CN114177309A - Self-oxygen-supply targeted bionic nuclear membrane structure nano-composite and preparation method and application thereof - Google Patents

Self-oxygen-supply targeted bionic nuclear membrane structure nano-composite and preparation method and application thereof Download PDF

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CN114177309A
CN114177309A CN202111293490.5A CN202111293490A CN114177309A CN 114177309 A CN114177309 A CN 114177309A CN 202111293490 A CN202111293490 A CN 202111293490A CN 114177309 A CN114177309 A CN 114177309A
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icg
liposome
bionic
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loaded
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伍满香
李强
陈天翔
吴爱国
王联芙
李建斌
史燕巧
房倩兰
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People's Hospital Affiliated To Ningbo University
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Abstract

The invention provides a self-oxygen supply targeting type bionic nuclear membrane structure nano compound and a preparation method and application thereof, wherein the nano compound comprises a bionic lipid membrane and hollow MnO2The nuclear membrane structure of (a); the bionic lipid membrane is a phospholipid bimolecular membrane with the surface containing tumor cell membrane protein modification; the hollow MnO2Is used for loading tumor therapeutic drugs, and the drugs are at least one of chemotherapeutic drugs and immunotherapy drugs; one of the purposes of the invention is to provide a self-oxygen-supply targeting type bionic nuclear membrane structure nano-composite which can be used for magnetic resonance imaging, fluorescence imaging, tumor chemotherapy and optical treatment; the second purpose of the invention is to provide a preparation method of the self-oxygen-supply targeting type bionic nuclear membrane structure nano-composite.

Description

Self-oxygen-supply targeted bionic nuclear membrane structure nano-composite and preparation method and application thereof
Technical Field
The invention relates to the technical field of drug carriers, in particular to a self-oxygen-supply targeting type bionic nuclear membrane structure nano-composite and a preparation method and application thereof.
Background
The traditional nano-medicine targeting strategy is divided into two steps: firstly, modifying the surface of the nano-particles with polyethylene glycol for escaping from the removal of a reticuloendothelial system; ligand modification is then performed to obtain targeting ability. However, recent studies show that the polyethylene glycol-modified nano-drug is rapidly cleared by the liver after continuous administration, and the ligand modification technology is complicated and has the problems of insufficient modification range, so that the actual in vivo targeting effect is often unsatisfactory.
The tumor hypoxia microenvironment is closely related to the occurrence and development of tumors, and is a great challenge in tumor treatment. Tumor hypoxia can cause various treatment resistances, and the curative effect of tumor treatment is greatly limited, so that the search of a proper strategy for improving the tumor hypoxia is very important for improving the curative effect of the tumor treatment. At present, the strategies for improving tumor hypoxia mainly comprise hypoxia improvement based on tumor blood vessel normalization, hypoxia improvement based on artificial oxygen supply carriers and hypoxia improvement based on tumor in-situ oxygen generation.
Disclosure of Invention
The invention aims to provide a self-oxygen-supply targeting type bionic nuclear membrane structure nano-composite which can be used for magnetic resonance imaging, fluorescence imaging, tumor chemotherapy and optical treatment.
In order to solve the above-mentioned object, the present invention provides a self-oxygen-supplying targeting type biomimetic nuclear membrane structure nanocomposite, which comprises a biomimetic lipid membrane and a hollow nuclear membrane structure of MnO 2; the bionic lipid membrane is a phospholipid bimolecular membrane with the surface containing tumor cell membrane protein modification; the hollow MnO2 is used for loading a tumor therapeutic drug, and the drug is at least one of a chemotherapeutic drug and an immunotherapy drug.
Optionally, the biomimetic lipid membrane contains a photosensitizer therein, wherein the photosensitizer is at least one of chlorin e6(Ce6), ICG-ODA, ICG-NH2, ICG-COOH, ICG-NHS, ICG-MAL, ICG-SH, ICG-N3, ICG-ALK, ICG-Biotin, IR780 and IR783 or IR 808.
Optionally, the hollow MnO2The mass percentage is 30-80%. Further optionally, the hollow MnO2The mass percentage is selected from 80%, 75%, 70%, 65%, 60%, 55%, 50%, 30%, 35%, 40% and 45%.
Optionally, the hollow MnO2The specific surface area of (A) is 60 to 200m2The average pore diameter is 1.5-10 nm, and the mesoporous surface is disordered.
Further optionally, the hollow MnO2Is selected from 200m in specific surface area2/g、180m2/g、160m2/g、140m2/g、60m2/g、80m2/g、100m2/g、120m2/g。
Optionally, the hollow MnO2The particle size is 50-150 nm, and the aperture is 2-10 nm; it has good drug loading capacity.
Optionally, the phospholipid bilayer membrane comprises lecithin, hydrogenated soya lecithin, hydrogenated egg yolk phospholipids, dipalmitoyl phosphatidylethanol, dipalmitoyl phosphatidylcholine, dioleoyl phosphatidylethanolamine, polyethylene glycol-distearoyl phosphatidylethanolamine, 1, 2-dipalmitoyl-sn-glycero-3-phosphatidic acid glycero-sodium salt, 1, 2-distearoyl-sn-glycero-3-phosphatidylcholine, 1, 2-dipalmitoyl-sn-glycero-3-phosphatidic acid-sodium salt, and 1, 2-dipalmitoyl-sn-glycero-3-phosphatidylcholine.
Optionally, the cancer comprises breast cancer, thyroid cancer, pancreatic cancer, lung cancer, kidney cancer, liver cancer, and melanoma.
Optionally, the photosensitizer is an indole cyanine dye comprising a mixture of two or more drugs selected from liposoluble derivatives of ICG such as ICG-ODA, ICG-NH2, ICG-COOH, ICG-NHS, ICG-MAL, ICG-SH, ICG-N3, ICG-ALK, ICG-Biotin, IR780, IR783, IR808, or chlorin e 6.
Optionally, the chemotherapeutic is selected from the group consisting of anthracycline anticancer drugs, paclitaxel anticancer drugs, vinca alkaloid anticancer drugs, platinum anticancer drugs, and at least one of 5-fluorouracil, methotrexate, cyclophosphamide, temozolomide, and pemetrexed.
Further optionally, the anthracycline anticancer drug is doxorubicin or epirubicin; the taxol anticancer drug is taxol or docetaxel; the vinca alkaloid anticancer drug is vinblastine, vincristine, vindesine or vinorelbine; the platinum anticancer drug is oxaliplatin or nedaplatin.
Optionally, the mass percentage of the photosensitizer is 2-5%; the mass percentage of the antitumor drug is 8-25%.
Optionally, the mass percentage of the ICG is 2-5%, and the mass percentage of the DOX is 8% -25%.
Optionally, the overall size of the self-oxygen-supply targeting type bionic nuclear membrane structure nano-composite is 100-400 nm.
Further optionally, the upper limit of the particle size range of the self-oxygen-supplying targeting type bionic nuclear membrane structure nano-composite is selected from 400nm, 350nm, 300nm, 250nm, 200nm, 100nm, 120nm and 150 nm.
Optionally, the self-oxygen-supplying targeting type bionic nuclear membrane structure nano-composite is dispersed in deionized water, PBS, physiological saline and other solutions.
Optionally, the self-oxygen-supplying targeting type biomimetic nuclear membrane structure nano-composite has a particle size change of less than 95% within 7 days, a particle size change of less than 92% within 14 days, a particle size change of less than 90% within 21 days, a particle size change of less than 88% within 28 days, a particle size change of less than 85% within 35 days, a particle size change of less than 80% within 42 days, a particle size change of less than 75% within 49 days, and a particle size change of less than 70% within 56 days.
Optionally, the hollow MnO2The material is prepared by any one of a template etching method, a coprecipitation method, a precipitation method, a hydrothermal reaction, a solvothermal reaction and an ultrasonic synthesis method.
Optionally, when the hollow MnO is prepared by the template etching method2Wherein the etching agent is selected from sodium carbonate, hydrofluoric acid, ammonium bifluoride, ammonium fluoride, tetrafluoromethane, buffered oxide etching solution (BOE), CHF3, CH2F2、CHF3/CH2F2、C4F8、C5F8、C4F8/C5F8Any one of them.
Optionally, the hollow MnO2The centrifugal speed of the centrifugal washing is selected from 4000 to 14000 g.
Further optionally, the centrifugation speed is selected from 14000g, 13000g, 12000g, 11000g, 10000g, 9000g, 4000g, 5000g, 6000g, 7000g, 8000 g.
Optionally, the nuclear membrane complex is prepared using a manual liposome extruder or an automatic liposome extruder.
Further optionally, the extrusion speed of the manual liposome extruder is 2-10 mL/s, and the extrusion speed is selected from 10mL/s, 9mL/s, 8mL/s, 7mL/s, 6mL/s, 2mL/s, 3mL/s, 4mL/s, and 5 mL/s. The extrusion force of the automatic liposome extruder is 2000-6000 psi.
Further optionally, the extrusion speed may have an upper limit selected from 5000psi, 4500psi, 4000psi, 3500psi, 3000psi and a lower limit selected from 1000psi, 1500psi, 2000psi, 2500 psi.
Optionally, the temperature control range of the extrusion of the nuclear membrane compound is 4-45 ℃. Further optionally, the extrusion temperature is selected from 45 deg.C, 42 deg.C, 40 deg.C, 37 deg.C, 35 deg.C, 32 deg.C, 30 deg.C, 27 deg.C, 25 deg.C, 22 deg.C, 20 deg.C, 5 deg.C, 7 deg.C, 10 deg.C, 12 deg.C, 15 deg.C, 17 deg.C.
The second purpose of the present invention is to provide a method for preparing the self-oxygen-supplying targeting type bionic nuclear membrane structure nano-composite, which comprises the following steps:
s1, preparing the photosensitizer-carrying bionic liposome with targeting performance: synthesizing a photosensitizer-carrying liposome modified by tumor cell membrane protein and membrane protein of tumor cells by a co-extrusion method;
s2 preparation of drug-loaded hollow MnO2: with SiO2Preparing hollow MnO for the template2(ii) a Co-stirring the obtained hollow manganese dioxide and the hydrophobic therapeutic drug to realize the loading of the hydrophobic drug in the hollow core;
s3, preparing bionic liposome/drug-loaded hollow MnO2Complex: mixing the bionic liposome prepared in the step S1 with the medicine-carrying hollow MnO in the step S22The finished product is prepared by co-extrusion of polycarbonate film, repeated extrusion, centrifugation, freeze drying.
Optionally, the biomimetic liposome hydration solution in the step S1 is selected from any one of deionized water, PBS buffer, Tris-HCl buffer, Tris buffer, physiological saline, 5% sucrose solution, and 5% glucose solution.
Optionally, the synthesized liposome carrying the photosensitizer is prepared by a thin film hydration method, a thin film ultrasonic dispersion method, a reverse evaporation method or a microemulsion ultrasonic dispersion method.
Optionally, the number of times of pressing in the co-pressing method in step S1 is between 20 and 60 times.
Further optionally, the number of squeezes is optionally from 60, 55, 50, 45, 40, 20, 25, 30, 35.
Alternatively, the synthetic photosensitizer-loaded liposome in step S1 is prepared by a thin film ultrasonic dispersion method, which includes the steps of: dissolving a phospholipid bimolecular membrane, DSPE-PEG2000, cholesterol and a photosensitizer with a molar ratio of 100:20:4:10 in a chloroform solution, fully mixing, performing rotary evaporation at 40 ℃ to form a membrane, and vacuumizing for 3 hours after the membrane is formed to completely volatilize an organic solvent; and adding PBS into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain the liposome carrying the photosensitizer.
Further optionally, the drug-loaded hollow MnO in step S22Is prepared by the following steps: preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4After continuing stirring, MnO was obtained2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano composite is placed in an etching solution to etch SiO2The inner core is sequentially subjected to centrifugation, washing and drying to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with the drug, and sequentially carrying out physical stirring, centrifugation, separation and freeze drying at normal temperature to obtain drug-loaded hollow MnO2。
Further optionally, the monodisperse SiO is prepared in the step S22When nano-microsphere, the SiO2The stirring speed of the preparation is 100-700 rpm, the reaction time is 12-36 h, and the reaction temperature is 25-60 ℃.
Further optionally, the ultrasonic condition in the step S2 includes that the ultrasonic power is 20-50W/cm2And the ultrasonic time is controlled to be 0.5-6 h.
Further optionally, a hollow MnO is obtained in the step S22In the process of (3), the drying method is any one of heating drying, heating vacuum drying and freezing vacuum drying.When vacuum drying is adopted, the heating temperature is selected from 60-300 ℃; the vacuum time is selected from 3 h-72 h; the freezing temperature is selected from-70 to-80 ℃.
Further optionally, in step S2, in preparing drug-loaded hollow MnO2In the process, the speed of physical stirring is 100-800 rpm, and the stirring time is selected from 12-48 h.
Further optionally, in step S2, drug-loaded hollow MnO is prepared2In the process, the centrifugation speed of the final product is selected from 4000 to 20000rpm, and the centrifugation temperature is selected from 4 to 25 ℃.
The third purpose of the invention is to provide an application of the self-oxygen-supplying targeted bionic nuclear membrane structure nano-composite in preparing a medicament for treating cancer, wherein the composite is used for precise targeting and combined synergistic treatment of various tumors in preparing the medicament for treating cancer.
Optionally, the cancer includes breast cancer, thyroid cancer, esophageal cancer, pancreatic cancer, lung cancer, kidney cancer, liver cancer, melanoma, and the like.
Optionally, the self-oxygen-supply targeting type bionic nuclear membrane structure nano-composite has T1 weighted MRI imaging performance, and the composite can be used for precise diagnosis and boundary identification of cancer in preparation of a medicament for treating cancer by virtue of the MRI imaging performance.
Optionally, the self-oxygen-supplying targeting type bionic nuclear membrane structure nano-composite has a fluorescence imaging performance, and can be used for accurate diagnosis and boundary identification of cancer in preparation of a medicament for treating cancer by virtue of the fluorescence imaging performance.
Optionally, the self-oxygen-supplying targeting type bionic nuclear membrane structure nano composite has an absorption peak at a wavelength of 808nm, can generate ROS and heat energy under irradiation, has good photo-thermal and photodynamic properties, and is used for photo-thermal and photodynamic treatment of tumors in preparation of drugs for treating cancers.
Optionally, the self-oxygen supply targeting type bionic nuclear membrane structure nano-composite has homologous targeting type energy.
Optionally, the self-oxygen-supply targeting type bionic nuclear membrane structure nano-composite has tumor microenvironment responsiveness, and can generate oxygen in situ at a tumor part for improving tumor hypoxia.
Optionally, the self-oxygenation targeting type bionic nuclear membrane structure nano compound carries chemotherapeutic drugs, and the compound is used for tumor chemotherapy in preparation of drugs for treating cancers.
Optionally, the self-oxygen-supplying targeting type bionic nuclear membrane structure nano-composite has good biological safety, and no obvious hemolytic reaction is seen.
The beneficial effects that this application can produce include:
1) according to the invention, the cell membrane is used as a carrier, so that the cell membrane has good biocompatibility, and long circulation and targeted delivery of the nanoparticle in the core in vivo can be promoted without considering the characteristics of the core nano material.
2) The self-oxygen-supply targeting type bionic nuclear membrane structure nano composite provided by the invention is MnO2As core, has pH and H2O2And glutathione responsiveness, can generate O in situ in tumors2And Mn2+So that the nano-composite has the responsive oxygen generation and MRI imaging performances, thereby realizing accurate diagnosis and enhancing the effect of combined treatment.
3) The raw materials of the invention are economical and easily available, the preparation method has mild conditions, low requirements on equipment, operation, environment and the like, is easy to implement, has good stability and reproducibility, and has the prospect of expanded production.
4) The cell membrane bionic technology is a simple top-down method, specifically targets homologous tumor cells by using the difference of protein expression on the surface of the tumor cell membrane, realizes accurate targeting of the medicine, and has higher biological safety. MnO2Is an inorganic material with tumor microenvironment responsiveness, and has PH and H2O2And glutathione responsiveness, can produce O2And Mn2 +The compound can improve local hypoxia of the tumor and enhance the curative effect of tumor treatment, and can also be used for MR (magnetic resonance) contrast imaging to monitor the growth of the tumor and the distribution and metabolism of the drug in the body.
Drawings
FIG. 1 is a schematic diagram of the composition of a self-oxygen-supplying targeted biomimetic nuclear membrane structured nanocomposite obtained in example 1;
FIG. 2 is a TEM image of the self-oxygen-supplying targeted biomimetic nuclear membrane structure nano-composite obtained in example 1;
FIG. 3 is a diagram showing the in vitro oxygen content production of the self-oxygen-supplying targeted biomimetic nuclear membrane structured nanocomposite in example 1;
FIG. 4 is a diagram showing the uptake of the self-oxygenating targeting biomimetic nuclear membrane structure nanocomposite in 4T1 tumor cells in example 1;
FIG. 5 is in vitro magnetic resonance imaging of the self-oxygenating targeting type biomimetic nuclear membrane structured nanocomposites in example 1;
FIG. 6 is a graph of in vitro photothermal performance of the self-oxygenating targeted biomimetic nuclear membrane structured nanocomposite in example 1;
FIG. 7 is a singlet oxygen detection diagram of the self-oxygen-supplying targeting biomimetic nuclear membrane structured nanocomposite in example 1;
FIG. 8 is a fluorescent image of a small animal of the self-oxygenating targeting biomimetic nuclear membrane structured nanocomposite obtained in example 1.
Description of reference numerals:
1. doxorubicin; 2. hollow manganese dioxide; 3. green indole; 4. 4T1 tumor cell membrane protein; 5. phospholipid bilayer membranes.
Detailed Description
For a more clear understanding of the technical features, objects and advantages of the present invention, reference is now made to the following detailed description of the embodiments of the present invention taken in conjunction with the accompanying drawings, which are included to illustrate and not to limit the scope of the present invention. In the examples, the respective starting reagents are commercially available, and the protocols without specifying the specific conditions are conventional methods and conditions well known in the art, or according to the conditions suggested by the manufacturer. The instruments and test conditions used in the examples were as follows:
the morphology of the material is observed on a JEOL-2100 transmission electron microscope, and the test conditions are as follows: 200KV, 101 μ A.
The particle size distribution was tested on a dynamic light scattering particle size analyzer of Zetasizer Nano ZS type.
Surface protein marker detection SDS-PAGE was performed on a Bio-Rad electrophoresis apparatus.
Cytotoxicity test cytotoxicity assays of materials were performed using the MTT method on a UV-2102PC type UV-vis spectrophotometer.
Magnetic resonance imaging analysis material imaging studies were performed on a 3.0T magnetic resonance imager, test condition T1: TR 650ms, TE 200 ms.
Example 1
Synthesizing the self-oxygen-supply targeting type bionic nuclear membrane structure nano compound according to the following steps:
s1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation at 40 ℃ to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding PBS into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the ice bath condition with a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4Stirring for 12h to obtain MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3Heating at 60 deg.C overnight to etch SiO2Centrifuging 8000g of inner core, washing, and freeze-drying for 48h to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at 300rpm at normal temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome at a concentration of 1mg/mL and 500 mu L of DOX-loaded hollow manganese dioxide at a concentration of 1mg/mL through a 200mm polycarbonate membrane under ice bath conditions, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain a nanocomposite product, wherein the nanocomposite product is shown in figure 1.
Example 2
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation at 40 ℃ to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding 5% of sucrose into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the ice bath condition with a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, freezing, drying and storing.
S2 preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 1h and then stirring overnight to obtain MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3Heating at 60 deg.C overnight to etch SiO2Centrifuging 8000g of inner core, washing, and freeze-drying for 48h to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at 400rpm at normal temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of 2mg/mL ICG-loaded bionic liposome and 500 mu L of 1mg/mL DOX-loaded hollow manganese dioxide through a 200mm polycarbonate membrane under the ice bath condition, repeatedly extruding for 20 times, centrifuging, freezing and drying to obtain the nano composite product.
Example 3
The self-oxygen-supply targeting type bionic nuclear membrane structure nano compound is synthesized according to the following steps
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation at 40 ℃ to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding a Tris solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder with a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4Stirring is continued overnight to obtain MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3Heating at 60 deg.C for 24h to etch SiO2Inner core, centrifuging at 9000g, washing, freeze drying for 48h to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at 400rpm at normal temperature, centrifuging, and separating to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of 1mg/mL ICG-loaded bionic liposome and 500 mu L of 2mg/mL DOX-loaded hollow manganese dioxide through a 200mm polycarbonate membrane under the ice bath condition, repeatedly extruding for 20 times, centrifuging, freezing and drying to obtain the nano composite product.
Example 4
The self-oxygen-supply targeting type bionic nuclear membrane structure nano compound is synthesized according to the following steps
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation at 40 ℃ to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding a Tris solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder with a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4Stirring is continued overnight to obtain MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid and stirred for 3 hours at normal temperature to etch SiO2Inner core, 7000g centrifugation, washing, freeze drying for 48h to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at 400rpm at normal temperature, centrifuging, and separating to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of 1mg/mL ICG-loaded bionic liposome and 500 mu L of 2mg/mL DOX-loaded hollow manganese dioxide through a 200mm polycarbonate membrane under the ice bath condition, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano composite product.
Example 5
The self-oxygen-supply targeting type bionic nuclear membrane structure nano compound is synthesized according to the following steps
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation at 40 ℃ to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding a Tris solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted MDA-MB-468 cell membrane protein by a liposome extruder with a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4Stirring for 24h to obtain MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid and stirred for 3 hours at normal temperature to etch SiO2Inner core, 7000g centrifugation, washing, freeze drying for 48h to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at 400rpm at normal temperature, centrifuging, and separating to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of 1mg/mL ICG-loaded bionic liposome and 500 mu L of 2mg/mL DOX-loaded hollow manganese dioxide through a 200mm polycarbonate membrane under the ice bath condition, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano composite product.
Example 6
The self-oxygen-supply targeting type bionic nuclear membrane structure nano compound is synthesized according to the following steps
S1, preparing a Ce 6-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and Ce6 in a molar ratio of 100:20:4:10 in a chloroform solution, fully mixing, performing rotary evaporation at 40 ℃ to form a film, and vacuumizing for 3h to completely volatilize an organic solvent after the film is formed; adding a Tris solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; the obtained Ce 6-loaded liposome and the extracted 4T1 tumor cell membrane protein are co-extruded by a liposome extruder through a 400nm polycarbonate membrane to obtain the Ce 6-loaded bionic liposome, and the bionic liposome is centrifugally cleaned and stored at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4Stirring is continued overnight to obtain MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid and stirred for 2 hours at normal temperature to etch SiO2Inner core, 7000g centrifugation, washing, freeze drying for 48h to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at 400rpm at normal temperature, centrifuging, and separating to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of 1mg/mL Ce 6-loaded bionic liposome and 500 mu L of 2mg/mL DOX-loaded hollow manganese dioxide through a 200mm polycarbonate membrane under the ice bath condition, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano composite product.
Example 7
The self-oxygen-supply targeting type bionic nuclear membrane structure nano compound is synthesized according to the following steps
S1, preparing the IR 780-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and IR780 in chloroform solution at a molar ratio of 100:20:4:10, mixing, performing rotary evaporation at 40 ℃ to form a film, and vacuumizing for 3h to completely volatilize the organic solvent; adding a Tris solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained IR 780-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder with a 400nm polycarbonate membrane to obtain the IR 780-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4Stirring is continued overnight to obtain MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid and stirred for 1 hour at normal temperature to etch SiO2Inner core, 7000g centrifugation, washing, freeze drying for 48h to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at 400rpm at normal temperature, centrifuging, and separating to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of 1mg/mL IR 780-loaded bionic liposome and 500 mu L of 1mg/mL DOX-loaded hollow manganese dioxide through a 200mm polycarbonate membrane under the ice bath condition, repeatedly extruding for 30 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 8
The self-oxygen-supply targeting type bionic nuclear membrane structure nano compound is synthesized according to the following steps
S1, preparing the IR 780-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and IR780 in chloroform solution at a molar ratio of 100:20:4:10, mixing, performing rotary evaporation at 40 ℃ to form a film, and vacuumizing for 4h to completely volatilize the organic solvent; adding a PBS solution into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained IR 780-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder with a 400nm polycarbonate membrane to obtain the IR 780-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation by sol-gel methodMonodisperse SiO2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4Stirring is continued overnight to obtain MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid and stirred for 2 hours at normal temperature to etch SiO2Inner core, 7000g centrifugation, washing, freeze drying for 48h to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at 400rpm at normal temperature, centrifuging, and separating to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome at a concentration of 1mg/mL and 500 mu L of DOX-loaded hollow manganese dioxide at a concentration of 1mg/mL through a 200mm polycarbonate membrane under ice bath, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 9
Preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:5:10, fully mixing, performing rotary evaporation at 40 ℃ to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding PBS into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 55 ℃, and centrifugally cleaning by deionized water to obtain ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the ice bath condition with a 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4Stirring is continued overnight for 12h to obtain MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3Heating at 65 deg.C overnight to etch SiO2Inner core, 7000g centrifugation, washing, freeze drying for 48h to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at 400rpm at normal temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome at a concentration of 1mg/mL and 500 mu L of DOX-loaded hollow manganese dioxide at a concentration of 1mg/mL through a 200mm polycarbonate membrane under ice bath, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 10
Preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation at 40 ℃ to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding PBS into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted MCF-7 cell membrane protein by a liposome extruder under the ice bath condition with a 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4Stirring is continued overnight for 12h to obtain MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3Heating at 60 deg.C overnight to etch SiO2Centrifuging 8000g of inner core, washing, and freeze-drying for 48h to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at 300rpm at normal temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide of 1mg/mL through a polycarbonate film of 100mm, repeatedly extruding for 20 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 11
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding PBS into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3To etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide with the concentration of 2mg/mL through a 100mm polycarbonate membrane, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 12
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding PBS into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted AKR tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain ICG-loaded bionic liposome, centrifuging, cleaning, and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3To etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, taking 500 mu L of 1mg/mL ICG-loaded bionic liposome and 500 mu L of 1mg/mL DOX-loaded hollow manganese dioxide, co-extruding the bionic liposome and the DOX-loaded hollow manganese dioxide through a 100mm polycarbonate membrane for ice bath, repeatedly extruding for 20 times, centrifuging, freezing and drying to obtain the nano composite product.
Example 13
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding PBS into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3To etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 14
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:10, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding a 5% sucrose solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3To etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide of 1mg/mL through a polycarbonate film of 200mm, repeatedly extruding for 20 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 15
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:5:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding a 5% sucrose solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, and freeze-drying for storage.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring for 24h to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3To etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide with the concentration of 2mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 16
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:5:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 4 hours to completely volatilize an organic solvent after forming the film; adding a PBS solution into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a polycarbonate membrane of 400nm to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring for 24h to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3To etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide with the concentration of 2mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 17
S1, preparing a Ce 6-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and Ce6 in a molar ratio of 100:20:5:20 in a chloroform solution, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 5 hours to completely volatilize an organic solvent after the film is formed; adding a PBS solution into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; the obtained Ce 6-loaded liposome and the extracted 4T1 tumor cell membrane protein are co-extruded by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the Ce 6-loaded bionic liposome, and the bionic liposome is centrifugally cleaned and stored at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring for 24h to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nanocomposite was placed in hydrofluoric acid to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of the Ce 6-loaded bionic liposome with 500 mu L of the manganese oxide with the concentration of 1mg/mL through a polycarbonate film with the thickness of 200mm, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano composite product.
Example 18
S1, preparing the IR 780-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and IR780 in a chloroform solution at a molar ratio of 100:20:5:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 5 hours to completely volatilize an organic solvent after forming the film; adding a PBS solution into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain the IR 780-loaded liposome; the obtained IR 780-loaded liposome and the extracted 4T1 tumor cell membrane protein are co-extruded by a liposome extruder under the ice bath condition by a polycarbonate membrane with the thickness of 400nm to obtain the IR 780-loaded bionic liposome, and the bionic liposome is centrifugally cleaned and stored at the temperature of 4 ℃.
S2 preparation by sol-gel methodPreparation of monodisperse SiO2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring for 24h to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nanocomposite was placed in hydrofluoric acid to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of the IR 780-loaded bionic liposome with 500 mu L of manganese oxide with the concentration of 2mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 19
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding a 5% glucose solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, and freeze-drying for storage.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid and stirred at normal temperature to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 20
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 4 hours to completely volatilize an organic solvent after the film is formed; adding a PBS solution into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, and freeze-drying for storage.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid and stirred at normal temperature to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 21
S1, preparing ICG-loaded bionic liposome: fully mixing lecithin, DSPE-PEG2000, cholesterol and ICG-ODA chloroform solution according to the molar ratio of 100:20:4:10, performing rotary evaporation to form a film, and vacuumizing for 3h to completely volatilize an organic solvent after the film is formed; adding a Tris solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a polycarbonate membrane of 400nm to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid and stirred at normal temperature to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide of 1mg/mL through a polycarbonate film of 200mm, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 22
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding a 5% glucose solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, and freeze-drying for storage.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3Stirring at 60 ℃ to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, stirring at room temperature, centrifuging, separating, and freeze dryingDrying to obtain hollow MnO with medicine2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 2mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 23
S1, preparing the IR 780-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and IR780 in a chloroform solution at a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3h to completely volatilize an organic solvent after forming the film; adding a 5% glucose solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an IR 780-loaded liposome; co-extruding the obtained IR 780-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the IR 780-loaded bionic liposome, centrifuging, cleaning, and freeze-drying for storage.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid and stirred at normal temperature to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of the IR 780-loaded bionic liposome with 500 mu L of manganese oxide of 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 24
S1, preparing a Ce 6-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and Ce6 in a chloroform solution according to a molar ratio of 100:20:4:10, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after the film is formed; adding a PBS solution into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain a Ce 6-loaded liposome; the obtained Ce 6-loaded liposome and the extracted 4T1 tumor cell membrane protein are co-extruded by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the Ce 6-loaded bionic liposome, and the bionic liposome is centrifugally cleaned and stored at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid and stirred at normal temperature to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of the Ce 6-loaded bionic liposome with 500 mu L of the manganese oxide with the concentration of 1mg/mL through a polycarbonate film with the thickness of 200mm, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano composite product.
Example 25
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:15, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 4 hours to completely volatilize an organic solvent after forming the film; adding a Tris solution into a round-bottom flask after film formation, hydrating under the ultrasonic condition of 55 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a polycarbonate membrane of 400nm to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring the mixture overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3Stirring overnight at 60 deg.C toEtching SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature for 24 hr, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide of 1mg/mL through a polycarbonate film of 200mm, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 26
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:15, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding a Tris solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a polycarbonate membrane of 400nm to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring at normal temperature for one night to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid solution and stirred for 1 hour at low speed to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide with the concentration of 2mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 27
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:15, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 4 hours to completely volatilize an organic solvent after forming the film; adding a 5% sucrose solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, freezing, drying and storing.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring at normal temperature for one night to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid solution and stirred for 2 hours at low speed to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 28
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:15, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 5 hours to completely volatilize an organic solvent after the film is formed; adding a 5% glucose solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a polycarbonate membrane of 400nm to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 3 hours and then continuing to stir at normal temperature overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 29
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:15, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 6 hours to completely volatilize an organic solvent after forming the film; adding a PBS solution into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, freezing, drying and storing.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring at normal temperature for one night to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid solution and stirred at low speed for 3 hours to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide with the concentration of 2mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 30
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:15, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding a Tris solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring at normal temperature for one night to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in hydrofluoric acid solution to vibrate at low speed for 1h to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 100mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 31
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:15, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 3 hours to completely volatilize an organic solvent after forming the film; adding a Tris solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, freezing, drying and storing.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring at normal temperature for one night to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid solution to vibrate at low speed for 2 hours so as to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide of 1mg/mL through a polycarbonate film of 200mm, repeatedly extruding for 20 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 32
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:15, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 4 hours to completely volatilize an organic solvent after forming the film; adding a Tris solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a polycarbonate membrane of 400nm to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring at normal temperature for one night to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, stirring at room temperature, centrifuging, separating, and freeze drying to obtain medicine carrierEmpty MnO2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 2mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 25 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 33
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 6 hours to completely volatilize an organic solvent after the film is formed; adding a Tris solution into the film-formed round-bottom flask, hydrating at 55 ℃ under an ultrasonic condition, and centrifugally cleaning with deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a polycarbonate membrane of 400nm to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring at normal temperature for one night to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 34
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 5 hours to completely volatilize an organic solvent after the film is formed; adding a Tris solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, freezing, drying and storing.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Stirring at normal temperature for one night to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 35
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 5 hours to completely volatilize an organic solvent after the film is formed; adding a Tris solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, freezing, drying and storing.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 30min, then stirring at normal temperature overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, stirring at room temperature, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide with the concentration of 2mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 36
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 5 hours to completely volatilize an organic solvent after the film is formed; adding a 5% glucose solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, freezing, drying and storing.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 1h, then stirring at normal temperature overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, physically stirring under ice bath condition, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 2mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 37
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:6:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 6 hours to completely volatilize an organic solvent after the film is formed; adding a 5% sucrose solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a 400nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, freezing, drying and storing.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 1h, then stirring at normal temperature overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, physically stirring under ice bath condition, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 38
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:5:10, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 6 hours to completely volatilize an organic solvent after forming the film; adding a 5% sucrose solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, freezing, drying and storing.
S2 preparation by sol-gel methodMonodisperse SiO2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 1h, then stirring at normal temperature overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, physically stirring under ice bath condition, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 100mm polycarbonate membrane, repeatedly extruding for 30 times, centrifuging, freezing and drying to obtain the nano-composite product.
Example 39
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:5:15, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 6 hours to completely volatilize an organic solvent after forming the film; adding a 5% sucrose solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning, freezing, drying and storing.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 1h, then stirring at normal temperature overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with paclitaxel, physically stirring under ice bath condition, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 100mm polycarbonate membrane, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 40
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:5:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 6 hours to completely volatilize an organic solvent after forming the film; adding a Tris solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a polycarbonate membrane of 400nm to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 1h, then stirring at normal temperature overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring under ice bath condition, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
EXAMPLE 41
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:15, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 6 hours to completely volatilize an organic solvent after forming the film; adding a Tris solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of a polycarbonate membrane of 400nm to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 1h, then stirring at normal temperature overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring under ice bath condition, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 200mm polycarbonate membrane, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Example 42
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 5 hours to completely volatilize an organic solvent after the film is formed; adding a Tris solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 1h, then stirring at normal temperature overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano-composite is placed in hydrofluoric acid solution to vibrate at low speed for 3 hours so as to etch SiO2Inner core, centrifuging, separating, freezingDrying to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring under ice bath condition, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, co-extruding 500 mu L of ICG-loaded bionic liposome with 500 mu L of manganese oxide in 1mg/mL through a 100mm polycarbonate membrane, repeatedly extruding for 20 times, centrifuging, and freeze-drying to obtain the nano-composite product.
Comparative example 1
S1, preparing ICG-loaded common liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 5 hours to completely volatilize an organic solvent after the film is formed; adding a Tris solution into the round-bottom flask after film formation, hydrating under the ultrasonic condition of 60 ℃, centrifugally cleaning by deionized water to obtain the ICG-loaded liposome, and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nanometer microsphere, washing, centrifuging, and adding KMnO under ultrasonic condition4Continuing to perform ultrasonic treatment for 1h, then stirring at normal temperature overnight to prepare MnO2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2Placing the nano-composite in Na2CO3In solution to etch SiO2Centrifuging, separating and freeze-drying the inner core to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with DOX, physically stirring under ice bath condition, centrifuging, separating, and freeze drying to obtain hollow MnO containing drug2。
S3, taking 500 mu L of ICG-loaded bionic liposome with 1mg/mL and 500 mu L of hollow MnO with 1mg/mL medicine2Co-extruding with 200mm polycarbonate film, extruding repeatedly for 20 times, centrifuging, and freeze drying to obtain the final product.
The nanocomposite product produced in this comparative example was a non-biomimetic modified nuclear membrane nanocomposite (HMD-I-L).
Comparative example 2
S1, preparing ICG-loaded bionic liposome: dissolving lecithin, DSPE-PEG2000, cholesterol and ICG-ODA in a chloroform solution according to a molar ratio of 100:20:4:20, fully mixing, performing rotary evaporation to form a film, and vacuumizing for 5 hours to completely volatilize an organic solvent after the film is formed; adding a Tris solution into the film-formed round-bottom flask, hydrating under the ultrasonic condition of 60 ℃, and centrifugally cleaning by deionized water to obtain an ICG-loaded liposome; co-extruding the obtained ICG-loaded liposome and the extracted 4T1 tumor cell membrane protein by a liposome extruder under the condition of 200nm polycarbonate membrane to obtain the ICG-loaded bionic liposome, centrifuging, cleaning and storing at 4 ℃.
S2 preparation of monodisperse SiO by sol-gel method2Nano microsphere, prepared by mixing the above materials2Adding CTAB solution, ammonia water and BTEB into the nano microsphere solution in sequence and continuing stirring to obtain solid SiO2The SiO2@ mSiO2 nano-particle takes a core and mesoporous SiO2 takes a shell. Placing the obtained core-shell structure compound in Na2CO3Removing template from the solution, centrifuging, separating, and freeze-drying to obtain hollow mesoporous SiO2(ii) a Hollow mesoporous SiO2Mixing with DOX, physically stirring, centrifuging, separating, and freeze drying to obtain DOX-loaded hollow mesoporous SiO2。
S3, taking 500 mu L of ICG-loaded bionic liposome with 1mg/mL and DOX-loaded hollow mesoporous SiO with 500 mu L2Co-extruding with 200mm polycarbonate film, extruding repeatedly for 20 times, centrifuging, and freeze drying to obtain the final product.
The nano composite product prepared by the comparative example is hollow mesoporous SiO2Drug-loaded nuclear membrane nanocomposites (HSD-I-BL).
Experimental example 1
Structural characterization of self-oxygen-supply targeting type bionic nuclear membrane structure nano-composite
Structural characterization of each composite prepared in example 1
The characterization method comprises the following steps: transmission electron micro-scanning instrument: FEITecnai F20
FIG. 2 shows a TEM image of the product obtained in example 1.
The same structural characterization was performed on each of the complexes prepared in the other examples of the present invention, which proves that the method of the present invention is indeed used to obtain the self-oxygen-supplying targeted biomimetic nuclear membrane structure nanocomposite.
Experimental example 2
Oxygen production experiment of self-oxygen supply targeting type bionic nuclear membrane structure nano compound
The self-oxygen-supplying targeting type bionic nuclear membrane structure nano composite (HMD-I-BL) obtained in example 1 and the hollow mesoporous SiO obtained in comparative example 2 are configured2Loading nuclear membrane nanocomposite (HSD-I-BL), adding each to a solution containing hydrogen peroxide (10mM H)2O2) And glutathione (10mM GSH) in aqueous solution to mimic the responsive oxygen production of the nanocomposite in the tumor microenvironment in vitro. Irradiating for 5 minutes (1W/cm) under 808nm laser2) And measuring the oxygen content in the solution. The oxygen content is shown in figure 3.
The results show that the hollow mesoporous SiO2The medicine-carrying nuclear membrane nano-composite can obviously produce oxygen in vitro, and the obtained self-oxygen-supply targeting bionic nuclear membrane structure nano-composite has better in vitro oxygen production capacity under laser irradiation and is used for improving tumor hypoxia in situ.
Experimental example 3
Self-oxygen-supply targeting type bionic nuclear membrane structure nano compound targeting experiment
5000 pieces of 4T1 breast cancer cells were inoculated in a 6-well plate at 37 ℃ in 5% CO2After overnight incubation, the self-oxygenating targeting type bionic nuclear membrane nano-composite (HMD-I-BL) prepared in example 1 and the non-bionic modified nuclear membrane nano-composite (HMD-I-L) prepared in comparative example 1 were added in an ICG amount of 20. mu.g/mL respectively, and after 2 hours of incubation, PBS was washed and fixed with 4% paraformaldehyde. The uptake of the complex by 4T1 cells under confocal laser microscopy is shown in FIG. 4.
The obtained self-oxygen-supplying targeted bionic nuclear membrane structure nano compound has the intake in 4T1 cells obviously higher than that of a non-bionic modified nano compound, has good targeting effect and is used for tumor targeted drug delivery.
Experimental example 4
In-vitro magnetic resonance imaging performance of self-oxygen-supply targeting type bionic nuclear membrane structure nano compound
A typical example of the magnetic resonance imaging is shown in fig. 5. It can be seen that when the surrounding water phase environment is black, example 1 presents a concentration-dependent bright signal (MRI T1 contrast function is enhanced), which indicates that the sample self-oxygen-supply targeted biomimetic nuclear membrane structure nanocomposite has an obvious T1 weighted magnetic resonance imaging function.
Experimental example 5
In-vitro photo-thermal performance of self-oxygen supply targeting type bionic nuclear membrane structure nano composite
The self-oxygen-supplying targeting type bionic nuclear membrane nano-composite prepared in the embodiment 1 and measured by 20 mu g/mL ICG is dissolved in deionized water, a 808nm laser is adopted for irradiation for 5min, and the temperature change condition of the solution under the laser irradiation observed by a near-infrared thermal imager is shown in figure 6.
The obtained self-oxygen-supply targeted bionic nuclear membrane structure nano composite has an obvious temperature rise effect under laser irradiation, and the temperature rise effect is similar to that of free ICG with the same concentration, so that the self-oxygen-supply targeted bionic nuclear membrane structure nano composite has good photo-thermal performance and is used for tumor photo-thermal treatment.
Experimental example 6
In vitro of self-oxygen-supply targeting type bionic nuclear membrane structure nano compound1O2Generating an assessment
The self-oxygen-supply targeting bionic nuclear membrane nano compound prepared in the embodiment 1 and measured by 20 mu g/mL ICG is dissolved in H-containing solution2O2And DPBF deionized water, irradiating for 5min by using a 808nm laser, and observing the change of the absorbance of the solution at different time points by using an ultraviolet spectrophotometer, wherein the change is shown in figure 7.
The absorbance of the obtained self-oxygen-supply targeted bionic nuclear membrane structure nano compound is obviously reduced under the irradiation of laser, which shows that the self-oxygen-supply targeted bionic nuclear membrane structure nano compound can effectively generate under the irradiation of the laser1O2Can be used for photodynamic therapy of tumor.
Experimental example 7
Small animal fluorescence imaging of self-oxygen-supply targeting type bionic nuclear membrane structure nano compound
The self-oxygen-supplying targeting type bionic nuclear membrane nano-composite prepared in the embodiment 1 and measured by 20 mug/mL ICG is dissolved in deionized water, injected into a mouse body through tail vein, and observed by a small animal living body imaging instrument to change the fluorescence intensity as shown in figure 8.
The obtained self-oxygen supply targeting type bionic nuclear membrane structure nano compound is gradually enriched at a tumor part, and the concentration of the self-oxygen supply targeting type bionic nuclear membrane structure nano compound is obviously higher than that of a non-bionic modified compound, so that the self-oxygen supply targeting type bionic nuclear membrane structure nano compound has good targeting performance and is used for accurate targeting treatment of tumors.
Although the present disclosure has been described above, the scope of the present disclosure is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present disclosure, and such changes and modifications will fall within the scope of the present invention.

Claims (10)

1. A self-oxygen supply targeting type bionic nuclear membrane structure nano-composite is characterized in that: the nano composite comprises a bionic lipid membrane and hollow MnO2The nuclear membrane structure of (a); the bionic lipid membrane is a phospholipid bimolecular membrane with the surface containing tumor cell membrane protein modification;
the hollow MnO2Is used for loading tumor therapeutic drugs, and the drugs are chemotherapeutic drugs or immunotherapy drugs.
2. The self-oxygen-supplying targeting type bionic nuclear membrane structure nano-composite according to claim 1, characterized in that: the bionic lipid membrane contains a photosensitizer, and the photosensitizer is selected from at least one of ICG-ODA, ICG-NH2, ICG-COOH, ICG-NHS, ICG-MAL, ICG-SH, ICG-N3, ICG-ALK, ICG-Biotin, IR780, IR783 and IR 808.
3. The self-oxygen-supplying targeting type bionic nuclear membrane structure nano-composite according to claim 1, characterized in that: the chemotherapeutic drug is selected from anthracycline anticancer drug, taxol anticancer drug, vinca alkaloid anticancer drug, platinum anticancer drug and at least one of 5-fluorouracil, methotrexate, cyclophosphamide, temozolomide and pemetrexed.
4. The self-oxygen-supplying targeting type bionic nuclear membrane structure nano-composite according to claim 1, characterized in that: the phospholipid bilayer membrane comprises lecithin, hydrogenated soybean phospholipid, hydrogenated yolk phospholipid, dipalmitoyl phosphatidyl ethanol, dipalmitoyl phosphatidyl choline, dioleoyl phosphatidyl ethanolamine, polyethylene glycol-distearoyl phosphatidyl ethanolamine, 1, 2-dipalmitoyl-sn-glyceryl-3-phosphatidic acid glyceryl-sodium salt, 1, 2-distearoyl-sn-glyceryl-3-phosphatidylcholine, 1, 2-dipalmitoyl-sn-glyceryl-3-phosphatidic acid-sodium salt and 1, 2-dipalmitoyl-sn-glyceryl-3-phosphatidylcholine.
5. A method for preparing the self-oxygen-supplying targeting type bionic nuclear membrane structure nano-composite according to any one of claims 1 to 4, which comprises the following steps:
s1, preparing the photosensitizer-carrying bionic liposome with targeting performance: synthesizing a photosensitizer-carrying liposome modified by tumor cell membrane protein and membrane protein of tumor cells by a co-extrusion method;
s2 preparation of drug-loaded hollow MnO2: with SiO2Preparing hollow MnO for the template2Co-stirring the obtained hollow manganese dioxide and the hydrophobic therapeutic drug to realize the loading of the hydrophobic drug in the hollow core;
s3, preparing bionic liposome/drug-loaded hollow MnO2Complex: mixing the bionic liposome prepared in the step S1 with the medicine-carrying hollow MnO in the step S22The finished product is prepared by co-extrusion of polycarbonate film, repeated extrusion, centrifugation, freeze drying.
6. The preparation method of the self-oxygen-supplying targeted biomimetic nuclear membrane structure nanocomposite as claimed in claim 5, wherein the preparation method comprises the following steps: the synthesized liposome carrying the photosensitizer is prepared by a film hydration method, a film ultrasonic dispersion method, a reverse evaporation method or a microemulsion ultrasonic dispersion method.
7. The method for preparing the self-oxygen-supplying targeted nano composite with the bionic nuclear membrane structure according to claim 6, wherein the synthesized liposome loaded with the photosensitizer in the step S1 is prepared by a thin-film ultrasonic dispersion method, and the method comprises the following steps: dissolving a phospholipid bimolecular membrane, DSPE-PEG2000, cholesterol and a photosensitizer with a molar ratio of 100:20:4:10 in a chloroform solution, fully mixing, performing rotary evaporation at 40 ℃ to form a membrane, vacuumizing for 3h after membrane formation to completely volatilize the organic solvent, adding PBS into a round-bottomed flask after membrane formation, hydrating under an ultrasonic condition at 60 ℃, and centrifugally cleaning with deionized water to obtain the liposome carrying the photosensitizer.
8. The preparation method of the self-oxygen-supplying targeted biomimetic nuclear membrane structure nanocomposite as claimed in claim 5, wherein the preparation method comprises the following steps: drug-loaded hollow MnO in step S22The preparation method specifically comprises the following steps: preparation of monodisperse SiO by sol-gel method2Washing nanometer microsphere with ethanol/water solution, centrifuging at room temperature, and adding KMnO under ultrasonic condition4After continuing stirring, MnO was obtained2Wrapped SiO2A nanocomposite; MnO to be obtained2Wrapped SiO2The nano composite is placed in an etching solution to etch SiO2The inner core is sequentially subjected to centrifugation, washing and drying to obtain hollow MnO2(ii) a To make hollow MnO2Mixing with the drug, and sequentially carrying out physical stirring, centrifugation, separation and freeze drying at normal temperature to obtain drug-loaded hollow MnO2。
9. Use of the self-oxygenating targeted biomimetic nuclear membrane structured nanocomposite as recited in any of claims 1-4 in the preparation of a medicament for treating cancer.
10. Use according to claim 9, characterized in that: the cancer includes breast cancer, thyroid cancer, esophageal cancer, pancreatic cancer, lung cancer, kidney cancer, liver cancer and melanoma.
CN202111293490.5A 2021-11-03 2021-11-03 Self-oxygen-supply targeted bionic nuclear membrane structure nano-composite and preparation method and application thereof Pending CN114177309A (en)

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Citations (3)

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Publication number Priority date Publication date Assignee Title
CN110179754A (en) * 2019-06-12 2019-08-30 福州大学 With redox responsiveness and the multi-functional liposome of tissue infiltration can be enhanced
CN112402379A (en) * 2020-12-04 2021-02-26 中南大学湘雅二医院 Bionic exosome and preparation and application thereof
CN113493223A (en) * 2020-04-03 2021-10-12 长春可研生物科技有限公司 Preparation method and application of hollow manganese dioxide nanospheres

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110179754A (en) * 2019-06-12 2019-08-30 福州大学 With redox responsiveness and the multi-functional liposome of tissue infiltration can be enhanced
CN113493223A (en) * 2020-04-03 2021-10-12 长春可研生物科技有限公司 Preparation method and application of hollow manganese dioxide nanospheres
CN112402379A (en) * 2020-12-04 2021-02-26 中南大学湘雅二医院 Bionic exosome and preparation and application thereof

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