Neutrophile granulocyte intercellular effect-mediated high-efficiency mononuclear phagocyte targeting siRNA (small interfering Ribonucleic Acid) nanoparticle as well as preparation method and application thereof

Abstract

The invention discloses siRNA (small interfering Ribonucleic Acid) nanoparticles capable of efficiently targeting mononuclear phagocytes and mediated by intercellular effect of neutrophils as well as a preparation method and application of the siRNA nanoparticles. The siRNA nanoparticles comprise mesoporous silica nanoparticles carrying siRNA, and functional cell membranes over-expressing ICAM-1 ligands are wrapped outside the mesoporous silica nanoparticles carrying siRNA. Key sentinel cells (mononuclear phagocytes, MPs) for targeted mitochondrial DNA release to treat inflammatory responses caused by mtDNA. The efficient delivery of MPs is realized by using a functionalized cell membrane of an overexpressed ICAM-1 ligand to wrap nano-particles and by virtue of infiltration and rapid apoptosis of a large number of neutrophils in an inflammatory microenvironment and a process (cell burial effect) of specific phagocytosis of the neutrophils by the MPs. Compared with a traditional targeting scheme, the in-vitro targeting efficiency and the in-vivo targeting efficiency of the FCM-coated siNPs scheme are improved by 360% and 120% respectively. The siRNA is guided to target MPs, and the expression of STING is inhibited, so that the inflammatory reaction caused by mtDNA release is regulated and controlled, and an excellent anti-inflammatory curative effect is shown in an inflammatory model mouse.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to a highly efficient siRNA nanoparticle targeting mononuclear phagocytes mediated by neutrophil burial, its preparation method, and its application. Background Technology

[0002] Mitochondria play a crucial role in the body's immunity, especially their powerful innate immune agonist—mitochondrial DNA (mtDNA). The key regulatory role of mtDNA in innate immunity is increasingly recognized. Various cellular stress conditions trigger the release of mtDNA from mitochondria into the cytoplasm, activating inflammatory responses through pathways such as cGAS-STING. Evidence of this phenomenon has been found in various inflammatory diseases such as nephritis, pneumonia, and periodontitis, and some mechanisms have been elucidated in fibroblasts, monocytes, and epithelial cells. Mononuclear macrophages (MPs) are key cells for mtDNA release; therefore, there is an urgent need to develop efficient targeted delivery strategies to precisely regulate the mtDNA release-mediated inflammatory response.

[0003] Precision targeted therapy is the core of precision medicine, and cell-targeted regulation is receiving increasing attention in diseases such as inflammation and tumors. In recent decades, many types of targeted delivery systems have been developed, including inorganic particles, polymers, and liposomes. These particles achieve targeted delivery to specific sites or target cells through passive targeting, responsive local accumulation, or active strategies mediated by targeting molecules. However, these traditional nanomaterials face challenges such as insufficient target site enrichment and rapid clearance by the bloodstream. To address these issues, cell membrane coating technology is considered one of the most prominent methods for functionalizing nanoparticles (NPs). Cell membrane coating technology is a simple top-down approach. Because the membrane coating is structurally and functionally similar to the host cell, it has many advantages, including good biocompatibility, low immunogenicity, good stability, and a wide range of ligands. Therefore, the significant advantages and abundant cell membrane sources of functionalized cell membrane biomimetic nanodelivery platforms provide a solid foundation for their industrial production and implementation in personalized precision medicine. However, for specific cells, precise targeted delivery often faces challenges of low efficiency and low specificity.

[0004] In recent years, the theory of intercellular material flow has been applied to the development of new targeting strategies to improve in vivo targeted delivery capabilities. Neutrophils, due to their high tumor local infiltration, bone marrow homing, and ability to cross the blood-brain barrier, have been used as novel delivery vehicles, achieving promising results in targeted delivery to specific sites. For example, neutrophils can penetrate inflamed brain tumors; neutrophils carrying liposomes containing paclitaxel (PTX) can penetrate the brain and inhibit glioma recurrence in mice whose tumors have been surgically removed. Utilizing the bone marrow homing effect of senescent neutrophils under CXCR4 / CXCL12 signaling, drug-loaded polylactic acid-glycolic acid nanoparticles are taken up by neutrophils and then transported across the bone marrow-blood barrier. However, the short half-life of neutrophils (approximately 12 hours) and their rapid apoptosis and clearance by phagocytes through cytotoxicity are considered significant limitations. Furthermore, current cell-mediated delivery systems can only achieve delivery to specific tissues and organs; achieving precise and efficient targeted delivery to target cells using in vivo "mediator" cells remains to be achieved. Summary of the Invention

[0005] To address the aforementioned technical problems, the first objective of this invention is to provide a highly efficient siRNA nanoparticle that targets mononuclear phagocytes in a manner mediated by neutrophil burial; the second objective is to provide a method for its preparation; and the third objective is to provide its applications.

[0006] To achieve the above objectives, the present invention is implemented through the following technical solution: a highly efficient siRNA nanoparticle targeting mononuclear phagocytes mediated by neutrophil burial, characterized in that: it comprises mesoporous silica nanoparticles carrying siRNA, and a functionalized cell membrane overexpressing ICAM-1 ligand is wrapped around the mesoporous silica nanoparticles carrying siRNA.

[0007] In the above scheme: the siRNA is a siRNA that is specific to the STING protein.

[0008] In the above scheme: the functionalized cell membrane overexpressing ICAM-1 ligand comes from the ICAM-1 overexpressing NIH3T3 cell membrane.

[0009] This invention presents a neutrophil-mediated siRNA delivery system designed to target key sentinel cells (monocytes, MPs) that release mitochondrial DNA (mtDNA) to treat mtDNA-induced inflammatory responses. The invention utilizes functionalized cell membrane-encapsulated nanoparticles (FCM@siNPs) overexpressing the ICAM-1 ligand. By leveraging the infiltration, rapid apoptosis, and specific phagocytosis of MPs by a large number of neutrophils in the inflammatory microenvironment (cell burial indirectly targets monocytes via neutrophil burial), highly efficient delivery of MPs is achieved. Compared to traditional targeting methods, the FCM@siNPs approach improves targeting efficiency by 360% in vitro and 120% in vivo. By guiding siRNA to target MPs and inhibiting STING expression, the inflammatory response induced by mtDNA release is regulated, demonstrating excellent anti-inflammatory efficacy in an inflammatory mouse model. This invention addresses the problems of limited delivery efficiency and poor cell targeting specificity of nanomaterials by developing a nanodelivery platform that utilizes the neutrophil burial mechanism in vivo to amplify targeting efficiency. This provides a new strategy for targeted delivery systems and has broad potential for clinical applications.

[0010] A method for preparing highly efficient siRNA nanoparticles targeting mononuclear phagocytes mediated by neutrophil burial, characterized by the following steps:

[0011] (1) Synthesize siNPs composite nanoparticles by loading siRNA into mesoporous silica using the calcium silicate sealing method to obtain siNPs composite nanoparticles;

[0012] (2) Extraction of ICAM-1 overexpressing NIH3T3 cell membrane;

[0013] (3) The fibroblast membrane obtained in step (2) is mixed with the siNPs nanoparticles and extruded through a polycarbonate membrane to obtain siRNA nanoparticles that are highly efficient at targeting mononuclear phagocytes mediated by neutrophil burial.

[0014] In the above scheme: the specific operation of step (1) is as follows: prepare an enzyme-free CaCl2 aqueous solution, filter the impurities in the solution, mix the CaCl2 solution, siRNA and mesoporous silica nanoparticle solution, sonicate in the dark, centrifuge at low temperature, remove the supernatant, wash once with anhydrous ethanol and sterile water, centrifuge at low temperature, remove the supernatant to obtain siNPs composite nanoparticles.

[0015] In the above scheme: the specific operation of step (2) is as follows: construct the NIH3T3 cell line overexpressing ICAM-1 by lentivirus infection, expand it in a culture dish, collect it, wash it once with PBS, digest the cells with 0.02% EDTA cell dissociation solution at 37°C, centrifuge to collect the cells, remove the supernatant, resuspend the cells in PBS, and then use sonication to destroy the cell membrane, centrifuge, collect the supernatant to remove the cell nucleus and unbroken cells, centrifuge to precipitate cell membrane fragments, remove the supernatant, resuspend the precipitate, and obtain cell membrane fragments.

[0016] In the above scheme: the operation of step (3) is as follows: mix siNPs with cell membrane at a mass ratio of 3:1, and pass them through nanoscale polycarbonate porous membranes with pore sizes of 400nm and 200nm 21 times by physical extrusion, and centrifuge at low temperature to purify and obtain siRNA nanoparticles that are highly efficient in targeting mononuclear phagocytes mediated by neutrophil burial.

[0017] The application of a highly efficient siRNA nanoparticle targeting mononuclear phagocytes mediated by neutrophil burial in the treatment of mtDNA-induced inflammation. The siRNA nanoparticle, capable of targeting mononuclear phagocytes via ICAM-1 / LFA-1 ligand-receptor binding, specifically targets and binds to neutrophils and mononuclear phagocytes (MPs). The inflammation is periodontitis.

[0018] This invention indirectly targets mononuclear phagocytes (MPs) by leveraging neutrophil burial, improving delivery efficiency both in vitro and in vivo. It provides a highly efficient and specific targeted delivery strategy using neutrophils as the primary "transporter" in vivo. Utilizing the characteristics of a large infiltration of neutrophils in the inflammatory microenvironment, their powerful phagocytic capacity, and rapid apoptosis, the invention efficiently and specifically targets MPs through burial. Mesoporous silica is used as the siRNA carrier, and a functionalized cell membrane modified with overexpression of ICAM-1 ligand is employed to target neutrophils and MPs via ICAM-1 / LFA-1 ligand-receptor binding. This significantly inhibits mtDNA release-mediated inflammatory responses.

[0019] Beneficial effects

[0020] (1) The biocompatible, multi-specific, and targeted nanodelivery platform for mononuclear phagocytes constructed in this invention can directly target MPs; on the other hand, it can first target a large number of neutrophils, utilizing the mechanism of rapid apoptosis of neutrophils and phagocytosis by MPs, thereby achieving more and more specific targeted delivery of MPs and precisely regulating the inflammatory response. The delivery system of this invention can precisely guide siRNA to target MPs, inhibit the expression of STING, thereby regulating the inflammatory response triggered by mtDNA release and significantly improving periodontitis symptoms.

[0021] (2) Compared with the prior art, the present invention uses the ICAM-1 overexpressing NIH3T3 cell membrane as the delivery carrier of siNPs nanoparticles. Compared with the traditional chemically synthesized carriers, it has great advantages. It is simpler to obtain, less expensive, has a higher yield, and has a simpler composition. Moreover, the present invention uses the method of preparing cancer cell membrane biomimetic nanoparticles by membrane extrusion, which can realize the large-scale and low-cost preparation of biomimetic nanoparticle carriers.

[0022] (3) The method for preparing biomimetic nanoparticles in this invention can adjust the carrier size by changing the pore size of the polycarbonate membrane, and the prepared biomimetic nanoparticles have uniform particle size. The equipment is inexpensive and easy to operate.

[0023] (4) The siSTING-based siRNA delivery vector prepared in this invention contains siSTING loaded within biomimetic nanoparticles, which effectively protects siSTING from degradation by ribozymes. The application of cell membrane biomimetic technology in this invention enables the nanodelivery system to have good immune evasion and in vitro and in vivo targeting capabilities, ensuring that the particles remain in the nanoscale and maintain stability.

[0024] (5) The ligands modified by genetic engineering in this invention have natural in vitro and in vivo activities, and through ligand-receptor interaction, they have better specific binding ability than other target molecules.

[0025] (6) These obvious advantages and abundant cell membrane sources of the present invention provide a solid foundation for its industrial production and translation into personalized precision medicine. Attached Figure Description

[0026] Figure 1 To enable TEM observation of the morphology and elemental distribution of nanoparticles (MSN, siNPs, and FCM@siNPs).

[0027] Figure 2 CLSM images and semi-quantitative fluorescence statistics of fluorescently labeled nanoparticles targeting MPs and neutrophils are shown. In A, fluorescently labeled nanoparticles targeting MPs are shown, and in B, fluorescently labeled nanoparticles targeting neutrophils are shown. C and D are semi-quantitative fluorescence statistics of Dil and FAM in MPs and neutrophils, respectively.

[0028] Figure 3This diagram illustrates the cellular experiment of neutrophil-mediated FCM@siNPs delivery to PDM and the CLSM image of the same process. Figures A and B show the cellular experiment, confocal fluorescence observation, and quantitative statistical analysis of the neutrophil-mediated FCM@siNPs delivery to PDM, respectively. Figure C shows the CLSM image of the neutrophil-mediated FCM@siNPs delivery to PDM, including the Dil and FAM fluorescence channels.

[0029] Figure 4 This document describes the fluorescence intensity and quantitative statistics of nanoparticle phagocytosis analyzed by flow cytometry. A and B represent the fluorescence intensity and quantitative statistics of nanoparticle phagocytosis analyzed by flow cytometry, respectively.

[0030] Figure 5 This diagram illustrates the in vivo transport of FCM@siNPs by neutrophils and the fluorescence intensity and quantitative statistics of macrophage phagocytosis of nanoparticles. A shows the in vivo transport of FCM@siNPs by neutrophils; B and C are the fluorescence intensity and quantitative statistics of macrophage phagocytosis of nanoparticles observed by CLSM, respectively.

[0031] Figure 6 Figure A shows the phagocytic activity of macrophages on nanoparticles after neutrophil depletion, as assessed by flow cytometry, and Figure B shows the quantitative analysis.

[0032] Figure 7 These are representative 3D reconstructed images of alveolar bone obtained from micro-CT scans before and after LIP treatment in mice. Detailed Implementation

[0033] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0034] Example 1: Construction and characterization of engineered membrane-coated nanoparticles FCM@siNPs

[0035] First, prepare a 2M CaCl2 solution (enzyme-free water) and filter out impurities. Mix the CaCl2 solution (2M), siSTING (siRNA specific to STING protein) solution (5nM), and mesoporous SiO2 solution (5mg / mL) at a volume ratio of 5:1:1. All solutions are enzyme-free aqueous solutions. Sonicate in the dark for 15 min, then centrifuge at 4℃ for 5 min. Remove the supernatant, wash once with anhydrous ethanol and once with sterile water, centrifuge at low temperature for 5 min, and discard the supernatant to obtain SiO2-si-STING (siNPs).

[0036] NIH3T3 cell lines overexpressing ICAM-1 were constructed using lentiviral infection. After amplification in culture dishes, cells were collected, washed once with PBS, and digested with 0.02% EDTA cell dissociation buffer at 37°C. Cells were collected by centrifugation, and the supernatant was aspirated. Cells were resuspended in PBS and then sonicated (20% power, 2s on, 3s off, 40 cycles) to disrupt the cell membrane. The cells were centrifuged at 4°C for 10 min, and the supernatant was carefully collected into a new centrifuge tube to remove nuclei and undisrupted cells. The cells were then centrifuged at 4°C for 30 min to precipitate cell membrane fragments. The supernatant was removed, and the precipitate was resuspended to obtain cell membrane fragments.

[0037] After obtaining cell membranes and siNPs using the above method, the siNPs (with 1 mg / mL aqueous solution added) were mixed with the cell membrane at a mass ratio of 3:1. The mixture was then passed through nanoscale polycarbonate porous membranes with pore sizes of 400 nm and 200 nm 21 times by physical extrusion. After centrifugation at low temperature for 10 min, the mixture was purified to obtain cell membrane-encapsulated nanoparticles FCM@siNPs.

[0038] As described above, FCM@siNPs consist of mesoporous silica loaded with siRNA and engineered cell membrane modifications. Figure 1 Transmission electron microscopy (TEM) images showed that MSNs were spherical particles with typical ordered porous characteristics. After loading siRNA (siNPs), their structure and size did not change significantly, while a clear cell membrane structure was visible on the outer layer of the particles after encapsulation. EDS-mapping results revealed the elemental distribution of silicon (Si), oxygen (O), and carbon (C) in FCM@siNPs. Si originated from the interior of the MSN nanoparticles, O was distributed in the MSN, siRNA, and cell membrane, and C mainly originated from the cell membrane and siRNA, thus being most densely distributed in the outermost layer and less abundant in the central region. These elemental distribution images are consistent with the expected structure of membrane-encapsulated nanoparticles, confirming the successful preparation of FCM@siNPs.

[0039] Application example: "Neutral granule transporter" delivers FCM@siNPs in vitro and in vivo via endocytosis-mediated targeted delivery.

[0040] The ability of FCM@siNPs to target macrophages and neutrophils was evaluated: siRNA was fluorescently labeled with FAM (green), and the cell membrane was labeled with Dil (red). FCM@siNPs and CM@siNPs (prepared according to Example 1, except that the modified cell membrane did not overexpress ICAM-1) were incubated with peritoneal derived macrophages (PDM) at 37°C in a 5% CO2 incubator for 4 hours. The uptake of nanoparticles by PDM was assessed by the fluorescence intensity of intracellular Dil and FAM. Figure 2CLSM results and semi-quantitative statistics showed that the fluorescence intensities of Dil and FAM in the FCM@siNPs group were significantly higher than those in the CM@siNPs group, approximately three times higher. Similar results were observed in polymorphonuclear leukocytes (PMNs). These results indicate that, under the same conditions, both PDM and PMNs exhibit significantly stronger phagocytic capacity for FCM@siNPs than CM@siNPs, suggesting that nanoparticles with the ICAM-1 ligand can better target macrophages and neutrophils.

[0041] Evaluation of FCM@siNPs delivery efficiency mediated by the in vitro "neutrophil transporter": siRNA was labeled with FAM fluorescent markers (green), and the cell membrane was labeled with Dil markers (red). FCM@siNPs and CM@siNPs were first taken up by PMNs. PMNs that had taken up the nanoparticles were induced to apoptosis and then co-incubated with PDM for 30 min. Control group nanoparticles followed the same treatment trajectory but did not undergo PMN uptake. Results are as follows... Figure 3 As shown, within 30 minutes, the fluorescence intensity of the FCM@siNPs+PMN group was significantly higher than that of other groups, including the CM@siNPs+PMN group, indicating that the ICAM-1 ligand plays an important role in the targeted delivery process. Importantly, the fluorescence intensity of the FCM@siNPs+PMN group was much higher than that of the FCM@siNPs group, and quantitative fluorescence statistics support this result. This indicates that PDM can rapidly engulf nanoparticles in apoptotic PMNs, while its engulfment ability for FCM@siNPs alone is weak. In addition, flow cytometry analysis yielded the same results. Figure 4 Statistical analysis showed that PMN mediated a 360% increase in the number of FCM@siNPs delivered to PDM. This result strongly supports the transport mechanism role of neutrophils in the phagocytosis of nanoparticles by macrophages.

[0042] Evaluation of in vivo neutrophil transport mechanism-mediated FCM@siNPs delivery efficiency: The effectiveness of this mechanism in vivo is crucial, but the in vivo microenvironment is more complex, and both nanoparticles and target cells face significantly increased interference. Therefore, we further validated the in vivo neutrophil transport mechanism-mediated targeted delivery of FCM@siNPs. First, peritoneal PMNs were extracted, and apoptotic PMNs that had engulfed nanoparticles were induced in vitro. These PMNs were then injected back into C57 mice that had induced PDM accumulation in the peritoneal cavity. Two hours later, PDM was extracted from the mouse peritoneal cavity, and the engulfment of nanoparticles by PDM was observed in vitro using fluorescence. Figure 5CLSM results showed that, compared to direct injection of FCM@siNPs into mice, FCM@siNPs pre-phagocytosed by PMNs were delivered to PDMs more rapidly and efficiently, with significantly stronger red fluorescence in PMN-mediated PDMs. Quantitative fluorescence analysis showed an improvement of approximately 120% compared to the control group. To further demonstrate the enhancing role of neutrophils in the in vivo delivery of FCM@siNPs, we constructed an anti-Ly6G depleted neutrophil mouse model and induced intraperitoneal neutrophil recruitment in normal mice and neutrophil-depleted mice. FCM@siNP nanoparticles were then injected into the peritoneal cavities of both groups of mice, followed by further induction of PDM recruitment. Finally, the PDMs in the peritoneal cavity were collected. Figure 6 Flow cytometry results showed that the fluorescence intensity in PDM of neutrophil-depleted mice was significantly reduced compared to normal mice, and quantitative statistical analysis showed that the delivery capacity of FCM@siNPs in neutrophil-depleted mice was inhibited by about half. These results strongly demonstrate that FCM@siNPs can more efficiently target macrophages in vivo by riding on neutrophil transport vehicles, laying the foundation for more efficient in vitro and in vivo treatment.

[0043] Application 3: The therapeutic effect of FCM@siNPs nanoparticles on periodontitis

[0044] Six-week-old male C57BL / 6 mice were anesthetized by isoflurane inhalation. A 5.0mm suture was wrapped around both second molars with microforceps and tied at the palatal side to establish an acute mouse model of periodontitis. The ligated periodontitis (LIP) mice were divided into four groups of eight mice each. For one week after modeling, mice were injected daily with saline, FCM@NPs, CM@siNPs, and FCM@siNPs (siRNA dose of 25 nmol / kg) into the periodontal tissues, while the control group received saline. Mouse weight was recorded daily. After one week, mice were sacrificed using CO2 inhalation, and periodontal and organ tissues were extracted for subsequent experiments. Changes in alveolar bone resorption were analyzed using Micro-CT scans. Figure 7 3D reconstructed images showed that LIP mice exhibited significantly greater alveolar bone resorption compared to control mice. In contrast, the alveolar bone height in the FCM@siNPs treatment group was significantly increased compared to the periodontitis group.

[0045] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A siRNA nanoparticle that efficiently targets mononuclear phagocytes mediated by neutrophil efferocytosis, characterized in that: The invention comprises mesoporous silica nanoparticles carrying siRNA, wherein a functional cell membrane over-expressing ICAM-1 ligand is wrapped outside the mesoporous silica nanoparticles carrying siRNA.

2. The siRNA nanoparticles mediated by neutrophil efferocytosis and highly efficient targeting to mononuclear phagocytes according to claim 1, characterized in that: The siRNA is specific to the STING protein.

3. The siRNA nanoparticles mediated by neutrophil efferocytosis and highly efficient targeting to mononuclear phagocytes according to claim 1 or 2, characterized in that: The functionalized cell membrane overexpressing ICAM-1 ligand comes from ICAM-1 overexpressing NIH3T3 cell membrane.

4. A method for preparing siRNA nanoparticles that efficiently target mononuclear phagocytes mediated by neutrophil efferocytosis according to any one of claims 1 to 3, characterized in that: Prepare according to the following steps: (1) synthesizing siNPs composite nanoparticles, and loading siRNA into mesoporous silica using a calcium silicate sealing method to obtain siNPs composite nanoparticles; (2) Extraction of ICAM-1 overexpressing NIH3T3 cell membranes; (3) Mixing the fibroblast membrane obtained in step (2) with the siNPs nanoparticles and extruding them through a polycarbonate membrane to obtain siRNA nanoparticles that are highly efficient in targeting mononuclear phagocytes and mediated by neutrophil efferocytosis.

5. The method for preparing the siRNA nanoparticles that efficiently target mononuclear phagocytes mediated by neutrophil efferocytosis according to claim 4, characterized in that: The specific operation of step (1) is as follows: preparing a CaCl2 enzyme-free aqueous solution, filtering impurities in the solution, mixing the CaCl2 solution, siRNA and mesoporous silica nanoparticle solution, performing light-protected ultrasound, performing low-temperature centrifugation, removing the supernatant solution, washing once with anhydrous ethanol and once with sterile water, performing low-temperature centrifugation, and removing the supernatant to obtain siNPs composite nanoparticles.

6. The method for preparing the siRNA nanoparticles that efficiently target mononuclear phagocytes mediated by neutrophil efferocytosis according to claim 5, characterized in that: The specific operation of step (2) is as follows: constructing an NIH3T3 cell line overexpressing ICAM-1 by lentiviral infection, amplifying it in a culture dish, collecting and washing it with PBS, digesting the cells with 0.02% EDTA cell dissociation solution at 37°C, collecting the cells by centrifugation, removing the supernatant, resuspending the cells in PBS, destroying the cell membrane by ultrasonic disruption, centrifuging, collecting the supernatant to remove the cell nucleus and unbroken cells, centrifuging to precipitate cell membrane fragments, removing the supernatant, resuspending the precipitate, and obtaining cell membrane fragments.

7. The method for preparing the siRNA nanoparticles that efficiently target mononuclear phagocytes mediated by neutrophil efferocytosis according to claim 6, characterized in that: The operation of step (3) is as follows: siNPs and cell membranes are mixed at a mass ratio of 3:1, and the mixture is passed through a nanoscale polycarbonate porous membrane with a pore size of 400 nm and 200 nm 21 times by physical extrusion, and centrifuged at low temperature to purify siRNA nanoparticles that are highly efficient in targeting mononuclear phagocytes and mediated by neutrophil efferocytosis.

8. A use of the siRNA nanoparticles mediated by neutrophil efferocytosis and efficiently targeting mononuclear phagocytes as described in any one of claims 1 to 3 as a drug for treating inflammation caused by mtDNA, wherein the siRNA nanoparticles mediated by neutrophil efferocytosis and efficiently targeting mononuclear phagocytes can target and bind to neutrophils and MPs through ICAM-1 / LFA-1 ligand-receptor binding.

9. The use according to claim 8, characterized in that: The inflammation is periodontitis.

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