CN113476603B - Magnetic nanoparticle wrapped by erythrocyte membrane as well as preparation method and application of magnetic nanoparticle - Google Patents
Magnetic nanoparticle wrapped by erythrocyte membrane as well as preparation method and application of magnetic nanoparticle Download PDFInfo
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- CN113476603B CN113476603B CN202110770716.XA CN202110770716A CN113476603B CN 113476603 B CN113476603 B CN 113476603B CN 202110770716 A CN202110770716 A CN 202110770716A CN 113476603 B CN113476603 B CN 113476603B
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Images
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- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
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- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
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Abstract
The invention relates to magnetic nanoparticles wrapped by erythrocyte membranes, a preparation method and application thereof, in particular to PEG-Fe prepared by a hydrothermal method3O4Extruding and wrapping with target-modified erythrocyte membrane fragments to prepare nanoparticle erythrocyte membrane-PEG-Fe with immune evasion and sensitive magnetic response3O4The folic acid-sensitive cancer cell can be adsorbed in a targeted manner, so that the folic acid-sensitive cancer cell can be conveniently killed by magnetic field control or photo-thermal stimulation. Erythrocyte membrane-PEG-Fe3O4Can be effectively adsorbed on folic acid sensitive cancer cell membranes in a targeted way, endows the cancer with magnetic responsiveness, and is favorable for controlling movement by a magnetic field and killing by photo-thermal stimulation. Therefore, the method is suitable for targeted therapy and industryHas potential application prospect in a wide range.
Description
Technical Field
The invention belongs to the field of nano biomedical materials, and relates to a preparation method of magnetic nanoparticles wrapped by erythrocyte membranes, and a product and application thereof.
Background
Cancer is life threatening to humansThe traditional Chinese medicine composition has limited curative effect on major diseases by conventional chemotherapy, radiotherapy and surgery, has high side effect, and urgently needs the development of new technology according to the requirement of a cancer market. Targeting magnetic nanoparticles is the focus of current research, magnetic Fe3O4The material has unique magnetic property and heat conduction capability, has good biocompatibility, is widely concerned in cancer treatment, can excite a large amount of heat with a small amount of infrared energy, is a good material for photothermal therapy, and has magnetic property which is favorable for endowing cancer cells with magnetic responsiveness and is convenient for magnetic field control. Fe3O4The disadvantages of (1) are that the magnetic effect makes the particles easy to agglomerate and precipitate, the oversize particles are not beneficial to human metabolism and easy to accumulate in liver, kidney and lymph node, which causes intolerable side effects for human body. Although the prior art currently passes through Fe in the magnetic nanoparticle preparation process3O4The surface modification of (2) can enhance the suspension property, but in order to achieve the purpose of reducing the particle size, surfactants with high toxicity such as sodium dodecyl benzene sulfonate, organic solvents such as tetrahydrofuran and DMSO are often adopted, and recognition and phagocytosis of immune cells cannot be avoided, and although the prepared particles have excellent biological effect, the potential toxicity hazard and the risk of phagocytosis by the immune cells also exist. The selection range is reduced and the risk of wrong selection is increased compared with the surfactants and organic solvents, and the chitosan and sodium alginate which have been used and have lower toxicity are taken as the surfactants and are finally failed.
Disclosure of Invention
Aiming at the defects of the prior art, the invention mainly aims to: provides a method for preparing simple, low-toxicity and immune evasive targeted nanoparticles. In particular to PEG-Fe prepared by a hydrothermal method3O4The nano-particles with immune evasion and sensitive magnetic responsiveness are prepared by extruding and wrapping red cell membrane fragments subjected to targeted modification, and due to the isolation effect of the red cell membrane, folic acid targeted molecules are easily obtained, so that more cancer cells sensitive to folic acid are obtained, the particles are prepared into the nano-particles capable of performing targeted adsorption on the cancer cells sensitive to folic acid, the cancer cells can be conveniently killed by magnetic field control or photo-thermal stimulation, the particle size of the particles can be effectively reduced, and besides a hydrothermal method, PEG6000-10000 mixed oleic acid is also usedThe dispersant prepared from sodium successfully reduces the particle size to 80nm, the erythrocyte membrane plays a role in stabilizing the particle size after the dispersant is removed, and the whole preparation process effectively avoids a surfactant and an organic solvent with high toxicity.
A preparation method of magnetic nanoparticles wrapped by erythrocyte membranes comprises the following steps:
(1)PEG-Fe3O4preparing;
(2) extracting and targeted modifying erythrocyte membrane fragments;
(3) erythrocyte membrane-PEG-Fe3O4The preparation of (1): mixing PEG-Fe3O4Adding a dispersion prepared by adding ultra-pure water into PEG6000-10000 and sodium oleate, performing ultrasonic treatment, standing, removing precipitates, taking an upper layer suspension, blending and stirring the target modified erythrocyte membrane obtained in the step (2) and the upper layer suspension, putting the mixture into a liposome extruder, and extruding for multiple times to obtain the liposome.
Further, in the step (3), the mass ratio of PEG6000-10000 (preferably PEG6000) to sodium oleate is 1: 2-2: 1, preferably 1:1, adding ultrapure water to prepare 6-8% dispersion, preferably 6% dispersion. PEG-Fe3O4The concentration in the dispersion is 100-300 mu g/mL; preferably 150. mu.g/mL, and more preferably 200. mu.g/mL.
Further, the power of the ultrasonic cell disruptor in the step (3) is 600- & lt 1000 & gt W, preferably 800W, and the frequency is 15-25kHz, preferably 20 kHz. Performing ultrasonic treatment for 30-40 min.
Further, standing for at least 12h after the ultrasonic treatment in the step (3) to remove precipitates.
Further, in the step (3), the target modified erythrocyte membrane obtained in the step (2) and the supernatant suspension are blended and stirred, heated to 38-45 ℃, preferably 40 ℃, and then extruded.
Further, in step (3), per ml of PEG-Fe3O4The ratio of the two in the mixed solution of the target modified erythrocyte membrane is 150-250 mu g:3mg, and the two are extruded from 800nm and 400nm by a liposome extruder and extruded from 200nm membrane for 3-7 times at 40-45 ℃. Preferably: PEG-Fe per ml3O4The ratio of the two in the mixed solution of the red cell membrane and the target modified red cell membrane is 180-230 mu g:3mg, and thenThe extrusion was carried out sequentially at 40 ℃ from 800nm to 400nm using a liposome extruder and 7 times under a 200nm membrane.
The membrane of step (3) is preferably a Whatman polycarbonate membrane.
Step (1) PEG-Fe3O4The preparation process comprises the following steps: FeCl2With FeCl3Mixing the raw materials in a ratio of 1: 1.5-1: distilled water is added according to the molar ratio of 2, and FeCl is added2PEG6000-10000 with the mass of 2-4 times, and ultrasonic treatment for 5-10min (the power of an ultrasonic cell disruptor is 600-1000W, and the frequency is 15-25kHz) until dissolution; stirring at 1400-2600r/min under inert atmosphere, heating to 45-60 ℃, and reacting for 30-40 min; adding FeCl2PEG6000-10000 solution with the mass of 2-4 times, transferring the reaction solution to a polytetrafluoroethylene reaction kettle, setting the temperature at 160-200 ℃, and reacting for 5-10 hours; to obtain Fe3O4Precipitating nanoparticles, washing the precipitate with pure water and ethanol, repeating the steps for several times, and drying in a vacuum oven at 45-60 deg.C for 10-24 hr to obtain PEG-Fe3O4And (3) nanoparticles.
Preferably: step (1) PEG-Fe3O4The preparation process comprises the following steps: FeCl2With FeCl3Mixing the raw materials in a ratio of 1: distilled water is added according to the molar ratio of 2, and FeCl is added2PEG6000 with the mass 3 times, and performing ultrasonic treatment for 10min (the power of an ultrasonic cell disruptor is 800W, and the frequency is 20kHz) until dissolution; stirring at 2600r/min under nitrogen atmosphere, heating to 60 deg.C, and reacting for 30 min; adding FeCl2Transferring the reaction solution to a polytetrafluoroethylene reaction kettle by using PEG6000 solution with the mass being 3 times that of the reaction solution, setting the temperature at 160 ℃, heating for 1 hour, and reacting for 5 hours; to obtain Fe3O4Precipitating nanoparticles, washing the precipitate with pure water and ethanol, repeating the steps for multiple times, and drying in a vacuum oven at 60 deg.C for 10 hr to obtain PEG-Fe3O4And (3) nanoparticles.
Step (2) extraction and targeted modification process of erythrocyte membrane fragments: centrifuging anticoagulated mammal blood to precipitate erythrocyte, removing supernatant, washing with ice PBS, soaking the crushed erythrocyte in hypotonic solution, centrifuging, leaving precipitate to remove supernatant, washing the precipitate with ice PBS, repeating for several times until the precipitate becomes light pink or white, and mixing erythrocyte membrane and sodium taurocholate solution according to the formula: red cell membrane 1: mixing at a mass ratio of 6-1:4, dissolving in ethanol with a volume of 4-6 times, ultrasonically mixing, loading into an injector, rapidly injecting PBS (phosphate buffer solution) with a volume of 8-12 times that of ethanol solution at 40-50 deg.C, and rotary steaming at 50-70 deg.C for 20-30min to remove ethanol;
and (3) targeted modification process: mixing DSPE-PEG and targeting molecules and/or fluorescent molecules in a PBS solution according to a molar ratio of 1.5-1:1 to form the DSPE-PEG-targeting molecules and/or fluorescent molecules, wherein the fluorescent molecules comprise at least one of FITC and rhodamine isothiocyanate, and the targeting molecules comprise at least one of folic acid and mannitol; adding DSPE-PEG-targeting molecules and/or fluorescent molecules into a very small amount of erythrocyte membranes diluted by PBS solution, and mixing by a shaking table; red cell membrane: DSPE-PEG-targeting and/or fluorescent molecules 4: 1-10: 1 mass ratio.
Preferably: step (2) extraction and targeted modification process of erythrocyte membrane fragments: centrifuging sheep blood at 5000r/min to precipitate red blood cells, removing supernatant, washing with 0 deg.C ice PBS, repeating for three to four times, soaking the crushed red blood cells in 1/4 times of PBS hypotonic solution for 1 hr, centrifuging at 13000r/min for 15min, removing supernatant, washing the precipitate with 0 deg.C ice PBS, repeating for multiple times until the precipitate becomes pale pink or white, determining standard curve with lecithin on ultraviolet to obtain erythrocyte membrane concentration, and adjusting to 3 mg/ml; preparing sodium taurocholate into a concentration of 1mg/ml, wherein the concentration of the erythrocyte membrane and the sodium taurocholate solution is as follows: red cell membrane 1: mixing at a mass ratio of 4, dissolving in 5 times volume of ethanol, ultrasonically mixing, loading into an injector, rapidly injecting PBS solution with a volume of 10 times that of the ethanol solution at 45 ℃, and rotary-steaming at 60 ℃ for 25min to remove ethanol.
And (3) targeted modification process: mixing DSPE-PEG and targeting molecules and/or fluorescent molecules in a PBS solution according to a molar ratio of 1:1 to form a DSPE-PEG-targeting molecule and/or fluorescent molecule solution (namely DSPE-PEG-X), adding DSPE-PEG-X into a very small amount of erythrocyte membranes diluted by the PBS solution, and mixing by a shaking table; red cell membrane: DSPE-PEG-X is 4:1 by mass.
The DSPE-PEG-targeting molecule (FA) is used for targeted adsorption of folate-sensitive cancer cells, so that the cancer cells can be conveniently and accurately killed by magnetic field control or photo-thermal stimulation.
The DSPE-PEG-fluorescent molecules are used for laboratory testing needs or computational pharmacokinetic testing needs in clinical procedures.
The invention can prepare DSPE-PEG-targeting molecule or DSPE-PEG-fluorescent molecule separately, and also can prepare DSPE-PEG to connect the targeting molecule and the fluorescent molecule at the same time.
The invention can also adopt PEG to be connected with different medicines and then coated with a targeted erythrocyte membrane for targeted therapy.
The DSPE-PEG-fluorescent molecule and/or FA (folic acid) can be prepared by self or directly adopts the products of Shanghai Peng Shuichi Biotech Co.
DSPE-PEG is distearoylphosphatidylethanolamine-polyethylene glycol, wherein the molecular weight of PEG is 2000-6000, preferably 2000.
Further, mixing erythrocyte membrane with DSPE-PEG-X, and shaking for 1-2 hr at 36-37 deg.C; then sequentially extruding at 40-45 deg.C from 800nm and 400nm with liposome extruder, and extruding under 200nm membrane for 3-7 times. Centrifuging at 12000-13000 r/min at 4 ℃, washing with ice PBS, re-dispersing for 30-60 seconds by using an ultrasonic cell disruptor, preserving with PBS at 4 ℃ at power of 600-1000W, preferably 800W and frequency of 15-25kHz, preferably 20 kHz.
Further, dialyzing the extruded erythrocyte membrane magnetic nanoparticle solution for 48h by using a 70nm dialysis bag, removing redundant DSPE-PEG-fluorescent molecules and/or targeting molecules, sodium oleate and PEG6000, standing for 48h at 4 ℃, removing precipitates, and taking upper-layer liquid.
The second purpose of the invention is to provide the magnetic nanoparticle wrapped by the erythrocyte membrane obtained by the preparation method.
The third purpose of the invention is to provide the application of the magnetic nano-particles wrapped by the erythrocyte membranes as a targeting material, in particular to the application as a medical targeting material.
The magnetic nano-particles wrapped by the erythrocyte membranes can reflect the magnetic responsiveness and the photothermal performance of the central core of the magnetic nano-particles, and the good biocompatibility and immune evasion of the liposome of the externally loaded erythrocyte membranes, and on the other hand, the use of PEG molecules effectively reduces the particle size of the magnetic cores, improves the suspension stability of the magnetic fluid, and the use of PEG and sodium oleate dispersant enables the preparation process to be avoidedToxic surfactants and organic solvents. After the magnetic nano-particles coated by the erythrocyte membrane are adsorbed on the folate sensitive cancer cells in a targeted manner, the magnetic responsiveness of the cancer cells can be effectively endowed, and Fe can be excited by a small amount of infrared rays3O4The heat is released, and the cancer cells are effectively killed. The erythrocyte membrane endows magnetic particle immunity evasion, greatly prolongs the blood circulation time, and enables the erythrocyte membrane to obtain sufficient time and accurately capture cancer cells. Therefore, the compound has excellent application prospect in the biomedical field and the cancer treatment aspect.
The invention has the advantages that:
(1) the nano material prepared by the invention has magnetic responsiveness.
(2) The nano material prepared by the invention has the function of specifically targeting tumors.
(3) The nano material prepared by the invention has the function of killing cancer cells under the excitation of infrared rays.
(4) The nano material prepared by the invention has good biocompatibility and long blood circulation function.
(5) The nano material prepared by the invention is wrapped by erythrocyte membranes, and the erythrocyte membranes are softened by an ethanol softening method, so that the deformability is increased, the nano material is easy to pass through a polycarbonate membrane, and the nano particles are easy to be adsorbed by tumor tissues by ESR effect.
(6) The nano material prepared by the invention has the advantages of target adsorption to the surface of a cancer cell membrane and endowing the cancer cell with a magnetic response function.
(7) The preparation process of the invention has no toxic material and organic solvent in the whole process, and has simple preparation, low cost, safety and reliability.
Drawings
FIG. 1 is a schematic cross-sectional view of a targeted magnetic nanoparticle prepared according to the present invention;
in the figure, (1) erythrocyte membrane; (2) PEG; (3) fe3O4A magnetic core; (4) DSPE-PEG-FA;
FIG. 2 is a comparison of the dispersion effect of PEG6000 and sodium sulfate dispersant in different proportions in example 1;
in the figure, (a) sodium oleate and PEG6000 is 1: 1; (b) sodium oleateAnd PEG6000 is 1: 2; (c) sodium oleate and PEG6000 is 2: 1; (d) oleic acid free and PEG6000 free Fe3O4All stand for 24 hours.
FIG. 3 is a comparison of the dispersing effect of different dispersants used in step (1) or step (3) of example 1 of the present invention;
in the figure: (a) PEG6000-Fe3O4Adding a sodium dodecyl benzene sulfonate dispersing agent; (b) chitosan Oligosaccharide (OCS) -Fe3O4Adding PEG-sodium oleate dispersant; (c) sodium alginate-Fe3O4Adding PEG-sodium oleate dispersant; (d) PEG6000-Fe3O4Adding a polyvinyl alcohol-sodium dodecanesulphonate dispersing agent; (e) PEG600 coated Fe3O4Adding PEG-sodium oleate dispersant; (f) OCS-Fe on the left3O4Extruding 200nm film, right side PEG6000-Fe3O4A 200nm film was extruded.
FIG. 4 shows PEG-Fe prepared in example 13O4Five-day DLS particle size monitoring graph;
in the figure (a) PEG-Fe after ultrasonication3O4The particle size distribution of the nanoparticles; (b) PEG-Fe after standing for 1 day3O4The particle size distribution of the nanoparticles; (c) PEG-Fe after standing for 3 days3O4The particle size distribution of the nanoparticles; (d) PEG-Fe after standing for 5 days3O4The particle size distribution of the nanoparticles.
FIG. 5 shows PEG-Fe prepared in example 13O4A magnetic property characterization map;
in the figure, a is PEG-Fe in suspension3O4Magnetic particles, b is PEG-Fe3O4The magnetic particles move with the magnetic field, and c is a hysteresis loop of the powdery magnetic particles.
FIG. 6 is a TEM image of the erythrocyte membrane prepared in example 1 after being dispersed by softening extrusion;
FIG. 7 is a graph showing the effect of UV-irradiation on erythrocyte membrane deformability of sodium taurocholate in different ratios and at different temperatures in example 1
In the figure a, the influence of different temperatures on the membrane flexibility of pure red blood cells;
b, the influence of different temperatures on the ultraviolet peak value of the extruded pure erythrocyte membrane;
c influence of different surfactant ratios on erythrocyte membrane softness;
d, influence of different surfactant ratios on ultraviolet peak values of the softened erythrocyte membranes after extrusion;
e the influence of different temperatures on the softness of the red cell membrane;
f influence of different temperatures on UV peak of softened erythrocyte membrane after extrusion.
FIG. 8 is a graph of the ratio of 1:4(m/m) research on the stability of erythrocyte membrane (RBC) modified by sodium taurocholate and unmodified erythrocyte membrane;
(a) pure RBC particle size distribution; (b)1/6RBC size distribution modified by sodium taurocholate; (c)1/5RBC size distribution modified by sodium taurocholate; (d)1/4 RBC-quality sodium taurocholate-modified RBC particle size distribution.
FIG. 9 shows modified RBC and PEG-Fe in example 13O4Particle size diagrams after 3 hours and 48 hours after co-extrusion through the membrane, and RBC-PEG-Fe3O4 Particle size distribution 5 days after passage through 200nm film.
FIG. 10 is a fluorescence spectrum of DSPE-PEG-Rho (Rho is rhodamine isothiocyanate) blended with red cell membranes at different volume ratios in example 1.
FIG. 11 is the magnetic attraction diagram and magnetic field movement diagram of the targeted magnetic nanoparticle-cancer cell prepared in example 1;
FIG. 12 shows magnetic nanoparticles (RBC-PEG-Fe) encapsulated in erythrocyte membrane prepared in example 1 of the present invention3O4) The precise targeting of the cancer cells and the evasion of immune cells are compared;
(a) a549 carcinoma cell location distribution; (b) a549 adsorbing RBC-PEG-Fe prepared by the invention3O4The fluorescence distribution of the particles; (c) PMBC cell location distribution; (d) PMBC co-incubated RBC-PEG-Fe prepared by the invention3O4Particle fluorescence distribution.
FIG. 13 is a CCK8 assay of RBC-PEG-Fe prepared by the present invention in example 13O4Cell activity of a549 by particle treatment and infrared irradiation;
(a) treating A549 cells by irradiation of medicine and infrared ray for 5 min; (b) treating A549 cells with medicine; (c) a549 cells have no medicine and no infrared radiation; (d)1 hour after adding the A549 cells into the CCK 8; (e) 3 hours after the addition of the A549 cells to the CCK 8; (f) a549 cells were added 12 hours after CCK 8.
FIG. 14 is a CCK8 assay of RBC-PEG-Fe prepared in example 1 of the present invention3O4Cellular activity of particle-treated and infrared-irradiated PMBC;
(a) adding pure medicines with different volumes into a culture medium; (b) adding medicine into PMBC, irradiating with infrared ray for 5min, and adding CCK8 for one hour; (c) PMBC add drug and CCK8 for one hour; (d) adding drugs into PMBC, irradiating with infrared ray for 5min, and adding CCK8 for twelve hours; (e) PMBC was added to the drug and CCK8 was added for twelve hours.
Detailed Description
The technical solution of the present invention is further described below by specific examples. The following examples are further illustrative of the present invention and do not limit the scope of the present invention.
Example 1
(1)PEG-Fe3O4Preparation:
2.00g(0.01mol)FeCl2·4H2O，4.725g(0.02mol)FeCl3·6H2and adding 20mL of distilled water into the O, adding 6g of PEG6000, and carrying out ultrasonic treatment for 10min (the power of an ultrasonic cell disruptor is 800W, and the frequency is 20kHz) until the O is dissolved. Stirring at 1400r/min under nitrogen atmosphere, heating to 60 deg.C, and reacting for 30 min. Then 6g of PEG6000 solution is added dropwise, the volume of the solution is about 1/5 of the reaction solution, the reaction solution is transferred to a polytetrafluoroethylene reaction kettle, the temperature is set to be 160 ℃, the temperature is raised for 1 hour, and the reaction is carried out for 5 hours. To obtain Fe3O4Precipitating nanoparticles, washing the precipitate with pure water and ethanol, repeating the steps for multiple times, and drying in a vacuum oven at 60 deg.C for 10 hr to obtain PEG-Fe3O4And (3) nanoparticles.
(2) Extraction and targeted modification of red cell membrane fragments:
subpackaging sheep blood into a centrifuge tube, centrifuging at 6000r/min for 15min, removing supernatant from the red blood cells, washing with ice PBS, repeating for three to four times, soaking and crushing the remaining red blood cells with ice 1/4 times of PBS hypotonic solution for 1 hour, centrifuging at 13000r/min for 15min, leaving the precipitate, removing supernatant, washing the precipitate with ice PBS, repeating for multiple times until the precipitate becomes light pink or white, and storing at 4 ℃. Determining a standard curve on ultraviolet by using lecithin to obtain the concentration of erythrocyte membranes, and adjusting the concentration to be 3 mg/ml; preparing 1mg/ml of sodium taurocholate, wherein the sodium taurocholate is prepared by the following steps: mixing erythrocyte membrane 1:4(m/m), dissolving in ethanol (10mL) with five times of volume, ultrasonically mixing, loading into a syringe, rapidly injecting into PBS solution at 45 ℃ of 100mL, and rotary-steaming at 60 ℃ for 25 min.
And (3) targeted modification process: mixing DSPE-PEG (DSPE-PEG2000) and targeting molecules and/or fluorescent molecules in a PBS (phosphate buffer solution) according to a molar ratio of 1:1 to form a DSPE-PEG-targeting molecule and/or fluorescent molecule solution (namely DSPE-PEG-X), adding DSPE-PEG-X into a very small amount of erythrocyte membranes diluted by the PBS solution, mixing the mixture in a shaker at 37 ℃, and stirring the mixture for 1 hour; red cell membrane: DSPE-PEG-X is 4:1 by mass. X in the DSPE-PEG-X is a fluorescent molecule (Rho) or a targeting molecule (FA), or the fluorescent molecule and the targeting molecule are mixed in a PBS solution according to a molar ratio of 1: 1.
(3) Erythrocyte membrane-PEG-Fe3O4The preparation of (1):
adding ultra-pure water into PEG6000 and sodium oleate at a mass ratio of 1:1 to prepare 6% dispersion, PEG-Fe3O4The concentration in the dispersion was 200. mu.g/mL; mixing PEG-Fe3O4Adding ultrasound for half an hour, controlling the power of an ultrasonic cell disruptor to be 800W and the frequency to be 20kHz, standing for 24 hours, removing sediment and taking an upper suspension. The target modified erythrocyte membrane is blended with the supernatant suspension, and each milliliter of PEG-Fe3O4The proportion of the two components in the mixed solution of the target modified erythrocyte membrane is 200 mug to 3mg, the mixed solution is stirred and heated to 40 ℃, an extruder is kept in water bath at 40 ℃, then the Whatman membrane is placed in a liposome extruder, large particles possibly blocking the microporous membrane are filtered out by sequentially passing through 800nm and 400nm, and then the mixed large particles are extruded for 7 times through the aperture of 200 nm. Dialyzing with 70nm dialysis bag at room temperature for 48 hr, standing at 4 deg.C for 48 hr, removing precipitate, and collecting upper layer liquid.
The following is a search for conditions based on example 1 (single factor change, other conditions are the same):
FIG. 2 is a comparison of the dispersion effect of PEG6000 sodium oleate dispersant in different proportions in example 1;
TABLE 1 dispersant Effect of different ratios of sodium oleate to PEG
Figure 2 and table 1 show different ratios of sodium oleate to PEG6000 as 1:1,1: 2,2: 1 and ultrapure water are prepared into 3 percent, 4 percent, 6 percent and 8 percent of dispersion liquid by mass ratio, and the same amount of PEG-Fe is added3O4Dropping into the solution, performing ultrasonic treatment for 30min, standing for 24 hr, and observing the suspension property after the liquid is stable. Due to PEG-Fe3O4When the concentration reaches hundreds of micrograms per milliliter, all the liquid is layered to different degrees, but after 24 hours, the liquid basically reaches the stability, the precipitation speed can be ignored, the upper suspension liquid is the thickest after the dispersion liquid with the mass ratio of 6 percent is stabilized, and the suspension property is the best. And PEG-Fe without dispersant treatment3O4After 24 hours of standing, the suspension property of the suspension was almost maintained.
FIG. 3 is a comparison of the dispersing effect of different dispersants used in step (1) or step (3) of example 1 of the present invention;
in the figure: (a) PEG6000-Fe3O4Adding a sodium dodecyl benzene sulfonate dispersing agent; (b) chitosan oligosaccharide OCS-Fe3O4Adding PEG-sodium oleate dispersant; (c) sodium alginate-Fe3O4Adding PEG-sodium oleate dispersant; (d) PEG6000-Fe3O4Adding a polyvinyl alcohol-sodium dodecanesulphonate dispersing agent; (e) PEG600 coated Fe3O4Adding PEG-sodium oleate dispersant; (f) OCS-Fe on the left3O4Extrusion of 200nm film, PEG-Fe on the right3O4A 200nm film was extruded. FIGS. (a) - (e) are schematic diagrams of PEG-Fe dispersed with different dispersants3O4In case of failure after 24 hours of standing, OCS-Fe can be seen in the graph (f)3O4The residue on the polycarbonate membrane is far larger than that of PEG-Fe3O4。
FIG. 4 shows PEG-Fe prepared in example 13O4Five-day DLS particle size monitoring graph;
in the figure (a) Fe after ultrasonication3O4The particle size distribution of the nanoparticles; (b) after standing for 1 day, Fe3O4The particle size distribution of the nanoparticles; (c) fe after 3 days of standing3O4The particle size distribution of the nanoparticles; (d) after standing for 5 days Fe3O4The particle size distribution of the nanoparticles.
As shown, the PEG-Fe pairs are continuously processed for five days3O4The particle size of the magnetic suspension is monitored, and the particle size is kept stable at about 80nm, which indicates that the magnetic fluid is convenient to be used for preparing RBC-PEG-Fe3O4Subsequent wrapping work.
FIG. 5 shows PEG-Fe prepared in example 13O4And (4) a magnetic property characterization chart.
The graphs (a) and (b) show the magnetic field responsiveness of nanoparticles, but the particle particles are too small and wait for more than 1 hour until the responsiveness appears, and the hysteresis loop of the graph (c) shows that the magnetic properties of the particle material are higher than the average value, and the magnetic response time is long in relation to the small particle diameter and in relation to the magnetic properties of the magnetic particle material.
FIG. 6 is a TEM image of the erythrocyte membrane prepared in example 1 after being dispersed by softening extrusion;
the flexibility of the erythrocyte membrane treated by the sodium taurocholate is enhanced, and the erythrocyte membrane enters an extruder under the slightly-hot environment of 45 ℃ and successfully passes through a 200nm membrane. The erythrocyte membrane solution extruded by the 200nm membrane is dropped on a copper net, and is dried in the shade for 72 hours and then is photographed into a TEM picture, so that the vesicle formed by the extruded cell membrane has relatively uniform grain diameter, a plurality of cell fragments exist around the vesicle, the dispersion degree is satisfactory, the size of the membrane cluster is stable, and only a very small amount of cell membrane diameter expansion agglomeration phenomenon exists.
FIG. 7 is a graph showing the effect of UV-irradiation on erythrocyte membrane deformability of sodium taurocholate in different ratios and at different temperatures in example 1
In the figure a, the influence of different temperatures on the membrane flexibility of pure red blood cells;
b, the influence of different temperatures on the ultraviolet peak value of the extruded pure erythrocyte membrane;
c influence of different surfactant ratios on erythrocyte membrane softness;
d, influence of different surfactant ratios on ultraviolet peak values of the softened erythrocyte membranes after extrusion;
e influence of different temperatures on the softness of the softened erythrocyte membranes;
f, influence of different temperatures on ultraviolet peak value of the softened erythrocyte membrane after extrusion;
extruding pure erythrocyte membranes which are not softened at different temperatures, erythrocyte membranes which are treated by taking sodium taurocholate as a surfactant in different proportions by using 100nm polycarbonate membranes, and fixing at different temperatures by using a fixing device 1:4, measuring ultraviolet peak of the liquid obtained by the red cell membrane softened by the sodium taurocholate to obtain red cell membrane liquid with different concentrations, wherein the red cell membrane liquid can reflect the deformability of the red cell membrane under different conditions, and the more red cell membranes passing through the micropores of 100nm under high pressure, the better the deformability of the membrane is. The effect of temperature on pure red blood cell membrane deformability is shown in FIG. a and FIG. b, and the best deformability was obtained for pure red blood cell membranes at 40 ℃. As shown in the graphs c and d, the effect of sodium taurocholate treatment on erythrocyte membrane with different mass ratios on the flexibility is obtained, and when the sodium taurocholate is 1/4 with the erythrocyte membrane mass, the erythrocyte membrane softening effect is best. As shown in the graphs e and f, the flexibility of the sodium taurocholate with fixed 1/4 erythrocyte membrane quality at different temperatures is obtained, and 40 ℃ is the optimal temperature for softening the erythrocyte membrane deformability, so that the maximum deformability of the erythrocyte membrane can be obtained by treating the erythrocyte membrane with the sodium taurocholate with 1/4 erythrocyte membrane quality and extruding at 40 ℃.
FIG. 8 is a graph of the ratio of 1:4(m/m) research on the stability of erythrocyte membranes modified by sodium taurocholate and unmodified erythrocyte membranes;
(a) pure RBC particle size distribution; (b)1/6 RBC-modified RBC particle size distribution of sodium taurocholate of RBC mass; (c)1/5RBC size distribution modified by sodium taurocholate; (d)1/4RBC size distribution modified by sodium taurocholate;
FIG. 8 shows the DLS particle size distribution measured 48 hours after the pure erythrocyte membrane and the erythrocyte membrane modified by sodium taurocholate with different proportions are extruded through a 100nm pore membrane, the just prepared erythrocyte membrane is gradually destabilized and fused from 100nm along with the time, the particle size is enlarged, the destabilization of the pure erythrocyte membrane is the largest, the cell size is increased to about 800nm from 100nm during the initial extrusion within 48 hours, and in FIG. 8, the sodium taurocholate has the function of stabilizing the erythrocyte membrane to a certain extent, the more the dosage is, the larger the stabilizing effect is, and the particle size of the erythrocyte membrane modified by the sodium taurocholate with the quality of 1/4 erythrocyte membrane is only increased to about 300nm within 48 hours.
FIG. 9 shows modified RBC (with both Rho and FA attached) and PEG-Fe as in example 13O4Particle size diagrams after 3 hours and 48 hours after co-extrusion through the membrane, and RBC-PEG-Fe3O4 Particle size distribution 5 days after passing 200nm film;
FIG. 9 shows RBC and prepared PEG-Fe modified by both targeting molecule (FA) and fluorescent molecule (Rho)3O4After passing through a 200nm polycarbonate film, the particle size is intensively distributed about 200nm in hours after preparation, no obvious particle size expansion is caused, and Fe less than 200nm can be presumed3O4Particles of more than 200nm Fe coated3O4The particles had been filtered off in a gradient extrusion of 800nm, 400nm and then 200 nm. OCS-Fe3O4Only 400nm film can be passed. Although the particle size of the powder is 145nm, each particle cannot correspondingly pass through each pore of the 200nm film in the high-pressure process, the powder cannot be fully dispersed in the extrusion compaction process due to the overlapping of a large number of particles, and some particles are adsorbed on the polycarbonate film, so that the powder fails to pass through the 200nm film. Sodium alginate-Fe3O4The membrane experiment could not be performed because of the total precipitation. The magnetic nano-particles with high dispersity and small particle size can more easily and fully utilize the adsorption space of folate receptors on cancer cell membranes, and are favorable for adsorbing a larger amount of magnetic particles, so that RBC-PEG-Fe passing through a 200nm membrane is selected3O4And (6) entering the next cell experiment. As can be seen in FIG. 9, RBC-PEG-Fe3O4The particle size distribution of the initially extruded 200nm membrane is almost unchanged in 48 hours, so that the membrane can play a role in preventing the particles from being instable and fused under the combined action of PEG and sodium taurocholate, and the erythrocyte membrane replaces PEG6000 and sodium oleate dispersing agent to a certain extent to play a role in stabilizing the particle size.
FIG. 10 is the fluorescence spectra of DSPE-PEG-Rho blended with erythrocyte membrane at different volume ratios in example 1;
the figure shows the fluorescence spectra obtained by adding 200 microliters of erythrocyte membranes with the concentration of 3mg/ml, adding 1mg/ml of DSPE-PEG-Rho with different volumes, mixing by a shaker, carrying out high-speed centrifugation and precipitation, and carrying out redispersion once, wherein the fluorescence of 120 microliters to 140 microliters is stable, and the optimal volume is 1:5(m/m) -1:4 (m/m). The further increase of the DSPE-PEG-Rho concentration is of little significance.
FIG. 11 shows the erythrocyte membrane-coated magnetic nanoparticles (RBC-PEG-Fe) prepared in example 13O4) -cancer cell sorption and magnetic field motility profiles;
magnetic nanoparticles (RBC-PEG-Fe) encapsulated by erythrocyte membrane3O4) The mesoerythrocyte membrane has been modified with targeting molecules (FA) and fluorescent molecules (Rho).
Graphs (a) and (b) show the relative distribution positions of the cancer cells and the fluorescent targeting magnetic nanoparticle molecules, so that the cancer cell membranes on which the fluorescent targeting magnetic nanoparticles are distributed can be clearly seen, and the positions of the cancer cells in the graph (a) and the positions of the fluorescence in the graph (b) can be in one-to-one correspondence; the graphs (c), (d), (e) and (f) show that the cancer cells treated with the fluorescent targeting magnetic nanoparticles are placed in a magnetic field and are directionally controlled to move by the magnetic field, which indicates that the cancer cells are endowed with magnetic responsiveness by the targeting magnetic nanoparticles.
FIG. 12 shows magnetic nanoparticles (RBC-PEG-Fe) encapsulated in erythrocyte membrane prepared in example 1 of the present invention3O4) The precise targeting of the cancer cells is compared with the evasion of immune cells;
(a) a549 carcinoma cell location distribution; (b) a549 adsorbing RBC-PEG-Fe prepared by the invention3O4The fluorescence distribution of the particles; (c) PMBC cell location distribution; (d) PMBC co-incubated RBC-PEG-Fe prepared by the invention3O4Particle fluorescence distribution;
magnetic nanoparticles (RBC-PEG-Fe) encapsulated by erythrocyte membrane3O4) The mesoerythrocyte membrane has been modified with targeting molecules (FA) and fluorescent molecules (Rho).
Respectively culturing A549 cell and sheep blood PMBC (peripheral blood mononuclear cell) with peripheral blood mononuclear cellPlates, incubation at 37 ℃ for 24 hours and addition of different concentrations of RBC-PEG-Fe3O4After adding medicine for several hours, high content photography is used for displaying the distribution situation of fluorescence, the edges of the obtained cancer cells in the graphs (a) and (b) are clearly displayed by Rhodamine6G, the positions of most of the cancer cells in the graphs (a) and (b) can be almost in one-to-one correspondence with the positions of the fluorescence, the graphs (c) and (d) display denser PMBC distribution, the distribution of the fluorescence particles does not strictly follow the position distribution of the original cells, the randomness is stronger, and RBC-PEG-Fe can be presumed3O4PMBC are not easy to adsorb, and phagocytosis is not obvious.
FIG. 13 is a CCK8 assay of RBC-PEG-Fe prepared in example 1 of the present invention3O4Cell activity of a549 by particle (i.e., drug) treatment and infrared irradiation;
(a) treating A549 cells by irradiation of medicine and infrared ray for 5 min; (b) treating A549 cells with medicine; (c) a549 cells have no medicine and no infrared radiation; (d)1 hour after adding the A549 cells into the CCK 8; (e) 3 hours after the addition of the A549 cells to the CCK 8; (f) 12 hours after the addition of the A549 cells to CCK 8;
magnetic nanoparticles (RBC-PEG-Fe) encapsulated by erythrocyte membrane3O4) The mesoerythrocyte membrane has been modified with targeting molecules (FA) and fluorescent molecules (Rho).
FIG. 13 shows RBC-PEG-Fe obtained from 450nm absorbance data obtained by microplate reader3O4Effect of particle and infrared irradiation on proliferation rate and activity of a549 cancer cells within 12 hours. As shown in FIG (a), A549 cells were treated with RBC-PEG-Fe3O4After the particles are treated by 5min of infrared irradiation, the proliferation speed of the plate holes with 20 mul is the maximum along with the time, the proliferation speed is slightly increased compared with that of the holes without medicine only irradiation, low dose heat probably from the excitation of a small amount of nanoparticles by infrared rays promotes the proliferation of cancer cells, the proliferation speed of the cells is obviously reduced along with the increase of the particle dose, compared with a graph (b), the absorbance of the graph (a) starts to be reduced from the 20 mul plate holes, the absorbance of the graph (b) starts to be reduced from the 40 mul plate holes, and the RBC-PEG-Fe is enhanced by visible light irradiation3O4The anti-cancer cellularity of the particle. As shown in FIG. c, the blank group without nanoparticles and infrared treatment exhibited the highest cancer cell proliferation rate. Such asAs shown in the graph (d), the absorbance of the cells treated differently was concentrated in a relatively narrow range of 0.3-0.5 within one hour of the addition of the cells to CCK8, and the difference was not easily seen, and after 3 hours, as shown in the graph (e), the proliferation rate difference was further increased after 12 hours, as shown in the graph (f), in the drug-and infrared-added cell group, which began to proliferate slower than the drug-added group, and slower than the non-treated group.
FIG. 14 is a CCK8 assay of RBC-PEG-Fe prepared in example 1 of the present invention3O4Cellular activity of particle (i.e., drug) treated and infrared irradiated PMBC;
(a) adding pure medicines with different volumes into a culture medium; (b) adding medicine into PMBC, irradiating with infrared ray for 5min, and adding CCK8 for one hour; (c) PMBC add drug and CCK8 for one hour; (d) adding drugs into PMBC, irradiating with infrared ray for 5min, and adding CCK8 for twelve hours; (e) PMBC was added to the drug and CCK8 was added for twelve hours.
Magnetic nanoparticles (RBC-PEG-Fe) encapsulated by erythrocyte membrane3O4) The mesoerythrocyte membrane has been modified with targeting molecules (FA) and fluorescent molecules (Rho).
FIG. 14 shows the absorbance data of PMBC at 1 and 12 hours after CCK8 addition, due to RBC-PEG-Fe3O4The particles have certain influence on absorbance, and PMBC has weak adherence, so that the nanoparticles cannot be directly poured off to remove the influence. As can be seen from each of fig. 14, the PMBC proliferated in the drug-and-infrared irradiation treatment for 1 hour and 12 hours was in a linear relationship with the increase in the drug concentration course, and the slope of the curve was almost parallel to the absorbance curve of the pure drug concentration regardless of whether the drug was irradiated with infrared light, and the PMBC proliferated in the drug-free and infrared-free conditions for 1 hour fluctuated around the average value of 0.35 and for 12 hours fluctuated around the average value of 0.66. The stability of the proliferation speed of the PMBC at 1-12 hours can be presumed from the absorbance curve which grows in parallel along the time, and the RBC-PEG-Fe is not used in the process3O4The particles and infrared radiation effect and a significant speed reduction or enhancement occurs.
Claims (10)
1. A preparation method of magnetic nanoparticles wrapped by erythrocyte membranes is characterized by comprising the following steps:
（1）PEG-Fe3O4preparing;
(2) extracting and targeted modifying erythrocyte membrane fragments;
(3) erythrocyte membrane-PEG-Fe3O4The preparation of (1): mixing PEG-Fe3O4Adding a dispersion prepared by adding ultra-pure water into PEG6000-10000 and sodium oleate, performing ultrasonic treatment, standing, removing precipitates, taking an upper layer suspension, blending and stirring the target modified erythrocyte membrane obtained in the step (2) and the upper layer suspension, putting the mixture into a liposome extruder, and extruding for multiple times to obtain the target modified erythrocyte membrane;
step (2) extraction process of erythrocyte membrane fragments: centrifuging anticoagulated mammal blood to precipitate erythrocyte, removing supernatant, washing with ice PBS, soaking the crushed erythrocyte in hypotonic solution, centrifuging, leaving precipitate to remove supernatant, washing the precipitate with ice PBS, repeating for several times until the precipitate becomes light pink or white, and mixing erythrocyte membrane and sodium taurocholate solution according to the formula: red cell membrane = 1: mixing at 6 or 1:4 mass ratio, dissolving in 4-6 times volume of ethanol, ultrasonically mixing, loading into an injector, rapidly injecting PBS (phosphate buffer solution) with volume 8-12 times of ethanol solution at 40-45 deg.C, and rotary steaming at 50-70 deg.C for 15-25 min;
in the step (3), the mass ratio of PEG6000-10000 to sodium oleate is 1: 2-2: 1, adding ultrapure water to prepare 6-8% dispersion liquid.
2. The production method according to claim 1,
in the step (3), the mass ratio of PEG6000-10000 to sodium oleate is 1:1, adding ultrapure water to prepare 6% dispersion liquid; PEG-Fe3O4The concentration in the dispersion was 100-300. mu.g/mL.
3. The preparation method according to claim 1, wherein the ultrasound in step (3) is performed by using an ultrasonic cell disruptor with a power of 600- "1000W and a frequency of 15-25 kHz.
4. The preparation method according to claim 1, wherein the targeted modified erythrocyte membrane obtained in step (2) and the supernatant suspension are blended and stirred in step (3), heated to 38-45 ℃, and then extruded.
5. The method according to claim 1, wherein the PEG-Fe is added per ml in the step (3)3O4The proportion of the two in the mixed solution of the red cell membrane and the target modified red cell membrane is 150-250 mug: 3 mg.
6. The method according to claim 1, wherein the step (3) comprises extruding the mixture sequentially from 800nm to 400nm using a liposome extruder, and extruding the mixture 3 to 7 times under a 200nm membrane at 40 to 45 ℃.
7. The production method according to claim 1,
step (1) PEG-Fe3O4The preparation process comprises the following steps: FeCl2With FeCl3Mixing the raw materials in a ratio of 1: 1.5-1: distilled water is added according to the molar ratio of 2, and FeCl is added2PEG6000-10000 with the mass 2-4 times, and ultrasonic processing for 5-10min until dissolution, wherein the ultrasonic processing adopts an ultrasonic cell disruption instrument, the power is 600-1000W, and the frequency is 15-25 kHz; stirring at 1400-2600r/min under inert atmosphere, heating to 45-60 ℃, and reacting for 30-40 min; adding FeCl2Transferring the PEG6000 solution with the mass 2-4 times of that of the reaction solution to a polytetrafluoroethylene reaction kettle, setting the temperature at 160-200 ℃, and reacting for 5-10 hours; to obtain Fe3O4Precipitating nanoparticles, washing the precipitate with pure water and ethanol, repeating the steps for several times, and drying in a vacuum oven at 45-60 deg.C for 10-24 hr to obtain PEG-Fe3O4And (3) nanoparticles.
8. The production method according to claim 1,
step (2) targeted modification process of erythrocyte membrane fragments: mixing DSPE-PEG and targeting molecules in a PBS solution according to the molar ratio =1.5-1:1 to form the DSPE-PEG-targeting molecules, wherein the targeting molecules comprise at least one of folic acid and mannitol; adding DSPE-PEG-targeting molecules into a very small amount of erythrocyte membranes diluted by PBS solution, and mixing by a shaking table; the mass ratio of the erythrocyte membrane to the DSPE-PEG-targeting molecule is 4: 1-10: 1.
9. a magnetic nanoparticle coated with erythrocyte membrane obtained by the preparation method of any one of claims 1 to 8.
10. Use of the erythrocyte membrane-coated magnetic nanoparticle of claim 9 in the preparation of a targeting material.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104637644A (en) * | 2015-03-06 | 2015-05-20 | 山东大学 | Particle coating method for preparing magnetic liquid |
Non-Patent Citations (3)
| Title |
|---|
| Ferrofluid based on polyethylene glycol-coated iron oxide nanoparticles:Characterization and properties;Sonia García-Jimeno,et al.;《Colloids and Surfaces A: Physicochemical and Engineering Aspects》;20121226;第420卷;第74-81页 * |
| 基于RBC-OCS-Fe3O4磁性纳米粒子自组装与磁控制癌细胞运动的研究;彭逸舟，等;《科技风》;20200630;第280页 * |
| 聚乙二醇化Fe3O4作为光热治疗剂对大鼠C6胶质瘤细胞光热治疗的可行性研究;袁罡;《中国优秀博硕士学位论文全文数据库(博士)医药卫生科技辑》;20150815(第08期);第E072-4页 * |
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