Mesoporous bioactive glass composite graphene oxide hemostatic aerogel and preparation method and application thereof

Abstract

The application discloses a mesoporous bioglass composite graphene oxide hemostatic aerogel and a preparation method and application thereof. The mesoporous bioglass composite graphene oxide hemostatic aerogel is an aerogel with a three-dimensionally connected porous structure and is composed of mesoporous bioglass, graphene oxide and a polymer matrix; the mesoporous bioglass is adhered to the surface of the graphene oxide sheet; and the mass ratio of the mesoporous bioglass, the graphene oxide and the polymer matrix is (1-3):(4-6):3. The mesoporous bioglass composite graphene oxide hemostatic aerogel has good water absorption and hydrophilicity, excellent hemostatic performance, photothermal antibacterial performance, good blood compatibility and cell compatibility, can promote the antibacterial and healing of an infectious skin wound, and can be used as a rapid hemostatic material and an infectious skin wound antibacterial and healing material.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biomaterials, and particularly relates to a mesoporous bioglass composite graphene oxide hemostatic aerogel and a preparation method and application thereof. BACKGROUND

[0002] Uncontrolled bleeding caused by traffic accidents, accidental injuries and surgical bleeding and other trauma is one of the main causes of death, and it is of great significance to research and develop excellent hemostatic materials for emergency trauma rescue. So far, a series of materials such as tourniquets, gauze, gelatin sponge, powder, cellulose composite material, hemostatic hydrogel and the like have been studied for rapid and effective hemostasis. Among them, medical gauze, gelatin sponge and tourniquet are commonly used for hemostasis of small, shallow wound bleeding. However, the hemostatic effect of these traditional materials is limited, and it is difficult to treat deep wound bleeding, puncture wound bleeding and large area bleeding. On the other hand, due to the damage of the protective barrier (skin, mucosa), trauma is more susceptible to bacterial infection, leading to increased inflammation and slow healing, and antibacterial and healing after hemostasis are particularly important, while traditional hemostatic materials generally do not have significant antibacterial and healing functions. Therefore, the rapid hemostasis of large hemorrhagic trauma after emergency trauma and antibacterial and healing still have very high challenges.

[0003] Inorganic materials have a long history of application in rapid hemostasis and antibacterial healing of trauma. Since the U.S. Food and Drug Administration (FDA) approved the zeolite-based topical hemostatic agent "QuikClot" in 2002, inorganic powder materials have attracted great interest for rapid hemostasis. Mesoporous bioglass is an artificially synthesized inorganic mesoporous material, which has a high specific surface area and an ordered mesoporous structure of nanometer size, exhibits good water absorption and hydrophilicity, and has been shown to be able to rapidly absorb blood and concentrate blood clotting factors such as red blood cells, platelets, and the like, and it can also release Ca 2+ activate the extrinsic coagulation pathway to accelerate blood clotting. However, mesoporous bioglass releases a large amount of ions during hemostasis, resulting in an alkaline microenvironment at the wound site, and is difficult to clean up after being left at the wound site, hindering tissue healing.

[0004] In recent years, researchers have found that using the photothermal properties of materials for antibacterial treatment is an excellent strategy, because the material can convert light energy into heat energy under irradiation of a specific wavelength of light, inactivating bacterial proteins and lysing and killing bacteria through a physical method, thereby playing an antibacterial role. However, in the past, wound dressings with photothermal therapy function have mostly incorporated inorganic materials with photothermal properties into organic matrices such as hydrogels, electrospun membranes and the like, and the antibacterial performance of these dressings is limited by the content of inorganic materials, and these antibacterial dressings generally do not have the function of rapid hemostasis. Therefore, an effective way to solve the above problems is to prepare a high-inorganic-content hemostatic material with excellent hemostatic activity and antibacterial performance. SUMMARY

[0005] In view of the problems of the prior art, the present application aims to provide a mesoporous bioglass composite graphene oxide hemostatic aerogel and a preparation method and application thereof, the aerogel having a highly connected three-dimensional porous structure, good water absorption and hydrophilicity, excellent hemostatic performance, photothermal antibacterial performance, good blood compatibility and cell compatibility, and being capable of promoting the antibacterial and healing of infectious skin wounds, and can be used as a rapid hemostatic material and an infectious skin wound antibacterial and healing material.

[0006] In a first aspect, the present application provides a mesoporous bioglass composite graphene oxide hemostatic aerogel. The mesoporous bioglass composite graphene oxide hemostatic aerogel is an aerogel having a three-dimensional connected porous structure composed of mesoporous bioglass, graphene oxide and a polymer matrix; the mesoporous bioglass adheres to the surface of the graphene oxide sheet; wherein the mass ratio of the mesoporous bioglass, graphene oxide and polymer matrix is (1-3):(4-6):3.

[0007] The hemostatic material needs excellent hydrophilicity and rapid absorption capacity for water and blood exudate, so that the blood coagulation factors, red blood cells and platelets can rapidly gather to form a blood clot and a blood clot to prevent further bleeding. Mesoporous bioglass (MBG) is an inorganic powder with ordered mesoporous structure, high specific surface area and mesoporous volume, good hydrophilicity, and can release blood coagulation factor Ca 2+ accelerate hemostasis. Graphene oxide (GO) is a carbon material with excellent photothermal performance, a large number of hydroxyl and carboxyl functional groups on the surface endowing it with good hydrophilicity, and also has the functions of activating platelets and promoting blood coagulation. The polymer matrix, such as sodium alginate (SA), is a biocompatible organic binder, which is compounded with graphene oxide to enhance the mechanical properties. In the present application, a mesoporous bioglass composite graphene oxide hemostatic aerogel with good biocompatibility, effective hemostatic activity and excellent photothermal antibacterial performance is prepared by using mesoporous bioglass, graphene oxide and polymer matrix, which is used for rapid hemostasis and antibacterial and healing of infectious skin wounds.

[0008] Preferably, the porosity of the mesoporous bioglass composite graphene oxide hemostatic aerogel is 98.0-98.6%, and the pore size distribution is 30-50 μm.

[0009] Preferably, the water absorption rate of the mesoporous bioglass composite graphene oxide hemostatic aerogel is 5000-6700%, and the Zeta potential is -25 to -15 mV.

[0010] Preferably, the 30-second BCI blood coagulation index of the mesoporous bioglass composite graphene oxide hemostatic aerogel is less than 10%.

[0011] Preferably, the polymer matrix is at least one of sodium alginate, polyvinyl alcohol, chitosan, and methacrylated gelatin.

[0012] Preferably, the mesoporous bioglass has a chemical composition of 80SiO2·15CaO·5P2O5, a specific surface area of 350-400 m2 / g, a mesopore size of 3-4 nm, and a mesopore volume of 0.3-0.4 cm3 / g. 2 3 Preferably, the mesoporous bioglass has a chemical composition of 80SiO2·15CaO·5P2O5, a specific surface area of 350-400 m2 / g, a mesopore size of 3-4 nm, and a mesopore volume of 0.3-0.4 cm3 / g.

[0013] Preferably, the graphene oxide has a size of 1-10 μm.

[0014] In a second aspect, the present application provides a method for preparing the mesoporous bioglass composite graphene oxide hemostatic aerogel as described in any of the above. The method comprises: mixing a polymer matrix with water to obtain a polymer matrix solution; mixing mesoporous bioglass and graphene oxide with water to obtain a mesoporous bioglass / graphene oxide aqueous dispersion; mixing the polymer matrix solution and the mesoporous bioglass / graphene oxide aqueous dispersion and homogenizing to obtain a suspension; freezing the suspension under low-temperature conditions to obtain an uncrosslinked green body; crosslinking the uncrosslinked green body by immersing it in a calcium chloride solution, washing with water, and freeze-drying under low-temperature conditions to obtain the mesoporous bioglass composite graphene oxide hemostatic aerogel; preferably, the mass ratio of the polymer matrix to water in the polymer matrix solution is 1-10:100; the mass ratio of the total mass of the mesoporous bioglass and graphene oxide to water in the mesoporous bioglass / graphene oxide aqueous dispersion is 0.1-1:100; more preferably, the pressure of the freeze-drying under low-temperature conditions is 10-70 Pa, the temperature is -30 to -10℃, and the time is 24-48 hours.

[0015] The present application utilizes freeze-drying technology to prepare a mesoporous bioglass composite graphene oxide hemostatic aerogel with a controllable three-dimensionally interconnected porous morphology. This method has the advantages of simple process and easy control of conditions. In addition, a three-dimensionally porous structure with high porosity is particularly crucial for enabling a hemostatic material to have stronger water absorption capacity, and freeze-drying technology is a suitable method for manufacturing a material with high porosity. The present application first prepares a mesoporous bioglass composite graphene oxide hemostatic aerogel with high porosity, water absorption, good hydrophilicity, excellent in-vitro and in-vivo hemostatic and blood-activating functions, and skin wound antibacterial and healing functions, which has a good clinical application prospect.

[0016] Preferably, the polymer matrix is selected from one of low-viscosity sodium alginate, medium-viscosity sodium alginate, and high-viscosity sodium alginate, and is preferably high-viscosity sodium alginate.

[0017] In a third aspect, the present application provides the use of the mesoporous bioglass composite graphene oxide hemostatic aerogel as described in any of the above in the preparation of a rapid hemostatic material for external injuries and an antibacterial and healing material for infected skin wounds.​

[0018] Advantages:

[0019] The mesoporous bioglass composite graphene oxide hemostatic aerogel has good water absorption, hydrophilicity, excellent hemostatic activity, photothermal performance and biocompatibility. Animal experiments prove that the mesoporous bioglass composite graphene oxide hemostatic aerogel has excellent in-vivo rapid hemostatic performance and the dual functions of anti-bacterial and healing of infectious skin wounds. Therefore, the mesoporous bioglass composite graphene oxide hemostatic aerogel is a new type of dual-function biomaterial, which has a wide prospect in the clinical application of rapid hemostasis and anti-bacterial and healing of infectious skin wounds. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 The figure is the phase and microstructure characterization of the synthesized mesoporous bioglass. (a) XRD pattern, (b) TEM image.

[0021] Figure 2 The figure is the BET test of the mesoporous bioglass. (a) Nitrogen adsorption-desorption curve, (b) Mesopore size distribution curve, (c) Mesopore structure parameters. Figure 1 and Figure 2 It is shown that the mesoporous bioglass is an amorphous glass phase structure, which has an ordered mesoporous structure, a nanoscale mesopore size, a high specific surface area and a mesopore volume.

[0022] Figure 3 The figure is the macroscopic morphology optical photo and microstructure SEM image of the mesoporous bioglass composite graphene oxide hemostatic aerogel. (a) The mesoporous bioglass composite graphene oxide hemostatic aerogel is prepared in the shape of a block, (b-d) The SEM images of the microstructure with different magnifications. It is shown in the figure that the mesoporous bioglass composite graphene oxide hemostatic aerogel prepared by the freeze-drying technology has good formability, has a three-dimensionally connected porous structure, and the mesoporous bioglass can be seen to adhere to the graphene oxide sheet.

[0023] Figure 4 The figure is the physicochemical property experiment of the mesoporous bioglass composite graphene oxide hemostatic aerogel. (a) Water absorption rate, (b) Zeta potential, (c) Hydrophilicity. It is shown in the figure that the mesoporous bioglass composite graphene oxide hemostatic aerogel has a high water absorption rate (5000-6700%) and a negative Zeta potential, because the mesoporous bioglass and the graphene oxide constituting the hemostatic aerogel have negative charged groups; the mesoporous bioglass composite graphene oxide hemostatic aerogel shows good hydrophilicity and can absorb water in a very short time. Figure 4(a) and (b) of FIG. 1 from left to right are 30SA / 70GO, 30SA / 60GO / 10MBG, 30SA / 50GO / 20MBG and 30SA / 40GO / 30MBG in order.

[0024] Figure 5 In vitro hemostatic performance experiment of mesoporous bioglass composite graphene oxide hemostatic hydrogel. (a) BCI blood clotting index, (b) partial thrombin activation time, (c) hemolysis rate, (d) SEM image of red blood cell and platelet adhesion. Figure 5 (a) of FIG. 2 from left to right are Blank, Gauze, Gelatin, 30SA / 70GO, 30SA / 60GO / 10MBG, 30SA / 50GO / 20MBG and 30SA / 40GO / 30MBG in order; (b) of FIG. 2 from left to right are Blank, Gelatin, 30SA / 70GO, 30SA / 60GO / 10MBG, 30SA / 50GO / 20MBG and 30SA / 40GO / 30MBG in order; (c) of FIG. 2 the same concentration of each group of column chart from left to right are 30SA / 70GO, 30SA / 60GO / 10MBG, 30SA / 50GO / 20MBG and 30SA / 40GO / 30MBG in order. The 30-second BCI blood clotting index of mesoporous bioglass composite graphene oxide hemostatic hydrogel can be lower than 10%, and can activate the endogenous coagulation pathway, the hemolysis rate is <5% at a concentration of 1-8 mg / mL, and the SEM image shows that a large number of red blood cells and platelets adhere to the surface of the hemostatic hydrogel, indicating that the mesoporous bioglass composite graphene oxide hemostatic hydrogel has excellent hemostatic activity and good blood compatibility.

[0025] Figure 6 Mesoporous bioglass composite graphene oxide hemostatic hydrogel is used for rat liver puncture injury hemostasis. (a) Schematic diagram of rat liver puncture injury hemostasis, (b) hemostatic time, (c) bleeding amount. Figure 6 (b) and (c) of FIG. 3 from left to right are Blank, Gelatin, 30SA / 70GO, 30SA / 60GO / 10MBG, 30SA / 50GO / 20MBG and 30SA / 40GO / 30MBG in order. It is shown in the figure that the mesoporous bioglass composite graphene oxide hemostatic hydrogel can significantly shorten the bleeding time of rat liver puncture injury and reduce the bleeding amount compared with the control group, and has excellent rapid hemostatic function.

[0026] Figure 7Photothermal performance experiments of mesoporous bioglass-graphene oxide hemostatic aerogels. (a) Dry temperature rise curve, (b) Wet temperature rise curve, and (c) Photothermal cycling stability. The figures demonstrate that the mesoporous bioglass-graphene oxide hemostatic aerogels exhibit excellent photothermal performance and stability, allowing for rapid temperature increases within a short period of time.

[0027] Figure 8 In vitro antibacterial experiments on mesoporous bioglass-graphene oxide hemostatic aerogels. (a-b) Optical images and antibacterial rates of Staphylococcus aureus antibacterial experiments, (c-d) Optical images and antibacterial rates of Escherichia coli antibacterial experiments. Figure 8 (b) and (d) show, from left to right, Blank, 30SA / 70GO, 30SA / 60GO / 10BMG, 30SA / 70GO+PTT, and 30SA / 60GO / 10BMG+PTT. The figures show that the mesoporous bioglass-graphene oxide hemostatic aerogel exhibits excellent photothermal antibacterial properties, achieving approximately 99% inhibition against both Staphylococcus aureus and Escherichia coli.

[0028] Figure 9 Figure 2 shows the cytocompatibility of mesoporous bioglass-graphene oxide hemostatic aerogels. (a) Proliferation results of human fibroblasts cultured in extracts of mesoporous bioglass-graphene oxide hemostatic aerogels (the bars in each group on the same days, from left to right, are blank, 30SA / 70GO, and 30SA / 60GO / 10MBG). (b) Proliferation results of human umbilical vein endothelial cells cultured in extracts of mesoporous bioglass-graphene oxide hemostatic aerogels (the bars in each group on the same days, from left to right, are blank, 30SA / 70GO, and 30SA / 60GO / 10MBG). The figure shows that the mesoporous bioglass-graphene oxide hemostatic aerogels have good cytocompatibility and have no inhibitory effect on cell proliferation.

[0029] Figure 10 This is an antibacterial and healing study of infected skin wounds in mice using mesoporous bioglass-graphene oxide hemostatic aerogels. (a) Optical photographs of infected skin wounds at different time points, (b) Relative wound area statistics of infected skin wounds at different time points (bar graphs for each group on the same day, from left to right, represent the blank group, 30SA / 70GO, and 30SA / 60GO / 10MBG), and (c) HE staining and (d) Masson trichrome staining of the wound site skin tissue on day 14. The figures demonstrate that the mesoporous bioglass-graphene oxide hemostatic aerogels exhibit excellent photothermal antibacterial properties, killing most bacteria in the early stages of infected skin wound formation, thereby promoting wound healing, collagen fiber deposition, and new epidermal formation. DETAILED DESCRIPTION

[0030] The present application is further illustrated by the following examples, which are intended to be illustrative only and not limiting of the present application. Unless otherwise specified, each percentage is by mass.

[0031] The mesoporous bioglass composite graphene oxide hemostatic aerogel is an aerogel with a three-dimensionally interconnected porous structure composed of mesoporous bioglass, graphene oxide and a polymer matrix. The mesoporous bioglass of the mesoporous bioglass composite graphene oxide hemostatic aerogel adheres to the surface of the graphene oxide sheets.

[0032] In the ternary aerogel system of mesoporous bioglass / graphene oxide / polymer matrix, the graphene oxide provides the main support performance. As the content of graphene oxide decreases, the self-supporting property of the aerogel material weakens, the volume shrinks slightly, and the porosity and water absorption rate also decrease. In addition, in the mesoporous bioglass composite graphene oxide hemostatic aerogel, a small amount of mesoporous bioglass added can enhance the ability of the hemostatic aerogel to activate the endogenous coagulation pathway, thereby reducing the partial thrombin activation time to achieve better performance in promoting coagulation and accelerating hemostasis. The mass ratio of mesoporous bioglass, graphene oxide and polymer matrix is (1-3):(4-6):3. At this time, the water absorption rate of the aerogel is the highest. Too high or too low mass ratio of the polymer matrix will result in a decrease in the water absorption rate of the aerogel. When the mass ratio of graphene oxide is too low, the aerogel has poor formability and cannot be prepared into a stable aerogel block. In some technical solutions, the mass ratio of the mesoporous bioglass composite graphene oxide hemostatic aerogel is 10-30% mesoporous bioglass, 40-60% graphene oxide and 30% polymer matrix. As an example, the chemical ratio is 60% GO / 10% MBG / 30% SA, 50% GO / 20% MBG / 30% SA, 40% GO / 30% MBG / 30% SA, etc.

[0033] The mesoporous bioglass composite graphene oxide hemostatic aerogel has a highly interconnected three-dimensional porous structure. In an alternative embodiment, the pore size of the mesoporous bioglass composite graphene oxide hemostatic aerogel is 30-50 μm, and the porosity is 98.0-98.6%.

[0034] The mesoporous bioglass composite graphene oxide hemostatic aerogel has good water absorption and hydrophilicity. In an alternative embodiment, the water absorption rate of the mesoporous bioglass composite graphene oxide hemostatic aerogel can reach 5000-6700%; and the Zeta potential is -25 to -15 mV.

[0035] The mesoporous bioglass composite graphene oxide hemostatic aerogel has excellent hemostatic performance. In an alternative embodiment, the mesoporous bioglass composite graphene oxide hemostatic aerogel has a 30-second BCI blood clotting index of less than 10%.

[0036] The above shows that the mesoporous bioglass composite graphene oxide hemostatic aerogel has good water absorption and hydrophilicity, and has excellent hemostatic performance and photothermal antibacterial performance, and also has good blood compatibility and cell compatibility, and can promote the antibacterial and healing of infected skin wounds. Because in the composite hemostatic aerogel system, graphene oxide constitutes the main porous structure, and mesoporous bioglass adheres to the surface of the graphene oxide sheet as a small amount of modifier. This promotes the substantial reduction of the amount of mesoporous bioglass on the one hand, and the mesoporous bioglass no longer remains in the wound and is difficult to clean up, which effectively avoids the wound microenvironment and the powder residue caused by the direct hemostasis of the mesoporous bioglass powder, which hinders wound healing.

[0037] Next, a method for preparing the mesoporous bioglass composite graphene oxide hemostatic aerogel according to the present application is shown. The method uses mesoporous bioglass, graphene oxide and a polymer matrix as raw materials to prepare the mesoporous bioglass composite graphene oxide hemostatic aerogel by freeze-drying technology.

[0038] In an alternative embodiment, a mixed solution is prepared using mesoporous bioglass, graphene oxide and a polymer matrix as raw materials. After stirring for a period of time, the solution is frozen at low temperature and vacuum dried to obtain an uncrosslinked green body. The uncrosslinked green body is soaked in a calcium chloride solution for crosslinking, and then washed with deionized water, and then frozen at low temperature and vacuum dried to obtain the mesoporous bioglass composite graphene oxide hemostatic aerogel. The hemostatic aerogel has a three-dimensionally interconnected porous structure, has the functions of rapid hemostasis, antibiosis and wound healing promotion, and can be used as a rapid hemostatic and skin wound repair material.

[0039] Specifically, the following steps are used.

[0040] The polymer matrix is mixed with water to obtain a polymer matrix solution. The polymer matrix acts as a high molecular binder to improve the overall strength of the composite aerogel. The polymer matrix is preferably sodium alginate. The sodium alginate can be selected from low viscosity sodium alginate, medium viscosity sodium alginate and high viscosity sodium alginate. Preferably, the sodium alginate is high viscosity sodium alginate, because the composite aerogel prepared from high viscosity sodium alginate has the highest strength. In an alternative embodiment, the mass ratio of the polymer matrix to water in the polymer matrix solution is 1-10:100. As an example, the polymer matrix is mixed with deionized water at a mass ratio of 1:100, and stirred at room temperature for 2 hours to obtain a 1 wt.% polymer matrix solution.

[0041] The sodium alginate can also be replaced by other high-molecular-matrix such as polyvinyl alcohol (PVA), chitosan, gelatin methacrylated (GelMA), etc.

[0042] The mesoporous bioglass and graphene oxide are mixed with water to obtain a mesoporous bioglass / graphene oxide water dispersion.

[0043] In some embodiments, the mesoporous bioglass has a chemical composition of 80SiO2·15CaO5·5P2O5, a specific surface area of 350-400 m2 / g, a mesopore size of 3-4 nm, and a mesopore volume of 0.3-0.4 cm3 / g. 2 / g, a mesopore size of 3-4 nm, and a mesopore volume of 0.3-0.4 cm 3 / g. It should be understood that other bioglasses or bioceramics with other chemical compositions are also applicable to the present application, such as 58S mesoporous bioglass, bioglass containing active elements such as Sr and Mn, etc. Similarly, bioglasses without the above mesoporous structure, such as conventional bioglass powder, are also applicable to the present application.

[0044] The mesoporous bioglass can be synthesized by a sol-gel-evaporation induced self-assembly method. The raw materials for synthesizing the mesoporous bioglass include anhydrous ethanol, polyether P123, hydrochloric acid, tetraethyl orthosilicate, calcium nitrate tetrahydrate, and triethyl phosphate. As an example, 20 g of polyether P123 is first completely dissolved in 300 g of anhydrous ethanol, and then 33.5 g of tetraethyl orthosilicate, 7.12 g of calcium nitrate tetrahydrate, 3.65 g of triethyl phosphate, and 5 g of hydrochloric acid (0.5 M) are sequentially added, and stirring is continued for 24 hours. The solution is then poured into a large glass container and placed in a fume hood to evaporate the solvent for 7 days to obtain a dry gel. The dry gel is calcined at 650°C for 6 hours with a heating rate of 2°C / min. After the calcination is completed, a white powder is collected, ground, and sieved through a 320-mesh sieve to obtain the mesoporous bioglass.

[0045] The size of the graphene oxide can be 1-10 μm.

[0046] The mass ratio of the total mass of the mesoporous bioglass and the graphene oxide to the mass of water in the mesoporous bioglass / graphene oxide water dispersion is 0.1-1:100. As an example, the mesoporous bioglass, the graphene oxide, and deionized water are mixed in a mass fraction ratio of (1-3):(4-7):700, and ultrasonic treatment is performed for 30 minutes to obtain a mesoporous bioglass / graphene oxide water dispersion.

[0047] The high-molecular-matrix solution and the mesoporous bioglass / graphene oxide water dispersion are mixed and homogenized to obtain a suspension. As an example, the high-molecular-matrix solution and the mesoporous bioglass / graphene oxide water dispersion are mixed in a mass ratio of 3:7 and stirred for 12 hours to completely homogenize the three components, thereby obtaining a black water suspension.

[0048] The suspension is frozen at low temperature to obtain uncrosslinked precursor. In an alternative embodiment, the suspension is poured into a silica gel mold and frozen in a low temperature refrigerator, and then demolded to obtain a frozen gel. For example, the freezing temperature is controlled at -80°C, and the freezing time is 0.5-1 hour. The obtained frozen gel is placed in a vacuum drying device for vacuum drying to obtain the uncrosslinked precursor. For example, the vacuum drying machine parameters are vacuum degree of 10-70 Pa, and drying time of 24-48 hours.

[0049] The uncrosslinked precursor is immersed in a calcium chloride solution for sufficient crosslinking, washed with water, and low-temperature freeze-dried to obtain the mesoporous bioglass composite graphene oxide hemostatic aerogel. The obtained uncrosslinked precursor is immersed in a calcium chloride solution to crosslink the polymer matrix. Deionized water is used for washing to remove excess calcium chloride solution. In an alternative embodiment, the low-temperature freeze-drying pressure is 10-70 Pa, the temperature is -30 to -10°C, and the time is 24-48 hours. In one example, the uncrosslinked precursor is immersed in a 5 wt.% calcium chloride solution for 1 hour for sufficient crosslinking, and then the aerogel is immersed in deionized water for 0.5 hour, repeated three times, and then placed in a low-temperature refrigerator at -80°C for freezing for 0.5-1 hour, and vacuum dried at 10-70 Pa for 24-48 hours to obtain the mesoporous bioglass composite graphene oxide hemostatic aerogel.

[0050] As a specific example, the detailed steps for preparing the mesoporous bioglass composite graphene oxide hemostatic aerogel by freeze-drying technology are as follows: (A) mixing the polymer matrix and deionized water at a mass ratio of 1:100, stirring at room temperature for 1-2 hours to obtain a 1 wt.% polymer matrix aqueous solution; (B) mixing the mesoporous bioglass, graphene oxide and deionized water at a mass fraction ratio of (1-3):(4-7):700, and ultrasonic treatment for 30 min to obtain a mesoporous bioglass / graphene oxide aqueous dispersion; preferably, the total solid content of the mesoporous bioglass, graphene oxide and polymer matrix accounts for 1 wt.% of the total mass of the black aqueous suspension, and the remaining 99 wt.% is the water content; (C) mixing the 1 wt.% polymer matrix aqueous solution and the mesoporous bioglass / graphene oxide aqueous dispersion at a mass ratio of 3:7, stirring for 12 h to make the three components fully homogeneous, to obtain a black aqueous suspension; (D) pouring the obtained black aqueous suspension into a silica gel mold, and placing the silica gel mold in a -80℃ low-temperature refrigerator for freezing for 0.5-1 h to obtain a frozen gel; (E) demolding the obtained frozen gel, and placing it in a vacuum drying machine for drying under a vacuum of 10-70 Pa for 24-48 h to obtain an uncrosslinked blank; (F) mixing anhydrous calcium chloride and deionized water at a mass ratio of 1:19 to obtain a 5 wt.% calcium chloride solution, immersing the obtained uncrosslinked blank in the 5 wt.% calcium chloride solution for 1 h to crosslink the polymer matrix, and then immersing it in deionized water for washing 3 times, each time for 30 min; (G) freezing the washed uncrosslinked blank again in a -80℃ low-temperature refrigerator for 0.5-1 h, and then placing it in a vacuum drying machine for drying under a vacuum of 10-70 Pa for 24-48 h to obtain the mesoporous bioglass composite graphene oxide hemostatic aerogel.

[0051] The hemostatic aerogel material prepared by the method of the present application is completely made of biocompatible mesoporous bioglass, graphene oxide and polymer matrix combined with freeze-drying technology, and the method is simple, low in cost and can be mass-produced, and the obtained mesoporous bioglass composite graphene oxide hemostatic aerogel can be used as a new type of clinical emergency hemostatic and antibacterial and healing dressing for infectious wounds.

[0052] The following examples are further listed to illustrate the present application in detail. It should also be understood that the following examples are only used to further illustrate the present application, and cannot be understood as limiting the protection scope of the present application, and some non-essential improvements and adjustments made by those skilled in the art according to the above content of the present application all belong to the protection scope of the present application. The specific process parameters and the like in the following examples are only one example in the appropriate range, i.e. those skilled in the art can make appropriate selection within the range through the description herein, and are not limited to the specific values of the following examples.

[0053] Example 1

[0054] Mesoporous bioglass was synthesized using sol-gel-evaporation induced self-assembly method. 20 g of polyether P123 was completely dissolved in 300 g of anhydrous ethanol, then 33.5 g of tetraethyl orthosilicate, 7.12 g of calcium nitrate tetrahydrate, 3.65 g of triethyl phosphate and 5 g of hydrochloric acid (0.5 M) were added in turn, and stirring was continued for 24 hours to obtain a mixed solution. The solution was poured into a large glass container and placed in a fume hood to evaporate the solvent for 7 days to obtain a dry gel. The dry gel was heated to 650℃ at a heating rate of 2℃ / min, and burned at 650℃ for 6 hours to remove the organic template. The white powder obtained after burning was ground and sieved through a 320 mesh sieve to obtain mesoporous bioglass.

[0055] Structure characterization of mesoporous bioglass: it can be known from X-ray diffraction (XRD), transmission electron microscopy (TEM) and BET test that the mesoporous bioglass synthesized in the application has an amorphous glass phase, an ordered mesoporous structure, a high specific surface area, a nanoscale mesoporous size and a high mesoporous volume (for example, see Figure 1 and Figure 2 ).

[0056] Sodium alginate solution was configured. Sodium alginate and deionized water were mixed at a mass ratio of 1:100, and stirred at room temperature for 2 hours to obtain a 1wt.% sodium alginate solution. The aqueous dispersion of mesoporous bioglass / graphene oxide was configured. Mesoporous bioglass, graphene oxide and deionized water were mixed at a mass ratio of 1:6:700, and ultrasonic treatment was carried out for 30 min to obtain the aqueous dispersion of mesoporous bioglass / graphene oxide. The sodium alginate solution and the aqueous dispersion of mesoporous bioglass / graphene oxide were mixed at a mass ratio of 3:7 and stirred for 12 hours to homogenize the three components, and an aqueous suspension was obtained. The aqueous suspension was poured into a silica gel mold, and placed in a-80℃ low-temperature refrigerator for freezing for 0.5 hours, then demolded to obtain a frozen gel, and continued to be dried at-20℃ under 10Pa for 24 hours to obtain an uncrosslinked blank. The uncrosslinked blank was soaked in a 5wt.% calcium chloride solution for 1 hour for sufficient crosslinking, and then taken out to soak in deionized water for 0.5 hours, repeated three times. After being frozen in a-80℃ low-temperature refrigerator for 0.5 hours again, it was continued to be dried at-20℃ under 10Pa for 24 hours to obtain a mesoporous bioglass composite graphene oxide hemostatic aerogel (30SA / 60GO / 10MBG).

[0057] Morphology and microstructure characterization of mesoporous bioglass composite graphene oxide hemostatic aerogel: it can be known from morphology observation and scanning electron microscopy (SEM) that the mesoporous bioglass composite graphene oxide hemostatic aerogel of the application has black color, light weight, highly interconnected porous structure (for example, see Figure 3For example, by adjusting the composition ratio of mesoporous bioglass composite graphene oxide hemostatic aerogel (graphene oxide 40-60 wt.%, mesoporous bioglass 10-30 wt.%, sodium alginate 30 wt.%), mesoporous bioglass composite graphene oxide hemostatic aerogel with a pore size of 30-50 μm and a porosity of 98.0-98.6% can be obtained.

[0058] The water absorption, hydrophilicity, Zeta potential and in vitro and in vivo hemostatic performance of the mesoporous bioglass composite graphene oxide hemostatic aerogel (30SA / 60GO / 10MBG) were studied, and the results are shown in Table 2. Figures 4 to 6 .

[0059] Example 2

[0060] In this example 2, the preparation process of the mesoporous bioglass composite graphene oxide hemostatic aerogel was the same as that of example 1, except that the mesoporous bioglass, graphene oxide and deionized water were mixed in a mass ratio of 2:5:700 to obtain a mesoporous bioglass / graphene oxide aqueous dispersion, and the mesoporous bioglass composite graphene oxide hemostatic aerogel (30SA / 50GO / 20MBG) was prepared.

[0061] The photothermal performance, in vitro antibacterial performance, cell compatibility and antibacterial and healing experiment of infected skin wounds of the mesoporous bioglass composite graphene oxide hemostatic aerogel (30SA / 50GO / 20MBG) were studied, and the results are shown in Table 4. Figures 7 to 10 .

[0062] Example 3

[0063] In this example 3, the preparation process of the mesoporous bioglass composite graphene oxide hemostatic aerogel was the same as that of example 1, except that the mesoporous bioglass, graphene oxide and deionized water were mixed in a mass ratio of 3:4:700 to obtain a mesoporous bioglass / graphene oxide aqueous dispersion, and the mesoporous bioglass composite graphene oxide hemostatic aerogel (30SA / 40GO / 30MBG) was prepared.

[0064] The water absorption, hydrophilicity, Zeta potential, in vitro and in vivo hemostatic performance, photothermal performance, in vitro antibacterial performance, cell compatibility and antibacterial and healing experiment of infected skin wounds of the mesoporous bioglass composite graphene oxide hemostatic aerogel (30SA / 40GO / 30MBG) were studied, and the results are shown in Table 5. Figures 4 to 10 .

[0065] Comparative Example 1

[0066] Graphene oxide hemostatic aerogel was prepared by freeze-drying technique as a control group. The preparation process of graphene oxide hemostatic aerogel in Comparative Example 1 was according to Example 1, except that no mesoporous bioglass was added, and only graphene oxide and deionized water were mixed in a mass ratio of 1:100 to obtain a graphene oxide water dispersion, and the graphene oxide hemostatic aerogel (30SA / 70GO) was prepared.

[0067] Comparative Example 2

[0068] Mesoporous bioglass composite graphene oxide hemostatic aerogels with different contents of mesoporous bioglass and graphene oxide were prepared by freeze-drying technique as a control group. The preparation process of mesoporous bioglass composite graphene oxide hemostatic aerogel in Comparative Example 2 was according to Example 1, except that the mass ratio of sodium alginate, graphene oxide and mesoporous bioglass was 2:1:1, and the chemical composition of the mesoporous bioglass composite graphene oxide hemostatic aerogel was (50SA / 25GO / 25MBG).

[0069] Table 1 is the composition and performance parameters of the hemostatic aerogels prepared in Examples 1-3 and Comparative Examples 1-2

[0070]

[0071] Physicochemical properties of mesoporous bioglass composite graphene oxide hemostatic aerogel

[0072] The mesoporous bioglass composite graphene oxide hemostatic aerogel (3 parallel samples) was soaked in deionized water for 1 day to fully absorb water, and the weight difference before and after water absorption was measured to represent the water absorption rate. The Zeta potential of the mesoporous bioglass composite graphene oxide hemostatic aerogel (3 parallel samples) was measured in deionized water. The wettability of the mesoporous bioglass composite graphene oxide hemostatic aerogel to deionized water was studied, and the water static contact angle was measured. The results showed that the mesoporous bioglass composite graphene oxide hemostatic aerogel had excellent water absorption performance, and the water absorption rate of the mesoporous bioglass composite graphene oxide hemostatic aerogel with different proportions could reach 5000-6700%, and the water absorption rate increased with the increase of the content of graphene oxide. The mesoporous bioglass composite graphene oxide hemostatic aerogel showed negative Zeta potential, and the Zeta potential range was -25 to -15 mV, and the 30SA / 60GO / 10MBG group had the highest negative Zeta potential. The static water contact angle test results showed that the mesoporous bioglass composite graphene oxide hemostatic aerogel with different proportions had good hydrophilicity, and could completely absorb water within 0.02s (see, for example, Figure 4 ).

[0073] In vitro hemostatic performance of mesoporous bioglass composite graphene oxide hemostatic aerogel

[0074] The BCI blood coagulation index, partial thrombin activation time, hemolysis rate and red blood cell and platelet adhesion of the mesoporous bioglass composite graphene oxide hemostatic aerogels with different composition ratios were studied. The results showed that the mesoporous bioglass composite graphene oxide hemostatic aerogels with different composition ratios all had good blood absorption, and could accelerate blood coagulation. The BCI blood coagulation index of 30 s was less than 10%, which was much lower than the BCI blood coagulation index of 60% of medical gauze and the BCI blood coagulation index of 85% of gelatin sponge. The partial thrombin activation time test results showed that the hemostatic aerogel of the 30SA / 60GO / 10MBG group had the shortest partial thrombin activation time, which was significantly different from that of the gelatin sponge, proving that it could activate the endogenous coagulation pathway. The hemolysis rate of the mesoporous bioglass composite graphene oxide hemostatic aerogel was lower than the international standard of 5%, proving that it had good blood compatibility. After the mesoporous bioglass composite graphene oxide hemostatic aerogel absorbed blood, a large number of red blood cells and platelets could be observed on the surface in the SEM image, proving that the mesoporous bioglass composite graphene oxide hemostatic aerogel could effectively absorb plasma and make red blood cells and platelets aggregate, thereby accelerating blood coagulation (see, for example, Figure 5 ).

[0075] In vivo hemostatic performance of mesoporous bioglass composite graphene oxide hemostatic aerogels

[0076] It was confirmed that the mesoporous bioglass composite graphene oxide hemostatic aerogels had excellent in vivo hemostatic performance. The mesoporous bioglass composite graphene oxide hemostatic aerogels with different composition ratios and control materials such as medical gauze and gelatin sponge were used to study their hemostatic effect in a rat liver puncture injury. The results showed that the mesoporous bioglass composite graphene oxide hemostatic aerogels had good blood absorption and blood coagulation function in vivo, and the hemostatic schematic diagram showed that the wound around the mesoporous bioglass composite graphene oxide hemostatic aerogel was clean and had few blood clots after hemostasis, while the control group had a large amount of unabsorbed blood coagulation around the wound, which indicated that the mesoporous bioglass composite graphene oxide hemostatic aerogel had good hemostatic performance. Quantitative analysis further showed that the hemostatic time and bleeding amount of the mesoporous bioglass composite graphene oxide hemostatic aerogel were greatly reduced compared with the control group. In the rat liver injury hemostatic model, the hemostatic time and bleeding amount were reduced by 50% and 60% compared with the control group, respectively, and the 30SA / 60GO / 10MBG group containing mesoporous bioglass had faster hemostatic time and less bleeding amount than the 30SA / 70GO group without mesoporous bioglass, which indicated that the incorporation of mesoporous bioglass improved the hemostatic performance of the aerogel. The mesoporous bioglass composite graphene oxide hemostatic aerogel had excellent rapid hemostatic function, could quickly absorb blood in the early stage of wound bleeding, promote blood coagulation, shorten the bleeding time and greatly reduce the bleeding amount (see, for example, Figure 6 ).

[0077] Photothermal properties of mesoporous bioglass composite graphene oxide hemostatic aerogels

[0078] The photothermal properties of mesoporous bioglass composite graphene oxide hemostatic aerogels with different compositions were studied. The photothermal properties and photothermal stability of the mesoporous bioglass composite graphene oxide hemostatic aerogels under 808 nm laser irradiation, dry conditions, wet conditions, and cycling conditions were tested, respectively. The results showed that the mesoporous bioglass composite graphene oxide hemostatic aerogels had good photothermal properties, which was due to the excellent photothermal properties of the main component, graphene oxide. Under 0.3 W / cm 2 laser power, the mesoporous bioglass composite graphene oxide hemostatic aerogels could be heated to near 100°C in a short time (less than 30 seconds) under dry conditions; under 0.9 W / cm 2 laser power, the mesoporous bioglass composite graphene oxide hemostatic aerogels could also be heated to near 60°C in about 10 minutes under wet conditions; in addition, the mesoporous bioglass composite graphene oxide hemostatic aerogels also had photothermal stability, and the photothermal properties did not attenuate after multiple cycles in the photothermal cycling test (for example, see Figure 7 ).

[0079] In vitro antibacterial properties of mesoporous bioglass composite graphene oxide hemostatic aerogels

[0080] The in vitro antibacterial effects of mesoporous bioglass composite graphene oxide hemostatic aerogels on two common bacteria, Staphylococcus aureus and Escherichia coli, were investigated by the plate counting method. The results showed that the mesoporous bioglass composite graphene oxide hemostatic aerogels combined with photothermal effects exhibited significant antibacterial properties, which was due to their excellent photothermal properties, which could raise the temperature of the bacterial solution to above 60°C and maintain it for 10-15 minutes, meeting the requirements for effectively killing Staphylococcus aureus and Escherichia coli, with an inhibition rate of up to 99%; in addition, the mesoporous bioglass composite graphene oxide hemostatic aerogels of the 30SA / 60GO / 10MBG group also showed a certain inhibition of the growth of Escherichia coli, which was because Escherichia coli was less tolerant to alkaline environments, and the degradation of mesoporous bioglass would make the bacterial solution environment alkaline, thereby inhibiting the growth of Escherichia coli to a certain extent (for example, see Figure 8 ).

[0081] Cell compatibility of mesoporous bioglass composite graphene oxide hemostatic aerogels

[0082] Human fibroblast and human umbilical vein endothelial cells were cultured with the 30SA / 70GO group and 30SA / 60GO / 10MBG group mesoporous bioglass composite graphene oxide hemostatic aerogel leaching solution, and the effect of the material on the proliferation of human fibroblast and human umbilical vein endothelial cells was studied, and CCK-8 cell counting box was used for quantitative characterization. The results of the study showed that the 30SA / 70GO group and 30SA / 60GO / 10MBG group mesoporous bioglass composite graphene oxide hemostatic aerogel had good biocompatibility, and the leaching solution had no inhibition effect on the proliferation and growth of human fibroblast and human umbilical vein endothelial cells (see, for example Figure 9 ).

[0083] Antibacterial and healing experiments of mesoporous bioglass composite graphene oxide hemostatic aerogel on infected skin wounds in mice

[0084] The 30SA / 60GO / 10MBG group and 30SA / 70GO group were used as dressings to treat infected skin wounds in mice, and the function of promoting skin wound healing was studied. The results showed that the mesoporous bioglass composite graphene oxide hemostatic aerogel combined with photothermal antibacterial treatment could efficiently and quickly kill bacteria in the early stage of infected skin wounds. As can be seen from the photographs of the appearance of the skin wounds, on the 4th day, the skin wounds of the 30SA / 70GO group and 30SA / 60GO / 10MBG group mice were relatively dry, and no infected pus was visible, while a large amount of yellow infected pus was still visible on the skin wounds of the blank group mice; relative wound area quantitative statistics further confirmed this point, and the mesoporous bioglass composite graphene oxide hemostatic aerogel could promote the healing of infected skin wounds, and the experimental group 30SA / 60GO / 10MBG showed better function of promoting wound healing than the control group 30SA / 70GO; the results of HE staining and Masson's trichrome staining of the skin tissue sections of the wounds on the 14th day showed that the 30SA / 60GO / 10MBG group mesoporous bioglass composite graphene oxide hemostatic aerogel had the function of promoting skin wound healing, promoting the deposition of collagen fibers and the generation of new epidermis (see, for example Figure 10 ).

Claims

1. A mesoporous bioglass composite graphene oxide hemostatic aerogel, characterized in that: The mesoporous bioglass composite graphene oxide hemostatic aerogel is an aerogel with a three-dimensional interconnected porous structure composed of mesoporous bioglass, graphene oxide and a polymer matrix; the mesoporous bioglass is adhered to the surface of the graphene oxide sheet; wherein the mass ratio of the mesoporous bioglass, graphene oxide and polymer matrix is ​​(1-3): (4-6): 3; The porosity of the mesoporous bioglass composite graphene oxide hemostatic aerogel is 98.0-98.6%, and the pore size distribution is 30-50 μm; The mesoporous bioglass composite graphene oxide hemostatic aerogel has a water absorption rate of 5000-6700% and a Zeta potential of -25 to -15 mV; The 30-second BCI coagulation index of the mesoporous bioglass composite graphene oxide hemostatic aerogel is lower than 10%.

2. The mesoporous bioglass composite graphene oxide hemostatic aerogel according to claim 1, characterized in that: The polymer matrix is ​​selected from one of low-viscosity sodium alginate, medium-viscosity sodium alginate and high-viscosity sodium alginate.

3. The mesoporous bioglass composite graphene oxide hemostatic aerogel according to claim 1, characterized in that: The chemical composition of the mesoporous bioglass is 80SiO2·15CaO5·5P2O5, and the specific surface area is 350~400m 2 / g, the mesopore size is 3~4nm, and the mesopore volume is 0.3~0.4cm 3 / g.

4. The mesoporous bioglass composite graphene oxide hemostatic aerogel according to claim 1, characterized in that: The size of the graphene oxide is 1-10 μm.

5. A method for preparing the mesoporous bioglass composite graphene oxide hemostatic aerogel according to any one of claims 1 to 4, characterized in that: include: Mixing a polymer matrix with water to obtain a polymer matrix solution; mixing mesoporous bioglass and graphene oxide with water to obtain a mesoporous bioglass / graphene oxide aqueous dispersion; mixing the polymer matrix solution and the mesoporous bioglass / graphene oxide aqueous dispersion and homogenizing to obtain a suspension; Freezing the suspension under low temperature conditions to obtain an uncrosslinked green body; The uncrosslinked green body is immersed in a calcium chloride solution for full crosslinking, washed with water, and freeze-dried at low temperature to obtain the mesoporous bioglass composite graphene oxide hemostatic aerogel.

6. The preparation method according to claim 5, characterized in that The mass ratio of the polymer matrix to water in the polymer matrix solution is 1-10:100; the mass ratio of the total mass of the mesoporous bioglass and graphene oxide to water in the mesoporous bioglass / graphene oxide aqueous dispersion is 0.1-1:

100.

7. The preparation method according to claim 5, characterized in that The pressure of low-temperature freeze drying is 10~70Pa, the temperature is -30 to -10℃, and the time is 24~48 hours.

8. Use of the mesoporous bioglass composite graphene oxide hemostatic aerogel according to any one of claims 1 to 4 in the preparation of a rapid hemostatic material for trauma and an antibacterial and healing material for infectious skin wounds.

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