Preparation method of dendritic mesoporous silicon bionic nanoparticles loaded with RNA (Ribonucleic Acid) molecules

Abstract

The invention provides a preparation method of dendritic mesoporous silicon bionic nanoparticles loaded with RNA (Ribonucleic Acid) molecules, which comprises the following steps: preparing dendritic mesoporous silicon nanoparticles DMSNs by adopting a double-phase layering method, dissolving a surfactant CTAC and a catalyst TEA in water to serve as a water phase solution, dissolving ethyl silicate TEOS in cyclohexane to serve as an oil phase, slowly dropwise adding the oil phase into the water phase, and reacting for 2-4 hours to obtain the dendritic mesoporous silicon bionic nanoparticles loaded with the RNA molecules. The DMSNs nanoparticles are obtained; amino groups are introduced to the surfaces of DMSNs through gamma-aminopropyltriethoxysilane APTES so as to enhance electrostatic attraction to nucleic acid, and after a surfactant template is removed, the DMSNs are mixed with RNA molecules in ribozyme-free water; meanwhile, a cytoplasmic membrane is harvested by using a hypotonic method, and the dendritic mesoporous silicon bionic nanoparticles loaded with the RNA molecules are prepared through ultrasonic treatment and polycarbonate film extrusion. The method is simple and convenient, and has a wide application prospect in RNA delivery.

Description

Technical Field

[0001] The invention relates to a method in the technical field of biomedical materials, in particular to a method for preparing dendritic mesoporous silicon bionic nanoparticles loaded with RNA molecules. Background Art

[0002] RNA therapy is an emerging therapy that promotes disease treatment or prevention by manipulating the genetic code RNA. In the past few decades, RNA therapy has shown great promise in biomedicine. It has many advantages. Compared with drugs targeting proteins (small molecule compounds or antibodies), RNA drugs can target a wider range of genetic molecules because most DNA sequences in the human genome are transcribed into non-coding transcripts and about 85% of proteins lack specific cracks for small molecules to bind. Secondly, RNA drugs are long-lasting and do not cause irreversible genomic changes, and do not cause potential genetic toxicity risks similar to DNA therapy. Although people have a strong interest in RNA therapy, the difficulties in RNA delivery have become a bottleneck for RNA therapy. First, RNA has a high molecular weight and is negatively charged, which may repel negatively charged cell membranes and make it difficult to enter cells and function. Second, naked RNA molecules are extremely susceptible to degradation by nucleases in cells or the environment and lose their function. Third, RNA is easily cleared by endosomes and lysosomes after entering cells through endocytosis. Fourth, the lack of targeting itself may cause serious side effects and limit the therapeutic effect. In view of the above problems, the design of the delivery system is particularly critical.

[0003] In recent years, the emergence and development of nanotechnology has provided a new idea for nucleic acid drug carriers. Various non-viral nanocarriers have shown broad application prospects in nucleic acid delivery, tumor diagnosis and treatment. Mesoporous silica has attracted widespread attention due to its uniform and adjustable particle size, pore size and shape, modular surface chemistry and excellent biocompatibility. The mesopore size of traditional mesoporous silica (MSNs) is usually below 2 nm. For biological macromolecules such as nucleic acids and enzymes, the use of DMSNs with larger pores can undoubtedly increase their cargo load. In addition, given the complexity of the biological environment, RNA carrier design considers effective ways to interact with the environment and reduce nonspecific interactions. Therefore, providing a cell membrane biomimetic mesoporous silica nanoparticle and a preparation method thereof has important scientific research and application value. Summary of the invention

[0004] The present invention aims to provide a method for preparing RNA molecule-loaded dendritic mesoporous silicon biomimetic nanoparticles.

[0005] The object of the present invention is achieved through the following scheme: a method for preparing dendritic mesoporous silicon biomimetic nanoparticles loaded with RNA molecules, wherein dendritic mesoporous silicon nanoparticles (DMSNs) are prepared by a two-phase layering method, amino groups are introduced on the surface of the DMSNs by using γ-aminopropyltriethoxysilane (APTES) to enhance the electrostatic attraction to nucleic acids, and after removing the surfactant template, the DMSNs are mixed with RNA molecules in nuclease-free water; at the same time, the cell plasma membrane is harvested by a hypotonic and cyclic freeze-thaw method, and the dendritic mesoporous silicon biomimetic nanoparticles loaded with RNA molecules are prepared by ultrasonic treatment and polycarbonate film extrusion, comprising the following steps: (1) Preparation of dendritic mesoporous silica nanoparticles: The surfactant hexadecyltrimethylammonium chloride (CTAC) and the catalyst triethanolamine (TEA) were dissolved in deionized water as the aqueous phase solution. At the same time, ethyl silicate (TEOS) was dissolved in cyclohexane as the oil phase. The oil phase was slowly dripped on top of the aqueous phase and stirred at 400 rpm at 40 °C for 18 h to obtain dendritic mesoporous silica nanoparticles DMSNs. (2) Amino modification of dendritic mesoporous silica nanoparticles: Add γ-aminopropyltriethoxysilane (APTES) ethanol solution to the aqueous phase and continue the reaction for 4 h. After the reaction is completed, centrifuge at 50,000 g for 15 min to harvest the amino-modified dendritic mesoporous silica nanoparticles. (3) Removal of surfactant CTAC: The nanoparticles were repeatedly washed with concentrated hydrochloric acid-ethanol solution, and the obtained dendritic mesoporous silica nanoparticles (DMSNs) were stored in anhydrous ethanol at 4°C; (4) RNA loading: Dissolve the RNA powder in nuclease-free water, add the DMSNs from step (3) at a mass ratio of RNA:DMSNs = 1:5-1:20, mix the DMSN nanoparticles and RNA thoroughly, incubate at 4°C for 15 min, adsorb the RNA on the DMSNs, and centrifuge at 20,000 g for 10 min to remove unloaded RNA molecules; (5) Coating the cell membrane on the surface of dendritic mesoporous silica nanoparticles: The collected cell membrane and DMSNs loaded with RNA molecules were mixed evenly in a mass ratio of 1:1-1:2, ultrasonicated for 1 min in an ice bath probe, and extruded through 1000 nm, 400 nm, and 200 nm polycarbonate films in sequence. The obtained dendritic mesoporous silica biomimetic nanoparticles were resuspended in PBS to obtain dendritic mesoporous silica biomimetic nanoparticles loaded with RNA molecules.

[0006] The advantages of the present invention are high-efficiency RNA molecule loading and cell biomimetic, relatively simple operation and low cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Attached Figure 1Effect of TEOS concentration on the size and morphology of dendritic mesoporous silica. DMSN-5, DMSN-10, and DMSN-20 represent 5%, 10%, and 20% TEOS concentrations in the oil phase during preparation, respectively. Attached Figure 2 TEM image of dendritic mesoporous silica erythrocyte membrane biomimetic nanoparticles loaded with PDL1-siRNA. DETAILED DESCRIPTION

[0008] The following examples are implemented based on the technical solutions of the invention, and provide detailed implementation methods and specific operation processes, but the protection scope of the present invention is not limited to the following examples.

[0009] Example 1 A dendritic mesoporous silica biomimetic nanoparticle loaded with RNA molecules, wherein the dendritic mesoporous silica nanoparticle DMSNs is prepared by a two-phase layering method, and amino groups are introduced on the surface of the dendritic mesoporous silica nanoparticle using γ-aminopropyltriethoxysilane APTES to enhance the electrostatic attraction to nucleic acids, and after removing the surfactant template, the dendritic mesoporous silica nanoparticle is mixed with RNA molecules in nuclease-free water; at the same time, the cell plasma membrane is harvested by a hypotonic method, and the dendritic mesoporous silica biomimetic nanoparticle loaded with RNA molecules is prepared by ultrasonic treatment and polycarbonate film extrusion, and the preparation is performed according to the following steps: (1) Preparation of dendritic mesoporous silica nanoparticles: 120 mL of 25 wt.% CTAC aqueous solution and 0.9 g of catalyst TEA were dissolved in 180 mL of deionized water and stirred thoroughly, which was recorded as aqueous solution A. At the same time, 2 mL of TEOS was dissolved in 18 mL of cyclohexane to prepare 20 mL (10% TEOS) oil phase solution B. 60 mL of solution A was added to a 100 mL round-bottom flask and stirred at 400 rpm for 30 min at 40 °C to allow CTAC to form lamellar micelles in water. The stirring speed was reduced to 200 rpm, and 20 mL of solution B was slowly added dropwise to solution A, paying attention to the formation of the interface between the aqueous phase and the oil phase. The stirring speed was increased to 400 rpm and the two-phase system was maintained at 40 °C for 18 h to obtain dendritic mesoporous silica nanoparticles DMSNs. (2) Amino modification of dendritic mesoporous silica nanoparticles: APTES was dissolved in 250 μL of anhydrous ethanol, mixed and dissolved, and APTES was added to the lower aqueous phase solution to ensure that the APTES / TEOS molar ratio was 0.5 / 10. The reaction was continued for 4 h. After the reaction was completed, the amino-modified dendritic mesoporous silica nanoparticles DMSNs were harvested by centrifugation at 50,000 g for 15 min. (3) Removal of surfactant CTAC: Prepare a 0.12 M hydrochloric acid-ethanol solution, wash the nanoparticles repeatedly with the concentrated hydrochloric acid-ethanol solution for 5 times, and store the obtained nanoparticles in anhydrous ethanol at 4°C; (4) Cytoplasmic membrane extraction: 20 mL of rat whole blood was collected and centrifuged three times at low speed to wash blood cells. Red blood cells were collected and placed in 0.25X PBS. Protease inhibitors were added and hypotonic treatment was performed for 1 h. Red blood cell membrane (RBCM) was obtained by centrifugation at 20,000 g. The RBCM was washed three times to remove soluble proteins and ultrasonicated for 5 min. The obtained RBCM was added to a liposome extruder and passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. Membrane vesicles were obtained by centrifugation at 20,000 g. The protein concentration was quantified using the BCA method and stored in aliquots at -80°C. (5) RNA loading: Wash the DMSNs with nuclease-free water for three times, add the DMSNs from step (3) at a mass ratio of RNA:DMSNs = 1:5-1:20, incubate at 4°C for 15 min to obtain DMSNs-RNA; centrifuge at 20,000 g for 10 min to remove unloaded RNA molecules; (6) Coating the plasma membrane on the surface of dendritic mesoporous silica nanoparticles: The collected red blood cell membrane and DMSNs-RNA were mixed evenly in a mass ratio of 1:1-1:2, ultrasonicated for 1 min in an ice bath, added to a liposome extruder, and passed through 1000 nm, 400 nm, and 200 nm polycarbonate nuclear pore membranes in sequence. The obtained 10,000 g dendritic mesoporous silica biomimetic nanoparticles were resuspended in PBS and frozen at -80°C.

[0010] Effect of TEOS concentration on the size and morphology of dendritic mesoporous silica. DMSN-5, DMSN-10, and DMSN-20 represent 5%, 10%, and 20% TEOS concentrations in the oil phase during preparation, respectively. Figure 1 shown.

[0011] TEM images of dendritic mesoporous silica erythrocyte membrane biomimetic nanoparticles loaded with PDL1-siRNA are shown in the attached Figure 2 .

[0012] Example 2 A dendritic mesoporous silica biomimetic nanoparticle loaded with RNA molecules is prepared by the following steps similar to those in Example 1: (1) Preparation of dendritic mesoporous silica nanoparticles: Accurately weigh 0.9 g of TEA, 180 mL of deionized water, and 120 mL of a 25 wt. % CTAC aqueous solution, and stir them thoroughly to mix them, which is recorded as aqueous solution A. Dissolve 1 mL of TEOS in 19 mL of cyclohexane to prepare 20 mL (5% TEOS) of oil phase solution B. Add 60 mL of solution A to a 100 mL round-bottom flask and stir at 400 rpm for 30 min at 40 °C to allow CTAC to form lamellar micelles in water. Reduce the stirring speed to 200 rpm and slowly add 20 mL of solution B dropwise into solution A, paying attention to the formation of the interface between the aqueous phase and the oil phase. Increase the stirring speed to 400 rpm and maintain the two-phase system at 40 °C for 18 h to obtain dendritic mesoporous silica nanoparticles DMSNs. (2) Amino modification of dendritic mesoporous silica nanoparticles: APTES was dissolved in 250 μL of anhydrous ethanol, mixed and dissolved, and APTES was added to the lower layer solution to ensure that the APTES / TEOS molar ratio was 0.5 / 10; the reaction was continued for 4 h. After the reaction was completed, DMSNs were collected by centrifugation at 50,000 g; (3) Removal of surfactant CTAC: Prepare 0.12 M hydrochloric acid-ethanol solution, wash DMSNs 5 times, and resuspend in ethanol for storage; Use concentrated hydrochloric acid-ethanol solution to repeatedly wash the nanoparticles, and store the obtained nanoparticles in anhydrous ethanol at 4°C; (4) Cytoplasmic membrane extraction: 20 mL of rat whole blood was collected and centrifuged three times at low speed to wash blood cells. Red blood cells were collected and placed in 0.25X PBS. Protease inhibitors were added and hypotonic treatment was performed for 1 h. Red blood cell membrane (RBCM) was obtained by centrifugation at 20,000 g. The RBCM was washed three times to remove soluble proteins and ultrasonicated for 5 min. The obtained RBCM was added to a liposome extruder and passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. Membrane vesicles were obtained by centrifugation at 20,000 g. The protein concentration was quantified using the BCA method and stored in aliquots at -80°C. (4) RNA loading: DMSNs were washed three times with nuclease-free water, and RNA and DMSNs were mixed at a mass ratio of RNA:DMSNs = 1:5-1:20. The mixture was incubated at 4°C for 15 min, and centrifuged at 20,000 g for 10 min to remove unloaded RNA molecules to obtain DMSNs-RNA.

[0013] (6) Coating the plasma membrane on the surface of dendritic mesoporous silica nanoparticles: The membrane vesicles and DMSNs-RNA were mixed in a mass ratio of 1:1-1:2 and evenly added to the liposome extruder. The mixture was ultrasonicated in an ice bath for 1 min and passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. The dendritic mesoporous silica biomimetic nanoparticles loaded with RNA molecules were obtained by centrifugation at 10,000 g. The mixture was resuspended in PBS and frozen at -80°C.

[0014] Effect of TEOS concentration on the size and morphology of dendritic mesoporous silica. DMSN-5, DMSN-10, and DMSN-20 represent 5%, 10%, and 20% TEOS concentrations in the oil phase during preparation, respectively. Figure 1 shown.

[0015] Example 3 A dendritic mesoporous silica biomimetic nanoparticle loaded with RNA molecules is prepared by the following steps similar to those in Example 1: (1) Preparation of dendritic mesoporous silica nanoparticles: Accurately weigh 0.9 g of TEA, 180 mL of deionized water, and 120 mL of 25 wt. % CTAC aqueous solution, and stir them thoroughly, which is recorded as aqueous solution A. Dissolve 4 mL of TEOS in 16 mL of cyclohexane to obtain 20 mL (20% TEOS) of oil phase solution B. Add 60 mL of solution A to a 100 mL round-bottom flask and stir at 400 rpm for 30 min at 40 °C to allow CTAC to form lamellar micelles in water. Reduce the stirring speed to 200 rpm and slowly add 20 mL of solution B dropwise into solution A, paying attention to the formation of the interface between the aqueous phase and the oil phase. Increase the stirring speed to 400 rpm and maintain the two-phase system at 40 °C for 18 h to obtain dendritic mesoporous silica nanoparticles DMSNs. (2) Amino modification of dendritic mesoporous silica nanoparticles: APTES was dissolved in 250 μL of anhydrous ethanol, mixed and dissolved, and APTES was added to the lower layer solution A to ensure that the APTES / TEOS molar ratio was 0.5 / 10; amino-modified DMSNs were collected by centrifugation at 50,000 g. (3) Removal of surfactant CTAC: Prepare 0.12 M hydrochloric acid-ethanol solution, wash DMSNs repeatedly with concentrated hydrochloric acid-ethanol solution 5 times, and resuspend in ethanol for storage; (4) Cytoplasmic membrane extraction: ID8 ovarian tumor cell line was cultured in DMEM complete medium at 37°C and 5% CO2 until the logarithmic phase. The cells were collected and placed in hypotonic TM buffer. Protease inhibitors were added and hypotonic treatment was performed overnight. After mechanical disruption of the cells, 1M sucrose solution was added to a final concentration of 0.25 M. Tumor cell membranes were obtained by centrifugation at 2000 g. The probe was sonicated for 5 min. The obtained cell membranes were added to a liposome extruder and passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. Membrane vesicles were obtained by centrifugation at 20,000 g. The protein concentration was quantified using the BCA method and the cells were aliquoted and stored at -80°C. (5) RNA loading: DMSNs were washed three times with nuclease-free water, RNA and DMSNs were mixed at a mass ratio of 1:5-1:20, incubated at 4°C for 15 min, and centrifuged at 20,000 g for 10 min to remove unloaded RNA molecules to obtain DMSNs-RNA; (6) Coating the plasma membrane on the surface of dendritic mesoporous silica nanoparticles: The membrane vesicles and DMSNs-RNA were mixed in a mass ratio of 1:1-1:2 and added to the liposome extruder, and passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in turn; centrifuged at 10,000 g to obtain dendritic mesoporous silica biomimetic nanoparticles loaded with RNA molecules, which were resuspended in PBS and frozen at -80°C.

[0016] Effect of TEOS concentration on the size and morphology of dendritic mesoporous silica. DMSN-5, DMSN-10, and DMSN-20 represent 5%, 10%, and 20% TEOS concentrations in the oil phase during preparation, respectively. Figure 1 shown.

[0017] Example 4 A dendritic mesoporous silica biomimetic nanoparticle loaded with RNA molecules is prepared by the following steps similar to those in Example 1: (1) Preparation of dendritic mesoporous silica nanoparticles: Accurately weigh 0.9 g of TEA, 180 mL of deionized water, and 120 mL of 25 wt. % CTAC aqueous solution, and stir them thoroughly, which is recorded as aqueous solution A. Dissolve 8 mL of TEOS in 12 mL of cyclohexane to prepare 20 mL (40% TEOS) of oil phase solution B. Add 60 mL of solution A to a 100 mL round-bottom flask and stir at 400 rpm for 30 min at 40 °C to allow CTAC to form lamellar micelles in water. Reduce the stirring speed to 200 rpm and slowly add 20 mL of solution B dropwise into solution A, paying attention to the formation of the interface between the aqueous phase and the oil phase. Increase the stirring speed to 400 rpm and maintain the two-phase system at 40 °C for 18 h to obtain dendritic mesoporous silica nanoparticles DMSNs. (2) Amino modification of dendritic mesoporous silica nanoparticles: APTES was dissolved in 250 μL of anhydrous ethanol, mixed and dissolved, and APTES was added to the lower solution to ensure that the APTES / TEOS molar ratio was 0.5 / 10. After the reaction was continued for 4 h, the amino-modified DMSNs were collected by centrifugation at 50,000 g. (3) Removal of surfactant CTAC: Prepare a 0.12 M hydrochloric acid-ethanol solution, wash the DMSNs repeatedly 5 times with the concentrated hydrochloric acid-ethanol solution, and resuspend the obtained nanoparticles in ethanol for storage; (4) Cytoplasmic membrane extraction: RAW264.7 macrophage cell line was cultured in DMEM complete culture medium at 37°C, 5% CO2 to the logarithmic phase. The cells were collected and placed in hypotonic TM buffer. Protease inhibitors were added and hypotonic treatment was performed overnight. After mechanical disruption of the cells, 1M sucrose solution was added to a final concentration of 0.25 M. The macrophage membrane was obtained by centrifugation at 2000 g. The macrophage membrane was washed three times and ultrasonicated for 5 min. The obtained cell membrane was added to a liposome extruder and passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. Membrane vesicles were obtained by centrifugation at 20,000 g. The protein concentration was quantified using the BCA method and the cells were aliquoted and stored at -80°C. (5) RNA loading: DMSNs were washed three times with nuclease-free water, and RNA and DMSNs were mixed at a mass ratio of 1:5-1:20, incubated at 4°C for 15 min, and centrifuged at 20,000 g for 10 min to remove unloaded RNA molecules to obtain RNA-loaded DMSNs, which were recorded as DMSNs-RNA. (6) Coating the plasma membrane on the surface of dendritic mesoporous silica nanoparticles: The membrane vesicles and DMSNs-RNA were mixed in a mass ratio of 1:1-1:2 and added to the liposome extruder. The mixture was passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. The dendritic mesoporous silica biomimetic nanoparticles were obtained by centrifugation at 10,000 g. The obtained dendritic mesoporous silica biomimetic nanoparticles loaded with RNA molecules were resuspended in PBS for storage.

[0018] Effect of TEOS concentration on the size and morphology of dendritic mesoporous silica. DMSN-5, DMSN-10, and DMSN-20 represent 5%, 10%, and 20% TEOS concentrations in the oil phase during preparation, respectively. Figure 1 shown.

Claims

1. A method for preparing RNA-loaded dendritic mesoporous silica biomimetic nanoparticles, characterized in that: Dendritic mesoporous silica nanoparticles DMSNs were prepared by a two-phase layering method. γ-aminopropyltriethoxysilane APTES was used to introduce amino groups on its surface to enhance the electrostatic attraction to nucleic acids. After removing the surfactant template, it was mixed with RNA molecules in nuclease-free water. At the same time, the cell plasma membrane was harvested by a hypotonic method, and dendritic mesoporous silica biomimetic nanoparticles loaded with RNA molecules were prepared by ultrasonic treatment and polycarbonate film extrusion. The steps include: (1) Preparation of dendritic mesoporous silicon nanoparticles: The surfactant CTAC and the catalyst TEA were dissolved in deionized water as the aqueous phase solution. At the same time, TEOS was dissolved in cyclohexane as the oil phase. The oil phase was slowly dripped on top of the aqueous phase and stirred at 400 rpm at 40 °C for 18 h to obtain dendritic mesoporous silicon nanoparticles DMSNs. (2) Amino modification of dendritic mesoporous silica nanoparticles: Add γ-aminopropyltriethoxysilane (APTES) ethanol solution to the aqueous phase and continue the reaction for 4 h. After the reaction is completed, centrifuge at 50,000 g to collect the amino-modified dendritic mesoporous silica nanoparticles. (3) Removal of surfactant CTAC: The nanoparticles were repeatedly washed with concentrated hydrochloric acid-ethanol solution, and the obtained nanoparticle DMSNs were resuspended in anhydrous ethanol for storage; (4) RNA loading: Dissolve DMSNs in nuclease-free water, add DMSNs from step (3) at a mass ratio of RNA:DMSNs of 1:5-1:20, incubate at 4°C for 15 min, and centrifuge at 20,000 g for 10 min to remove unloaded RNA molecules; (5) Coating the plasma membrane on the surface of dendritic mesoporous silica nanoparticles: The collected plasma membrane and DMSNs loaded with RNA molecules were mixed evenly in a mass ratio of 1:1-1:2, ultrasonicated for 1 min in an ice bath probe, and then extruded through polycarbonate films of 1000 nm, 400 nm, and 200 nm in sequence. The obtained dendritic mesoporous silica biomimetic nanoparticles were resuspended in PBS to obtain dendritic mesoporous silica biomimetic nanoparticles loaded with RNA molecules.

2. The method for preparing a dendritic mesoporous silicon biomimetic nanoparticle loaded with RNA molecules according to claim 1, characterized in that: The structure of dendritic mesoporous silica nanoparticles is a flower-like structure, and the dendritic subunits are a layered structure.

3. The method for preparing a dendritic mesoporous silicon biomimetic nanoparticle loaded with RNA molecules according to claim 1, characterized in that: The size range of nanoparticles is 30-100 nm, and the size range of mesopores is 5-15 nm.

4. The method for preparing a dendritic mesoporous silicon biomimetic nanoparticle loaded with RNA molecules according to claim 1, characterized in that: The loaded RNA molecules include at least one of antisense oligonucleotides (ASO), small interfering RNA (siRNA), micro RNA (miRNA), messenger RNA (mRNA), ribozymes, and CRISPR guide RNAs (gRNA).

5. The method for preparing RNA-loaded dendritic mesoporous silica biomimetic nanoparticles according to claim 1, characterized in that: The membrane sources of cell membrane biomimetic nanoparticles include eukaryotic and prokaryotic cells.

6. The method for preparing RNA-loaded dendritic mesoporous silica biomimetic nanoparticles according to claim 1, characterized in that: The particle size of the RNA-loaded dendritic mesoporous silica biomimetic nanoparticles is 100-200 nm, with a typical core-shell structure.

7. The method for preparing a dendritic mesoporous silicon biomimetic nanoparticle loaded with RNA molecules according to any one of claims 1 to 6, characterized in that: Prepare as follows: (1) Preparation of dendritic mesoporous silica nanoparticles: 120 mL of 25 wt.% CTAC aqueous solution and 0.9 g of catalyst TEA were dissolved in 180 mL of deionized water and stirred thoroughly to obtain aqueous solution A. At the same time, 2 mL of TEOS was dissolved in 18 mL of cyclohexane to obtain 20 mL of oil solution B. 60 mL of solution A was added to a 100 mL round-bottom flask and stirred at 400 rpm for 30 min at 40 °C to allow CTAC to form lamellar micelles in water. The stirring speed was reduced to 200 rpm, and 20 mL of solution B was slowly added dropwise to solution A, paying attention to the formation of the interface between the aqueous phase and the oil phase. The stirring speed was increased to 400 rpm and the two-phase system was maintained at 40 °C for 18 h to obtain dendritic mesoporous silica nanoparticles DMSNs. (2) Amino modification of dendritic mesoporous silica nanoparticles: APTES was dissolved in 250 μL of anhydrous ethanol, mixed and dissolved, and APTES was added to the lower aqueous phase solution to ensure that the APTES / TEOS molar ratio was 0.5 / 10. The reaction was continued for 4 h. After the reaction was completed, the amino-modified dendritic mesoporous silica nanoparticles DMSNs were harvested by centrifugation at 50,000 g for 15 min. (3) Removal of surfactant CTAC: Prepare a 0.12 M hydrochloric acid-ethanol solution, wash the nanoparticles repeatedly with the concentrated hydrochloric acid-ethanol solution for 5 times, and store the obtained nanoparticles in anhydrous ethanol at 4°C; (4) Cytoplasmic membrane extraction: Take 20 mL of rat whole blood, centrifuge at low speed three times to wash blood cells, collect red blood cells and place them in 0.25X PBS, add protease inhibitors and treat hypotonic for 1 h, centrifuge at 20,000 g to obtain red blood cell membrane (RBCM), wash three times to remove soluble proteins, sonicate with the probe for 5 min, add the obtained RBCM to the liposome extruder, pass through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence; centrifuge at 20,000 g to obtain membrane vesicles, quantify the protein concentration using the BCA method, and store them in aliquots at -80°C; (5) RNA loading: Wash the DMSNs with nuclease-free water for three times, add the DMSNs from step (3) at a mass ratio of RNA:DMSNs = 1:5-1:20, incubate at 4°C for 15 min to obtain DMSNs-RNA; centrifuge at 20,000 g for 10 min to remove unloaded RNA molecules; (6) Coating the plasma membrane on the surface of dendritic mesoporous silica nanoparticles: The collected red blood cell membrane and DMSNs-RNA were mixed evenly in a mass ratio of 1:1-1:2, ultrasonicated for 1 min in an ice bath, added to a liposome extruder, and passed through 1000 nm, 400 nm, and 200 nm polycarbonate nuclear pore membranes in sequence. The obtained 10,000 g dendritic mesoporous silica biomimetic nanoparticles were resuspended in PBS and frozen at -80°C.

8. The method for preparing a dendritic mesoporous silica biomimetic nanoparticle loaded with RNA molecules according to any one of claims 1 to 6, characterized in that: Prepare as follows: (1) Preparation of dendritic mesoporous silica nanoparticles: Accurately weigh 0.9 g of TEA, 180 mL of deionized water, and 120 mL of a 25 wt. % CTAC aqueous solution, and stir them thoroughly to mix them, which is recorded as aqueous solution A. Dissolve 1 mL of TEOS in 19 mL of cyclohexane to prepare 20 mL of oily solution B. Add 60 mL of solution A to a 100 mL round-bottom flask and stir at 400 rpm for 30 min at 40 °C to allow CTAC to form lamellar micelles in water. Reduce the stirring speed to 200 rpm and slowly add 20 mL of solution B dropwise into solution A, paying attention to the formation of the interface between the aqueous phase and the oil phase. Increase the stirring speed to 400 rpm and maintain the two-phase system at 40 °C for 18 h to obtain dendritic mesoporous silica nanoparticles DMSNs. (2) Amino modification of dendritic mesoporous silica nanoparticles: APTES was dissolved in 250 μL of anhydrous ethanol, mixed and dissolved, and APTES was added to the lower layer solution to ensure that the APTES / TEOS molar ratio was 0.5 / 10; the reaction was continued for 4 h. After the reaction was completed, DMSNs were collected by centrifugation at 50,000 g; (3) Removal of surfactant CTAC: Prepare 0.12 M hydrochloric acid-ethanol solution, wash DMSNs 5 times, and resuspend in ethanol for storage; Use concentrated hydrochloric acid-ethanol solution to repeatedly wash the nanoparticles, and store the obtained nanoparticles in anhydrous ethanol at 4°C; (4) Cytoplasmic membrane extraction: 20 mL of rat whole blood was collected and centrifuged three times at low speed to wash blood cells. Red blood cells were collected and placed in 0.25X PBS. Protease inhibitors were added and hypotonic treatment was performed for 1 h. Red blood cell membrane (RBCM) was obtained by centrifugation at 20,000 g. The RBCM was washed three times to remove soluble proteins and ultrasonicated for 5 min. The obtained RBCM was added to a liposome extruder and passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. Membrane vesicles were obtained by centrifugation at 20,000 g. The protein concentration was quantified using the BCA method and stored in aliquots at -80°C. (4) RNA loading: DMSNs were washed three times with nuclease-free water, and RNA and DMSNs were mixed at a mass ratio of RNA:DMSNs = 1:5-1:

20. The mixture was incubated at 4°C for 15 min, and centrifuged at 20,000 g for 10 min to remove unloaded RNA molecules to obtain DMSNs-RNA. (6) Coating the plasma membrane on the surface of dendritic mesoporous silica nanoparticles: The membrane vesicles and DMSNs-RNA were mixed in a mass ratio of 1:1-1:2 and evenly added to the liposome extruder. The mixture was ultrasonicated in an ice bath for 1 min and passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. The dendritic mesoporous silica biomimetic nanoparticles loaded with RNA molecules were obtained by centrifugation at 10,000 g. The mixture was resuspended in PBS and frozen at -80°C.

9. The method for preparing a dendritic mesoporous silicon biomimetic nanoparticle loaded with RNA molecules according to any one of claims 1 to 6, characterized in that: Prepare as follows: (1) Preparation of dendritic mesoporous silica nanoparticles: Accurately weigh 0.9 g of TEA, 180 mL of deionized water, and 120 mL of 25 wt. % CTAC aqueous solution, and stir them thoroughly, which is recorded as aqueous solution A. Dissolve 4 mL of TEOS in 16 mL of cyclohexane to obtain 20 mL of oil phase solution B. Add 60 mL of solution A to a 100 mL round-bottom flask and stir at 400 rpm for 30 min at 40 °C to allow CTAC to form lamellar micelles in water. Reduce the stirring speed to 200 rpm and slowly add 20 mL of solution B dropwise into solution A, paying attention to the formation of the interface between the aqueous phase and the oil phase. Increase the stirring speed to 400 rpm and maintain the two-phase system at 40 °C for 18 h to obtain dendritic mesoporous silica nanoparticles DMSNs. (2) Amino modification of dendritic mesoporous silica nanoparticles: APTES was dissolved in 250 μL of anhydrous ethanol, mixed and dissolved, and APTES was added to the lower layer solution A to ensure that the APTES / TEOS molar ratio was 0.5 / 10; amino-modified DMSNs were collected by centrifugation at 50,000 g. (3) Removal of surfactant CTAC: Prepare 0.12 M hydrochloric acid-ethanol solution, wash DMSNs repeatedly with concentrated hydrochloric acid-ethanol solution 5 times, and resuspend in ethanol for storage; (4) Cytoplasmic membrane extraction: ID8 ovarian tumor cell line was cultured in DMEM complete medium at 37°C and 5% CO2 until the logarithmic phase. The cells were collected and placed in hypotonic TM buffer. Protease inhibitors were added and hypotonic treatment was performed overnight. After mechanical disruption of the cells, 1M sucrose solution was added to a final concentration of 0.25 M. Tumor cell membranes were obtained by centrifugation at 2000 g. The probe was sonicated for 5 min, and the obtained cell membranes were added to a liposome extruder and passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. Membrane vesicles were obtained by centrifugation at 20,000 g. The protein concentration was quantified using the BCA method and the cells were aliquoted and stored at -80°C. (5) RNA loading: DMSNs were washed three times with nuclease-free water, RNA and DMSNs were mixed at a mass ratio of 1:5-1:20, incubated at 4°C for 15 min, and centrifuged at 20,000 g for 10 min to remove unloaded RNA molecules to obtain DMSNs-RNA; (6) Coating the plasma membrane on the surface of dendritic mesoporous silica nanoparticles: The membrane vesicles and DMSNs-RNA were mixed in a mass ratio of 1:1-1:2 and added to the liposome extruder, and then passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in turn; centrifuged at 10,000 g to obtain dendritic mesoporous silica biomimetic nanoparticles loaded with RNA molecules, which were resuspended in PBS and frozen at -80°C.

10. The method for preparing a dendritic mesoporous silicon biomimetic nanoparticle loaded with RNA molecules according to any one of claims 1 to 6, characterized in that: Prepare as follows: (1) Preparation of dendritic mesoporous silica nanoparticles: Accurately weigh 0.9 g of TEA, 180 mL of deionized water, and 120 mL of 25 wt. % CTAC aqueous solution, and stir thoroughly to mix, which is recorded as aqueous solution A; 8 mL of TEOS was dissolved in 12 mL of cyclohexane to prepare 20 mL of oil phase solution B; 60 mL of solution A was added to a 100 mL round-bottom flask and stirred at 400 rpm for 30 min at 40 °C to allow CTAC to form lamellar micelles in water; the stirring speed was reduced to 200 rpm, and 20 mL of solution B was slowly added dropwise to solution A, while paying attention to the formation of the interface between the aqueous phase and the oil phase; the stirring speed was increased to 400 rpm and the two-phase system was maintained at 40 °C for 18 h to obtain dendritic mesoporous silica nanoparticles DMSNs; (2) Amino modification of dendritic mesoporous silica nanoparticles: APTES was dissolved in 250 μL of anhydrous ethanol, mixed and dissolved, and APTES was added to the lower solution to ensure that the APTES / TEOS molar ratio was 0.5 / 10. After the reaction was continued for 4 h, the amino-modified DMSNs were collected by centrifugation at 50,000 g. (3) Removal of surfactant CTAC: Prepare a 0.12 M hydrochloric acid-ethanol solution, wash the DMSNs repeatedly 5 times with the concentrated hydrochloric acid-ethanol solution, and resuspend the obtained nanoparticles in ethanol for storage; (4) Cytoplasmic membrane extraction: RAW264.7 macrophage cell line was cultured in DMEM complete culture medium at 37°C, 5% CO2 to the logarithmic phase. The cells were collected and placed in hypotonic TM buffer. Protease inhibitors were added and hypotonic treatment was performed overnight. After mechanical disruption of the cells, 1M sucrose solution was added to a final concentration of 0.25 M. The macrophage membrane was obtained by centrifugation at 2000 g. The macrophage membrane was washed three times and probe sonicated for 5 min. The obtained cell membrane was added to a liposome extruder and passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. Membrane vesicles were obtained by centrifugation at 20,000 g. The protein concentration was quantified using the BCA method and the cells were aliquoted and stored at -80°C. (5) RNA loading: DMSNs were washed three times with nuclease-free water, and RNA and DMSNs were mixed at a mass ratio of 1:5-1:20, incubated at 4°C for 15 min, and centrifuged at 20,000 g for 10 min to remove unloaded RNA molecules to obtain RNA-loaded DMSNs, which were recorded as DMSNs-RNA. (6) Coating the plasma membrane on the surface of dendritic mesoporous silica nanoparticles: The membrane vesicles and DMSNs-RNA were mixed evenly at a mass ratio of 1:1-1:2 and added to the liposome extruder. The mixture was passed through 1000 nm, 400 nm and 200 nm polycarbonate nuclear pore membranes in sequence. The dendritic mesoporous silica biomimetic nanoparticles were obtained by centrifugation at 10,000 g. The obtained dendritic mesoporous silica biomimetic nanoparticles loaded with RNA molecules were resuspended in PBS for storage.

Images