Hydrogel-modified heart valve prosthesis and preparation method and application thereof

Abstract

The invention discloses a hydrogel modified artificial heart valve as well as a preparation method and application thereof, and belongs to the technical field of biomedical materials. Comprising the following steps: S1, constructing a hyaluronic acid and folic acid dual-modified loaded CY-09-coated FA-HA-MSN; and S2, preparing the decellularized artificial valve CY-09 (at) MSN (at) HDAV modified by the ROS responsive hydrogel loaded with the targeted macrophage nanoparticles. The active oxygen ROS responsive hydrogel is modified on the decellularized valve stent, so that collagen in the decellularized valve stent is prevented from being in direct contact with blood, the chronic inflammatory reaction induced by the valve stent is reduced, and the antithrombotic performance of the artificial valve is improved; meanwhile, the hydrogel contains targeted macrophage drug-loaded nanoparticles, and the nanoparticles are made of a mesoporous silica (MSN) material and are loaded with a small molecule drug CY-09; and through the modification of hyaluronic acid and folic acid, the function of targeting macrophages is realized.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedical materials, and particularly relates to a hydrogel-modified artificial heart valve, a preparation method thereof, and an application thereof. Background Art

[0002] Heart valve disease seriously endangers human health. With the continuous deepening of the aging of the population, its incidence rate will further increase. Artificial heart valve replacement is the main treatment method for heart valve disease. At present, the commonly used artificial heart valves in clinical practice include mechanical valves and biological valves: among them, mechanical valves require patients to take anticoagulant drugs for life, and are prone to problems such as bleeding and embolism, and the noise during operation is relatively obvious; while biological valves do not require lifelong anticoagulation and have a high degree of integration with interventional techniques. With the technological innovation in recent decades, they have now surpassed mechanical valves and become the mainstream choice in clinical practice. However, the existing biological valves are mainly cross-linked with glutaraldehyde. As the implantation time prolongs, obvious calcification will occur in the biological valves, resulting in limited opening and closing of the valves, and then leading to valve stenosis or regurgitation. Therefore, the research and development of a new type of artificial valve that can resist calcification for life has great scientific value and practical significance.

[0003] At present, the tissue engineering in-situ regeneration technology is considered to be the most promising method to break through the limitations of existing artificial valves. This technology can realize the recellularization of artificial valves, thereby breaking through the technical limitations of valve calcification and decay, and maximizing the service life of artificial valves. The decellularization technology is currently the most potential artificial valve treatment technology. The biological valve scaffold obtained through this technology has significant advantages. It can retain the original shape and original three-dimensional structure of the valve, and minimize its immunogenicity; however, due to the lack of protection of endothelial cells, the collagen of the scaffold itself is completely exposed to the blood flow environment, which is extremely likely to induce chronic inflammatory reactions and thrombosis. The immune response is deeply involved in the valve regeneration process, and macrophages play an important role in it. The immune cells infiltrated in the valve scaffold can secrete a variety of cytokines, which can then induce the migration, proliferation, and differentiation of adjacent endothelial cells and endothelial progenitor cells, thereby realizing the tissue remodeling of the valve. Different subtypes of macrophages play different roles in the immune response; according to the different functions of macrophages, they are usually divided into M1-type macrophages mainly with pro-inflammatory effects and M2-type macrophages mainly with anti-inflammatory effects. Two different types of macrophages play important roles in different periods of valve remodeling. Therefore, regulating the proportion distribution of different types of macrophages at different times is of great significance for the regeneration of tissue engineering valves. Summary of the Invention

[0004] The present invention adopts the following technical solutions: In the first aspect, the present invention provides a preparation method of a hydrogel-modified artificial heart valve, comprising the following steps: S1. Construct CY-09@FA-HA-MSN: Dissolve 0.1 - 0.5 g of cetyltrimethylammonium bromide (CTAB) in an aqueous sodium hydroxide solution, stir and heat to 60 - 90 °C, add 0.1 - 0.5 mL of absolute ethanol and stir; dropwise add 1 - 5 mL of tetraethyl orthosilicate (TEOS) solution, and react at 60 - 100 °C for 1 - 5 h; centrifuge the reaction product at 10000 - 11000 rpm for 10 - 20 min; wash and dry overnight, and calcine at 500 - 600 °C for 2 - 7 h to obtain white powder, i.e., mesoporous silica nanoparticles (MSN); Mix 10 - 50 mg of MSN and 1 - 5 mg of CY-09 in 1 - 5 mL of PBS, and stir at 1 - 5 °C for 12 - 36 h to obtain CY-09@MSN; dissolve 10 - 50 mg of CY-09@MSN in 1 - 5 mL of phosphate buffer solution (PBS), then add 50 - 150 μL of a polyethyleneimine (PEI) solution with a concentration of 70 - 80 mg / mL, stir at room temperature for 20 - 40 min, then centrifuge at 5000 - 15000 rpm for 10 - 20 min, repeat centrifugation, wash and freeze-dry to obtain CY-09@PEI-MSN; dissolve 20 - 40 mg of CY-09@PEI-MSN, 10 - 20 mg of hyaluronic acid (HA), 10 - 50 mg of N-hydroxysuccinimide (NHS) and 10 - 50 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) in water, stir for 1 - 2 h to obtain CY-09@HA-PEI-MSN; add 10 - 20 mg of folic acid (FA), stir at room temperature for 12 - 36 h, centrifuge, wash and freeze-dry overnight to obtain CY-09@FA-HA-MSN; S2. Prepare CY-09@MSN@HDAV: Take 0.05 - 0.15 g of N,N,N′,N-tetramethyl-1,3-propanediamine and 0.1 - 1.0 g of 4-(bromomethyl)phenylboronic acid and dissolve them in 5 - 15 mL of N,N-dimethylformamide (DMF), stir at 50 - 70 °C for 10 - 20 h; add it to 100 - 300 mL of pre-cooled tetrahydrofuran (THF), let it stand for precipitation, discard the supernatant, then wash with THF and dry to obtain N1-(4-boronophenyl)-N3-(4-boronophenyl)-N1,N1,N3,N3-tetramethylpropane-1,3-diamine (TPA) product, and prepare a TPA solution with a concentration of 20 - 30 mg / mL; Add 1 - 6 mg of CY-09@FA-HA-MSN to 1 - 6 mL of a polyvinyl alcohol (PVA) solution with a concentration of 20 - 30 mg / mL. Take the decellularized aortic valve (DAV) and place it in this mixture. Shake it on a shaker at 1 - 5 °C and 50 - 100 rpm for 12 - 36 h, wash it, and then put it into the prepared TPA solution. Shake it on a shaker at 10 - 50 rpm for 1 - 2 h, and wash it to obtain CY-09@MSN@HDAV.

[0005] Further, in step S1, dissolve 0.4 g of CTAB in water, stir and heat it to 80 °C, add 0.4 mL of absolute ethanol and stir; dropwise add 2 mL of TEOS solution and react at 80 °C for 2 h; centrifuge the reaction product at 10200 rpm for 15 min; wash and dry it overnight, and calcine it at 550 °C for 5 h to obtain a white powder, which is MSN.

[0006] Further, the CTAB is dissolved in 1.44 mL of an aqueous sodium hydroxide solution with a concentration of 2 M.

[0007] Further, in step S1, mix 30 mg of MSN and 2 mg of CY-09 in 4 mL of PBS and stir at 4 °C for 24 h to obtain CY-09@MSN; dissolve 30 mg of CY-09@MSN in 4 mL of PBS, then add 100 μL of a PEI solution with a concentration of 75 mg / mL, stir at room temperature for 30 min, then centrifuge at 10000 rpm for 15 min, repeat centrifugation, wash, and freeze-dry to obtain CY-09@PEI-MSN; dissolve 30 mg of CY-09@PEI-MSN, 15 mg of HA, 40 mg of NHS, and 30 mg of EDC in water and stir for 1 h to obtain CY-09@HA-PEI-MSN; add 15 mg of FA and stir at room temperature for 24 h, centrifuge, wash, and freeze-dry overnight to obtain CY-09@FA-HA-MSN.

[0008] Further, in step S2, dissolve 0.1 g of N,N,N′,N-tetramethyl-1,3-propanediamine and 0.5 g of 4-(bromomethyl)phenylboronic acid in 10 mL of DMF and stir at 60 °C for 16 h; add it to 200 mL of pre-cooled THF, let it stand for precipitation, discard the supernatant, then wash it with THF and dry it to obtain the TPA product, and prepare a TPA solution with a concentration of 25 mg / mL.

[0009] Further, the TPA is prepared into a TPA solution using PBS as the solvent.

[0010] Further, in step S2, 5 mg of CY-09@FA-HA-MSN was added to 5 mL of a PVA solution with a concentration of 25 mg / mL. The decellularized valve DAV was placed in this mixture, and the mixture was shaken on a shaker at 4°C and 60 rpm for 24 h. After washing, it was placed in the prepared TPA solution and shaken on a shaker at 30 rpm for 1 h, and then washed to obtain CY-09@MSN@HDAV.

[0011] Further, PBS was used as a solvent to prepare the PVA solution for the PVA.

[0012] In a second aspect, the present invention also provides a hydrogel-modified artificial heart valve, which is obtained according to the described preparation method.

[0013] In a third aspect, the present invention also provides the application of the described artificial heart valve in the preparation of anti-thrombotic, endothelialization-promoting, and anti-calcification functional materials.

[0014] Mechanism of action: 1. The reactive oxygen species-responsive hydrogel can react with the ROS generated on the valve surface, thereby reducing the ROS concentration on the valve surface, further inhibiting the polarization of macrophages into the M1 type, reducing the M1 / M2 ratio, and promoting valve re-endothelialization; 2. The MSN nanoparticles loaded with CY-09, after being modified with folic acid and hyaluronic acid, significantly increase their targeting ability to macrophages; the engulfed MSN can achieve the gradual release of CY-09 in macrophages, further interfering with the formation of inflammasomes NLRP3 and AIM2, and then inhibiting the polarization of macrophages into M1-type macrophages and reducing the M1 / M2 ratio, promoting valve re-endothelialization; 3. The decellularized valve modified with the hydrogel can significantly enhance its mechanical properties, which is more in line with the actual requirements; 4. The decellularized valve modified with the hydrogel can prevent the collagen of the valve from being directly exposed to the blood environment, thereby inhibiting the formation of thrombus; 5. The decellularized valve modified with the hydrogel can enhance its anti-enzymatic degradation performance while inhibiting the polarization of macrophages into M1-type macrophages.

[0015] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention modifies a reactive oxygen species ROS-responsive hydrogel on the decellularized valve scaffold, avoiding the direct contact of the collagen in the decellularized valve scaffold with the blood, reducing the chronic inflammatory response induced by the valve scaffold, and improving the anti-thrombotic performance of the artificial valve; 2. The ROS-responsive hydrogel in the present invention can react rapidly with the ROS generated in the body, reducing the content of ROS and promoting the polarization of macrophages into the M2 type; 3. The hydrogel in the present invention contains macrophage-targeted drug-loaded nanoparticles: the nanoparticles are made of mesoporous silica (MSN), loaded with the small molecule drug CY-09; modified with hyaluronic acid and folic acid to achieve the function of targeting macrophages. Description of the Drawings

[0016] Figure 1 It is the SEM scanning result diagram of the nanoparticles in Example 1; Figure 2 It is the effect diagram of CY-09@FA-HA-MSN nanoparticles on macrophage ROS in Example 1; Figure 3 It is the SEM and EDS scanning result diagrams of different tissue valves in Example 1; Figure 4 It is the determination result of the mechanical properties of different tissue valves in Example 1; Figure 5 It is the result diagram of subcutaneous embedding in rats in Example 2; Figure 6 It is the fluorescence result of macrophage polarization of valve embedding in Example 2; Figure 7 It is the endothelial fluorescence result of valve abdominal aorta transplantation in Example 2; Detailed Embodiment

[0017] For better illustration of the present invention, the following examples are specifically listed. Obviously, the described examples are only a part of the present invention, rather than all the examples. Based on the examples in the present invention, other examples obtained by those skilled in the art without creative efforts all fall within the scope of protection of the present invention.

[0018] The technical solution of the present invention will be further described below with reference to the drawings and examples.

[0019] Example 1

[0020] 1. Construction of hyaluronic acid and folic acid dual-modified CY-09@FA-HA-MSN (1) Dissolve 0.4 g of CTAB (N-hexadecyltrimethylammonium bromide) in 200 mL of ultrapure water containing 1.44 mL of sodium hydroxide aqueous solution (2M), and heat it to 80 °C with stirring. Then add 0.4 mL of absolute ethanol and stir for another 5 min. Then, dropwise add 2 mL of tetraethyl orthosilicate (TEOS) solution and keep the reaction at 80 °C for 2 h. Centrifuge the product (10,200 revolutions, 15 min), and wash it three times with ethanol and deionized water respectively to remove residual impurities. To remove CTAB, dry the product overnight, calcine it at 550 °C for 5 h, and finally obtain a white powder denoted as MSN; (2) Mix 30 mg of MSN and 2 mg of CY-09 (dissolved in 200 μL of DMSO) in 4 mL of phosphate-buffered saline (PBS, 10 mM, pH = 7.4), and stir for 24 h at 4 °C to obtain CY-09@MSN. Take 30 mg of CY-09@MSN and dissolve it in 4 ml of PBS solution, then add 100 μl of PEI solution (75 mg / mL), and stir the mixture at room temperature for 30 min. Then, centrifuge the product (10,000 rpm, 15 min); repeat the centrifugation operation three times and wash it once with ultrapure water each time, and freeze-dry to obtain CY-09@PEI-MSN. Take 30 mg of freeze-dried CY-09@PEI-MSN, 15 mg of HA, 40 mg of NHS, and 30 mg of EDC and dissolve them in 10 mL of distilled water, and stir for 1 h to obtain CY-09@HA-PEI-MSN. Subsequently, add 15 mg of FA (dissolved in 5 ml of DMSO), and then continue to stir at room temperature for 24 h. Finally, obtain CY-09@FA-HA-MSN through centrifugation, washing, and overnight freeze-drying.

[0021] The obtained nanoparticles were scanned by scanning electron microscopy (SEM), and their Zeta potential and particle size were detected (the detection results are shown in Figure 1 ), and the results showed that after the MSN was modified with HA and FA, its surface morphology changed significantly.

[0022] 2. Preparation of acellular artificial valve CY-09@MSN@HDAV modified with ROS hydrogel loaded with nanoparticles (1) Dissolve 0.1 g of N,N,N′,N-tetramethyl-1,3-propanediamine and 0.5 g of 4-(bromomethyl)phenylboronic acid in 10 mL of dimethylformamide (DMF), and stir at 60 °C for 16 h. Add the reacted mixed solution to 200 ml of tetrahydrofuran (THF) pre-cooled to 4 °C, let it stand for the product to precipitate, and discard the supernatant; then add 100 mL of pre-cooled tetrahydrofuran again and wash three times repeatedly; then dry to obtain the product N1-(4-boronophenyl)-N3-(4-boronophenyl)-N1,N1,N3,N3-tetramethylpropane-1,3-diamine (TPA), and use PBS solution as the solvent to prepare a TPA solution with a concentration of 25 mg / ml; (2)Using PBS as a solvent, prepare 5 ml of a 25 mg / mL PVA (polyvinyl alcohol, with an average molar mass of 30,000 - 70,000) solution, and then add 5 mg of CY-09@FA-HA-MSN to it; Place 10 decellularized porcine valves (DAV) into this mixed solution, and shake it on a horizontal shaker at 4 °C for 24 h (60 rpm / min); Then wash the decellularized valves 3 times with PBS solution; Then place them into 5 ml of the prepared TPA solution and shake it on a horizontal shaker at a low speed for 1 h (30 rpm / min); Then wash the valves 3 times with PBS solution to obtain a decellularized artificial valve CY-09@MSN@HDAV modified with ROS hydrogel loaded with targeted macrophages nanoparticles.

[0023] Verify the effect of reducing macrophage ROS using the CY-09@FA-HA-MSN nanoparticles prepared in the above example of this embodiment. Use the fluorescent probe DCFH-DA to detect the content of reactive oxygen species in macrophages induced by the THP-1 cell line. The results show that CY-09@FA-HA-MSN can significantly reduce the reactive oxygen species in macrophages.

[0024] At the same time, use the decellularized valve DAV and the glutaraldehyde cross-linked valve GAV as control groups, and compare them with the artificial valve CY-09@MSN@HDAV prepared by the present invention.

[0025] Among them, the preparation method of the decellularized valve (DAV) is to place the porcine aortic valve in a TRIS-HCI buffer solution (40 mM, pH 7.8) containing 2% 3-[3-(cholamidopropyl)dimethylammonio]propanesulfonate (CHAPS) and 2 mmol / l tributylphosphine (TnBP), and continuously shake and decellularize at room temperature for 24 h. Rinse it 6 times with sterile water, 10 min each time. Then place it in a TRIS-HCI buffer solution (40 mM, pH 7.8) containing 2% CHAPS, 2 mmol / l TnBP, 1% amidinosulfobetaine (ASB-14), and 2% sulfobetaine 10 (SB 3-10), and continue to shake and decellularize at room temperature for 24 h to obtain DAV; The preparation method of the glutaraldehyde cross-linked valve GAV is to add 30 porcine aortic valves to 50 mL of a 0.625% glutaraldehyde solution, cross-link them in the dark at room temperature for 48 hours, and obtain GAV after repeated rinsing with PBS.

[0026] Perform SEM scanning on the three obtained valve materials, and detect the Si element using an energy dispersive spectrometer (EDS, Energy Dispersive Spectrometer). The results are shown in Figure 3Results showed that compared with DAV, the surface of CY-09@MSN@HDAV was smoother and the exposed collagen fibers were significantly reduced. By detecting the Si element on the valve surface with EDS, it was found that CY-09@MSN@HDAV was significantly higher than the DAV and GAV groups, proving the attachment of nanoparticles on its surface, that is, this experiment proved that the nanoparticles were successfully modified onto the valve.

[0027] Uniaxial tensile tests were performed on the three valve samples to compare the changes in mechanical properties after hydrogel modification: The valve samples were cut into strips with a length of 10 mm and a width of 4 mm, and the strips were uniaxially stretched at a speed of 15 mm / min. The results are shown in Figure 4 : It can be observed that the mechanical properties of the valves after hydrogel modification were significantly improved compared with ordinary DAV, and were basically equivalent to those of GVA valves.

[0028] Example 2

[0029] Two in vivo animal experiments, subcutaneous embedding and abdominal aorta transplantation in rats, were carried out on the valve samples obtained in Example 1. Sprague-Dawley (SD) rats were purchased from Liaoning Changsheng Biotechnology Co., Ltd. and were housed in the SPF barrier breeding room of the Animal Experiment Center of Huazhong University of Science and Technology. SD rats are generally housed in a barrier environment with a temperature of 20°C - 26°C, a humidity of 40% - 70%, a light cycle of 12 hours of light / 12 hours of darkness, and sufficient supply of feed and drinking water.

[0030] (1) Subcutaneous embedding experiment: CY-09@MSN@HDAV and DAV obtained in Example 1, as well as the glutaraldehyde-crosslinked bioprosthetic valve GAV, were respectively implanted under the back skin of 6-week-old SD rats. After 28 days, the rats were sacrificed, and the subcutaneous valve tissues were taken out, fixed with paraformaldehyde, embedded in paraffin, and sectioned. The sections were stained with silver nitrate and stained with CD163 (M2 macrophages, green) and iNOS (M1 macrophages, red). (The test results are shown in Figure 5 、 6 ); The results of 28-day subcutaneous embedding in rats showed that compared with GAV, neither DAV nor CY-09@MSN@HDAV showed obvious calcification.

[0031] Due to the large cytotoxicity of the valves in the GAV group, no macrophage attachment was observed; compared with DAV, the macrophages colonized on the surface of CY-09@MSN@HDAV were mainly M2 macrophages (CD163, green); the surface of DAV was mainly colonized by M1 macrophages (iNOS, red).

[0032] (2)Abdominal aorta transplantation experiment: The valve samples were trimmed into 5 mm × 5 mm square pieces and sewn into a pipeline with an 8-0 surgical suture needle. Male SD rats weighing about 200 g were anesthetized by intraperitoneal injection of 0.3% sodium pentobarbital at a dose of 0.01 - 0.02 mL / g. The abdomen of the SD rats was opened, and the aortic blood vessels were dissected to expose the abdominal aorta. The abdominal aorta was blocked and transected using a vascular clamp, and then the sewn pipeline was connected to the abdominal aorta by end-to-end anastomosis. After 7 days, samples were taken, fixed with paraformaldehyde, embedded in paraffin, and sectioned. The sections were stained with CD31 (endothelial cells, red) fluorescence.

[0033] The results of the 7-day abdominal aorta transplantation model in rats showed that compared with DAV and GAV, CY-09@MSN@HDAV had more obvious infiltration of endothelial cells (CD31, red), indicating its good potential for re-endothelialization.

[0034] The embodiments described above are only for describing the preferred mode of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A preparation method of a hydrogel-modified artificial heart valve, characterized in that, It includes the following steps: S1. Construct CY-09@FA-HA-MSN: Dissolve 0.1 - 0.5 g of cetyltrimethylammonium bromide (CTAB) in an aqueous sodium hydroxide solution, stir and heat to 60 - 90 °C, add 0.1 - 0.5 mL of absolute ethanol and stir; dropwise add 1 - 5 mL of tetraethyl orthosilicate (TEOS) solution and react at 60 - 100 °C for 1 - 5 h; centrifuge the reaction product at 10000 - 11000 rpm for 10 - 20 min; wash and dry overnight, and calcine at 500 - 600 °C for 2 - 7 h to obtain white powder, namely mesoporous silica nanoparticles (MSN); Mix 10 - 50 mg of MSN and 1 - 5 mg of CY-09 in 1 - 5 mL of PBS, stir at 1 - 5 °C for 12 - 36 h to obtain CY-09@MSN; dissolve 10 - 50 mg of CY-09@MSN in 1 - 5 mL of phosphate buffer solution (PBS), then add 50 - 150 μL of a polyethyleneimine (PEI) solution with a concentration of 70 - 80 mg / mL, stir at room temperature for 20 - 40 min, then centrifuge at 5000 - 15000 rpm for 10 - 20 min, repeat centrifugation, wash and freeze-dry to obtain CY-09@PEI-MSN; dissolve 20 - 40 mg of CY-09@PEI-MSN, 10 - 20 mg of hyaluronic acid (HA), 10 - 50 mg of N-hydroxysuccinimide (NHS) and 10 - 50 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) in water, stir for 1 - 2 h to obtain CY-09@HA-PEI-MSN; add 10 - 20 mg of folic acid (FA), stir at room temperature for 12 - 36 h, centrifuge, wash and freeze-dry overnight to obtain CY-09@FA-HA-MSN; S2. Prepare CY-09@MSN@HDAV: Dissolve 0.05 - 0.15 g of N,N,N′,N-tetramethyl-1,3-propanediamine and 0.1 - 1.0 g of 4-(bromomethyl)phenylboronic acid in 5 - 15 mL of N,N-dimethylformamide (DMF), stir at 50 - 70 °C for 10 - 20 h; add it to 100 - 300 mL of pre-cooled THF, let it stand for precipitation, discard the supernatant, then wash with THF and dry to obtain the product of N1-(4-boronophenyl)-N3-(4-boronophenyl)-N1,N1,N3,N3-tetramethylpropane-1,3-diamine (TPA), and configure TPA into a TPA solution with a concentration of 20 - 30 mg / mL; Add 1 - 6 mg of CY-09@FA-HA-MSN to 1 - 6 mL of a polyvinyl alcohol (PVA) solution with a concentration of 20 - 30 mg / mL. Take the decellularized aortic valve (DAV) and place it in this mixture. Shake it on a shaker at 1 - 5 °C and 50 - 100 rpm for 12 - 36 h, wash it, then put it into the prepared TPA solution and shake it on a shaker at 10 - 50 rpm for 1 - 2 h, and wash it to obtain the decellularized artificial valve modified with ROS hydrogel loaded with nanoparticles, CY-09@MSN@HDAV.

2. The preparation method according to claim 1, characterized in that, In step S1, take 0.4 g of CTAB and dissolve it in an aqueous sodium hydroxide solution. Stir and heat it to 80 °C, add 0.4 mL of absolute ethanol and stir; dropwise add 2 mL of TEOS solution and react at 80 °C for 2 h; centrifuge the reaction product at 10200 rpm for 15 min; wash and dry it overnight, and calcine it at 550 °C for 5 h to obtain a white powder, which is MSN.

3. The preparation method according to claim 2, wherein The aqueous sodium hydroxide solution is prepared by the following method: Add 1.44 mL of 2 M aqueous sodium hydroxide solution to 200 mL of ultrapure water.

4. The preparation method according to claim 1, characterized in that, In step S1, mix 30 mg of MSN and 2 mg of CY-09 in 4 mL of PBS and stir at 4 °C for 24 h to obtain CY-09@MSN; take 30 mg of CY-09@MSN and dissolve it in 4 mL of PBS, then add 100 μL of a PEI solution with a concentration of 75 mg / mL, stir at room temperature for 30 min, then centrifuge at 10000 rpm for 15 min, repeat centrifugation, wash and freeze-dry to obtain CY-09@PEI-MSN; take 30 mg of CY-09@PEI-MSN, 15 mg of HA, 40 mg of NHS and 30 mg of EDC and dissolve them in water, stir for 1 h to obtain CY-09@HA-PEI-MSN; add 15 mg of FA and stir at room temperature for 24 h, centrifuge, wash and freeze-dry overnight to obtain CY-09@FA-HA-MSN.

5. The preparation method according to claim 1, characterized in that, In step S2, take 0.1 g of N,N,N′,N-tetramethyl-1,3-propanediamine and 0.5 g of 4-(bromomethyl)phenylboronic acid and dissolve them in 10 mL of DMF, stir at 60 °C for 16 h; add it to 200 mL of pre-cooled THF, let it stand for precipitation, discard the supernatant, then wash it with THF and dry to obtain the TPA product, and prepare a TPA solution with a concentration of 25 mg / mL.

6. The preparation method according to claim 5, characterized in that, The TPA is used to prepare a TPA solution using PBS as a solvent.

7. The preparation method according to claim 1, characterized in that, In step S2, add 5 mg of CY-09@FA-HA-MSN to 5 mL of a PVA solution with a concentration of 25 mg / mL. Take the decellularized valve DAV and place it in this mixture. Shake it on a shaker at 4 °C and 60 rpm for 24 h, wash it, then put it into the prepared TPA solution and shake it on a shaker at 30 rpm for 1 h, and wash it to obtain CY-09@MSN@HDAV.

8. The preparation method according to claim 7, characterized in that, The PVA is used to prepare a PVA solution using PBS as a solvent.

9. A hydrogel-modified artificial heart valve, characterized in that, Prepared by the preparation method according to any one of claims 1 - 8.

10. Use of the artificial heart valve according to claim 9 in the preparation of anti-thrombosis, endothelialization-promoting and anti-calcification functional materials.

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