CN109078183A - A kind of multifunctional bionic nanometer formulation and its preparation method and application based on cancer cell membrane - Google Patents
A kind of multifunctional bionic nanometer formulation and its preparation method and application based on cancer cell membrane Download PDFInfo
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Abstract
The invention discloses a kind of multifunctional bionic nanometer formulation and its preparation method and application based on cancer cell membrane, the nanometer formulation forms kernel by chemotherapeutics desalination adriamycin and photo-thermal therapy drug indocyanine green, outer layer forms shell structure by human cervical carcinoma cell film, when preparation, it is prepared for one kind first by two kinds of anticancer drug (DOX, ICG) the nanoparticle kernel that self assembly obtains, then, human cervical carcinoma cell film, which is superscribed, in outer layer carries out bionic camouflage, finally obtain a kind of multifunctional nano preparation for being provided simultaneously with chemotherapy and photo-thermal therapy and there is cancer cell targeting.The nanometer formulation biological safety is high, and preparation is simple, causes the disturbance of cell membrane in the stimulation of near infrared light, the final quick release for realizing drug has very big application prospect.
Description
Technical field
The present invention relates to medical science technical fields, are provided simultaneously with chemotherapy and photo-thermal therapy effect more particularly to a kind of
Multifunctional bionic nanometer formulation based on cancer cell membrane and its preparation method and application.
Background technique
Cancer causes millions of people dead every year, is a big killer of human health.To the drug therapy of cancer
Research has important clinical meaning.However monotherapy significantly limits it and leads in clinic there are its intrinsic limitation
The application in domain.For example, a kind of conventional means of the chemotherapy as treating cancer, but its therapeutic effect receives dose dependent and causes
Toxic effect.Therefore, many studies and clinical applications show multiple therapy methods combination therapy, can play cooperative compensating
Effect, shows efficient tumor suppression, and a variety of advantages such as low side effect and delay drug resistance have good development prospect.
In a variety of synergistic treatment modes, the treatment mode of " photo-thermal therapy+chemotherapy " can effectively kill cancer cell, reach
Synergistic therapeutic effect.Currently, being usually loaded into chemotherapeutics and photosensitizer altogether in synthesis nano-carrier, to obtain Nano medication defeated
Send system.More this Nano medication carrier systems usually have certain toxic side effect in blood circulation, are easy to by netted interior
Skin is removed, and targeted therapy difficult to realize, therapeutic effect are poor.In recent years, bionic camouflage Nano medication transportation system
Gradually people is caused to pay close attention to.Using cell membrane packaging medicine, biocompatibility is not only increased, extends its blood circulation time,
Also improve aggregation of the delivery system near cancer cell.It, can be with since cell membrane surface has specific proteins molecule
By cancer cell identification, to achieve the purpose that targeted therapy.However, realizing that cell membrane loads drug quick release is still one
Challenge.
Summary of the invention
It is imitative based on cancer cell membrane that in view of the technical drawbacks of the prior art, it is an object of the present invention to provide a kind of
Raw multifunctional nano preparation and its preparation method and application.
The technical solution adopted to achieve the purpose of the present invention is:
A kind of multifunctional bionic nanometer formulation based on cancer cell membrane, by the cancer cell of Nano medication particle kernel and outer layer
Film is constituted, in which: the Nano medication particle kernel is by chemotherapeutics and the photo-thermal therapy drug structure being coated on outside chemotherapeutics
At, in which: the chemotherapeutics is with hydrophobic adriamycin, and the photo-thermal therapy drug is with amphiphilic indoles cyanines
Green, the cancer cell membrane is the cell membrane fragments of the cervical cancer cell of logarithmic phase growth.
Preferably, the nanometer formulation has regular spherical design, average grain diameter 220-240nm.
Preferably, the adriamycin is de- doxorubicin hydrochloride.
Another aspect of the present invention further includes a kind of preparation method of nanometer formulation, comprising the following steps:
(1) chemotherapeutics micro syringe is slowly dropped to shape in the pure water of magnetic agitation dropwise under the conditions of being protected from light
At the nanoparticle of chemotherapeutics, 20-40min is stirred；
(2) photo-thermal therapy drug is dissolved in water, and is added dropwise to step (1) institute dropwise under the conditions of being protected from light with micro syringe
It is stirred in the nano-particle solution stated, stirs 10-15h, photo-thermal therapy drug is coated on the outside of chemotherapeutics, obtains nanoparticle
Sub- kernel, wherein the molar ratio of the photo-thermal therapy drug and chemotherapeutics is 1:(2-3)；
(3) the nanoparticle kernel for obtaining step (2) and the classification of cancer cell membrane fragment squeeze out, wherein the nano particle
Kernel and cancer cell membrane fragment are mixed according to the ratio of mass ratio 1:(3-4)；
Wherein step (1)-(3) carry out under light protected environment.
Preferably, the chemotherapeutics in the step (1) is the hydrophobic adriamycin of desalination, the photo-thermal in the step (2)
Therapeutic agent is indocyanine green, and the cancer cell membrane in the step (3) is the cell membrane of the human cervical carcinoma cell of logarithmic phase growth.
Preferably, the classification in the step (3) squeezes out the following steps are included: nano particle kernel and cancer cell membrane is broken
Piece according to mass ratio 1:3 ratio mix, vortex mixed is uniform, stand 30-40 minute after successively pass through 1 μm, 0.85 μm with
0.45 μm of miillpore filter distinguishes filter 23-5 times to get the multifunctional bionic nanoparticle formulations wrapped up to cancer cell membrane.
Preferably, the preparation method of the cancer cell membrane fragment in the step (3) is the following steps are included: through hypotonic extraction method
Broken cell membrane removal content obtains cell membrane fragments, is then sonicated to obtain uniform nanoscale cancer cell membrane fragment, entirely
Journey ice bath is placed.
Preferably, cancer cell membrane uses hypotonic extraction method in the step (3), comprising the following steps: logarithmic phase is taken to grow
Human cervical carcinoma cell, using cell scraper by under cell scraper, cell, the PBS washing of isotonic pre-cooling, centrifugation is collected by centrifugation in 700g
Afterwards, abandon supernatant, be added hypotonic solution ice bath 15-20 minute, by the cell solution after ice bath in room temperature and liquid nitrogen multigelation,
Until cell is completely broken, then, 700g is centrifuged to obtain supernatant, and 14000g is centrifuged to obtain precipitating to be cell membrane fragments, on
State -80 DEG C of cell membrane fragments -- 90 DEG C of freeze-dryings save.
Preferably, the multifunctional bionic nanometer formulation the preparation method is as follows:
5mg doxorubicin hydrochloride is added in 10mL sample bottle and is protected from light processing, 4.5mL dimethyl sulphoxide solution, which is added, makes salt
Sour adriamycin sufficiently dissolves, and 0.5mL triethylamine is added under the conditions of magnetic agitation, and adjustment magneton revolving speed is 600rpm, reaction overnight.
Products therefrom is desalination adriamycin.
The above-mentioned desalination adriamycin (DOX) of 100 μ L is slowly dropped to magnetic force with micro syringe dropwise under the conditions of being protected from light
Adriamycin nano particle is formed in the 5mL pure water of stirring.After 30 minutes, 200 μ L indocyanine green solution (1mg/ of water will be dissolved in
ML) (ICG) is added dropwise in above-mentioned nano-particle solution is dropwise stirred overnight by the same method, obtains nanoparticle kernel
(DINPs)。
The hypotonic extraction method of cancer cell membrane is the following steps are included: extract reagent using epicyte protein and suppressor proteins
Box extracts cell membrane.The human cervical carcinoma cell for taking logarithmic phase to grow, using cell scraper by under cell scraper, 700g is centrifuged 5 minutes
Cell is collected, is washed using the PBS (pH 7.4) of isotonic pre-cooling, after centrifugation, abandons supernatant, is added quiet under hypotonic solution condition of ice bath
It sets 15 minutes.By above-mentioned cell solution in room temperature and multigelation in liquid nitrogen, until cell is completely broken.Then, 700g is centrifuged
Supernatant is carefully obtained within 10 minutes, it is cell membrane fragments (CMs) that 14000g, which is centrifuged and obtains within 30 minutes precipitating,.Above-mentioned cell membrane is broken
- 80 DEG C of freeze-dryings of piece save.
The nano particle is merged with the package of cancer cell membrane fragment the following steps are included: first that the cancer cell membrane of extraction is broken
Piece is dissolved in pure water, obtains finely dispersed nanoscale cell film fragment through ultrasonic cell disruption instrument ice-bath ultrasonic process.So
It is afterwards that nano particle and cancer cell membrane fragment is uniform according to mass ratio 1:3 ratio vortex mixed, 30 minutes are stood, is then successively passed through
Crossing 1 μm, 0.85 μm and 0.45 μm miillpore filter, filter 23 time obtains cancer cell membrane package multifunctional bionic nano particle respectively
(DICNPs)。
Another aspect of the present invention further includes the multifunctional bionic nanometer formulation, is preparing the application on anticancer drug.
In above scheme, it is due to hydrophobic effect power drive that DOX, which is added dropwise in water and forms nano particle,.ICG is wrapped in
It is amphipathic nature material that DOX nanoparticle outer layer, which is due to ICG, and in hydrophobic layer is wrapped in, hydrophilic layer is wrapped in outer.
Compared with prior art, the beneficial effects of the present invention are:
1, the cancer cell membrane multifunctional bionic nanometer formulation prepared by the present invention has chemotherapy and photo-thermal therapy synergistic treatment
Effect there is good biocompatibility, degradability, can effectively avoid reticular endothelium due to outer layer covers cancer cell membrane
The phagocytosis of system, while there is height targeting to cancer cell of the same race.
2, internal layer is two kinds of medicinal compositions, and DOX is chemotherapeutics, and ICG is photo-thermal therapy drug, it can be achieved that chemotherapy and light
Heat cure synergistic therapeutic effect, while the photothermal response of ICG can accelerate the rate of release of DOX drug.
3, the nanometer formulation not only has good biocompatibility, and the ability with active targeting cancer cell, realizes
The ability of chemotherapy and photo-thermal therapy synergistic treatment, and near infrared light (NIR) stimulating drug discharges process, makes drug in cancer cell
Position burst release, utmostly improves therapeutic effect, is of great significance during treatment of cancer.
Detailed description of the invention
The present invention is based on the TEM of cancer cell membrane multifunctional bionic nanometer formulation DICNPs figures by Fig. 1.
The present invention is based on the partial sizes of cancer cell membrane multifunctional bionic nanometer formulation DICNPs by Fig. 2.
The present invention is based on the targetings of cancer cell membrane multifunctional bionic nanometer formulation DICNPs to analyze (scale: 25 μm) by Fig. 3.
The present invention is based on the tablets in vitro figures of cancer cell membrane multifunctional bionic nanometer formulation DICNPs by Fig. 4.
The present invention is based on the analyses of the cell dark toxicity of cancer cell membrane multifunctional bionic nanometer formulation DICNPs by Fig. 5.
The present invention is based on the analyses of the cell phototoxicity of cancer cell membrane multifunctional bionic nanometer formulation DICNPs by Fig. 6.
Specific embodiment
The present invention is described in further detail below in conjunction with the drawings and specific embodiments.It should be appreciated that described herein
Specific embodiment be only used to explain the present invention, be not intended to limit the present invention.
Doxorubicin hydrochloride and indocyanine green are bought in Dalian U.S. logical sequence Technology Co., Ltd., epicyte protein and cell
Protein Extraction Reagent kit purchase is starched in green skies Bioisystech Co., Ltd.
(1) doxorubicin hydrochloride takes off HCl treatment: it weighs 5mg doxorubicin hydrochloride and is added in 10mL sample bottle and be protected from light processing,
4.5mL dimethyl sulfoxide (DMSO) solution, which is added, dissolves doxorubicin hydrochloride sufficiently, under the conditions of magnetic agitation (600rpm, 37 DEG C)
0.5mL triethylamine, reaction overnight is added.Products therefrom is desalination adriamycin.
(2) prepared by nano particle kernel: the above-mentioned desalination adriamycin (DOX) of 100 μ L is being protected from light condition with micro syringe
Under be slowly dropped in the 5mL pure water of magnetic agitation formation adriamycin nano particle (DOX NPs) dropwise.It, will be molten after 30 minutes
Above-mentioned receive is added dropwise to dropwise under the conditions of being protected from light with micro syringe in 200 μ L indocyanine green solution (1mg/mL) (ICG) of water
It is stirred overnight in rice corpuscles solution, obtains nanoparticle kernel (DINPs).
(3) cancer cell membrane extracts: being extracted using Low Osmotic Method, is mentioned using epicyte protein and suppressor proteins extracts kit
Take cell membrane.Protease to avoid containing on film or adsorb makes memebrane protein change, and operation should be under 4 DEG C of low temperature below
It carries out.The human cervical carcinoma cell for taking logarithmic phase to grow, using cell scraper by under cell scraper, it is heavy that 700g is centrifuged 5min collection cell
It forms sediment, the PBS (pH 7.4) that isotonic pre-cooling is added is resuspended, and after 600g is centrifuged 5min, abandons supernatant, subsequent 600g is centrifuged 1min, with heavy
Residual liquid and the further sedimentation cell to form sediment on centrifugation tube wall, exhaustion residual of trying one's best.Hypotonic solution ice bath item is added
15 minutes are stood under part.By above-mentioned cell solution in room temperature and liquid nitrogen multigelation for several times until cell is completely broken.Then,
700g is centrifuged 10 minutes and carefully obtains supernatant, to remove nucleus, organelle and unbroken cell.Finally, 14000g is centrifuged
Obtaining within 30 minutes precipitating is cell membrane fragments (CMs).- 80 DEG C of freeze-dryings of above-mentioned cell membrane fragments save.
(4) preparation based on cancer cell membrane multifunctional bionic nanometer formulation DICNPs: first by the cancer cell membrane fragment of extraction
It is dissolved in pure water, obtains finely dispersed nanoscale cell film fragment through ultrasonic cell disruption instrument ice-bath ultrasonic process.Then
Nano particle and cancer cell membrane fragment is uniform according to mass ratio 1:3 ratio vortex mixed, 30 minutes are stood, is then successively passed through
Filter 23 time obtains cancer cell membrane package multifunctional bionic nano particle to 1 μm, 0.85 μm and 0.45 μm miillpore filter respectively
(DICNPs).Its form such as Fig. 1 is characterized with TEM.The observable bionical carrier has regular spherical structure in figure, and size is equal
One, partial size is in 220nm or so.The partial size and Zeta potential result such as Fig. 2 of the laser nano particle size analyzer determination nanoparticle.
Targeting characterization based on cancer cell membrane multifunctional bionic nanometer formulation DICNPs, it is thin with monkey source nephrocyte COS 7
Born of the same parents and 929 cell of smooth muscle cell L are control, and operating procedure is as follows: by HeLa cell, 7 cell of COS and 929 cell of L point
It is not inoculated in the burnt ware of copolymerization, each ware 1 × 105A cell, each ware culture medium 1mL.It is added after cell culture is adherent for 24 hours
DICNPs continues to cultivate 2h, then absorbs liquid and clean cell with PBS, use 33342 dyes core of Hochest
15min is added paraformaldehyde and fixes 30min, cleans, be eventually adding 1mL PBS solution after being cleaned up with PBS.It will be above-mentioned total
Focusing culture dish, which is placed under laser co-focusing, to be observed and shoots photo.
For tumor-targeting result as shown in figure 3, blue represents the fluorescence of nucleus Hochest 33342, red represents DOX
Fluorescence, green represents the fluorescence of ICG, and since DICNPs has the shell of cancer cell membrane to coat, this nanoparticle can be effective
Raising to the targeting of same tumor cell, specific binding (fluorescence has occurred in DICNPs and HeLa cell as can be seen from Figure
Intensity is stronger), and the associativity of DICNPs and 929 cell of 7 cell of COS and L is poor (fluorescence intensity is weak), in addition, DICNPs exists
Near infrared light is better than the experimental group (DICNPs) for not providing near infrared light according to the fluorescence intensity of lower experimental group (DICNPs+NIR)
Fluorescence intensity illustrates that near infrared light (NIR) has the function of stimulating drug release.
Tablets in vitro behavior based on cancer cell membrane multifunctional bionic nanometer formulation DICNPs: should using dialysis process evaluation
The tablets in vitro behavior of nanometer system, dissolution medium are the PBS phosphate buffer of pH 7.4.In 37 DEG C of thermostat water baths,
160rpm oscillation carries out the drug release behavior of DINPs, DINPs+NIR, DICNPs and DICNPs+NIR, respectively at 0,1,2,4,6,8,
12,24,36,48,60,72h time point exchanges equivalent PBS solution for, by the medium of above-mentioned taking-up with sepectrophotofluorometer (EX:
480nm, EM:560nm) it measures and calculates accumulative release rate.As can be seen from Figure 4, the nanoparticle drug for wrapping up cancer cell membrane is released
It has been placed with and has obviously slowed down, and wrapped up cell membrane and the drug release of near infrared light photograph has been added to have and be remarkably reinforced.
Cytotoxicity analysis based on cancer cell membrane multifunctional bionic nanometer formulation DICNPs: cytotoxicity is to pass through MTT
Method measurement.HeLa cell is with 6 × 103The density of a cell per well is seeded in 96 orifice plates.Before material is added, in 100 μ
L, which contains in the DMEM culture medium of 10%FBS, to be cultivated 24 hours.Be separately added into later various concentration DOX, ICG, DINPs and
DICNPs.One piece of plank adds illumination, and another piece of plank is protected from light processing, fresh with 200 μ L after it is co-cultured 24 hours with cell
The DMEM culture medium containing 10%FBS substitute original culture medium, and 20 μ L MTT (5mg/mL in are added in every hole
PBS buffer) solution, continue culture 4 hours in 37 DEG C of incubators, discards culture medium, 150 μ L DMSO are added in every hole,
Photon absorbing intensity of the solution at 570nm is detected in microplate reader.The relative survival rate of cell is calculated as follows: Cell
Viability (%)=(OD570 (sample)/OD570 (control)) × 100%, wherein OD570 (Control) and
OD570 (sample) respectively represents no sample processing and has the solution light absorption value after sample treatment.Resulting value is parallel real three times
The average value of resulting value is tested, value is shown as average value ± standard deviation (SD).Comparison diagram 5-6, it can be seen that not plus when NIR
DICNPs all has lesser cytotoxicity, and after NIR light is shone, the cell survival rate of DICNPs group has obvious drop
It is low, illustrate that cell membrane has preferable coating function to DINPs, under near-infrared light stimulation, drug has apparent release,
Cytotoxicity, which has, to be obviously increased.
Illustrative description is done to the present invention above, the description thereof is more specific and detailed, but can not therefore understand
For limitations on the scope of the patent of the present invention.It should be pointed out that for those of ordinary skill in the art, not departing from this
Under the premise of inventive concept, any simple deformation, modification or other skilled in the art can not spend creative labor
Dynamic equivalent replacement all belongs to the scope of protection of the present invention.Therefore, the scope of protection of the patent of the present invention should advise you to require with appended
Subject to.
Claims (10)
1. a kind of multifunctional bionic nanometer formulation based on cancer cell membrane, which is characterized in that by Nano medication particle kernel and outside
The cancer cell membrane of layer is constituted, in which: the Nano medication particle kernel is by chemotherapeutics and the photo-thermal being coated on outside chemotherapeutics
Therapeutic agent is constituted, in which: the chemotherapeutics is with hydrophobic adriamycin, and the photo-thermal therapy drug is with amphiphilic
Property indocyanine green, the cancer cell membrane be logarithmic phase growth cervical cancer cell cell membrane fragments.
2. a kind of multifunctional bionic nanometer formulation based on cancer cell membrane as described in claim 1, which is characterized in that described to receive
Metric system agent has regular spherical design, average grain diameter 220-240nm.
3. a kind of multifunctional bionic nanometer formulation based on cancer cell membrane as described in claim 1, which is characterized in that Ah
Mycin is de- doxorubicin hydrochloride.
4. a kind of preparation method of the multifunctional bionic nanometer formulation based on cancer cell membrane as described in claim 1, feature
It is, comprising the following steps:
(1) chemotherapeutics micro syringe is slowly dropped to formationization in the pure water of magnetic agitation dropwise under the conditions of being protected from light
The nanoparticle of drug is treated, 20-40min is stirred；
(2) photo-thermal therapy drug is dissolved in water, and is added dropwise to dropwise under the conditions of being protected from light described in step (1) with micro syringe
It is stirred in nano-particle solution, stirs 10-15h, photo-thermal therapy drug is coated on the outside of chemotherapeutics, obtains in nanoparticle
Core, wherein the molar ratio of the photo-thermal therapy drug and chemotherapeutics is 1:(2-3)；
(3) the nanoparticle kernel for obtaining step (2) and the classification of cancer cell membrane fragment squeeze out, wherein the nano particle kernel
It is mixed with cancer cell membrane fragment according to the ratio of mass ratio 1:(3-4)；
Wherein step (1)-(3) carry out under light protected environment.
5. a kind of preparation method of the multifunctional bionic nanometer formulation based on cancer cell membrane as claimed in claim 4, feature
It is, the chemotherapeutics in the step (1) is the hydrophobic adriamycin of desalination, and the photo-thermal therapy drug in the step (2) is
Indocyanine green, the cancer cell membrane in the step (3) are the cell membrane of the human cervical carcinoma cell of logarithmic phase growth.
6. a kind of preparation method of the multifunctional bionic nanometer formulation based on cancer cell membrane as claimed in claim 4, feature
It is, the classification in the step (3) squeezes out the following steps are included: by nano particle kernel and cancer cell membrane fragment according to quality
Ratio mixing than 1:3, vortex mixed is uniform, successively by 1 μm, 0.85 μm and 0.45 μm micropore filter after standing 30-40 minutes
Film distinguishes filter 23-5 times to get the multifunctional bionic nanoparticle formulations wrapped up to cancer cell membrane.
7. a kind of preparation method of the multifunctional bionic nanometer formulation based on cancer cell membrane as claimed in claim 4, feature
It is, the preparation method of the cancer cell membrane fragment in the step (3) is the following steps are included: through hypotonic extraction method broken cell membrane
Removal content obtains cell membrane fragments, is then sonicated to obtain uniform nanoscale cancer cell membrane fragment, whole ice bath is placed.
8. a kind of preparation method of the multifunctional bionic nanometer formulation based on cancer cell membrane as claimed in claim 4, feature
It is, cancer cell membrane uses hypotonic extraction method in the step (3), comprising the following steps: the human cervical carcinoma for taking logarithmic phase to grow
Cell, using cell scraper by under cell scraper, cell is collected by centrifugation in 700g, and the PBS washing of isotonic pre-cooling after centrifugation, abandons supernatant,
It is added hypotonic solution ice bath 15-20 minutes, by the cell solution after ice bath in room temperature and multigelation in liquid nitrogen, until cell is complete
Complete broken, then, 700g is centrifuged to obtain supernatant, and 14000g is centrifuged to obtain precipitating to be cell membrane fragments, and above-mentioned cell membrane is broken
- 80 DEG C of piece -- 90 DEG C of freeze-dryings save.
9. a kind of preparation method of the multifunctional bionic nanometer formulation based on cancer cell membrane as claimed in claim 4, feature
Be, the multifunctional bionic nanometer formulation the preparation method is as follows:
5mg doxorubicin hydrochloride is added in 10mL sample bottle and is protected from light processing, be added 4.5mL dimethyl sulphoxide solution make hydrochloric acid Ah
Mycin sufficiently dissolves, and 0.5mL triethylamine is added under the conditions of magnetic agitation, and adjustment magneton revolving speed is 600rpm, reaction overnight, gained
Product is desalination adriamycin；
The above-mentioned desalination adriamycin micro syringe of 100 μ L is slowly dropped to the 5mL of magnetic agitation dropwise under the conditions of being protected from light
Adriamycin nano particle is formed in pure water, after 30 minutes, will be dissolved in 200 μ L indocyanine green solution of water by the same method by
It is added dropwise in above-mentioned nano-particle solution and is stirred overnight, wherein the concentration of indocyanine green solution is 1mg/mL, obtains nanoparticle
Sub- kernel；
The hypotonic extraction method of cancer cell membrane using epicyte protein and suppressor proteins extracts kit the following steps are included: mentioned
Cell membrane is taken, the human cervical carcinoma cell for taking logarithmic phase to grow, using cell scraper by under cell scraper, 700g is centrifuged 5 minutes and collects
Cell is washed using the PBS of isotonic pre-cooling, and the pH of PBS is 7.4, after centrifugation, abandons supernatant, is added under hypotonic solution condition of ice bath
15 points are stood, by above-mentioned cell solution in room temperature and multigelation in liquid nitrogen, until cell is completely broken, then, 700g is centrifuged
Supernatant is carefully obtained within 10 minutes, it is cell membrane fragments, above-mentioned cell membrane fragments -80 that 14000g, which is centrifuged and obtains within 30 minutes precipitating,
DEG C freeze-drying save；
The nano particle is merged with the package of cancer cell membrane fragment the following steps are included: first that the cancer cell membrane fragment of extraction is molten
In pure water, finely dispersed nanoscale cell film fragment is obtained through ultrasonic cell disruption instrument ice-bath ultrasonic process, then will
Nano particle and cancer cell membrane fragment are uniform according to mass ratio 1:3 ratio vortex mixed, stand 30 minutes, then successively pass through 1 μ
M, filter 23 time obtains cancer cell membrane package multifunctional bionic nano particle respectively for 0.85 μm and 0.45 μm of miillpore filter.
10. a kind of multifunctional bionic nanometer formulation based on cancer cell membrane as described in claim 1 is on preparing anticancer drug
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103099784A (en) * | 2012-10-29 | 2013-05-15 | 深圳先进技术研究院 | Nanometer medicine particle, preparation method and application thereof |
| CN104922671A (en) * | 2015-06-09 | 2015-09-23 | 深圳先进技术研究院 | Indocyanine green composite nano-particles and preparation method and application thereof |
| CN105288622A (en) * | 2015-11-03 | 2016-02-03 | 浙江大学 | Preparation method of cell membrane microcapsule simultaneously loaded with chemotherapeutic drug and photodynamic therapeutic drug |
-
2017
- 2017-06-13 CN CN201710443034.1A patent/CN109078183B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103099784A (en) * | 2012-10-29 | 2013-05-15 | 深圳先进技术研究院 | Nanometer medicine particle, preparation method and application thereof |
| CN104922671A (en) * | 2015-06-09 | 2015-09-23 | 深圳先进技术研究院 | Indocyanine green composite nano-particles and preparation method and application thereof |
| CN105288622A (en) * | 2015-11-03 | 2016-02-03 | 浙江大学 | Preparation method of cell membrane microcapsule simultaneously loaded with chemotherapeutic drug and photodynamic therapeutic drug |
Non-Patent Citations (1)
| Title |
|---|
| CAITONG YU ET AL: ""Smart doxorubicin nanoparticles with high drug payload for enhanced chemotherapy against drug resistance and cancer diagnosis"", 《NANOSCALE》 * |
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