A hydrogel-modified artificial heart valve and its preparation method and application

Abstract

The present invention discloses a hydrogel-modified artificial heart valve, its preparation method, and application, belonging to the field of biomedical materials technology. The method comprises the following steps: S1, constructing a hyaluronic acid and folic acid dual-modified loaded CY‑09@FA‑HA‑MSN; S2, preparing a ROS-responsive hydrogel-modified decellularized artificial valve CY‑09@MSN@HDAV loaded with macrophage-targeting nanoparticles. The present invention modifies a decellularized valve stent with a reactive oxygen species (ROS)-responsive hydrogel to prevent direct contact of the collagen in the decellularized valve stent with blood, thereby reducing the chronic inflammatory response induced by the valve stent and improving the anti-thrombotic properties of the artificial valve. Simultaneously, the hydrogel contains macrophage-targeted drug-loaded nanoparticles made of mesoporous silica (MSN) and loaded with the small molecule drug CY‑09. The hydrogel is modified with hyaluronic acid and folic acid to achieve the function of targeting macrophages.

Description

Technical Field

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

[0002] Heart valve disease seriously endangers human health, and its incidence will further increase as the aging population continues to deepen. Artificial heart valve replacement is the main treatment for heart valve disease. At present, the artificial heart valves commonly used in clinical practice include mechanical valves and biological valves: among them, mechanical valves require patients to take anticoagulants for life, are prone to bleeding, embolism and other problems, and are more noisy during operation; while biological valves do not require lifelong anticoagulation and are highly integrated with interventional technology. With the technological innovations in recent decades, they have now surpassed mechanical valves to become the mainstream choice in clinical practice. However, the existing biological valves are mainly cross-linked with glutaraldehyde. As the implantation time increases, the biological valve will undergo significant calcification, resulting in limited valve opening and closing, and then leading to valve stenosis or regurgitation. Therefore, the research and development of new artificial valves that can resist calcification for life has great scientific value and practical significance.

[0003] Currently, tissue engineering in situ regeneration is considered the most promising approach to overcome the limitations of existing prosthetic valves. This technology enables recellularization of the prosthetic valve, thereby overcoming the technical limitations of valve calcification and decay, and maximizing the lifespan of the prosthetic valve. Decellularization is currently the most promising prosthetic valve processing technology. The bioprosthetic valve scaffolds obtained using this technology offer significant advantages, preserving the original morphology and three-dimensional structure of the valve while minimizing its immunogenicity. However, due to the lack of endothelial cell protection, the collagen in the scaffold is completely exposed to the bloodstream, making it highly susceptible to chronic inflammation and thrombosis. The immune response is deeply involved in the valve regeneration process, with macrophages playing a crucial role. Immune cells infiltrating the valve scaffold secrete a variety of cytokines, which in turn induce the migration, proliferation, and differentiation of adjacent endothelial cells and endothelial progenitor cells, thereby achieving valve tissue remodeling. Different macrophage subtypes play distinct roles in the immune response; based on their functional differences, macrophages are generally divided into M1 macrophages, which primarily promote inflammation, and M2 macrophages, which primarily suppress inflammation. Both types of macrophages play important roles in different stages of valve remodeling. Therefore, regulating the distribution of different types of macrophages at different stages is of great significance for the regeneration of tissue-engineered valves. Summary of the Invention

[0004] The present invention adopts the following technical solutions:

[0005] In a first aspect, the present invention provides a method for preparing a hydrogel-modified artificial heart valve, comprising the following steps:

[0006] S1. Build CY-09@FA-HA-MSN:

[0007] Dissolve 0.1-0.5g of hexadecyltrimethylammonium bromide (CTAB) in a sodium hydroxide aqueous solution, stir and heat to 60-90°C, then add 0.1-0.5mL of anhydrous ethanol and stir. Add 1-5mL of tetraethyl orthosilicate (TEOS) solution dropwise, and react at 60-100°C for 1-5 hours. Centrifuge the reaction product at 10,000-11,000 rpm for 10-20 minutes, wash and dry overnight, and calcinate at 500-600°C for 2-7 hours to obtain a white powder, namely mesoporous silica nanoparticles (MSNs).

[0008] 10-50 mg of MSN and 1-5 mg of CY-09 were mixed in 1-5 mL of PBS and stirred at 1-5 ° C for 12-36 h to obtain CY-09@MSN; 10-50 mg of CY-09@MSN was dissolved in 1-5 mL of phosphate buffer PBS, and then 50-150 μL of polyethyleneimine PEI solution with a concentration of 70-80 mg / mL was added, stirred at room temperature for 20-40 min, and then centrifuged at 5000-15000 rpm for 10-20 min. After repeated centrifugation, washing and freeze-drying were obtained to obtain CY-09@PEI-MSN; 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 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC) was dissolved in water and stirred for 1-2 hours to obtain CY-09@HA-PEI-MSN. 10-20 mg of folic acid (FA) was added and stirred at room temperature for 12-36 hours. The mixture was centrifuged, washed, and freeze-dried overnight to obtain CY-09@FA-HA-MSN.

[0009] S2. Preparation of CY-09@MSN@HDAV:

[0010] 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 are dissolved in 5-15 mL of N,N-dimethylformamide (DMF) and stirred at 50-70° C. for 10-20 h. The mixture is added to 100-300 mL of pre-cooled tetrahydrofuran (THF), allowed to settle, the supernatant discarded, and the product washed with THF and dried to obtain N1-(4-borylphenyl)-N3-(4-borylphenyl)-N1,N1,N3,N3-tetramethylpropane-1,3-diamine (TPA). TPA is prepared into a TPA solution with a concentration of 20-30 mg / mL.

[0011] Add 1-6 mg of CY-09@FA-HA-MSN to 1-6 mL of 20-30 mg / mL polyvinyl alcohol (PVA) solution, take the decellularized aortic valve (DAV) and place it in the mixture. Shake at 1-5°C and 50-100 rpm for 12-36 hours, wash, and place it in the prepared TPA solution. Shake at 10-50 rpm for 1-2 hours and wash to obtain CY-09@MSN@HDAV.

[0012] Furthermore, in step S1, 0.4 g of CTAB was dissolved in water, heated to 80° C. with stirring, and 0.4 mL of anhydrous ethanol was added with stirring; 2 mL of TEOS solution was added dropwise, and the mixture was reacted at 80° C. for 2 h; the reaction product was centrifuged at 10,200 rpm for 15 min; washed and dried overnight, and calcined at 550° C. for 5 h to obtain a white powder, namely MSN.

[0013] Furthermore, the CTAB was dissolved in 1.44 mL of a 2M sodium hydroxide aqueous solution.

[0014] Furthermore, in step S1, 30 mg of MSN and 2 mg of CY-09 were mixed into 4 mL of PBS and stirred at 4°C for 24 h to obtain CY-09@MSN; 30 mg of CY-09@MSN was dissolved in 4 mL of PBS, and then 100 μL of 75 mg / mL PEI solution was added, stirred at room temperature for 30 min, and then centrifuged at 10,000 rpm for 15 min. After repeated centrifugation, the mixture was washed and freeze-dried to obtain CY-09@PEI-MSN; 30 mg of CY-09@PEI-MSN, 15 mg of HA, 40 mg of NHS and 30 mg of EDC were dissolved in water and stirred for 1 h to obtain CY-09@HA-PEI-MSN; 15 mg of FA was added, stirred at room temperature for 24 h, centrifuged, washed, and freeze-dried overnight to obtain CY-09@FA-HA-MSN.

[0015] Furthermore, in step S2, 0.1 g of N,N,N′,N-tetramethyl-1,3-propanediamine and 0.5 g of 4-(bromomethyl)phenylboronic acid were dissolved in 10 mL of DMF and stirred at 60° C. for 16 h; the mixture was added to 200 mL of pre-cooled THF, allowed to stand for precipitation, the supernatant was discarded, and the product was washed with THF and dried to obtain a TPA product, and the TPA was prepared into a TPA solution with a concentration of 25 mg / mL.

[0016] Furthermore, the TPA is prepared into a TPA solution using PBS as a solvent.

[0017] Furthermore, in step S2, 5 mg of CY-09@FA-HA-MSN was added to 5 mL of a 25 mg / mL PVA solution, and the decellularized valve DAV was placed in the mixture, shaken at 4°C and 60 rpm for 24 h, washed, and placed in the prepared TPA solution, shaken at 30 rpm for 1 h, and washed to obtain CY-09@MSN@HDAV.

[0018] Furthermore, the PVA is prepared into a PVA solution using PBS as a solvent.

[0019] In a second aspect, the present invention also provides a hydrogel-modified artificial heart valve prepared according to the preparation method.

[0020] In a third aspect, the present invention further provides the use of the artificial heart valve in the preparation of anti-thrombotic, pro-endothelialization and anti-calcification functional materials.

[0021] Mechanism of action:

[0022] 1. The reactive oxygen species-responsive hydrogel can neutralize ROS generated on the valve surface, thereby reducing the ROS concentration on the valve surface, inhibiting macrophage polarization to M1, reducing the M1 / M2 ratio, and promoting valve re-endothelialization;

[0023] 2. CY-09-loaded MSN nanoparticles, modified with folic acid and hyaluronic acid, significantly enhance their targeting ability to macrophages. The engulfed MSNs gradually release CY-09 within macrophages, intervening in the formation of inflammasomes NLRP3 and AIM2, thereby inhibiting macrophage polarization toward M1 macrophages, reducing the M1 / M2 ratio, and promoting valve re-endothelialization.

[0024] 3. The hydrogel-modified acellular flap can significantly enhance its mechanical properties, which is more in line with actual needs;

[0025] 4. The hydrogel-modified acellular valve can prevent the valve collagen from being directly exposed to the blood environment, thereby inhibiting the formation of blood clots;

[0026] 5. The hydrogel-modified acellular flap can inhibit the polarization of macrophages into M1 macrophages while enhancing their anti-enzymatic properties.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] 1. The present invention modifies the decellularized valve stent with a reactive oxygen species (ROS)-responsive hydrogel to prevent direct contact of the collagen in the decellularized valve stent with blood, thereby reducing the chronic inflammatory response induced by the valve stent and improving the anti-thrombotic properties of the artificial valve.

[0029] 2. The ROS-responsive hydrogel of the present invention can rapidly react with ROS generated in the body, reduce the content of ROS, and promote the polarization of macrophages to M2 type;

[0030] 3. The hydrogel of the present invention contains macrophage-targeted drug-loaded nanoparticles: the nanoparticles are made of mesoporous silica (MSN) and loaded with the small molecule drug CY-09; they are modified with hyaluronic acid and folic acid to achieve the function of targeting macrophages. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is the SEM scanning result of the nanoparticles in Example 1;

[0032] Figure 2 This is a diagram showing the effect of CY-09@FA-HA-MSN nanoparticles on macrophage ROS in Example 1;

[0033] Figure 3 The SEM and EDS scanning results of different tissue valves in Example 1;

[0034] Figure 4 The results of mechanical properties measurement of valves with different tissues in Example 1 are as follows;

[0035] Figure 5 This is a diagram showing the subcutaneous embedding results of rats in Example 2;

[0036] Figure 6 This is the polarized fluorescence result of valve-embedded macrophages in Example 2;

[0037] Figure 7 This is the endothelial fluorescence result of the valve abdominal aorta transplantation in Example 2; DETAILED DESCRIPTION

[0038] To better illustrate the present invention, the following embodiments are listed. Obviously, the embodiments described are only part of the present invention, not all of the embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without making any creative efforts are also within the scope of protection of the present invention.

[0039] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0040] Example 1

[0041] 1. Construction of hyaluronic acid and folic acid dual-modified loaded CY-09@FA-HA-MSN

[0042] (1) 0.4 g of CTAB (N-hexadecyltrimethylammonium bromide) was dissolved in 200 mL of ultrapure water containing 1.44 mL of sodium hydroxide aqueous solution (2 M) and heated to 80 °C under stirring. Subsequently, 0.4 mL of anhydrous ethanol was added and stirred for 5 min. Then, 2 mL of tetraethyl orthosilicate (TEOS) solution was added dropwise and the reaction was maintained at 80 °C for 2 h. The product was centrifuged (10,200 rpm, 15 min) and washed three times with ethanol and deionized water to remove residual impurities. In order to remove CTAB, the product was dried overnight and calcined at 550 °C for 5 h to obtain a white powder and was designated as MSN.

[0043] (2) 30 mg of MSN and 2 mg of CY-09 (dissolved in 200 μL of DMSO) were mixed in 4 mL of phosphate buffered saline (PBS, 10 mM, pH = 7.4) and stirred at 4 °C for 24 h to obtain CY-09@MSN. 30 mg of CY-09@MSN was dissolved in 4 mL of PBS solution, and then 100 μL of PEI solution (75 mg / mL) was added. The mixture was stirred at room temperature for 30 min. Then, the product was centrifuged (10,000 rpm, 15 min); the centrifugation operation was repeated 3 times, each time washed with ultrapure water, and lyophilized to obtain CY-09@PEI-MSN. 30 mg of lyophilized CY-09@PEI-MSN, 15 mg of HA, 40 mg of NHS, and 30 mg of EDC were dissolved in 10 mL of distilled water and stirred for 1 h to obtain CY-09@HA-PEI-MSN. Subsequently, 15 mg of FA (dissolved in 5 ml of DMSO) was added, and stirring was continued at room temperature for 24 h. Finally, CY-09@FA-HA-MSNs were obtained after centrifugation, washing, and overnight lyophilization.

[0044] The obtained nanoparticles were scanned by scanning electron microscope (SEM) and their Zeta potential and particle size were detected (see the test results in Figure 1 ), the results showed that the surface morphology of MSN changed significantly after being modified with HA and FA.

[0045] 2. Preparation of Nanoparticle-Loaded ROS Hydrogel Modified Acellular Artificial Valve CY-09@MSN@HDAV

[0046] (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 reaction mixture to 200 mL of tetrahydrofuran (THF) pre-cooled at 4°C, let it stand until the product precipitates, and discard the supernatant. Then add 100 mL of pre-cooled tetrahydrofuran again and wash repeatedly three times. Then dry to obtain the product N1-(4-borylphenyl)-N3-(4-borylphenyl)-N1,N1,N3,N3-tetramethylpropane-1,3-diamine (TPA). Use PBS solution as solvent to prepare a TPA solution with a concentration of 25 mg / ml.

[0047] (2) Using PBS as a solvent, 5 ml of 25 mg / mL PVA (polyvinyl alcohol, average molar mass of 30,000-70,000) solution was prepared, and then 5 mg of CY-09@FA-HA-MSN was added to it; 10 decellularized porcine valves (DAVs) were placed in the mixed solution and shaken on a horizontal shaker at 4°C for 24 h (60 rpm / min); the decellularized valves were then washed three times with PBS solution; then they were placed in 5 ml of the prepared TPA solution and shaken on a horizontal shaker at low speed for 1 h (30 rpm / min); the valves were then washed three times with PBS solution to obtain the ROS hydrogel-modified decellularized artificial valve CY-09@MSN@HDAV loaded with macrophage-targeted nanoparticles.

[0048] The CY-09@FA-HA-MSN nanoparticles prepared in this example were used to verify their ability to reduce ROS in macrophages. The fluorescent probe DCFH-DA was used to detect the levels of reactive oxygen species in macrophages induced by the THP-1 cell line. The results showed that CY-09@FA-HA-MSN significantly reduced the levels of reactive oxygen species in macrophages.

[0049] At the same time, the decellularized valve DAV and the glutaraldehyde-crosslinked valve GAV were used as control groups for comparison with the artificial valve CY-09@MSN@HDAV prepared in the present invention.

[0050] The decellularized valve (DAV) was prepared by placing the porcine aortic valve in TRIS-HCl buffer (40 mM, pH 7.8) containing 2% 3-[3-(cholamidopropyl)dimethylamino]propanesulfonic acid inner salt (CHAPS) and 2 mmol / l tributylphosphine (TnBP) and continuously shaking at room temperature for 24 hours to decellularize. The valve was then rinsed with sterile water six times for 10 minutes each. The valve was then placed in TRIS-HCl buffer (40 mM, pH 7.8) containing 2% CHAPS, 2 mmol / l TnBP, 1% amidinosulfobetaine (ASB-14), and 2% sulfobetaine 10 (SB 3-10) and continued to shake at room temperature for 24 hours to obtain the DAV.

[0051] The preparation method of glutaraldehyde cross-linked valve GAV is as follows: 30 pieces of porcine aortic valves are added to 50 mL of 0.625% glutaraldehyde solution, cross-linked at room temperature in the dark for 48 hours, and then repeatedly rinsed with PBS to obtain GAV.

[0052] The three valve materials were scanned by SEM and the Si element was detected by EDS (Energy Dispersive Spectrometer). Figure 3 The results showed that compared to DAV, the surface of CY-09@MSN@HDAV was smoother, with significantly fewer exposed collagen fibers. EDS analysis of the Si element on the valve surface revealed a significant increase in the CY-09@MSN@HDAV compared to both the DAV and GAV groups, demonstrating the presence of nanoparticles on the surface. This experiment confirmed the successful nanoparticle modification of the valve.

[0053] Uniaxial tensile tests were performed on three valve samples to compare the changes in mechanical properties after hydrogel modification: the valve samples were cut into strips 10 mm long and 4 mm wide, and the strips were uniaxially stretched at a speed of 15 mm / min. Figure 4 : It can be observed that the mechanical properties of the valve after hydrogel modification are significantly improved compared with ordinary DAV, and are basically equivalent to the mechanical properties of the GVA valve.

[0054] Example 2

[0055] The valve samples obtained in Example 1 were used for two in vivo experiments in rats: subcutaneous embedding and abdominal aorta transplantation. Sprague-Dawley (SD) rats were purchased from Liaoning Changsheng Biotechnology Co., Ltd. and maintained in an SPF-shielded enclosure at the Animal Experimental Center of Huazhong University of Science and Technology. SD rats are typically housed in a barrier environment with a temperature of 20°C-26°C, a humidity of 40%-70%, a 12-hour light / 12-hour dark cycle, and ample feed and drinking water.

[0056] (1) Subcutaneous embedding experiment: CY-09@MSN@HDAV and DAV obtained in Example 1, as well as the glutaraldehyde-crosslinked bioprosthetic valve GAV, were implanted under the back skin of 6-week-old SD rats. After 28 days, the rats were sacrificed, and the subcutaneous valve tissue was removed and fixed with paraformaldehyde, embedded in paraffin, and sliced. The slices were stained with silver nitrate and stained for CD163 (M2 macrophages, green) and iNOS (M1 macrophages, red). (See the test results.) Figure 5 、 6 );

[0057] The results of subcutaneous embedding in rats at 28 days showed that compared with GAV, DAV and CY-09@MSN@HDAV did not undergo obvious calcification.

[0058] Due to the greater cytotoxicity of the valves in the GAV group, no macrophage attachment was observed. Compared with DAV, the macrophages colonizing 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).

[0059] (2) Abdominal aorta transplantation experiment: The valve samples were trimmed into 5mm×5mm square pieces and sewn into tubes using 8-0 surgical suture needles. Male SD rats weighing approximately 200g were anesthetized with 0.3% sodium pentobarbital 0.01-0.02mL / g intraperitoneal injection. The SD rats were opened and the aorta was freed to expose the abdominal aorta. The abdominal aorta was blocked and severed using a vascular clamp. The sewn tube was then connected to the abdominal aorta using an end-to-end anastomosis method. Samples were taken 7 days later and fixed with paraformaldehyde, embedded in paraffin, and sliced. The slices were stained with CD31 (endothelial cells, red) fluorescence.

[0060] The results of the 7-day abdominal aorta transplantation model in rats showed that the infiltration of endothelial cells (CD31, red) of CY-09@MSN@HDAV was more obvious than that of DAV and GAV, demonstrating its good re-endothelialization potential.

[0061] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.

Claims

1. A method for preparing a hydrogel-modified artificial heart valve, characterized in that: The following steps are involved: S1. Build CY-09@FA-HA-MSN: Dissolve 0.1-0.5g of hexadecyltrimethylammonium bromide (CTAB) in a sodium hydroxide aqueous solution, stir and heat to 60-90°C, then add 0.1-0.5mL of anhydrous ethanol and stir. Add 1-5mL of tetraethyl orthosilicate (TEOS) solution dropwise, and react at 60-100°C for 1-5 hours. Centrifuge the reaction product at 10,000-11,000 rpm for 10-20 minutes, wash and dry overnight, and calcinate at 500-600°C for 2-7 hours to obtain a white powder, namely mesoporous silica nanoparticles (MSNs). 10-50 mg of MSN and 1-5 mg of CY-09 were mixed in 1-5 mL of PBS and stirred at 1-5°C for 12-36 h to obtain CY-09@MSN. 10-50 mg of CY-09@MSN was dissolved in 1-5 mL of phosphate buffered saline (PBS), and then 50-150 μL of a 70-80 mg / mL polyethyleneimine (PEI) solution was added. The mixture was stirred at room temperature for 20-40 min, and then centrifuged at 5000-15000 rpm for 10-20 min. After repeated centrifugation, the mixture was washed and freeze-dried to obtain CY-09@PEI-MSN. 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) were taken. Dissolve in water and stir for 1-2 hours to obtain CY-09@HA-PEI-MSN; add 10-20 mg of folic acid FA, stir at room temperature for 12-36 hours, centrifuge, wash, and freeze-dry overnight to obtain CY-09@FA-HA-MSN; S2. Preparation of 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) and stir at 50-70°C for 10-20 hours. Add the resulting mixture to 100-300 mL of pre-cooled THF, allow to settle, discard the supernatant, wash with tetrahydrofuran (THF), and dry to obtain the product N1-(4-borylphenyl)-N3-(4-borylphenyl)-N1,N1,N3,N3-tetramethylpropane-1,3-diamine (TPA). 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 polyvinyl alcohol (PVA) solution with a concentration of 20-30 mg / mL, take the decellularized aortic valve (DAV) and place it in the mixture. Shake it at 1-5°C and 50-100 rpm for 12-36 hours, wash it, and place it in the prepared TPA solution. Shake it at 10-50 rpm for 1-2 hours and wash it to obtain the ROS hydrogel-modified decellularized artificial valve CY-09@MSN@HDAV loaded with nanoparticles.

2. The preparation method according to claim 1, characterized in that In step S1, 0.4 g of CTAB was dissolved in a sodium hydroxide aqueous solution, heated to 80° C. with stirring, and 0.4 mL of anhydrous ethanol was added with stirring; 2 mL of TEOS solution was added dropwise, and the mixture was reacted at 80° C. for 2 h; the reaction product was centrifuged at 10,200 rpm for 15 min; the product was washed and dried overnight, and calcined at 550° C. for 5 h to obtain a white powder, namely MSN.

3. The preparation method according to claim 2, characterized in that The sodium hydroxide aqueous solution was prepared by the following method: 1.44 mL of 2M sodium hydroxide aqueous solution was added to 200 mL of ultrapure water.

4. The preparation method according to claim 1, characterized in that In step S1, 30 mg of MSN and 2 mg of CY-09 were mixed in 4 mL of PBS and stirred at 4°C for 24 h to obtain CY-09@MSN; 30 mg of CY-09@MSN was dissolved in 4 mL of PBS, and then 100 μL of 75 mg / mL PEI solution was added, stirred at room temperature for 30 min, and then centrifuged at 10,000 rpm for 15 min. After repeated centrifugation, the mixture was washed and freeze-dried to obtain CY-09@PEI-MSN; 30 mg of CY-09@PEI-MSN, 15 mg of HA, 40 mg of NHS, and 30 mg of EDC were dissolved in water and stirred for 1 h to obtain CY-09@HA-PEI-MSN; 15 mg of FA was added, stirred at room temperature for 24 h, centrifuged, washed, and freeze-dried overnight to obtain CY-09@FA-HA-MSN.

5. The preparation method according to claim 1, characterized in that In step S2, 0.1 g of N,N,N′,N′-tetramethyl-1,3-propylenediamine and 0.5 g of 4-(bromomethyl)phenylboronic acid were dissolved in 10 mL of DMF and stirred at 60° C. for 16 h. The mixture was added to 200 mL of pre-cooled THF, allowed to settle, the supernatant was discarded, and the product was washed with THF and dried to obtain a TPA product. The TPA solution was prepared to a concentration of 25 mg / mL.

6. The preparation method according to claim 5, characterized in that The TPA solution was prepared by using PBS as a solvent.

7. The preparation method according to claim 1, characterized in that In step S2, 5 mg of CY-09@FA-HA-MSN was added to 5 mL of a 25 mg / mL PVA solution, and the decellularized valve DAV was placed in the mixture, shaken at 4°C and 60 rpm for 24 hours, washed, and placed in the prepared TPA solution, shaken at 30 rpm for 1 hour, and washed to obtain CY-09@MSN@HDAV.

8. The preparation method according to claim 7, characterized in that The PVA was prepared into a PVA solution using PBS as a solvent.

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

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

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