A multi-substrate-adhesion hydrogel capable of efficiently delivering nucleic acid molecules and its application in tumor treatment

Abstract

The present invention discloses a multi-substrate adhesive hydrogel capable of efficiently delivering nucleic acid molecules and its application in tumor treatment. The preparation steps of the hydrogel are as follows: preparing a MXene nanosheet solution by hydrofluoric acid etching; preparing a disulfide solid powder by recrystallization; mixing the disulfide solid powder with the MXene nanosheet solution and stirring at room temperature to obtain solution A; mixing CpG oligonucleotide with solution A and dialyzing to obtain an ONMs nanoparticle solution; grafting sodium alginate and amino-modified ATP aptamer to obtain DNA SA solution; ONMs nanoparticle solution and DNA SA solution was mixed and incubated at room temperature to obtain DNA SA@ONMs solution; DNA The SA@ONMs solution was mixed with acrylamide, and then an ionic crosslinker, a crosslinker, a crosslinking accelerator, and an initiator were added in sequence. The mixture was stirred at room temperature and allowed to stand to solidify into a gel, thereby obtaining a multi-substrate adhesion hydrogel.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biomaterials, and particularly relates to a multi-substrate adhesive hydrogel capable of efficiently delivering nucleic acid molecules and application thereof in tumor treatment. BACKGROUND

[0002] Malignant tumors seriously endanger human health, among which primary malignant tumors usually occur in childhood and adolescence, and about 30% of primary bone tumor patients die within 5 years due to poor response to treatment. Bone tumor is a primary malignant tumor, which is prone to recurrence and has a high early metastasis rate, and has an invasive biological characteristic, and its clinical treatment has always been a difficulty in orthopedic and oncology research. Consolidation chemotherapy or radiotherapy is very important for controlling tumor metastasis and improving patient survival rate, however, many problems such as treatment efficiency, pathogenesis, adjuvant therapy mode and evaluation method have not been well solved. With the use of chemotherapy drugs and repeated illness, the toxic side effects of normal tissues, poor prognosis caused by multidrug resistance mechanism and radioresistance tendency make it difficult to completely eliminate bone tumor cells, which are prone to recurrence, and at the same time cause damage to normal cells in the bone tumor site, which is not conducive to the repair of bone defects. Therefore, designing a new type of tissue engineering scaffold material with the dual functions of promoting bone regeneration and preventing tumor recurrence provides a new idea for bone tumor treatment.

[0003] Deoxyribonucleic acid (DNA) as a carrier of genetic information is also considered as a block copolymer for constructing biomaterials. DNA hydrogel is a three-dimensional network biomaterial composed of DNA chains, which has a series of advantages including sequence designability, functional sequence responsiveness, good stability, flexibility and biodegradability. By assembling various functional sequences containing DNA modules, DNA hydrogels with specific functions can be prepared. In recent years, DNA hydrogels have been widely used for delivering cancer treatment drugs, which benefits from the sequence programmability and molecular recognition ability of DNA molecules, so that DNA hydrogels can achieve efficient loading of anticancer drugs, integrate specific DNA sequences with cancer treatment effects, realize targeted drug delivery and drug controlled release, and are beneficial to cancer treatment. The future development direction of DNA hydrogel in cancer treatment can also be combined with two-dimensional nanomaterials to form three-dimensional DNA nanomaterial composite gels, so that the DNA gel has new functions. The combined application of composite materials of polymers and nucleic acids carrying nucleic acid drugs with different action mechanisms is expected to not only realize in-situ regeneration induction of bone defects after tumor resection, but also further enhance the treatment effect and reduce the recurrence and metastasis of bone tumors.

[0004] With the development of material science, biomaterials have attracted attention due to their unique biological properties, excellent tumor specificity and high drug loading capacity, and suitable biomaterials can provide multiple functions such as tumor treatment and promotion of bone regeneration. Through screening of various materials, it is found that the application of polymer and nucleic acid composite materials can simultaneously enhance the therapeutic effect and reduce the recurrence and metastasis of bone tumors, and in order to prevent the problem that nano drugs are easily eliminated in the body, sodium alginate / polyacrylamide (SA / PAAm) double network hydrogel and Aapt nucleic acid aptamer are introduced as the hydrogel base, and finally a DNA SA / PAAm@ONMs polymer-nucleic acid composite hydrogel. The composite hydrogel carries CpG oligonucleotide polymer composite drugs that can act on tumor cells and dendritic cells, and is released under the synergistic action of heat and ATP to realize the integration of treatment, prognosis and promotion of bone repair. The physical and chemical properties of the nucleic acid / polymer composite hydrogel and the hydrogel are characterized by various means such as morphology analysis, particle size distribution and adhesion test, and the successful preparation of the nucleic acid / polymer composite hydrogel and the hydrogel is confirmed, and the experimental results such as photothermal conversion effect, active oxygen generation capacity and nucleic acid drug release rate are well verified. The composite hydrogel can respond and release in multiple modes, and has shown good bone tumor treatment effect. SUMMARY

[0005] Based on this, the technical problem to be solved by the present application is to provide a multi-substrate adhesive hydrogel capable of efficiently delivering nucleic acid molecules and its application in tumor treatment.

[0006] To achieve the above-mentioned application purposes, the technical scheme adopted by the present application is as follows:

[0007] A preparation method of a multi-substrate adhesive hydrogel capable of efficiently delivering nucleic acid molecules, comprising the following steps:

[0008] S1: preparing a MXene nanosheet solution by using a hydrofluoric acid etching method;

[0009] S2: preparing a disulfide solid powder by using a recrystallization method;

[0010] S3: mixing the disulfide solid powder and the MXene nanosheet solution, stirring at room temperature for 2h, to prepare a solution A;

[0011] S4: mixing and dialyzing CpG oligonucleotides with the solution A to prepare an ONMs nanoparticle solution;

[0012] S5: grafting sodium alginate and amino-modified ATP aptamer to prepare DNA a SA solution;

[0013] S6: ONMs nanoparticle solution and DNA The SA solution was mixed and incubated at room temperature for 2 h to prepare DNA SA@ONMs solution;

[0014] S7: DNA The SA@ONMs solution was mixed with acrylamide, and then an ionic crosslinker, a crosslinker, a crosslinking accelerator, and an initiator were added in sequence. The mixture was stirred at room temperature for 8 to 24 hours and allowed to stand to solidify into a gel, resulting in a multi-substrate adhesion hydrogel that can efficiently deliver nucleic acid molecules.

[0015] The concentration of the MXene nanosheet solution is 1 mg / mL;

[0016] The ratio of the disulfide solid powder to the MXene nanosheet solution is 10:100 w / v;

[0017] The step S4 is specifically: mixing 1-3 mL of solution A and 0.5 mL of 50-200 nM CpG oligonucleotide in 10 mL of Tris-HCl buffer at 37° C. for 15 minutes, then transferring to a dialysis bag with a molecular weight cutoff of 3,500 Da, and dialyzing with deionized water for 2 days, and the resulting dialysate is the ONMs nanoparticle solution; preferably, the step S4 is specifically: mixing 2 mL of solution A and 0.5 mL of 100 nM CpG oligonucleotide in 10 mL of Tris-HCl buffer at 37° C. for 15 minutes, then transferring to a dialysis bag with a molecular weight cutoff of 3,500 Da, and dialyzing with deionized water for 2 days, and the resulting dialysate is the ONMs nanoparticle solution;

[0018] The step S5 is specifically as follows: 1 mL of 100-400 nM amino-modified ATP aptamer and 10 mL of 2.8 wt% sodium alginate were mixed in 20 mL of MES buffer at 37°C for 2 h, followed by the addition of 1 mL of 1 M EDC·HCl and shaking at 37°C for 2 h, followed by the addition of 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol, the system was transferred to -80°C for 10 min, and then centrifuged at 10,000 rpm for 10 min, the precipitate was collected, washed with anhydrous ethanol, and redissolved with 1 mL of deionized water to obtain DNASA solution; preferably, the step S5 is specifically as follows: 1 mL of 200 nM amino-modified ATP aptamer and 10 mL of 2.8 wt% sodium alginate are mixed in 20 mL of MES buffer at 37°C for 2 h, followed by adding 1 mL of 1 M EDC·HCl, shaking at 37°C for 2 h, and then adding 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol in sequence, transferring the system to -80°C for 10 min, and then centrifuging at 10,000 rpm for 10 min, collecting the precipitate, washing it with anhydrous ethanol, and then redissolving it with 1 mL of deionized water to obtain DNA SA solution;

[0019] The step S6 is specifically as follows: 1 mL DNA SA solution was mixed with 0.5 mL ONMs nanoparticle solution and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution;

[0020] The specific step S7 is: DNA The SA@ONMs solution was mixed with 1 mL of 15 wt% acrylamide solution, and an ionic crosslinker with a final concentration of 0.1 wt%-0.4 wt% was added, a crosslinker with a final concentration of 0.01 wt%-0.05 wt% was added, a crosslinking accelerator with a final concentration of 0.02 wt%-0.10 wt% was added, and an initiator with a final concentration of 0.1 wt%-0.3 wt% was added. The mixture was stirred at room temperature for 8 h and then allowed to stand for 30 min to solidify into a gel. DNA SA / PAAm@ONMs hydrogel is a multi-substrate adhesion hydrogel that can efficiently deliver nucleic acid molecules;

[0021] The ionic crosslinking agent is calcium chloride, the crosslinking agent is N,N'-methylenebisacrylamide, the crosslinking accelerator is tetramethylethylenediamine, and the initiator is ammonium persulfate;

[0022] The sequence of the CpG oligonucleotide is 5'-Cy5-TCCATGACGTTCCTGACGTTACCTTCCTCCGCAA-3', and the sequence of the amino-modified ATP aptamer is 5'-NH2-ACCTGGGGGAGTATTGCGGAGGAAGGT-3'.

[0023] A multi-substrate adhesion hydrogel capable of efficiently delivering nucleic acid molecules is prepared by the above-mentioned preparation method.

[0024] The use of the above-mentioned multi-substrate adhesion hydrogel in the preparation of tumor treatment drugs, wherein the tumor is osteosarcoma.

[0025] Compared with the prior art, the innovation of the present invention lies in:

[0026] (1) The present application DNA The SA / PAAm@ONMs hydrogel based on the metal complexation of PAAm, SA and different substrates, electrostatic interaction, hydrogen bonding of nucleic acid, etc. presents multi-substrate adhesion characteristics. The adhesion to different substrates endows the hydrogel with the ability to adhere to the tumor site for a long time, so as to realize long-term release of drugs and provide a performance basis for further use in combination with medical devices;

[0027] (2) DNA The SA / PAAm@ONMs hydrogel as a polymer-nucleic acid composite hydrogel for eliminating tumors can realize multi-stage stimulus-responsive release of nucleic acid nanomedicine and promote lysosome escape, improve the drug uptake efficiency of cells, and under near-infrared light and ultrasonic conditions, the ONMs nanoparticles in the hydrogel can generate active oxygen and produce heat to eliminate tumor cells, the immune adjuvant CpG can activate immune response to kill in situ tumors and inhibit distant tumor metastasis / recurrence, and improve the treatment efficiency;

[0028] (3) DNA The ONMs nanoparticles in the SA / PAAm@ONMs hydrogel have excellent photothermal conversion capability, can realize photoacoustic imaging, and have good photoacoustic imaging capability, can realize deep monitoring of the tumor site, the nucleic acid modified fluorescent group realizes real-time monitoring of the drug, realizes fluorescence / photoacoustic / magnetic resonance multi-mode imaging real-time monitoring of the tumor area. BRIEF DESCRIPTION OF DRAWINGS

[0029] Figure 1 For the present application DNA Preparation process and mechanism diagram of SA / PAAm@ONMs hydrogel.

[0030] Figure 2 : A, B are SEM photos of MXene nanosheets and ONMs nanoparticles respectively; C, D are TEM photos of MXene nanosheets and ONMs nanoparticles respectively; E, F are particle size distribution column charts of MXene nanosheets and ONMs nanoparticles respectively; G, H are energy dispersive X-ray spectroscopy test charts of MXene nanosheets and ONMs nanoparticles respectively.

[0031] Figure 3 : A is the Zeta potential analysis chart of MXene nanosheets and ONMs nanoparticles; B is the X-ray diffraction analysis chart of MXene nanosheets and ONMs nanoparticles; C is the ultrasonic (US: 0.5 W / cm2, 1 MHz, 3 min) and laser (Laser: 808 nm, 1.0 W / cm 2, 5min) stimulation; D is the thermal imaging of MXene nanosheets and ONMs nanoparticles under near-infrared light (808nm); E is the GSH (5mM) response test result of MXene nanosheets and ONMs nanoparticles; F is the GSH (5mM) response test result of MXene nanosheets and ONMs nanoparticles under 808nm laser (1.0W / cm 2 ) stimulation; G is the photothermal performance analysis diagram of MXene nanosheets and nanoparticles ONMs under 808nm laser (1.0W / cm 2 ) Analysis diagram of cyclic photothermal performance under stimulation.

[0032] Figure 4 : AB is the SEM image of SA / PAAm hydrogel; CD is DNA SEM images of SA / PAAm@ONMs hydrogel; E is the SEM images of SA / PAAm hydrogel and DNA Infrared spectrum test diagram of SA / PAAm@ONMs hydrogel; F is SA / PAAm hydrogel and DNA Swelling behavior and contact angle test diagram of SA / PAAm@ONMs hydrogel; G is SA / PAAm hydrogel and DNA Degradation behavior test diagram of SA / PAAm@ONMs hydrogel; H is SA / PAAm hydrogel and DNA Compressive strength test diagram of SA / PAAm@ONMs hydrogel.

[0033] Figure 5 :A is DNA Schematic diagram of the adhesion mechanism of SA / PAAm@ONMs hydrogel; B is the adhesion mechanism of SA / PAAm hydrogel and DNA Adhesion ability test diagram of SA / PAAm@ONMs hydrogel; C is DNA Adhesion of SA / PAAm@ONMs hydrogel on different material surfaces; DG is DNA Responsive release test of SA / PAAm@ONMs hydrogel under different conditions.

[0034] Figure 6 : Example 1 preparation DNA Qualitative and quantitative analysis of the biocompatibility and anticancer properties of SA / PAAm@ONMs hydrogel. A is the biocompatibility of SA / PAAm@ONMs with different concentrations. DNA SA / PAAm@ONMs hydrogel co-cultured for different time periods of umbilical cord mesenchymal stem cells (MSCs) live and dead staining data. B is with different concentrations DNA Figure 2 shows the CCK8 test data of MSCs cells co-cultured with SA / PAAm@ONMs hydrogel for 1 day. DNASA / PAAm@ONMs hydrogel co-cultured 1d human osteosarcoma cells (143B cells) CCK8 test data graph. D is the ROS production of 143B cells co-cultured with different stimulation conditions DNA SA / PAAm@ONMs hydrogel co-cultured 2h 143B cells ROS test data graph. E is the ROS production of 143B cells co-cultured with different stimulation conditions DNA SA / PAAm@ONMs hydrogel co-cultured 2h 143B cells live and dead staining data graph. 1 represents no co-culture with hydrogel; 2 represents co-culture with SA / PAAm hydrogel; 3 represents co-culture with SA / PAAm hydrogel, and given ultrasound (US: 0.5 W / cm 2 , 1 MHz) and laser (Laser: 808 nm, 1.0 W / cm 2 ) stimulation; 4 represents co-culture with DNA SA / PAAm@ONMs hydrogel; 5 represents co-culture with DNA SA / PAAm@ONMs hydrogel, and given ultrasound (US: 0.5 W / cm 2 , 1 MHz) and laser (Laser: 808 nm, 1.0 W / cm 2 ) stimulation.

[0035] Figure 7 : SA / PAAm@ONMs hydrogel prepared in Example 1 DNA SA / PAAm@ONMs hydrogel released ONMs nanoparticle uptake ability analysis graph. DETAILED DESCRIPTION

[0036] In order to further understand the present application, the preferred embodiments of the present application are described below in conjunction with examples, but it should be understood that these descriptions are only for further illustrating the features and advantages of the present application, and are not limitations on the claims of the present application.

[0037] In the following examples, the concentration of Tris-HCl buffer is 20 mM, and pH = 7.5.

[0038] In the following examples, the concentration of MES buffer is 0.05 M, and pH = 6.7.

[0039] In the following examples, the nucleotide sequence of the CpG oligonucleotide is:

[0040] 5'-Cy5-TCCATGACGTTCCTGACGTTACCTTCCTCCGCAA-3'.

[0041] In the following examples, the nucleotide sequence of the amino-modified aptamer Aapt is: 5'-NH2-ACCTGGGGGAGTATTGCGGAGGAAGGT-3'.

[0042] Example 1

[0043] DNA The preparation method of the SA / PAAm@ONMs hydrogel is carried out in the following steps:

[0044] S1: 5.0 g of carbon aluminized titanium is soaked into 40 mL of 40 wt% aqueous hydrofluoric acid solution at room temperature for etching for 3 days, filtered, washed with anhydrous ethanol and deionized water until the filtrate is neutral, to obtain pretreated carbon aluminized titanium; the obtained pretreated carbon aluminized titanium is added into 30 mL of 25 wt% aqueous tetrapropylammonium hydroxide solution, continuously stirred at room temperature for 72 h, the supernatant is removed after centrifugation at 3,000 rpm, the lower layer precipitate is dispersed in 50 mL of deionized water and ultrasonicated for 2 h, then centrifuged at 10,000 rpm for 15 min, and the supernatant is collected to obtain a Mxene nanosheet solution with a concentration of 1 mg / mL.

[0045] S2: 0.784 g of a-lipoic acid and 0.812 g of 1,1'-carbonyldiimidazole are dissolved in 25 mL of anhydrous dichloromethane, stirred at 0°C and 30°C for 40 min respectively, dried and concentrated after washing with 0.9 wt% saline, redissolved in 30 mL of anhydrous dichloromethane, 2.5 g of 1H-pyrazole-1-carboxamide hydrochloride is added after stirring at room temperature for 4 h, dissolved in 1 mL of methanol after removing the solvent, then 10 mL of diethyl ether is added to induce precipitation, the precipitate is collected, washed with diethyl ether, and naturally volatilized and dried at room temperature to obtain a light yellow disulfide solid powder.

[0046] S3: the disulfide solid powder obtained in step S2 is mixed with the MXene nanosheet solution obtained in step S1 at a ratio of 10:100 w / v, stirred at room temperature for 2 h to obtain solution A.

[0047] S4: 2 mL of solution A obtained in step S3 and 0.5 mL of 100 nM CpG oligonucleotide are mixed in 10 mL of Tris-HCl buffer at 37°C for 15 min, then transferred to a dialysis bag with a molecular weight cut-off of 3,500 Da, dialyzed with deionized water for 2 days, and the obtained dialysate is the ONMs nanoparticle solution, which is stored at 4°C for subsequent use.

[0048] S5: 1 mL of 200 nM amino-modified ATP aptamer Aapt was mixed with 10 mL of 2.8 wt% sodium alginate (SA) in 20 mL MES buffer at 37°C for 2 h, followed by the addition of 1 mL of 1 M EDC·HCl and the mixture was shaken at 37°C for 2 h. 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol were then added in sequence. The mixture was transferred to -80°C for 10 min and then centrifuged at 10,000 rpm for 10 min. The precipitate was collected, washed with anhydrous ethanol, and redissolved in 1 mL of deionized water to obtain DNA SA solution.

[0049] S6: 1 mL of the product from step S5 DNA The SA solution was mixed with 0.5 mL of the ONMs nanoparticle solution obtained in step S4 and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution.

[0050] S7: 1 mL of the product from step S6 DNA The SA@ONMs solution was mixed with 1 mL of 15 wt% acrylamide (AAm) solution, and the ionic crosslinker calcium chloride, the crosslinker N,N'-methylenebisacrylamide, the crosslinking accelerator tetramethylethylenediamine, and the initiator ammonium persulfate were added in sequence. The mixture was stirred at room temperature for 8 h and then allowed to stand for 30 min to solidify into a gel. DNA SA / PAAm@ONMs hydrogel. DNA In the SA / PAAm@ONMs hydrogel, the final concentration of calcium chloride was 0.2 wt %, the final concentration of N,N'-methylenebisacrylamide was 0.03 wt %, the final concentration of tetramethylethylenediamine was 0.05 wt %, and the final concentration of ammonium persulfate was 0.2 wt %.

[0051] Example 2

[0052] DNA The preparation method of SA / PAAm@ONMs hydrogel is carried out according to the following steps:

[0053] S1: Same as step S1 in Example 1.

[0054] S2: Same as step S2 in Example 1.

[0055] S3: Same as step S3 in Example 1.

[0056] S4: Mix 1 mL of solution A obtained in step S3 and 0.5 mL of 50 nM CpG oligonucleotide in 10 mL of Tris-HCl buffer at 37°C for 15 minutes, then transfer to a dialysis bag with a molecular weight cutoff of 3,500 Da and dialyze against deionized water for 2 days. The resulting dialysate is the ONMs nanoparticle solution, which is stored at 4°C for subsequent use.

[0057] S5: 1 mL of 100 nM amino-modified ATP aptamer Aapt was mixed with 10 mL of 2.8 wt% sodium alginate (SA) in 20 mL MES buffer at 37°C for 2 h, followed by the addition of 1 mL of 1 M EDC·HCl and the mixture was shaken at 37°C for 2 h. 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol were then added in sequence. The mixture was transferred to -80°C for 10 min and then centrifuged at 10,000 rpm for 10 min. The precipitate was collected, washed with anhydrous ethanol, and redissolved in 1 mL of deionized water to obtain DNA SA solution.

[0058] S6: 1 mL of the product from step S5 DNA The SA solution was mixed with 0.5 mL of the ONMs nanoparticle solution obtained in step S4 and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution.

[0059] S7: 1 mL of the product from step S6 DNA The SA@ONMs solution was mixed with 1 mL of 15 wt% acrylamide (AAm) solution, and the ionic crosslinker calcium chloride, the crosslinker N,N'-methylenebisacrylamide, the crosslinking accelerator tetramethylethylenediamine, and the initiator ammonium persulfate were added in sequence. The mixture was stirred at room temperature for 8 h and then allowed to stand for 30 min to solidify into a gel. DNA SA / PAAm@ONMs hydrogel. DNA In the SA / PAAm@ONMs hydrogel, the final concentration of calcium chloride was 0.2 wt %, the final concentration of N,N'-methylenebisacrylamide was 0.03 wt %, the final concentration of tetramethylethylenediamine was 0.05 wt %, and the final concentration of ammonium persulfate was 0.2 wt %.

[0060] Example 3

[0061] DNA The preparation method of SA / PAAm@ONMs hydrogel is carried out according to the following steps:

[0062] S1: Same as step S1 in Example 1.

[0063] S2: Same as step S2 in Example 1.

[0064] S3: Same as step S3 in Example 1.

[0065] S4: Mix 1 mL of solution A obtained in step S3 and 0.5 mL of 50 nM CpG oligonucleotide in 10 mL of Tris-HCl buffer at 37°C for 15 minutes, then transfer to a dialysis bag with a molecular weight cutoff of 3,500 Da and dialyze against deionized water for 2 days. The resulting dialysate is the ONMs nanoparticle solution, which is stored at 4°C for subsequent use.

[0066] S5: 1 mL of 200 nM amino-modified ATP aptamer Aapt was mixed with 10 mL of 2.8 wt% sodium alginate (SA) in 20 mL MES buffer at 37°C for 2 h, followed by the addition of 1 mL of 1 M EDC·HCl and the mixture was shaken at 37°C for 2 h. 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol were then added in sequence. The mixture was transferred to -80°C for 10 min and then centrifuged at 10,000 rpm for 10 min. The precipitate was collected, washed with anhydrous ethanol, and redissolved in 1 mL of deionized water to obtain DNA SA solution.

[0067] S6: 1 mL of the product from step S5 DNA The SA solution was mixed with 0.5 mL of the ONMs nanoparticle solution obtained in step S4 and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution.

[0068] S7: 1 mL of the product from step S6 DNA The SA@ONMs solution was mixed with 1 mL of 15 wt% acrylamide (AAm) solution, and the ionic crosslinker calcium chloride, the crosslinker N,N'-methylenebisacrylamide, the crosslinking accelerator tetramethylethylenediamine, and the initiator ammonium persulfate were added in sequence. The mixture was stirred at room temperature for 8 h and then allowed to stand for 30 min to solidify into a gel. DNA SA / PAAm@ONMs hydrogel. DNA In the SA / PAAm@ONMs hydrogel, the final concentration of calcium chloride was 0.2 wt %, the final concentration of N,N'-methylenebisacrylamide was 0.03 wt %, the final concentration of tetramethylethylenediamine was 0.05 wt %, and the final concentration of ammonium persulfate was 0.2 wt %.

[0069] Example 4

[0070] DNA The preparation method of SA / PAAm@ONMs hydrogel is carried out according to the following steps:

[0071] S1: Same as step S1 in Example 1.

[0072] S2: Same as step S2 in Example 1.

[0073] S3: Same as step S3 in Example 1.

[0074] S4: 2 mL of solution A obtained in step S3 and 0.5 mL of 100 nM CpG oligonucleotide were mixed in 10 mL of Tris-HCl buffer at 37 °C for 15 min, and then transferred to a dialysis bag with a molecular weight cut-off of 3,500 Da, and dialyzed with deionized water for 2 days. The dialysate was the ONMs nanoparticle solution, which was stored at 4 °C for subsequent use.

[0075] S5: 1 mL of 200 nM amino-modified ATP aptamer Aapt was mixed with 10 mL of 2.8 wt% sodium alginate (SA) in 20 mL of MES buffer at 37 °C for 2 h, followed by the addition of 1 mL of 1 M EDC-HCl, and then stirred at 37 °C for 2 h. Subsequently, 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol were added in sequence, and the system was transferred to -80 °C for 10 min, followed by centrifugation at 10,000 rpm for 10 min. The precipitate was washed with anhydrous ethanol and then redissolved with 1 mL of deionized water to obtain DNA SA solution.

[0076] S6: 1 mL of the DNA SA solution obtained in step S5 was mixed with 0.5 mL of the ONMs nanoparticle solution obtained in step S4, and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution.

[0077] S7: 1 mL of the DNA SA@ONMs solution obtained in step S6 was mixed with 1 mL of 15 wt% acrylamide (AAm) solution, and ion crosslinking agent calcium chloride, crosslinking agent N,N'-methylene bisacrylamide, crosslinking promoter tetramethyl ethylenediamine, and initiator ammonium persulfate were added in sequence. The reaction was stirred at room temperature for 8 h, and then left to stand for 30 min to facilitate curing into a gel to obtain DNA SA / PAAm@ONMs hydrogel. In the DNA SA / PAAm@ONMs hydrogel, the final concentration of calcium chloride was 0.2 wt%, the final concentration of N,N'-methylene bisacrylamide was 0.03 wt%, the final concentration of tetramethyl ethylenediamine was 0.05 wt%, and the final concentration of ammonium persulfate was 0.2 wt%.

[0078] Example 5

[0079] DNA The preparation method of the SA / PAAm@ONMs hydrogel was as follows:

[0080] S1: The same as step S1 of Example 1.

[0081] S2: The same as step S2 of Example 1.

[0082] S3: Same as step S3 in Example 1.

[0083] S4: Mix 1 mL of solution A obtained in step S3 and 0.5 mL of 200 nM CpG oligonucleotide in 10 mL of Tris-HCl buffer at 37°C for 15 minutes, then transfer to a dialysis bag with a molecular weight cutoff of 3,500 Da and dialyze against deionized water for 2 days. The resulting dialysate is the ONMs nanoparticle solution, which is stored at 4°C for subsequent use.

[0084] S5: 1 mL of 200 nM amino-modified ATP aptamer Aapt was mixed with 10 mL of 2.8 wt% sodium alginate (SA) in 20 mL MES buffer at 37°C for 2 h, followed by the addition of 1 mL of 1 M EDC·HCl and the mixture was shaken at 37°C for 2 h. 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol were then added in sequence. The mixture was transferred to -80°C for 10 min and then centrifuged at 10,000 rpm for 10 min. The precipitate was collected, washed with anhydrous ethanol, and redissolved in 1 mL of deionized water to obtain DNA SA solution.

[0085] S6: 1 mL of the product from step S5 DNA The SA solution was mixed with 0.5 mL of the ONMs nanoparticle solution obtained in step S4 and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution.

[0086] S7: 1 mL of the product from step S6 DNA The SA@ONMs solution was mixed with 1 mL of 15 wt% acrylamide (AAm) solution, and the ionic crosslinker calcium chloride, the crosslinker N,N'-methylenebisacrylamide, the crosslinking accelerator tetramethylethylenediamine, and the initiator ammonium persulfate were added in sequence. The mixture was stirred at room temperature for 8 h and then allowed to stand for 30 min to solidify into a gel. DNA SA / PAAm@ONMs hydrogel. DNA In the SA / PAAm@ONMs hydrogel, the final concentration of calcium chloride was 0.2 wt %, the final concentration of N,N'-methylenebisacrylamide was 0.03 wt %, the final concentration of tetramethylethylenediamine was 0.05 wt %, and the final concentration of ammonium persulfate was 0.2 wt %.

[0087] Example 6

[0088] DNA The preparation method of SA / PAAm@ONMs hydrogel is carried out according to the following steps:

[0089] S1: Same as step S1 in Example 1.

[0090] S2: Same as step S2 in Example 1.

[0091] S3: Same as step S3 in Example 1.

[0092] S4: Mix 2 mL of solution A obtained in step S3 and 0.5 mL of 100 nM CpG oligonucleotide in 10 mL of Tris-HCl buffer at 37°C for 15 minutes, then transfer to a dialysis bag with a molecular weight cutoff of 3,500 Da and dialyze against deionized water for 2 days. The resulting dialysate is the ONMs nanoparticle solution, which is stored at 4°C for subsequent use.

[0093] S5: 1 mL of 100 nM amino-modified ATP aptamer Aapt was mixed with 10 mL of 2.8 wt% sodium alginate (SA) in 20 mL MES buffer at 37°C for 2 h, followed by the addition of 1 mL of 1 M EDC·HCl and the mixture was shaken at 37°C for 2 h. 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol were then added in sequence. The mixture was transferred to -80°C for 10 min and then centrifuged at 10,000 rpm for 10 min. The precipitate was collected, washed with anhydrous ethanol, and redissolved in 1 mL of deionized water to obtain DNA SA solution.

[0094] S6: 1 mL of the product from step S5 DNA The SA solution was mixed with 0.5 mL of the ONMs nanoparticle solution obtained in step S4 and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution.

[0095] S7: 1 mL of the product from step S6 DNA The SA@ONMs solution was mixed with 1 mL of 15 wt% acrylamide (AAm) solution, and the ionic crosslinker calcium chloride, the crosslinker N,N'-methylenebisacrylamide, the crosslinking accelerator tetramethylethylenediamine, and the initiator ammonium persulfate were added in sequence. The mixture was stirred at room temperature for 8 h and then allowed to stand for 30 min to solidify into a gel. DNA SA / PAAm@ONMs hydrogel. DNA In the SA / PAAm@ONMs hydrogel, the final concentration of calcium chloride was 0.2 wt %, the final concentration of N,N'-methylenebisacrylamide was 0.03 wt %, the final concentration of tetramethylethylenediamine was 0.05 wt %, and the final concentration of ammonium persulfate was 0.2 wt %.

[0096] Example 7

[0097] DNA The preparation method of SA / PAAm@ONMs hydrogel is carried out according to the following steps:

[0098] S1: Same as step S1 in Example 1.

[0099] S2: Same as step S2 in Example 1.

[0100] S3: Same as step S3 in Example 1.

[0101] S4: Mix 2 mL of solution A obtained in step S3 and 0.5 mL of 50 nM CpG oligonucleotide in 10 mL of Tris-HCl buffer at 37°C for 15 minutes, then transfer to a dialysis bag with a molecular weight cutoff of 3,500 Da and dialyze against deionized water for 2 days. The resulting dialysate is the ONMs nanoparticle solution, which is stored at 4°C for subsequent use.

[0102] S5: 1 mL of 200 nM amino-modified ATP aptamer Aapt was mixed with 10 mL of 2.8 wt% sodium alginate (SA) in 20 mL MES buffer at 37°C for 2 h, followed by the addition of 1 mL of 1 M EDC·HCl and the mixture was shaken at 37°C for 2 h. 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol were then added in sequence. The mixture was transferred to -80°C for 10 min and then centrifuged at 10,000 rpm for 10 min. The precipitate was collected, washed with anhydrous ethanol, and redissolved in 1 mL of deionized water to obtain DNA SA solution.

[0103] S6: 1 mL of the product from step S5 DNA The SA solution was mixed with 0.5 mL of the ONMs nanoparticle solution obtained in step S4 and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution.

[0104] S7: 1 mL of the product from step S6 DNA The SA@ONMs solution was mixed with 1 mL of 15 wt% acrylamide (AAm) solution, and the ionic crosslinker calcium chloride, the crosslinker N,N'-methylenebisacrylamide, the crosslinking accelerator tetramethylethylenediamine, and the initiator ammonium persulfate were added in sequence. The mixture was stirred at room temperature for 8 h and then allowed to stand for 30 min to solidify into a gel. DNA SA / PAAm@ONMs hydrogel. DNA In the SA / PAAm@ONMs hydrogel, the final concentration of calcium chloride was 0.2 wt %, the final concentration of N,N'-methylenebisacrylamide was 0.03 wt %, the final concentration of tetramethylethylenediamine was 0.05 wt %, and the final concentration of ammonium persulfate was 0.2 wt %.

[0105] Comparative Example 1

[0106] DNA The preparation method of SA / PAAm@ONMs hydrogel is carried out according to the following steps:

[0107] S1: Same as step S1 of Example 1.

[0108] S2: Same as step S2 of Example 1.

[0109] S3: Same as step S3 of Example 1.

[0110] S4: 3 mL of solution A obtained in step S3 and 0.5 mL of 100 nM CpG oligonucleotide were mixed in 10 mL of Tris-HCl buffer at 37°C for 15 min, and then transferred to a dialysis bag with a molecular weight cut-off of 3,500 Da, and dialyzed against deionized water for 2 days. The dialysate was the ONMs nanoparticle solution, which was stored at 4°C for subsequent use.

[0111] S5: 1 mL of 200 nM amino-modified ATP aptamer Aapt was mixed with 10 mL of 2.8 wt% sodium alginate (SA) in 20 mL of MES buffer at 37°C for 2 h, followed by the addition of 1 mL of 1 M EDC-HCl, and the system was uniformly shaken at 37°C for 2 h. Then, 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol were added in sequence, and the system was transferred to -80°C for 10 min, followed by centrifugation at 10,000 rpm for 10 min. The precipitate was washed with anhydrous ethanol and redissolved with 1 mL of deionized water to obtain DNA SA solution.

[0112] S6: 1 mL of the DNA SA solution obtained in step S5 was mixed with 0.5 mL of the ONMs nanoparticle solution obtained in step S4, and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution.

[0113] S7: 1 mL of the DNA SA@ONMs solution obtained in step S6 was mixed with 1 mL of 15 wt% acrylamide (AAm) solution, and ion crosslinking agent calcium chloride, crosslinking agent N,N'-methylene bisacrylamide, crosslinking promoter tetramethyl ethylenediamine, and initiator ammonium persulfate were added in sequence. The system was stirred at room temperature for 8 h, and then left to stand for 30 min to facilitate gelation to obtain DNA SA / PAAm@ONMs hydrogel. In the DNA SA / PAAm@ONMs hydrogel, the final concentration of calcium chloride was 0.2 wt%, the final concentration of N,N'-methylene bisacrylamide was 0.03 wt%, the final concentration of tetramethyl ethylenediamine was 0.05 wt%, and the final concentration of ammonium persulfate was 0.2 wt%.

[0114] Comparative Example 2

[0115] DNAThe preparation method of SA / PAAm@ONMs hydrogel is carried out according to the following steps:

[0116] S1: Same as step S1 in Example 1.

[0117] S2: Same as step S2 in Example 1.

[0118] S3: Same as step S3 in Example 1.

[0119] S4: Mix 2 mL of solution A obtained in step S3 and 0.5 mL of 100 nM CpG oligonucleotide in 10 mL of Tris-HCl buffer at 37°C for 15 minutes, then transfer to a dialysis bag with a molecular weight cutoff of 3,500 Da and dialyze against deionized water for 2 days. The resulting dialysate is the ONMs nanoparticle solution, which is stored at 4°C for subsequent use.

[0120] S5: 1 mL of 100 nM amino-modified ATP aptamer Aapt was mixed with 10 mL of 2.8 wt% sodium alginate (SA) in 20 mL MES buffer at 37°C for 2 h, followed by the addition of 1 mL of 1 M EDC·HCl and the mixture was shaken at 37°C for 2 h. 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol were then added in sequence. The mixture was transferred to -80°C for 10 min and then centrifuged at 10,000 rpm for 10 min. The precipitate was collected, washed with anhydrous ethanol, and redissolved in 1 mL of deionized water to obtain DNA SA solution.

[0121] S6: 1 mL of the product from step S5 DNA The SA solution was mixed with 0.5 mL of the ONMs nanoparticle solution obtained in step S4 and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution.

[0122] S7: 1 mL of the product from step S6 DNA The SA@ONMs solution was mixed with 1 mL of 15 wt% acrylamide (AAm) solution, and the ionic crosslinker calcium chloride, the crosslinker N,N'-methylenebisacrylamide, the crosslinking accelerator tetramethylethylenediamine, and the initiator ammonium persulfate were added in sequence. The mixture was stirred at room temperature for 8 h and then allowed to stand for 30 min to solidify into a gel. DNA SA / PAAm@ONMs hydrogel. DNA In the SA / PAAm@ONMs hydrogel, the final concentration of calcium chloride was 0.2 wt %, the final concentration of N,N'-methylenebisacrylamide was 0.03 wt %, the final concentration of tetramethylethylenediamine was 0.05 wt %, and the final concentration of ammonium persulfate was 0.2 wt %.

[0123] Comparative Example 3

[0124] The preparation method of the SA / PAAm hydrogel is carried out in the following steps:

[0125] 1 mL of 2.8 wt% sodium alginate (SA) is mixed with 1 mL of 15 wt% acrylamide (AAm) solution, and then ion crosslinking agent calcium chloride, crosslinking agent N,N'-methylene bisacrylamide, crosslinking promoter tetramethyl ethylenediamine and initiator ammonium persulfate are added in sequence, and the reaction is stirred at room temperature for 8 h, and then left for 30 min for solidification into glue, to obtain the SA / PAAm hydrogel. In the SA / PAAm hydrogel, the final concentration of calcium chloride is 0.2 wt%, the final concentration of N,N'-methylene bisacrylamide is 0.03 wt%, the final concentration of tetramethyl ethylenediamine is 0.05 wt%, and the final concentration of ammonium persulfate is 0.2 wt%.

[0126] Figure 1 For the present application DNA The preparation process and mechanism diagram of the SA / PAAm@ONMs hydrogel.

[0127] Figures 2-3 For the preparation of Example 1 DNA The basic physicochemical performance characterization diagram of the ONMs nanoparticles in the SA / PAAm@ONMs hydrogel. Figure 2 A to Figure 2 D By SEM, TEM and other morphology characterization, it is proved that after loading the polymer, the Mxene nanosheet becomes more rough; and Figure 2 E to Figure 2 The particle size analysis of F shows that the size of the nanoparticles before and after modification is still stable and uniform; Figure 2 G to Figure 3 B By elemental analysis zeta potential analysis and XRD analysis, it is found that the modified nanoparticles appear disulfide and characteristic elements on the CpG oligonucleotide, the potential changes, the characteristic peak of aluminum element disappears, and the wide peak of the polymer appears, proving that the prepared ONMs nanoparticles successfully carry disulfide and CpG oligonucleotide; Figure 3 C By using DCFH probe fluorescence characterization of the ROS produced by the nanoparticles under the stimulation of ultrasound (0.5 W / cm 2 , 1 MHz, 3 min) and 808 nm laser (1.0 W / cm 2 , 5 min), it is found that the ONMs nanoparticles can effectively produce ROS under the stimulation of both; Figure 3 E By characterizing the fluorescence intensity before and after adding GSH, it is proved that the ONMs nanoparticles have good GSH response function; Figure 3 D, Figure 3 F to Figure 3The nanoparticle solution was characterized by thermal imaging under the influence of 808 nm laser (1.0 W / cm 2 The results show that ONM nanoparticles have excellent photothermal conversion capabilities. Therefore, ONM nanoparticles successfully carry disulfide and CpG oligonucleotides, and have good photothermal performance, ROS generation performance, and GSH response capacity.

[0128] Figures 4-5 Prepared in Example 1 DNA Morphological structure and performance characterization diagram of SA / PAAm@ONMs hydrogel and SA / PAAm hydrogel prepared in Comparative Example 3. Figure 4 A to Figure 4 D The morphology of ONMs nanoparticles was characterized by SEM. DNA SA / PAAm@ONMs hydrogel; Figure 4 The PO characteristic peaks in the infrared test of E indicate that the ATP aptamer has been successfully integrated. DNA SA / PAAm@ONMs hydrogel substrate; Figure 4 F to Figure 4 G characterizes the swelling and degradation properties by measuring the mass change of the hydrogel and performing contact angle analysis on the hydrogel. DNA SA / PAAm@ONMs hydrogel has good hydrophilicity; Figure 4 H to Figure 5 C The mechanical properties of hydrogel compression and adhesion are tested to show that DNA SA / PAAm@ONMs hydrogel has better mechanical properties and adhesion ability than SA / PAAm hydrogel, and the digital image shows DNA SA / PAAm@ONMs hydrogel can effectively adhere to various substrates; Figure 5 D to Figure 5 G is illustrated by characterizing the fluorescent groups modified on CpG. DNA SA / PAAm@ONMs hydrogel was stimulated by 808 nm laser (1.0 W / cm 2 ) conditions, ONMs nanoparticles can be effectively released, and they have multiple stimulus response capabilities. DNA SA / PAAm@ONMs hydrogel successfully carried ONMs nanoparticles and had good adhesion properties and the ability to respond to release under near-infrared and ATP stimulation.

[0129] Figure 6 Prepared in Example 1 DNA Qualitative and quantitative analysis of the biocompatibility and anticancer properties of SA / PAAm@ONMs hydrogel and SA / PAAm hydrogel prepared in Comparative Example 3.Figure 6 A to Figure 6 B is to culture MSCs cells with different concentrations of DNA SA / PAAm@ONMs hydrogel co-culture, the results show that, DNA SA / PAAm@ONMs hydrogel has no obvious difference in the proliferation activity of MSCs cells; Figure 6 C to Figure 6 E is to culture 143B cells with DNA SA / PAAm@ONMs hydrogel, SA / PAAm hydrogel co-culture under different stimulation conditions, the results show that, DNA SA / PAAm@ONMs hydrogel generates the most active oxygen under near-infrared light and ultrasonic stimulation, and the ONMs nanoparticles inside have excellent light response and ultrasonic response ROS generation ability, DNA SA / PAAm@ONMs hydrogel has the strongest killing effect on 143B cells under near-infrared light and ultrasonic stimulation. Therefore, DNA SA / PAAm@ONMs hydrogel shows good biocompatibility and anticancer performance.

[0130] Figure 7 SA / PAAm@ONMs hydrogel prepared in Example 1 DNA SA / PAAm@ONMs hydrogel releases ONMs nanoparticles, and the uptake ability analysis diagram is shown. CpG oligonucleotide, DNA SA / PAAm@ONMs composite hydrogel (VAC-CpG) was co-cultured with 143B cells in transwell for 4h, and its uptake was tested by fluorescence labeling. From Figure 7 It can be seen that compared with free CpG oligonucleotide, 143B cells have stronger uptake performance for DNA SA / PAAm@ONMs composite hydrogel has strong uptake performance for ONMs.

[0131] SA / PAAm@ONMs hydrogel prepared in Example 1 and Comparative Example 1 DNA The anti-tumor performance of SA / PAAm@ONMs hydrogel was tested, and normal cells (MSCs cells) and tumor cells (143B cells) were co-cultured with the prepared hydrogel under ultrasonic (0.5W / cm 2 , 1MHz) and near-infrared light (808nm, 1.0W / cm 2 ) stimulation conditions for 2h, and tested by CCK8. The results show that when the concentrations of other components remain unchanged, the group with high MXene concentration has stronger anti-tumor ability. However, it should be noted that when the concentration of MXene is high, DNA The cytotoxicity of SA / PAAm@ONMs hydrogel will increase (Table 1).

[0132] Table 1 Cell survival test results

[0133] Serial Number Normal cell survival rate Tumor cell killing rate Example 1 95％ 75％ Comparative Example 1 90％ 80％

[0134] The drug release performance of the SA / PAAm@ONMs hydrogel prepared in Example 1 and Comparative Example 2 was tested. The prepared hydrogel was tested for drug release after 2 h under the condition of whether near-infrared light (808 nm, 1.0 W / cm2) stimulation was given. The results showed that, when the concentrations of other components were unchanged, the response release ability of ONMs would decrease when the concentration of aptamer Aapt was reduced, that is, the drug release rate of the SA / PAAm@ONMs hydrogel would decrease (Table 2). DNA 2 ) stimulation. The results showed that, when the concentrations of other components were unchanged, the response release ability of ONMs would decrease when the concentration of aptamer Aapt was reduced, that is, the drug release rate of the SA / PAAm@ONMs hydrogel would decrease (Table 2). DNA

[0135] Table 2. Drug release test results

[0136] Serial Number Release rate without administration of a stimulating drug Release rate with administration of a stimulating drug Example 1 20％ 80％ Comparative Example 2 50％ 70％

[0137] The above examples are only used to help understand the method of the present application and its core idea. It should be pointed out that, for those skilled in the art, without departing from the principles of the present application, the present application can be improved and modified in several ways, and these improvements and modifications also fall within the protection scope of the claims of the present application.​​

Claims

1. A method for preparing a multi-substrate adhesive hydrogel capable of efficiently delivering nucleic acid molecules, characterized by: The following steps are involved: S1: MXene nanosheet solution was prepared by hydrofluoric acid etching; S2: preparing disulfide solid powder by recrystallization; S3: Mixing the disulfide solid powder with the MXene nanosheet solution and stirring at room temperature for 2 h to prepare solution A; S4: 0.5 mL of 50-200 nM CpG oligonucleotide was mixed with 1-2 mL of solution A and dialyzed to prepare an ONM nanoparticle solution. S5: 10 mL of 2.8 wt% sodium alginate and 1 mL of 200-400 nM amino-modified ATP aptamer were grafted to prepare DNA SA solution; S6: ONMs nanoparticle solution and DNA The SA solution was mixed and incubated at room temperature for 2 h to prepare DNA SA@ONMs solution; S7: DNA The SA@ONMs solution was mixed with acrylamide, and then an ionic crosslinker, a crosslinker, a crosslinking accelerator, and an initiator were added in sequence. The mixture was stirred at room temperature for 8 to 24 hours and allowed to stand to solidify into a gel, resulting in a multi-substrate adhesion hydrogel that can efficiently deliver nucleic acid molecules. The concentration of the MXene nanosheet solution is 1 mg / mL; The ratio of the disulfide solid powder to the MXene nanosheet solution is 10:100 w / v; The sequence of the CpG oligonucleotide is 5'-Cy5-TCCATGACGTTCCTGACGTTACCTTCCTCCGCAA-3', and the sequence of the amino-modified ATP aptamer is 5'-NH2-ACCTGGGGGAGTATTGCGGAGGAAGGT-3'.

2. The preparation method according to claim 1, wherein: Specifically, step S4 comprises mixing solution A and CpG oligonucleotide in 10 mL of Tris-HCl buffer at 37° C. for 15 minutes, then transferring the mixture to a dialysis bag with a molecular weight cutoff of 3,500 Da, and dialyzing the mixture with deionized water for 2 days. The resulting dialysate is the ONMs nanoparticle solution.

3. The preparation method according to claim 1, wherein: The step S5 is specifically as follows: the amino-modified ATP aptamer and sodium alginate are mixed in 20 mL of MES buffer at 37°C for 2 h, 1 mL of 1 M EDC·HCl is added, and the mixture is shaken at 37°C for 2 h. Then, 5 mL of 3 M sodium acetate and 30 mL of anhydrous ethanol are added in sequence. The mixture is transferred to -80°C and placed for 10 min, followed by centrifugation at 10,000 rpm for 10 min, the precipitate is collected, washed with anhydrous ethanol, and redissolved with 1 mL of deionized water to obtain DNA SA solution.

4. The preparation method according to claim 1, wherein: The step S6 is specifically as follows: DNA SA solution was mixed with 0.5 mL ONMs nanoparticle solution and incubated at room temperature for 2 h to obtain DNA SA@ONMs solution.

5. The preparation method according to claim 1, wherein: The specific step S7 is: DNA The SA@ONMs solution was mixed with 1 mL of 15 wt% acrylamide solution, and an ionic crosslinker with a final concentration of 0.1 wt% to 0.4 wt% was added, a crosslinker with a final concentration of 0.01 wt% to 0.05 wt% was added, a crosslinking accelerator with a final concentration of 0.02 wt% to 0.10 wt% was added, and an initiator with a final concentration of 0.1 wt% to 0.3 wt% was added. The mixture was stirred at room temperature for 8 h and then allowed to stand for 30 min to solidify into a gel. DNA SA / PAAm@ONMs hydrogel is a multi-substrate adhesion hydrogel that can efficiently deliver nucleic acid molecules.

6. The preparation method according to claim 1, wherein: The ionic crosslinking agent is calcium chloride, the crosslinking agent is N,N-methylenebisacrylamide, the crosslinking accelerator is tetramethylethylenediamine, and the initiator is ammonium persulfate.

7. A multi-substrate adhesive hydrogel capable of efficiently delivering nucleic acid molecules, characterized by: The compound is prepared by the preparation method according to any one of claims 1 to 6.

8. Use of the multi-substrate adhesive hydrogel according to claim 7 in preparing a drug for treating tumors, characterized in that: The tumor is osteosarcoma.

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