CN110152019A - Bionical bimodal inversion of phases nanoscale ultrasound contrast agents and preparation method thereof - Google Patents
Bionical bimodal inversion of phases nanoscale ultrasound contrast agents and preparation method thereof Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0002—General or multifunctional contrast agents, e.g. chelated agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- 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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
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- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Acoustics & Sound (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Radiology & Medical Imaging (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a kind of bionical bimodal inversion of phases nanoscale ultrasound contrast agents, preparation is spherical core-shell structure, and 200~300nm of average grain diameter, shell membrane is interior to coat liquid fluorocarbon, and shell membrane is made of the erythrocyte membrane for carrying near infrared fluorescent dye IR-780.Open preparation method simultaneously, acquisition animal's whole blood simultaneously obtains erythrocyte membrane through separation and Extraction, the latter mixes the erythrocyte membrane for standing and obtain carrying IR-780 after high speed rotary-cut with IR-780, obtains the novel type radiographic contrast through ultrasonic emulsification after it is mixed with liquid fluorocarbon.Contrast agent prepared by the present invention is the nanoscale of uniform particle diameter as the result is shown, property is stablized, the function of erythrocyte membrane is remained simultaneously, it can be realized efficient immunologic escape, it is expected to that more acoustic contrast agent targetings is made to be gathered in tumor tissues, carry out tumor cells targeting bimodal to have complementary advantages imaging, effective cancer target photo-thermal further mediated to melt, for realize tumour efficient noninvasive precisely detecting and target photo-thermal therapy opens new thinking.
Description
Technical field
The invention belongs to contrast agent and its preparation technical fields, and in particular to bionical bimodal inversion of phases nanoscale ultrasound is made
Shadow agent and preparation method thereof.
Background technique
Malignant tumour seriously threatens the life and health of contemporary people, carries out early diagnosis and efficient targeting to malignant tumour
Treatment, can improve the prognosis and survival rate of patient.Currently, the research of ultrasound molecular image rapidly develops, the Clinics and Practices of tumour
Stride into from traditional mode to molecule field.That Ultrasonography has is convenient, "dead", can Real-time and Dynamic Detection etc. it is excellent
Point.The acoustic contrast agent of function admirable is the key that implement this technology.However, since conventional ultrasound contrast agent partial size is all micro-
It more than meter level, is not easy to penetrate tumor vascular endothelium gap and directly reaches tumor tissues, exploration develops that a kind of partial size is smaller, has height
The acoustic contrast agent for imitating tumor-targeting is extremely urgent.
Recent study discovery, partial size are less than advantage of the nanometer microvesicle due to its size of conventional ultrasound contrast agent, can benefit
Tumor locus is gathered in by the tumor vascular endothelium gap of 380-780nm with high-permeability and retention effect (EPR effect) to send out
The effect of waving.But this nanometer microvesicle stability is poor, and with the reduction of its partial size, echo enhancement effect weakens, ultrasonic imaging
Ability reduces, it is difficult to obtain satisfied imaging effect.Therefore, several recently using liquid fluorocarbon as the nanoscale ultrasound contrast agents of core
Year is developed, and stability is improved compared with the former, and can penetrate tumor vessel, occur under ultrasound or laser action
Liquid-gas phase transformation generates a large amount of microbubble, to enhance the clarity of ultrasonic contrast image.But form nano-liquid droplet not
Enter with shell membrane ingredient and easily identified and swallowed by reticuloendothelial system (RES) in vivo, then really reaches the amount meeting of tumor tissues
Greatly reduce, seriously affects it and play a role in diseased region.Erythrocyte membrane (RBCM) wraps up nanoparticle drug-loading system and escapes
Immune phagocytosis extends circulation time in vivo to implement the research of the high target administration of tumour in achievement in recent years and protrude.But it is red thin
After birth package liquid fluorocarbon there is no as acoustic contrast agent clearly to be reported.
Further, near infrared fluorescent dye IR-780 is because of the spy with tumor tissues specific target tropism and low bio-toxicity
Point is concerned as molecular probe in tumor research field in recent years.IR-780 can carry out near infrared imaging, make in conjunction with ultrasound
The acoustics visualization capabilities of shadow agent realize the bimodal diagnosis of diseased region；Simultaneously, moreover it is possible to by mediating photo-thermal, photodynamic effect real
Existing neoplasm targeted therapy.But IR-780 is insoluble in water, it is difficult to individually play a role, being carried on effective nano-carrier is solution
The certainly key of this problem.
In conclusion preparing a kind of erythrocyte membrane package liquid fluorocarbon, carrying the bionical of near infrared fluorescent dye IR-780
Bimodal inversion of phases nano-liquid droplet there is no correlative study to report at present as tumor cells targeted ultrasound contrast agent.
Summary of the invention
In order to solve problems of the prior art, the present invention discloses bionical bimodal inversion of phases nanoscale ultrasound and makes
Shadow agent and preparation method thereof can make erythrocyte membrane form shell membrane and carry IR-780, coat liquid fluorocarbon in shell membrane, can be used in
The early stage of tumour precisely detects and targeted therapy.
To achieve the goals above, the technical solution adopted by the present invention is that, bionical bimodal inversion of phases nanoscale ultrasound is made
Shadow agent, preparation are core-shell structure, and in sphere shape, average grain diameter is 200~300nm；Coat liquid fluorocarbon in shell membrane, shell membrane by
Mature erythrocyte membrane forms, and carries near infrared fluorescent dye IR-780 on the shell membrane.
Shell membrane raw material is the erythrocyte membrane of mammal, and the erythrocyte membrane uses Mice red cell film, rabbit erythrocyte film, dog
Erythrocyte membrane, swine erythrocyte film or human erythrocyte membrane.
Liquid fluorocarbon is perflexane, perflenapent, PF 5070, perfluorooctane, perfluoro bromide octane or perfluorodecalin.
Preferably, shell membrane raw material uses Mice red cell film；Liquid fluorocarbon uses perflexane.
The preparation method of bionical bimodal inversion of phases nanoscale ultrasound contrast agents, comprising the following steps:
S1 acquires mature animal's whole blood and is stored in the closed container equipped with anti-coagulants；
S2 after the animal's whole blood centrifugal treating for being stored S1, is washed after removing supernatant therein with buffer；
S3 repeats S2 for several times, and upper plasma and remaining haemocyte are removed, and collects pure red cell；
S4, distilled water and 10 × PBS after pre-cooling is sequentially added into pure red cell obtained by S3, carry out after mixing from
Heart processing, then remove supernatant, obtain erythrocyte membrane；
S5 repeats S4 for several times, collects erythrocyte membrane, erythrocyte membrane is suspended with 1 × PBS buffer solution of pre-cooling, is obtained red
The suspension of cell membrane；
S6 is added glycerol into the suspension of erythrocyte membrane obtained by S5 and is configured to aqua liquid；
S7 shears S6 products therefrom under the conditions of ice-water bath, obtains nanoscale erythrocyte membrane suspension；
S8 takes near infrared fluorescent dye IR-780 under the conditions of being protected from light, and prepares near infrared fluorescent dye IR-780 solution；
S9 mixes nanoscale erythrocyte membrane suspension obtained by S7 near infrared fluorescent dye IR-780 solution obtained by S8,
It is then allowed to stand to obtain the nanoscale erythrocyte membrane suspension for carrying near infrared fluorescent dye IR-780；
S10 mixes S9 products therefrom with liquid fluorocarbon, under the conditions of ice-water bath, carries out ultrasonic emulsification to emulsus and is suspended
Liquid obtains bionical bimodal inversion of phases nanoscale ultrasound contrast agents.
In S8, the concentration of near infrared fluorescent dye IR-780 solution is 1.0mg/mL~2.0mg/mL, near-infrared fluorescent dye
The solvent for expecting IR-780 solution is dimethyl sulfoxide or chloroform.
The taken animal's whole blood of S1, near infrared fluorescent dye IR-780 solution and liquid fluorocarbon obtained by S8 volume ratio be
24:0.6:(3-4)。
Compared with prior art, the present invention at least has the advantages that
Blood cell source needed for present invention preparation is in the self mature erythrocyte of mammal, and red blood cell contains in blood
Amount is the abundantest, and convenient for extracting during the experiment, red blood cell biological can drop after entering human body as the acoustic contrast agent of shell membrane
Solution, can reduce toxic side effect；
Bionical bimodal inversion of phases nanoscale ultrasound contrast agents shell membrane of the present invention is erythrocyte membrane, can be reduced netted
The intake of endothelial system inhibits the phagocytosis of macrophage, extends the circulation time of acoustic contrast agent in vivo, increases swollen
The aggregation at tumor position, while this acoustic contrast agent is also equipped with the sound of acoustic contrast agent while retaining erythrocyte membrane function
Characteristic is learned, the ability of diagnosing tumor and treatment is effectively improved；
Bionical bimodal inversion of phases nanoscale ultrasound contrast agents core of the present invention is liquid fluorocarbon, liquid fluorocarbon property
Stablize, after liquid fluorocarbon mutually becomes bubble, the ultrasonic development effect of tumor-infiltrated edge and metastatic lesion can be enhanced, can also be led to
It crosses cavitation effect and mechanical injuries is carried out to tumour；
The near infrared fluorescent dye IR- that bionical bimodal inversion of phases nanoscale ultrasound contrast agents surface of the present invention carries
780 have near infrared imaging ability, ultrasonic contrast can be cooperateed with to carry out bimodal imaging, while can target again and be gathered in tumour
Tissue realizes the efficient targeting ablation of tumour, the reduction pair while improving curative effect by mediating photo-thermal and optical dynamic therapy
The damage of human normal tissue.
Detailed description of the invention
Fig. 1 is the structural schematic diagram for preparing IR780-RBCM@Nds；
Fig. 2 is the grain size distribution of IR780-RBCM@Nds；
Fig. 3 is to observe IR780-RBCM@Nds (× 100 oil mirror) under laser confocal fluorescence microscope；
Fig. 4 is the scanning electron microscope (SEM) photograph of IR780-RBCM@Nds；
Fig. 5 is the stability of observation in vitro IR780-RBCM Nds at 37 DEG C；
Fig. 6 is the phase transformation and ultrasonic contrast effect of IR780-RBCM@Nds before and after observation in vitro ultrasound wave irradiation；
Fig. 7 is that laser confocal fluorescence microscope observes IR780-RBCM@Nds surface immune evasion PROTEIN C D47 by different sulphur
The anti-mouse CD47 labeling of monoclonal antibody of cyanic acid fluorescein coupling；
Fig. 8 is that real-time dynamic living cells imager observation in vitro macrophage swallows IR780-RBCM@Nds.
Specific embodiment
The present invention is illustrated in detail below with reference to embodiment and attached drawing.
Embodiment 1
The preparation of IR780-RBCM@Nds
S1, takes 8 nude mices, and nude mice comes from army medical university, PLA Air Force Experimental Animal Center, and weight 20~
25g, 6~8 week old carry out intraperitoneal anesthesia with 1% yellow Jackets (50mg/kg) that dosage is 100 μ L respectively, through arteria carotis
Blood is taken, the 4mL whole blood acquired in total is simultaneously stored in the heparin tube containing heparin sodium；
S2, the whole blood that S1 is stored uses centrifuge (German Ai Bende) with 3000r/min, under the conditions of 4 DEG C of temperature
Supernatant is abandoned in centrifugation 10 minutes, is then washed repeatedly with 1 × PBS (AM General) buffer of pre-cooling,
S3 is repeated S2 five times, until removal upper plasma and remaining haemocyte, collect 2mL pure red cell；
The distilled water that 80mL is pre-chilled is added into pure red cell obtained by S3 by S4, adds 10 × PBS of 80mL pre-cooling, and 10
It with distilled water is that 1:9 is mixed according to volume ratio that × PBS, which is 1 × PBS, is uniformly mixed, with the centrifugal rotational speed of 12000r/min at 4 DEG C
It is centrifuged 15 minutes under the conditions of temperature, abandons supernatant, obtain erythrocyte membrane；
S5 is repeated S4 five times, collects erythrocyte membrane, the 1 × PBS buffer solution suspension erythrocyte membrane being pre-chilled with 3.6mL,
Obtain erythrocyte membrane suspension；
400 μ L glycerols (Xi'an section sky) are added into erythrocyte membrane suspension obtained by S5 and are configured to 4mL Red Blood Cells Suspension by S6
The aqua liquid (1 × PBS: glycerol=9:1, volume ratio) of film；
Hand-held high-speed homogenization machine (believing only in Shanghai) is adjusted to 10000r/min under the conditions of ice-water bath by S7, to obtained by S6
Product carries out high speed rotary-cut, and time 20min obtains nanoscale erythrocyte membrane suspension；
S8 is weighed 2.0mg near infrared fluorescent dye IR-780 (U.S.'s Sigma-Aldrich) under the conditions of being protected from light, molten
In dimethyl sulfoxide (Xi'an section sky), it is configured to the IR-780 solution of 2mg/mL；
IR-780 solution obtained by the S8 of 100 μ L is added into nanoscale erythrocyte membrane obtained by S7, stands 30 points under room temperature by S9
Clock obtains the nanoscale erythrocyte membrane suspension for carrying near infrared fluorescent dye IR-780；
The perflexane (PFH, the auspicious happiness in Xi'an) of 667 μ L is added into S9 products therefrom by S10, under the conditions of ice-water bath, uses
Ultrasonic cell smash (silent 4710 series of Cole's pa, the U.S.) vibrates 130s with peak power output, obtains IR780-RBCM@
Nds: bionical bimodal inversion of phases nanoscale ultrasound contrast agents.
In the present embodiment, precooling temperature and ice water temperature are 4 DEG C.
Embodiment 2
The preparation of IR780-RBCM@Nds
S1, takes 8 nude mices, and nude mice comes from army medical university, PLA Air Force Experimental Animal Center, and weight 20~
25g, 6~8 week old carry out intraperitoneal anesthesia with 1% yellow Jackets (50mg/kg) that dosage is 100 μ L respectively, through arteria carotis
Blood is taken, the 4mL whole blood acquired in total is simultaneously stored in the heparin tube containing heparin sodium；
S2, the whole blood that S1 is stored, with 3000r/min, are centrifuged 10 minutes using centrifuge under the conditions of 4 DEG C of temperature, are abandoned
Then supernatant is washed repeatedly with 1 × PBS buffer solution of pre-cooling,
S3 is repeated S2 five times, until removal upper plasma and remaining haemocyte, collect 2mL pure red cell；
The distilled water that 80mL is pre-chilled is added into pure red cell obtained by S3 by S4, adds 10 × PBS of 80mL pre-cooling, and 10
It with distilled water is that 1:9 is mixed according to volume ratio that × PBS, which is 1 × PBS, is uniformly mixed, with the centrifugal rotational speed of 12000r/min at 4 DEG C
It is centrifuged 15 minutes under the conditions of temperature, abandons supernatant, obtain erythrocyte membrane；
S5 is repeated S4 five times, collects erythrocyte membrane, the 1 × PBS buffer solution suspension erythrocyte membrane being pre-chilled with 3.6mL,
Obtain erythrocyte membrane suspension；
400 μ L glycerols (Xi'an section sky) are added into erythrocyte membrane suspension obtained by S5 and are configured to 4mL Red Blood Cells Suspension by S6
The aqua liquid (1 × PBS: glycerol=9:1, volume ratio) of film；
Hand-held high-speed homogenization machine (believing only in Shanghai) is adjusted to 10000r/min under the conditions of ice-water bath by S7, to obtained by S6
Product carries out high speed rotary-cut, and time 20min obtains nanoscale erythrocyte membrane suspension；
S8 is weighed 1.5mg near infrared fluorescent dye IR-780 (U.S.'s Sigma-Aldrich) under the conditions of being protected from light, molten
In dimethyl sulfoxide (Xi'an section sky), it is configured to the IR-780 solution of 1.5mg/mL；
IR-780 solution obtained by the S8 of 100 μ L is added into nanoscale erythrocyte membrane obtained by S7, stands 30 points under room temperature by S9
Clock obtains the nanoscale erythrocyte membrane suspension for carrying near infrared fluorescent dye IR-780；
The perfluor pentane (PFP, the auspicious happiness in Xi'an) of 571 μ L is added into S9 products therefrom by S10, under the conditions of ice-water bath,
130s is vibrated with peak power output with Ultrasonic cell smash (silent 4710 series of Cole's pa, the U.S.), obtains IR780-
RBCM@Nds: bionical bimodal inversion of phases nanoscale ultrasound contrast agents.
In the present embodiment, precooling temperature and ice water temperature are 4 DEG C.
Embodiment 3
The preparation of IR780-RBCM@Nds
S1, takes 8 nude mices, and nude mice comes from army medical university, PLA Air Force Experimental Animal Center, and weight 20~
25g, 6~8 week old carry out intraperitoneal anesthesia with 1% yellow Jackets (50mg/kg) that dosage is 100 μ L respectively, through arteria carotis
Blood is taken, the 4mL whole blood acquired in total is simultaneously stored in the heparin tube containing heparin sodium；
S2, the whole blood that S1 is stored, with 3000r/min, are centrifuged 10 minutes using centrifuge under the conditions of 4 DEG C of temperature, are abandoned
Then supernatant is washed repeatedly with 1 × PBS buffer solution of pre-cooling,
S3 is repeated S2 five times, until removal upper plasma and remaining haemocyte, collect 2mL pure red cell；
The distilled water that 80mL is pre-chilled is added into pure red cell obtained by S3 by S4, adds 10 × PBS of 80mL pre-cooling, and 10
It with distilled water is that 1:9 is mixed according to volume ratio that × PBS, which is 1 × PBS, is uniformly mixed, with the centrifugal rotational speed of 12000r/min at 4 DEG C
It is centrifuged 15 minutes under the conditions of temperature, abandons supernatant, obtain erythrocyte membrane；
S5 is repeated S4 five times, collects erythrocyte membrane, the 1 × PBS buffer solution suspension erythrocyte membrane being pre-chilled with 3.6mL,
Obtain erythrocyte membrane suspension；
400 μ L glycerols (Xi'an section sky) are added into erythrocyte membrane suspension obtained by S5 and are configured to 4mL Red Blood Cells Suspension by S6
The aqua liquid (1 × PBS: glycerol=9:1, volume ratio) of film；
Hand-held high-speed homogenization machine (believing only in Shanghai) is adjusted to 10000r/min under the conditions of ice-water bath by S7, to obtained by S6
Product carries out high speed rotary-cut, and time 20min obtains nanoscale erythrocyte membrane suspension；
S8 is weighed 1.0mg near infrared fluorescent dye IR-780 (U.S.'s Sigma-Aldrich) under the conditions of being protected from light, molten
In chloroform (Xi'an section sky), it is configured to the IR-780 solution of 1.0mg/mL；
IR-780 solution obtained by the S8 of 100 μ L is added into nanoscale erythrocyte membrane obtained by S7, stands 30 points under room temperature by S9
Clock obtains the nanoscale erythrocyte membrane suspension for carrying near infrared fluorescent dye IR-780；
The PF 5070 (PFH, the auspicious happiness in Xi'an) of 500 μ L is added into S9 products therefrom by S10, under the conditions of ice-water bath, uses
Ultrasonic cell smash (silent 4710 series of Cole's pa, the U.S.) vibrates 130s with peak power output, obtains IR780-RBCM@
Nds: bionical bimodal inversion of phases nanoscale ultrasound contrast agents.
In the present embodiment, precooling temperature and ice water temperature are 4 DEG C.
The structure of IR780-RBCM@Nds
The structural schematic diagram of IR780-RBCM@Nds of the present invention, as shown in Figure 1, erythrocyte membrane is as shell membrane, package
Liquid fluorocarbon, surface carry near infrared fluorescent dye IR-780.
The IR780-RBCM@Nds core substance is liquid fluorocarbon；Such as: perflexane (Perfluorohexane,
PFH), perflenapent (Perfluoropentane, PFP), PF 5070, perfluorooctane, perfluoro bromide octane or perfluorodecalin.
The shell membrane of the IR780-RBCM@Nds is mammalian erythropoietin film；Such as: Mice red cell film, rabbit erythrocyte
Film, dog red blood cell film, swine erythrocyte film, human erythrocyte membrane；The preferred Mice red cell film of the present invention.
The label of the IR780-RBCM@Nds, can be unlimitedly using can be marked on erythrocyte membrane, and have swollen
The substance of tumor targeting and Photothermal characterisation；Such as near infrared fluorescent dye IR-780.
The IR780-RBCM@Nds: the partial size of bionical bimodal inversion of phases nanoscale ultrasound contrast agents is less than 780nm, excellent
Select 200~300nm.
The characteristic of IR780-RBCM@Nds
1. partial size, current potential and monodispersity index
At room temperature at 25 DEG C, with Beckman zeta current potential/nano-particle size analysis instrument detection IR780-RBCM@Nds partial size point
Cloth, surface potential and monodispersity index, each sample is in triplicate.As a result as shown in Fig. 2, IR780-RBCM@Nds partial size is in single
Crest, average grain diameter are 261.4 ± 58.6nm, its average potential is 8.51 ± 7.00mV, monodispersity index is 0.285 ±
0.010。
2. laser confocal fluorescence microscope phenetic analysis
Concentration is 1.2 × 10 after taking a drop to be diluted with 1 × PBS buffer solution6The IR780-RBCM@Nds of a/mL is dripped in load
On slide, (excitation wavelength 680nm, transmitted wave are observed with laser confocal fluorescence microscope high power oil mirror after covered
A length of 780nm), as a result the spheroidal as shown in figure 3, IR780-RBCM@Nds takes on a red color, is evenly distributed, size is uniform.
3. scanning electron microscope phenetic analysis
After IR780-RBCM@Nds suspension prepared by the embodiment of the present invention 1 is diluted with 1 × PBS buffer solution, drop to
It on copper mesh, is dried at room temperature, electron microscopic sample is made, by scanning electron microscope (Hitachi S-4800, Japan) observation, as a result such as
Shown in Fig. 4, IR780-RBCM@Nds is in sphere structure, and form rule, uniform in size, average grain diameter is 200~300nm, with grain
The result of degree analyzer detection matches.
The stability of IR780-RBCM@Nds in vitro
37 DEG C of body temperature in analogue body, the partial size change with the time by observing IR780-RBCM@Nds assess its stabilization
Property.Concentration is 1.2 × 10 after six groups of each 1mL are diluted with 1 × PBS buffer solution6The IR780-RBCM@Nds suspension of a/mL is put
Enter in centrifuge tube, be placed in 37 DEG C of water-bath (Changzhou state China, China), respectively 0min, 10min, 20min, 30min,
When 60min, 120min, its partial size is detected with Beckman zeta current potential/nano-particle size analysis instrument；The above experiment in triplicate, is tied
Fruit is as shown in figure 5, the partial size of IR780-RBCM@Nds corresponds in 0min, 10min, 20min, 30min, 60min, 120min and divides
Not Wei 213.4 ± 55.32nm, 247.6 ± 65.81nm, 247.3 ± 61.22nm, 306.4 ± 72.43nm, 382.6 ±
87.52nm, 553.8 ± 162.04nm, in 120min, the partial size of IR780-RBCM@Nds can smoothly lead to still less than 780nm
Tumor vessel gap is crossed, research shows that the stability of IR780-RBCM Nds can satisfy carries out diagnosing tumor and treatment in vivo
Requirement.
The external phase transformation of IR780-RBCM@Nds and ultrasonic contrast imaging
In vitro experiment, taking 2mL concentration is 2.2 × 106The IR780-RBCM@Nds suspension of a/mL is packed into latex
In emulsion capsule made of gloves finger tip, as a control group by isometric de aerated water；External application low-strength focusing ultrasonic instrument
(LIFU) ultrasonic imaging device, is adjusted to the twin display mode of two dimension, radiography by the emulsion capsule of vertical irradiation experiment group and control group
(Parkson, Italy), observation external supersonic develops and acquires image, as a result as shown in fig. 6, low-strength focusing ultrasonic is with 2.5W/
cm25s is irradiated, IR780-RBCM Nds phase becomes bubble, the enhancing of ultrasonoscopy ability.
Detection of the laser confocal fluorescence microscope to IR780-RBCM@Nds surface immune evasion PROTEIN C D47
Taking 200 μ L concentration is 1.8 × 106IR780-RBCM@Nds as experimental group, isometric isoconcentration lipid is received
Rice drop as a control group, is separately added into the anti-mouse that the fluorescein isothiocynate (FITC) that 4 μ L concentration are 0.5mg/mL is coupled
CD47 monoclonal antibody is observed after combining 12h at 4 DEG C with laser confocal fluorescence microscope；The excitation wavelength of IR-780 is
680nm, launch wavelength 780nm；The excitation wavelength of fluorescein isothiocynate is 488nm, launch wavelength 525nm；As a result such as
Shown in Fig. 7, the surface@Nds IR780-RBCM shows green fluorescence, and control group lipid shell membrane nano-liquid droplet surface redgreen is glimmering
Light, research shows that IR780-RBCM@Nds remains the characteristic of erythrocyte membrane, surface has immune evasion PROTEIN C D47.
The macrophage of IR780-RBCM@Nds swallows experiment
Under the conditions of being protected from light, 1mg cell membrane green fluorescence probe (DiO) (the green skies in Hangzhou) is dissolved in 1mL dimethyl
The DiO solution of 1mg/mL is prepared into sulfoxide (DMSO)；Taking following two concentration is 4.8 × 106Each 400 μ of the drop of a/ml
L is mixed respectively with DiO solution with the volume ratio of 40:1:
(1) IR780-RBCM@Nds, (2) lipid shell membrane nano-liquid droplet.
It is added in 400 μ L PBS and the DiO solution of same volume is added as a control group with same ratio.In cell incubator
It is middle to be incubated for 12 hours in 96 orifice plates of cell incubator with 264.7 macrophage of RAW respectively for every group with 100 holes μ L/, often
5000, hole, 264.7 macrophage of RAW；Image is shot using real-time dynamic living cells imager (gloomy west, the U.S.), DiO's swashs
Hair wavelength is 484nm, launch wavelength 501nm, as a result as shown in figure 8, the green fluorescence that IR780-RBCM@Nds group is shown
Few (figure), does not have notable difference with control group；And macrophage actively absorbs lipid shell membrane nano-liquid droplet, so that macrophage gulps down
A large amount of green fluorescences are generated after lipophagia mass shell film nano-liquid droplet；Erythrocyte membrane is by immunologic escape function from red blood cell transposition to novel
The removing of reticuloendothelial system can be effectively reduced in acoustic contrast agent surface, IR780-RBCM@Nds.
Claims (8)
1. bionical bimodal inversion of phases nanoscale ultrasound contrast agents, which is characterized in that preparation is core-shell structure, is in sphere shape,
Average grain diameter is 200~300nm；Liquid fluorocarbon is coated in shell membrane, shell membrane is made of mature erythrocyte membrane, takes on the shell membrane
Carry near infrared fluorescent dye IR-780.
2. bionical bimodal inversion of phases nanoscale ultrasound contrast agents according to claim 1, which is characterized in that shell membrane raw material
For the erythrocyte membrane of mammal, the erythrocyte membrane uses Mice red cell film, rabbit erythrocyte film, dog red blood cell film, pig red thin
After birth or human erythrocyte membrane.
3. bionical bimodal inversion of phases nanoscale ultrasound contrast agents according to claim 2, which is characterized in that shell membrane raw material
Using Mice red cell film.
4. bionical bimodal inversion of phases nanoscale ultrasound contrast agents according to claim 1, which is characterized in that liquid fluorocarbon
For perflexane, perflenapent, PF 5070, perfluorooctane, perfluoro bromide octane or perfluorodecalin.
5. bionical bimodal inversion of phases nanoscale ultrasound contrast agents according to claim 4, which is characterized in that liquid fluorocarbon
Using perflexane.
6. the preparation method of bionical bimodal inversion of phases nanoscale ultrasound contrast agents, which comprises the following steps:
S1 acquires mature animal's whole blood and is stored in the closed container equipped with anti-coagulants；
S2 after the animal's whole blood centrifugal treating for being stored S1, is washed after removing supernatant therein with buffer；
S3 repeats S2 for several times, and upper plasma and remaining haemocyte are removed, and collects pure red cell；
S4, distilled water and 10 × PBS after pre-cooling is sequentially added into pure red cell obtained by S3, carries out at centrifugation after mixing
Reason, then remove supernatant, obtain erythrocyte membrane；
S5 repeats S4 for several times, collects erythrocyte membrane, erythrocyte membrane is suspended with 1 × PBS buffer solution of pre-cooling, obtains red blood cell
The suspension of film；
S6 is added glycerol into the suspension of erythrocyte membrane obtained by S5 and is configured to aqua liquid；
S7 shears S6 products therefrom under the conditions of ice-water bath, obtains nanoscale erythrocyte membrane suspension；
S8 takes near infrared fluorescent dye IR-780 under the conditions of being protected from light, and prepares near infrared fluorescent dye IR-780 solution；
S9 mixes nanoscale erythrocyte membrane suspension obtained by S7 near infrared fluorescent dye IR-780 solution obtained by S8, then
Stand the nanoscale erythrocyte membrane suspension for obtaining carrying near infrared fluorescent dye IR-780；
S10 mixes S9 products therefrom with liquid fluorocarbon, under the conditions of ice-water bath, carries out ultrasonic emulsification to milky suspension, obtains
To bionical bimodal inversion of phases nanoscale ultrasound contrast agents.
7. the preparation method of bionical bimodal inversion of phases nanoscale ultrasound contrast agents according to claim 6, feature exist
In in S8, the concentration of near infrared fluorescent dye IR-780 solution is 1.0mg/mL~2.0mg/mL, near infrared fluorescent dye IR-
The solvent of 780 solution is dimethyl sulfoxide or chloroform.
8. the preparation method of bionical bimodal inversion of phases nanoscale ultrasound contrast agents according to claim 6, feature exist
In the volume ratio of near infrared fluorescent dye IR-780 solution obtained by the taken animal's whole blood of S1, S8 and liquid fluorocarbon is 24:0.6:
(3-4)。
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110772649A (en) * | 2019-11-21 | 2020-02-11 | 华中科技大学同济医学院附属协和医院 | Porous polymer ultrasonic contrast agent and preparation method thereof |
| CN111330025A (en) * | 2020-03-03 | 2020-06-26 | 中山大学附属第三医院 | Bionic microbubble ultrasound contrast agent and preparation method thereof |
| CN111330026A (en) * | 2020-03-19 | 2020-06-26 | 中南大学湘雅二医院 | Bionic phase-change ultrasonic contrast agent and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014041150A1 (en) * | 2012-09-14 | 2014-03-20 | Stichting Katholieke Universiteit | Contrast agent and its use for imaging |
| US20160317441A1 (en) * | 2015-04-30 | 2016-11-03 | National Tsing Hua University | Biomembrane phase-change droplets (pcd), drug carrier and use thereof |
| CN106267199A (en) * | 2016-08-25 | 2017-01-04 | 郑州大学 | A kind of preparation method and application of ultrasonic regulation type antitumor drug delivery system |
-
2019
- 2019-06-10 CN CN201910498246.9A patent/CN110152019A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014041150A1 (en) * | 2012-09-14 | 2014-03-20 | Stichting Katholieke Universiteit | Contrast agent and its use for imaging |
| US20160317441A1 (en) * | 2015-04-30 | 2016-11-03 | National Tsing Hua University | Biomembrane phase-change droplets (pcd), drug carrier and use thereof |
| CN106267199A (en) * | 2016-08-25 | 2017-01-04 | 郑州大学 | A kind of preparation method and application of ultrasonic regulation type antitumor drug delivery system |
Non-Patent Citations (2)
| Title |
|---|
| CHEN-CHAN HSIEH, ET AL.: "Biomimetic Acoustically-Responsive Vesicles for Theranostic Applications", 《THERANOSTICS》 * |
| 易衡静等: "靶向肿瘤细胞的相变型纳米粒多模态显像及其声动力治疗的体外实验研究", 《解放军医学杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110772649A (en) * | 2019-11-21 | 2020-02-11 | 华中科技大学同济医学院附属协和医院 | Porous polymer ultrasonic contrast agent and preparation method thereof |
| CN111330025A (en) * | 2020-03-03 | 2020-06-26 | 中山大学附属第三医院 | Bionic microbubble ultrasound contrast agent and preparation method thereof |
| CN111330025B (en) * | 2020-03-03 | 2022-07-26 | 中山大学附属第三医院 | Bionic microbubble ultrasound contrast agent and preparation method thereof |
| CN111330026A (en) * | 2020-03-19 | 2020-06-26 | 中南大学湘雅二医院 | Bionic phase-change ultrasonic contrast agent and preparation method thereof |
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