemostatic material (TA / NZ-2) of Example 2 of the present invention, the nano zeolite (NZ) of Comparative Example 1, and the natural zeolite (Z) of Comparative Example 3.

[0042] Figure 22 This is a comparison chart of the in vivo hemostatic time and blood loss of the composite hemostatic material (TA / NZ-2) of Example 2 of the present invention, the nano zeolite (NZ) of Comparative Example 1, and the natural zeolite (Z) of Comparative Example 3.

[0043] Figure 23 The figure shows the comparison of the antibacterial properties of Z, NZ and TA / NZ against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). DETAILED DESCRIPTION

[0044] The present invention is further described below with reference to specific embodiments. Based on these descriptions, a person skilled in the art will be able to implement the present invention. Furthermore, the embodiments of the present invention described below generally represent only a portion of the present invention, rather than all of the embodiments. Therefore, all other embodiments derived by a person skilled in the art without inventive effort based on the embodiments of the present invention should fall within the scope of protection of the present invention. The following abbreviations are used below: zeolite (Z), nanozeolite (NZ), and tannic acid (TA), with TA and NZ being used interchangeably in the accompanying drawings.

[0045] Embodiment 1: A composite hemostatic material, the preparation process of which is as follows: S1. Natural zeolite was mechanically ball-milled for 20 h to obtain zeolite powder with uniform particle size. The ball-milling parameters were as follows: ball-to-material mass ratio of 4:1, bead number ratio (2 mm: 4 mm: 6 mm) of 1:4:16, ball mill speed of 600 rpm, ball milling time of 10-25 h, and alternating forward and reverse rotations every 10 min with an interval of 5 min. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 2 weeks, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 1.0 mg / mL, and stirring at 30°C for 24 hours to obtain a tannic acid / zeolite composite colloid; S4. Freeze-drying the composite colloid to obtain a tannic acid / zeolite nanocomposite hemostatic material (TA / NZ-1).

[0046] Example 2: A composite hemostatic material, the preparation process of which is as follows: S1. Mechanically ball milling the natural zeolite for 20 h to obtain zeolite powder with uniform particle size. The ball milling parameters are the same as those described in Example 1. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 2 weeks, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 1.0 mg / mL, and stirring at 45°C for 24 hours to obtain a tannic acid / zeolite composite colloid; S4. Freeze-dry the composite colloid to obtain a tannic acid / zeolite nanocomposite hemostatic material (TA / NZ-2). Unless otherwise specified, "TA / NZ" in the accompanying drawings refers to the composite hemostatic material prepared in this example.

[0047] Example 3: A composite hemostatic material, the preparation process of which is as follows: S1. Mechanically ball milling the natural zeolite for 20 h to obtain zeolite powder with uniform particle size. The ball milling parameters are the same as those described in Example 1. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 2 weeks, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 1.0 mg / mL, and stirring at 60°C for 24 hours to obtain a tannic acid / zeolite composite colloid; S4. Freeze-drying the composite colloid to obtain a tannic acid / zeolite nanocomposite hemostatic material (TA / NZ-3).

[0048] Example 4: A composite hemostatic material, the preparation process of which is as follows: S1. Mechanically ball milling the natural zeolite for 20 h to obtain zeolite powder with uniform particle size. The ball milling parameters are the same as those described in Example 1. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 2 weeks, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 1.0 mg / mL, and stirring at 75°C for 24 h to obtain a tannic acid / zeolite composite colloid; S4. Freeze-drying the composite colloid to obtain a tannic acid / zeolite nanocomposite hemostatic material (TA / NZ-4).

[0049] Example 5: A composite hemostatic material, the preparation process of which is as follows: S1. Mechanically ball milling the natural zeolite for 20 h to obtain zeolite powder with uniform particle size. The ball milling parameters are the same as those described in Example 1. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 1 week, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 1.0 mg / mL, and stirring at 60°C for 12 h to obtain a tannic acid / zeolite composite colloid; S4. Freeze-drying the composite colloid to obtain a tannic acid / zeolite nanocomposite hemostatic material (TA / NZ-5).

[0050] Example 6: A composite hemostatic material, the preparation process of which is as follows: S1. Mechanically ball milling the natural zeolite for 20 h to obtain zeolite powder with uniform particle size. The ball milling parameters are the same as those described in Example 1. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 2 weeks, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 1.0 mg / mL, and stirring at 60°C for 36 h to obtain a tannic acid / zeolite composite colloid; S4. Freeze-drying the composite colloid to obtain a tannic acid / zeolite nanocomposite hemostatic material (TA / NZ-6).

[0051] Example 7: A composite hemostatic material, the preparation process of which is as follows: S1. Mechanically ball milling the natural zeolite for 20 h to obtain zeolite powder with uniform particle size. The ball milling parameters are the same as those described in Example 1. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 2 weeks, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 1.0 mg / mL, and stirring at 60°C for 48 h to obtain a tannic acid / zeolite composite colloid; S4. Freeze-drying the composite colloid to obtain a tannic acid / zeolite nanocomposite hemostatic material (TA / NZ-7).

[0052] Example 8: A composite hemostatic material, the preparation process of which is as follows: S1. Mechanically ball milling the natural zeolite for 20 h to obtain zeolite powder with uniform particle size. The ball milling parameters are the same as those described in Example 1. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 4 weeks, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 0.5 mg / mL, and stirring at 60°C for 12 h to obtain a tannic acid / zeolite composite colloid; S4. Freeze-drying the composite colloid to obtain a tannic acid / zeolite nanocomposite hemostatic material (TA / NZ-8).

[0053] Example 9: A composite hemostatic material, the preparation process of which is as follows: S1. Mechanically ball milling the natural zeolite for 20 h to obtain zeolite powder with uniform particle size. The ball milling parameters are the same as those described in Example 1. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 2 weeks, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 1.5 mg / mL, and stirring at 60°C for 12 h to obtain a tannic acid / zeolite composite colloid; S4. Freeze-drying the composite colloid to obtain a tannic acid / zeolite nanocomposite hemostatic material (TA / NZ-9).

[0054] Example 10: A composite hemostatic material, the preparation process of which is as follows: S1. Mechanically ball milling the natural zeolite for 20 h to obtain zeolite powder with uniform particle size. The ball milling parameters are the same as those described in Example 1. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 2 weeks, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 2.0 mg / mL, and stirring at 60°C for 12 h to obtain a tannic acid / zeolite composite colloid; S4. Freeze-drying the composite colloid to obtain a tannic acid / zeolite nanocomposite hemostatic material (TA / NZ-10).

[0055] Comparative Example 1: A hemostatic material, the preparation process of which is as follows: S1. Mechanically ball milling the natural zeolite for 20 h to obtain zeolite powder with uniform particle size. The ball milling parameters are the same as those described in Example 1. S2. Deionized water was added to the zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 2 weeks, and zeolite colloid was obtained after gravity sedimentation. S3. Zeolite colloid is freeze-dried to obtain nano zeolite (NZ).

[0056] Comparative Example 2: A hemostatic material, the preparation process of which is as follows: S1. Deionized water was added to natural zeolite powder, and the mixture was stirred under ultrasonic conditions for 2 h. The resulting suspension was allowed to stand for 2 weeks, and zeolite colloid was obtained after gravity sedimentation. S3, adding tannic acid to the zeolite colloid so that the concentration of tannic acid (TA) in the zeolite colloid is 1.0 mg / mL, and stirring at 45°C for 24 h to obtain a tannic acid / zeolite composite colloid; S4. Freeze-drying the composite colloid to obtain a hemostatic material (TA / Z).

[0057] Comparative Example 3: A hemostatic material, which is natural zeolite powder (Z).

[0058] from Figure 2 It can be seen that NZ is composed of zeolite agglomerates with diameters of about 50–200 nm, introducing a large number of mesopores and macropores, whose pore sizes range from tens to hundreds of nanometers. Figures 3 to 13 This is a scanning electron microscope image of the composite hemostatic material according to an embodiment of the present invention, especially from Figure 8 It can be seen that due to the introduction of tannic acid, TA / NZ is mainly composed of primary particles of about 10 nm, and the microporous structure of the zeolite itself is more exposed. These primary particles are evenly dispersed and construct a large number of mesoporous structures; there are a small amount of agglomerates on the surface, introducing mesoporous and macroporous structures.

[0059] Figures 14 to 19 The microstructure of the hemostatic materials of Example 2, Comparative Example 1 and Comparative Example 3 was further analyzed. After the material specific surface area and pore size analysis (BET), taking Example 2 as an example, the results showed that the specific surface area of NZ was 35.19 m 2 / g, of which the specific surface area in micropores is 5.37 m 2 / g, and the total pore volume is 0.2240 m 3 / g, and the micropore volume is 0.0024 m 3 / g; the specific surface area of TA / NZ is 28.33 m 2 / g, and the specific surface area in the micropores is 25.73 m 2 / g, and the total pore volume is 0.0566 m 3 / g, and the micropore volume is 0.0131 m3 / g. TA / NZ has a smaller specific surface area than NZ, but its micropore specific surface area is 4.62 times that of NZ. Its total pore volume is only one-fourth that of NZ, while its micropore volume is 5.46 times that of NZ. These data indicate that the introduction of tannic acid imparts a similar microporous structure to TA / NZ. The average pore diameter of NZ is 254.59 Å, while that of TA / NZ is 79.93 Å. The smaller pore size of TA / NZ further indicates that the particles in TA / NZ are smaller and more uniformly dispersed.

[0060] In vitro coagulation tests were performed on some of the above embodiments and comparative examples: At 37°C, 50 mg of each hemostatic material from the Examples and Comparative Examples was added to a 2 mL polystyrene tube. Then, 500 μL of activated blood was added. The tubes were inverted every 5 seconds to observe blood coagulation until the blood in the tubes stopped flowing. The corresponding clotting time was recorded. 100 mg of each hemostatic material from the Examples and Comparative Examples was placed in a 10 mL polystyrene tube. 200 μL of activated blood was added to each tube and incubated at 37°C for 5 min. After incubation, 5 mL of deionized water was added to the tubes. The tubes were incubated at 60 rpm for another 10 min. The supernatant was centrifuged and collected. The absorbance (OD) at 540 nm was measured using a microplate reader. 200 μL of blood was added to an empty tube and the above steps were repeated to obtain the OD value of the blank control group. The blood coagulation index (BCI) was calculated according to the following equation.

[0061] During the coagulation test, the temperature of the blood in the test tube is recorded simultaneously.

[0062] The representative embodiment 2 was selected for comparison. The coagulation test results of embodiment 2 and comparative examples 1 and 3 are shown as follows: Figure 20 As shown in FIG, the coagulation time of Example 2 is only 5 s, which is significantly improved compared to the 103 s of the natural zeolite hemostatic material. It can be seen that the coagulation effect of the nano zeolite with the addition of tannic acid is significantly improved. Figure 21 As shown in the figure, the maximum temperature of each group and the time exceeding 40°C during the coagulation test are marked. It can be seen that the natural zeolite material heats up significantly during the coagulation process and the high temperature time is long, which can easily cause thermal damage during use; while the maximum temperature of the composite hemostatic material in Example 2 during the coagulation process is only 48.9°C, and it only stays above 40°C for 1 second, which is enough to prove that tannic acid can produce a significant inhibitory effect on heat production.

[0063] In vivo hemostasis tests were performed on the hemostatic materials of some embodiments and comparative examples: Before the in vivo experiment, SD rats were fed for one week to allow them to adapt to the surrounding environment. Subsequently, the rats were randomly divided into 5 groups (n=5), namely the gauze group, Yunnan Baiyao (YNBY) group, Z group, NZ group, and TA / NZ (prepared in Example 2) group. All rats were fasted for 12 hours before surgery and deprived of water for 6 hours before surgery. Each rat was weighed and recorded, and anesthetized. No pain response was considered successful. After anesthesia, the rat's body temperature may drop, and the rat needs to be placed on a foam board to prevent hypothermia. Before the operation, the skin near the left thigh and groin of the rat was depilated and the skin was disinfected with iodine.

[0064] A longitudinal incision was made along the medial thigh using a scalpel. The skin was then dissected, followed by blunt dissection of the fascia and muscle layers to expose the vagus nerve, femoral artery, and femoral vein. The femoral artery was then isolated using surgical microforceps. The proximal end of the vessel was clamped with a hemostatic forceps. A 1 mL disposable syringe needle was then carefully used to create a rupture in the vessel. Scissors were then used to create a 5 mm long wound along the length of the vessel, starting from the rupture. Blood exuded from the wound was wiped away with a cotton ball. The hemostatic forceps were removed, and 0.4 g of hemostatic material was quickly applied to the wound. The wound was covered with a weighed medical gauze and manually applied. Pressure was applied continuously for 1 minute to stop bleeding. The gauze was then removed every 30 seconds to observe the progress of bleeding. The time to hemostasis was recorded after the wound ceased bleeding. The gauze group used only medical gauze for pressure to eliminate any influence on hemostasis. The gauze and hemostatic material were collected and weighed after hemostasis, and the increase in weight was used as blood loss.

[0065] The test results are as follows Figure 22 As shown, it can be seen that the composite hemostatic material of the present invention has a better coagulation effect than the traditional hemostatic material and can effectively reduce blood loss.

[0066] Antibacterial tests were performed on the hemostatic materials prepared in Example 2, Comparative Example 1, and Comparative Example 3: Sterilize the equipment required for the experiment in advance. Heat the sterilized LB solid culture medium until it is completely melted, transfer it to a sterile clean bench, and pour it into a plate when the LB solid culture medium cools to 50-60 °C. When the culture medium cools to completely solidify, close the culture dish lid to prevent the generation of water vapor. Measure 200 μL of a bacterial suspension with a concentration of 0.5 McFarland concentration and add it dropwise to the outer circle and center of the culture medium. Use a spreading rod to spread it until there is no obvious moisture on the surface to ensure that the bacteria are evenly dispersed. Use an agar puncher (D6 mm) to punch a hole in the culture dish and take another culture medium to seal the bottom of the hole. The hole should be free of cracks and set aside for later use. Use PBS to prepare the sample to 102.4 mg·mL -150 μL of sample dispersion was added to each well and the cells were incubated in a 37°C constant temperature incubator for 24 h. The inhibition zone was observed and the experiment was repeated three times for each sample.

[0067] The test results are as follows Figure 22 As shown, it can be seen that Z and NZ have no inhibition zone, while the composite hemostatic material prepared in Example 2 produces a relatively obvious inhibition zone, which can prove that the composite antibacterial material of the present invention has a good antibacterial effect.

[0068] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing a nanocomposite hemostatic material, characterized in that: The steps include: S1. ball milling the natural zeolite to obtain zeolite powder; S2. Stirring the zeolite powder in water under ultrasonic conditions to obtain a suspension, and then subjecting the suspension to gravity sedimentation to obtain nano-zeolite colloid; S3, adding polyphenol compounds to the nano-zeolite colloid, heating and stirring to obtain a nano-composite colloid; S4. Freeze-drying the nanocomposite colloid to obtain the nanocomposite hemostatic material.

2. The method for preparing a nanocomposite hemostatic material according to claim 1, characterized in that: In step S3, the polyphenolic compounds include one or more of tannic acid, catechin, proanthocyanidin, and quercetin.

3. The method for preparing a nanocomposite hemostatic material according to claim 1, wherein: In step S3, the concentration of the polyphenol compound in the zeolite colloid is 0.5-2.0 mg / mL.

4. The method for preparing a nanocomposite hemostatic material according to claim 1, wherein: The ball milling parameters in step S1 are as follows: ball-to-material mass ratio of 4:1, ball milling bead number ratio (2 mm: 4 mm: 6 mm) of 1:4:16, ball mill speed of 600 rpm, ball milling time of 10-25 h, alternating time of 1 time every 10 min, and interval of 5 min.

5. The method for preparing a nanocomposite hemostatic material according to claim 1, wherein: In step S3, the heating temperature is 30-75°C.

6. The method for preparing a nanocomposite hemostatic material according to claim 1, wherein: In step S3, the heating time is 12 to 48 hours.

7. A nanocomposite hemostatic material, characterized by: It is prepared by the preparation method according to any one of claims 1 to 6; the nanocomposite material comprises nano zeolite particles and polyphenol compounds grafted on the surface of the nano zeolite particles through hydrogen bonds and covalent bonds.

8. The nanocomposite hemostatic material according to claim 7, characterized in that: The primary particle size of the nanocomposite hemostatic material is 5-10 nm.

9. The nanocomposite hemostatic material according to claim 7, characterized in that: The volume of micropores in the composite hemostatic material accounts for 20-40% of the total pore volume.

10. Use of the composite hemostatic material according to any one of claims 7 to 9 in the preparation of medical hemostatic products.

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