CN111632154A - Phase-transition nanobubble, preparation method and application thereof - Google Patents
Phase-transition nanobubble, preparation method and application thereof Download PDFInfo
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- CN111632154A CN111632154A CN202010483519.5A CN202010483519A CN111632154A CN 111632154 A CN111632154 A CN 111632154A CN 202010483519 A CN202010483519 A CN 202010483519A CN 111632154 A CN111632154 A CN 111632154A
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- phase
- transition
- nanobubble
- nanobubbles
- polylactic acid
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
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Abstract
The invention discloses a phase-transition nanobubble, a preparation method and application thereof. The nanobubble is prepared by taking amphiphilic chitosan-polylactic acid graft copolymer as a coating material, taking perfluoropentane capable of undergoing liquid-gas phase transition under the action of temperature as a bubble core filling material and adopting an emulsion solvent volatilization method. The prepared nanobubble is cream yellow, has the particle size of 101.1 +/-2.7 nm, has small change of the particle size in 2 months in vitro, has good stability, can enhance the ultrasonic imaging effect under the action of ultrasonic waves, and can be broken by the ultrasonic waves. The chitosan-polylactic acid graft copolymer nano-bubbles prepared by the emulsion solvent volatilization method have good external stability and can enhance ultrasonic imaging. The nanobubbles can be loaded with MRI contrast agents, and the MRI imaging effect of tumor lesions is improved. The nanobubble can also load antitumor drugs, is used for the targeted therapy of tumors, is a novel multifunctional imaging nano contrast agent integrating diagnosis and treatment, and has great application value.
Description
Technical Field
The invention belongs to the technical field of medicine preparation, and particularly relates to a phase-transition nanobubble, a preparation method and application thereof.
Background
Chitosan is a basic polysaccharide with good biocompatibility and biodegradability. The chitosan has-OH and-NH on the main chain2The reactive group provides a bonding site for the grafting reaction, and is an excellent material for preparing the graft polymer. Has been widely used in the research of various drug carriers such as controlled release, targeting, intelligent drug delivery and the like.
The polylactic acid contains a large amount of ester bonds in the molecular structure, and has poor hydrophilicity, so that the polylactic acid has lower compatibility with other substances; polylactic acid molecules have no electric charge and no active groups, and are difficult to modify; the degradation period is difficult to control, etc. Aiming at the defects of the polylactic acid, the modification and modification of the polylactic acid focus on improving the mechanical property and the hydrophilicity of the polylactic acid while maintaining the degradability of the polylactic acid.
In recent years, the combination of polysaccharide and derivatives thereof with polylactic acid to prepare polylactic acid biomimetic materials with good biological performance has become a hotspot of research, and the synthetic methods of polylactic acid copolymers mainly comprise two methods: direct polymerization and ring-opening polymerization. Wherein the structure of the product synthesized by the ring-opening polymerization method is easy to control, so that the polymer with easy control of both the performance and the degradation period can be obtained.
The microbubble contrast agent is an ultrasonic imaging contrast agent, is a gas-containing vesicle formed by an outer envelope and an inner gas, and has the diameter of 1-8 mu m. The bubbles have strong scattering effect on the ultrasonic waves, and the tissue echo can be enhanced, so that the developing definition and resolution are improved. The microbubble ultrasound contrast agent goes through three development stages, the first generation is a free bubble type contrast agent which is physiological saline or emulsion containing carbon dioxide or air and other gases, and due to the large size and no membrane coating, the contrast agent in blood has short duration and loses acoustic reflectivity quickly, only right heart imaging can be carried out, and the use is limited to a certain extent. The second generation is a wrapped contrast agent, the shell membrane is prepared from albumin, a polymer, a surfactant or lipid by an ultrasonic vibration method, the contrast agent has certain stability, and the ultrasonic development of a left heart system can be realized by intravenous injection, so that the contrast agent has clinical practical value and is commercialized. The third generation is a wrapped contrast agent represented by a liquid fluorocarbon emulsion, the core is mainly perfluoropentane (PFP), Perfluorohexane (PFH), Perfluorodecalin (PFD), perfluorooctane bromide (PFOB) and the like, the shell membrane is a high molecular polymer or a lipid, the liquid fluorocarbon emulsion has better stability, the stability of microbubbles which are not easy to break is improved to a certain extent compared with the stability of a blood circulation system of a second generation microbubble product, the ultrasonic sound wave reflection performance is stronger, and myocardial ultrasonic development can be realized through coronary circulation.
Ultrasound contrast agents were discovered by Gramiak and Shah at rochester university as early as 1968. Sonoweiwei is the only ultrasound microbubble contrast agent approved to be on the market in China at present, has the particle size of 1-4 mu m, and can pass through the pulmonary circulation but can not pass through the vascular endothelium. The third-generation microbubbles marketed above are also available from Molecular Biosystems
Of Xian Ling Bao ya corporation
Sono of Bracco corporation
And ImaRx Pharmaceutical Co
From Sonus Pharmaceutical Co
Of Alliance Inc
And the like. The ultrasound contrast agent at this stage has a more stable gas core and a flexible shell and is therefore also more efficient and stable in scattering ultrasound signals. However, the particle size of the microbubbles is usually 1 to 10 μm, and after intravenous injection into the body, the microbubbles can only pass through the pulmonary circulation and cannot penetrate through the wall of the blood vessel, so that only blood pool imaging can be performed.
In recent years, the research of nanoscale gas-containing microbubbles-nanobubbles as novel ultrasonic imaging contrast agents draws attention. The nanobubbles have nanometer-scale particle size and can penetrate through vascular endothelium to enter tissue gaps, so that imaging of extravascular target tissues (such as tumor tissues) becomes possible. The Chinese invention patent 'ultrasonic sensitive drug-loaded nanobubble, 200810166862.6' discloses an ultrasonic sensitive drug-loaded nanobubble, but the fluorocarbon-loaded polymer micelle prepared by combining a dialysis method and an ultrasonic resonance method is not a real envelope nanobubble, and no animal in vivo test or cell test data proves the action effect. The Chinese invention patent 'a multifunctional ultrasonic contrast agent and a preparation method thereof, 201010505467.3' discloses a lipid emulsion containing shell-core structure nanoparticles, which can enhance ultrasonic, CT and MRI imaging, but the lipid emulsion is essentially a lipid emulsion carrying liquid fluorocarbon and nano magnetic spheres, is not a coating nanobubble in the true sense, and has no targeting property. The lipid emulsion adopts the traditional film hydration-dispersion emulsification-ultrasonic preparation process, the particle size is difficult to accurately control, the particle size distribution is wide, and the reproducibility of the preparation process is poor. The invention discloses an ultrasonic magnetic resonance combined contrast agent and a preparation method thereof, and 200610097375.X discloses a polymer microsphere for encapsulating a conventional magnetic resonance contrast agent and fluorocarbon gas, which is prepared by an ultrasonic acoustic vibration cavitation method, wherein the particle size of the polymer microsphere is 1-5 μm, and particle size distribution data is not given. The targeted modification is a simple physical mixing process, and the action effect of the targeted modification is proved by animal in vivo test or cell test data. At present, the disclosed ultrasound microbubbles are mostly prepared from natural materials such as albumin, phospholipid, starch, cellulose, and the like, or biodegradable high polymer materials such as Polylactide (PLA), glycolide-lactide copolymer (PLGA), Polycaprolactone (PCL), and the like. The microbubble prepared by the former has the defects of poor in-vivo pressure resistance, obvious sound attenuation, poor stability and the like, and the latter has the problems of poor hydrophilicity, long in-vivo degradation time, lack of active sites in molecules, difficulty in targeted modification and the like. The disclosed methods for preparing ultrasound microbubbles mainly include acoustic vibration cavitation, mechanical homogenization, thin film hydration, freeze drying, spray drying, and emulsion polymerization (dieshifei, et al, biomimetic membrane materials and technologies, science publishers, 2010). However, these methods have many problems in practical applications, such as: the method has the disadvantages that the nano-scale microbubbles are difficult to obtain, the prepared microbubbles have large particle size and poor particle size uniformity, the preparation process has poor reproducibility, and the severe mechanical action and/or high temperature in the preparation process can cause targeted factors such as polypeptides, proteins, antibodies and the like to lose biological activity and the like. Meanwhile, with the development of medical imaging, higher requirements are provided for an image contrast agent, the nano-contrast agent can be simultaneously applied to different imaging technologies such as ultrasound and MRI, and particularly applied to a novel multifunctional imaging nano-contrast agent integrating diagnosis and treatment of early diagnosis and targeted treatment of imaging of serious diseases such as tumors, and the nano-contrast agent has good application prospect clinically. Meanwhile, there is a need to develop new nano-contrast preparation technology.
Disclosure of Invention
In order to solve the technical problems, the invention provides the phase-transition nanobubble prepared by adopting a novel functional polymer material and an emulsifying solvent volatilization method, the nanobubble has uniform and controllable particle size and good in-vivo and in-vitro stability, and can enhance ultrasonic imaging. The invention also provides a preparation method and properties of the nanobubble, the preparation conditions are mild, the preparation process is good in reproducibility, and the nanobubble can be used as a novel nano contrast agent for imaging diagnosis.
The technical scheme provided by the invention is as follows:
one of the purposes of the invention is to provide a preparation method of phase-transition nanobubbles, which comprises the following steps:
(1) dissolving chitosan-polylactic acid graft copolymer as coating material in dichloromethane to obtain oil 1, namely O1Phase, liquid perfluoropentane as oil 2, O2Phase, mixing the two phases in an ice bath to obtain O2Phase is uniformly dispersed in O1Formation of O in phase2/O1Phase (1);
(2) mixing O with2/O1Gradually dripping the phase into the water phase under magnetic stirring in an ice bath to obtain pre-compound emulsion O2/O1A phase of/W;
(3) subjecting the pre-emulsion phase to ultrasonic treatment until O with uniform particle size is obtained2/O1a/W compound emulsion;
(4) mixing O with2/O1And the/W multiple emulsion is magnetically stirred under the ice bath condition, is solidified to form nano bubbles, is centrifuged to obtain precipitates, and is re-dispersed to obtain the nano bubble water dispersion body carrying liquid perfluoropentane.
Further, the structural formula of the chitosan-polylactic acid graft copolymer in the step (1) is as follows:
the structural formula of PLA is:
further, the number average molecular weight of the chitosan-polylactic acid graft copolymer is 3000-60000.
Furthermore, the number average molecular weight of the chitosan-polylactic acid graft copolymer is 5000-30000.
Further, the step (2) is pre-emulsion O2/O1The weight percentage of the chitosan-polylactic acid graft copolymer in the/W phase is 0.1-5.0 wt%, the weight percentage of the liquid perfluoropentane is 1.0-30.0 wt%, and the balance is ultrapure water; the water phase can also be an aqueous solution of surfactant with hydrophilic groups, including SDS, poloxamer, Span20, Tween80, and PEG.
Further, in the step (1), O1Phase sum of O2The phase mixing is high shear mixing, and the shear rate is 5000-30000 rpm. Preferably, the shear rate is 5000 to 20000 rpm.
Further, the ultrasonic treatment method in the step (3) comprises the following steps: carrying out ultrasonic treatment for multiple times, setting the ultrasonic power to be 30-300W, carrying out ultrasonic treatment for 1-10 s every time at intervals of 1-10 s, and repeating for 1-20 times until O with uniform particle size is obtained2/O1and/W compound emulsion. Preferably, the ultrasonic power is 50-200W, the ultrasonic treatment time is 2-8 s each time, the interval is 2-8 s, and the ultrasonic treatment is repeated for 5-15 times.
It is another object of the present invention to provide a phase-transition nanobubble prepared by the above method, which can realize liquid-gas phase transition by temperature change.
Furthermore, the particle size range of the nano-bubbles is 30-1000 nm, and the polydispersity index PDI is less than or equal to 0.35. The preferable particle size is 100-400 nm, and PDI is less than or equal to 0.15.
The third object of the present invention is to provide the use of the above-mentioned phase-transition nanobubbles: adding an MRI contrast agent to the phase-transition nanobubble for MRI imaging. The mass percentage of the MRI contrast agent is 0.01-3.0 wt%, preferably 0.1-1.5 wt%. The fourth object of the present invention is to provide another use of the above-mentioned phase-transition nanobubbles: the nanobubble is used for targeted delivery of antitumor drugs, and the loaded antitumor drugs comprise clinical antitumor drugs including paclitaxel, docetaxel, hydroxycamptothecin, adriamycin, mitomycin, tamoxifen, 5-fluorouracil, methotrexate, cytarabine, cyclophosphamide or platinum drugs.
The invention has the beneficial effects that:
(1) the chitosan-polylactic acid graft copolymer has the advantages of good biocompatibility, hydrophilicity and hydrophobicity, easy coupling with targeting factors and the like, and the mechanical property, the degradation time and the like can be regulated and controlled by selecting proper molecular weight. Thereby preparing the nano-bubble contrast agent with proper mechanical property (good toughness, moderate pressure resistance), good stability and proper degradation time.
(2) Compared with the conventional ultrasound microbubble contrast agent, the nanobubble can pass through the endothelium of tumor blood vessels and can be specifically targeted to a tumor focus under the action of a targeting factor. The liquid perfluoropentane undergoes liquid-gas phase conversion at the body temperature to form gas-containing nano bubbles, and the gas-containing nano bubbles are gathered and combined into micro bubbles under the action of ultrasound, so that the ultrasound imaging effect of the tumor focus is enhanced. And the kit can also load a conventional MRI contrast agent, improve the accuracy and sensitivity of MRI imaging of the tiny tumor focus, and improve the early diagnosis effect of tumor imaging.
(3) The emulsion solvent volatilization method can realize the accurate regulation and control of the particle size of the nanobubble, the prepared nanobubble has high uniformity of particle size, good monodispersity, good stability in vitro and in vivo, long blood circulation time, and can realize the repeated examination, dynamic monitoring and curative effect evaluation of tumor focus.
(4) The emulsifying solvent volatilization method has mild preparation conditions, and can avoid destroying the biological activity of targeted factors such as polypeptide, protein, antibody and the like in the preparation process.
(5) The preparation process has good reproducibility by controlling the ultrasonic condition and the emulsifying solvent volatilization method, the fluctuation of the particle size and the PDI value of the nanobubbles in different batches is very small, the preparation process can be amplified in proportion, and the large-scale preparation is easy to realize.
Drawings
FIG. 1 is an appearance diagram of a nanobubble;
FIG. 2 is a graph of the distribution of the nanobubble particle size;
FIG. 3 shows the results of the stability test of nanobubbles at 4 deg.C, wherein FIG. 3(a) is a particle size stability curve and FIG. 3(b) is a potential stability curve;
FIG. 4 is an image of the nanobubbles in a latex glove under 37 deg.C water bath condition, wherein 4a is physiological saline, 4b is coupling agent, and 4c is chitosan-polylactic acid nanobubbles.
Detailed Description
The present invention is explained in more detail below by means of examples, which are only illustrative and the scope of protection of the present invention is not limited by these examples.
A method for preparing phase-transition nanobubbles comprises the following steps:
(1) dissolving chitosan-polylactic acid graft copolymer as coating material in dichloromethane to obtain oil 1, namely O1Phase, liquid perfluoropentane as oil 2, O2Phase, mixing the two phases in an ice bath to obtain O2Phase is uniformly dispersed in O1Formation of O in phase2/O1Phase (1);
(2) mixing O with2/O1Gradually dripping the phase into the water phase under magnetic stirring in an ice bath to obtain pre-compound emulsion O2/O1A phase of/W;
(3) subjecting the pre-emulsion phase to ultrasonic treatment until O with uniform particle size is obtained2/O1a/W compound emulsion;
(4) mixing O with2/O1And the/W multiple emulsion is magnetically stirred under the ice bath condition, is solidified to form nano bubbles, is centrifuged to obtain precipitates, and is re-dispersed to obtain the nano bubble water dispersion body carrying liquid perfluoropentane.
Further, the structural formula of the chitosan-polylactic acid graft copolymer in the step (1) is as follows:
the structural formula of PLA is:
further, the number average molecular weight of the chitosan-polylactic acid graft copolymer is 3000-60000.
Furthermore, the number average molecular weight of the chitosan-polylactic acid graft copolymer is 5000-30000.
Further, the step (2) is pre-emulsion O2/O1The weight percentage of the chitosan-polylactic acid graft copolymer in the/W phase is 0.1-5.0 wt%, the weight percentage of the liquid perfluoropentane is 1.0-30.0 wt%, and the balance is ultrapure water; the water phase can also be an aqueous solution of surfactant with hydrophilic groups, including SDS, poloxamer, Span20, Tween80, and PEG.
Further, in the step (1), O1Phase sum of O2The phase mixing is high shear mixing, and the shear rate is 5000-30000 rpm. Preferably, the shear rate is 5000 to 20000 rpm.
Further, the ultrasonic treatment method in the step (3) comprises the following steps: carrying out ultrasonic treatment for multiple times, setting the ultrasonic power to be 30-300W, carrying out ultrasonic treatment for 1-10 s every time at intervals of 1-10 s, and repeating for 1-20 times until O with uniform particle size is obtained2/O1and/W compound emulsion. Preferably, the ultrasonic power is 50-200W, the ultrasonic treatment time is 2-8 s each time, the interval is 2-8 s, and the ultrasonic treatment is repeated for 5-15 times.
The nano-bubbles prepared by the invention can realize liquid-gas phase conversion through temperature change. The particle size range of the nanobubble is 30-1000 nm, and the polydispersity index PDI is less than or equal to 0.35. The preferable particle size is 100-400 nm, and PDI is less than or equal to 0.15.
After the prepared nanobubble is injected into a body, liquid-gas phase conversion of liquid fluorocarbon occurs at the body temperature to form gas-containing nanobubbles, and the nanobubbles are enriched at the tumor focus part through the specific combination of the targeting factors and tumor cells, so that the ultrasonic imaging effect of the tumor focus is improved. The nanobubble can load a conventional MRI contrast agent, improve the accuracy and sensitivity of MRI imaging of the tiny tumor focus, and improve the early diagnosis effect of tumor imaging. The nanobubble can also load antitumor drugs, is used for the targeted therapy of tumors, is used as a targeted delivery material of the antitumor drugs, and is a novel diagnosis and treatment integrated multifunctional imaging nano contrast agent.
The chitosan-polylactic acid graft copolymer can realize tumor targeting by chemically coupling tumor specific targeting factors, wherein the tumor specific targeting factors are folic acid, lactoferrin receptor single-chain antibody, transferrin receptor single-chain antibody, alpha-fetoprotein (AFP) receptor monoclonal antibody, RGD peptide or monoclonal antibodies of various cancer cells and the like.
The nano bubble core filling material is perfluoropentane (C5F12), which is liquid at normal temperature and has a boiling point of 29.5 ℃, and liquid-gas phase conversion occurs in vivo to gas.
The nano bubble loaded MRI contrast agent is a conventional MRI contrast agent used in clinic and comprises superparamagnetic nano Fe3O4Superparamagnetic nano Fe2O3Gadolinium compounds (e.g., Gd-DTPA, Gd-DOTA, Gd-BOPTA, etc.) and manganese compounds (e.g., Mn-DPDP, manganese porphyrin, etc.). The MRI contrast agent can account for 0.05-3.0 wt% of the total mass of the nano bubble water dispersion, and the optimal dosage is 0.1-1.5 wt%. The nano bubble loaded antitumor drug comprises paclitaxel, docetaxel, hydroxycamptothecin, adriamycin, mitomycin, tamoxifen, 5-fluorouracil, methotrexate, cytarabine, cyclophosphamide, platinum drugs (cisplatin, carboplatin or oxaliplatin) and other clinical common antitumor drugs.
The nano bubble water dispersoid can be added with common additives of injection, such as preservative sodium azide, thimerosal, phenol and the like, and the additives can account for 0-2.0 wt% of the total mass of the nano bubble water dispersoid.
The phase transition nanobubbles of the present invention, and the method of volatilizing the emulsion solvent and the use thereof will be described below by way of examples. The present disclosure is to be accorded the widest scope possible within the terms of persons skilled in the art and, therefore, the preferred embodiments of the present invention are to be understood as illustrative only and not as limiting the present invention in any way.
Example 1
(1) Preparation of Low molecular weight Chitosan
According to a literature method (Guokawa. the process for preparing the chitosan oligomer by degrading through a hydrogen peroxide method is discussed in [ J ]. Si Gu, 2010(19):25+34.), 5g of chitosan is taken into 100ml of 2% acetic acid solution, after the chitosan is completely dissolved, H2O2 with a certain concentration is slowly dropped into the solution, the pH value of the solution is adjusted to be neutral by 2mol/L of NaoH after the reaction is carried out for 5 hours at the temperature of 60 ℃, impurities are removed by filtration, the solution is precipitated and separated out by 3-5 times of absolute ethyl alcohol, low molecular weight chitosan is separated out, and the solution is refrigerated for 24 hours, filtered, and dried in vacuum.
(2) Preparation of Chitosan-polylactic acid graft copolymer
According to a literature method (Liangyanhui preparation of an amphiphilic chitosan-polylactic acid graft copolymer as an agricultural bactericide carrier and performance research thereof [ D ]. China academy of agricultural sciences, 2012), 1g of low-molecular-weight chitosan and 10g of lactide are put into a 50ml round-bottom flask, 15ml of DMSO is added, magnetic stirring is carried out for 1h in a vacuum state to enable the chitosan and the lactide to be fully swelled, a proper amount of triethylamine is slowly dropped into the flask by using an injector under the protection of nitrogen, the reaction temperature is slowly increased to 83 ℃, stirring is continuously carried out for reaction for 10h, at the moment, a reaction solution becomes clear, a crude product is obtained after the reaction is finished, the crude product is precipitated by using ice water and then filtered, deionized water is washed by using toluene at 85 ℃ to remove byproducts, and the product is obtained by.
(3) Preparing nano-bubbles by adopting emulsion solvent volatilization method
Weighing 0.0500g of chitosan-polylactic acid, placing in a test tube, adding dichloromethane, stirring for dissolving, placing the test tube in an ice bath container, then quickly adding 0.5mL perfluoropentane (PFP) into the solution, and adding 1 mL perfluoropentane (PFP)The mixed solution was high sheared for 3min at 0000rpm to form O1/O2Colostrum type; the colostrum formed is then added dropwise to 5mL of a 1% aqueous solution of Sodium Dodecyl Sulfate (SDS) with stirring to form O1/O2Type V/W pre-compound emulsion; and finally, carrying out ultrasonic treatment on the formed pre-compound emulsion to form the nano-emulsion, wherein the ultrasonic condition is that the time is 3s, the interval is 3s, the process is repeated for 10 times, and the ultrasonic power is set to be 100W. Under the ice bath condition, the nano-emulsion is stirred overnight and solidified to form nano-bubbles; centrifuging the obtained nanobubble solution, removing supernatant, dispersing the precipitate in physiological saline again, and storing in a refrigerator at 4 deg.C for use.
The nanobubble particle size was measured at 4 deg.C using a laser particle sizer (Zetasizer/Nano ZS90, Malvern) and found to be 101.1. + -. 2.7nm, as shown in FIG. 2. The results of the laser particle size analyzer tests show that the monodispersity is good and the particle size is uniform.
Example 2
Nanobubble standing stability test
The nanobubbles prepared in example 1 were stored in a refrigerator at 4 deg.C, sampled at regular intervals and measured for particle size using a laser particle sizer, and the stability of the nanobubbles at 4 deg.C was evaluated, the test results are shown in FIG. 3. As can be seen from the test, the particle size and potential change of the nanobubble is very small when the nanobubble is placed at 4 ℃ for 2 months, which indicates that the nanobubble has good stability at 4 ℃.
Example 3
Ultrasonic imaging test of nanobubbles at 37 deg.C
The nanobubbles prepared in example 1 and the control physiological saline and couplant are placed in a 37 ℃ constant temperature water bath, and a transmission electron microscope (Tecnai G220, Fei, the Netherlands) is used for respectively sampling to characterize the microscopic morphology of the nanobubbles and the control physiological saline and couplant under the condition of 37 ℃, as a result, as shown in FIG. 4, the nanobubbles show bright spots in the glove under the irradiation of ultrasonic waves (as shown in FIG. 4 c), the uniform and dense bright spots form ultrasonic high signals and can enhance ultrasonic imaging, and no bright spots are seen in the control physiological saline and couplant (as shown in FIGS. 4a and 4 b).
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any modification, equivalent replacement, and improvement made by those skilled in the art within the technical scope of the present invention should be included in the scope of the present invention.
Claims (10)
1. A method for preparing phase-transition nanobubbles is characterized by comprising the following steps:
(1) dissolving chitosan-polylactic acid graft copolymer as coating material in dichloromethane to obtain oil 1, namely O1Phase, liquid perfluoropentane as oil 2, O2Phase, mixing the two phases in an ice bath to obtain O2Phase is uniformly dispersed in O1Formation of O in phase2/O1Phase (1);
(2) mixing O with2/O1Gradually dripping the phase into the water phase under magnetic stirring in an ice bath to obtain pre-compound emulsion O2/O1A phase of/W;
(3) subjecting the pre-emulsion phase to ultrasonic treatment until O with uniform particle size is obtained2/O1a/W compound emulsion;
(4) mixing O with2/O1And the/W multiple emulsion is magnetically stirred under the ice bath condition, is solidified to form nano bubbles, is centrifuged to obtain precipitates, and is re-dispersed to obtain the nano bubble water dispersion body carrying liquid perfluoropentane.
2. The method for preparing phase-transition nanobubbles according to claim 1 wherein: the structural formula of the chitosan-polylactic acid graft copolymer in the step (1) is as follows:
the structural formula of PLA is:
3. the method for preparing phase-transition nanobubbles according to claim 1 wherein: the number average molecular weight of the chitosan-polylactic acid graft copolymer is 3000-60000.
4. The method for preparing phase-transition nanobubbles according to claim 3 wherein: the number average molecular weight of the chitosan-polylactic acid graft copolymer is 5000-30000.
5. The method for preparing phase-transition nanobubbles according to claim 1 wherein: the step (2) of pre-emulsion O2/O1The weight percentage of the chitosan-polylactic acid graft copolymer in the/W phase is 0.1-5.0 wt%, the weight percentage of the liquid perfluoropentane is 1.0-30.0 wt%, and the balance is ultrapure water; in addition, the aqueous phase may also be an aqueous solution of a surfactant with hydrophilic groups, including SDS, poloxamers, Span20, Tween80, PEG.
6. The method for preparing phase-transition nanobubbles according to claim 1 wherein: o in the step (1)1Phase sum of O2The phase mixing is high shear mixing, and the shear rate is 5000-30000 rpm.
7. The method for preparing phase-transition nanobubbles according to claim 1 wherein the ultrasonic treatment in the step (3) is: carrying out ultrasonic treatment for multiple times, setting the ultrasonic power to be 30-300W, carrying out ultrasonic treatment for 1-10 s every time at intervals of 1-10 s, and repeating for 1-20 times until O with uniform particle size is obtained2/O1and/W compound emulsion.
8. A phase-transition nanobubble prepared according to any of claims 1 to 7, characterized by: the nanobubbles can realize liquid-gas phase transition through temperature change.
9. The phase-shifting nanobubble of claim 8, characterized by: the particle size range of the nanobubble is 30-1000 nm, and the polydispersity index PDI is less than or equal to 0.35.
10. Use of the phase-transition nanobubbles prepared according to any of claims 1 to 7, characterized by: adding an MRI contrast agent to the phase-transition nanobubble for MRI imaging.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113706810A (en) * | 2021-07-22 | 2021-11-26 | 宁波讯强电子科技有限公司 | Acoustic-magnetic anti-theft label, label shell, label sealing cover and preparation method thereof |
| CN115006555A (en) * | 2022-05-07 | 2022-09-06 | 湖北科技学院 | Nanoscale ultrasound/magnetic resonance bimodal contrast agent and preparation method and application thereof |
| CN115887762A (en) * | 2022-10-11 | 2023-04-04 | 苏州大学 | Microfluidic electrostatically spun fibrous membrane microsphere, preparation method thereof and application thereof in vascular repair |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102552946A (en) * | 2012-03-01 | 2012-07-11 | 中国科学院苏州纳米技术与纳米仿生研究所 | Double-mode imaging and medicine-loading integrated nano medicine carrier and preparation method thereof |
| CN109966514A (en) * | 2019-04-26 | 2019-07-05 | 湖北科技学院 | A kind of phase transition targeted nano bubble, preparation method and application |
-
2020
- 2020-06-01 CN CN202010483519.5A patent/CN111632154A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102552946A (en) * | 2012-03-01 | 2012-07-11 | 中国科学院苏州纳米技术与纳米仿生研究所 | Double-mode imaging and medicine-loading integrated nano medicine carrier and preparation method thereof |
| CN109966514A (en) * | 2019-04-26 | 2019-07-05 | 湖北科技学院 | A kind of phase transition targeted nano bubble, preparation method and application |
Non-Patent Citations (1)
| Title |
|---|
| YAN WU ET AL: "Synthesis and characterization of a novel amphiphilic chitosan–polylactide graft copolymer", 《CARBOHYDRATE POLYMERS》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113706810A (en) * | 2021-07-22 | 2021-11-26 | 宁波讯强电子科技有限公司 | Acoustic-magnetic anti-theft label, label shell, label sealing cover and preparation method thereof |
| CN115006555A (en) * | 2022-05-07 | 2022-09-06 | 湖北科技学院 | Nanoscale ultrasound/magnetic resonance bimodal contrast agent and preparation method and application thereof |
| CN115006555B (en) * | 2022-05-07 | 2024-01-12 | 湖北科技学院 | Nanoscale ultrasound/magnetic resonance bimodal contrast agent, and preparation method and application thereof |
| CN115887762A (en) * | 2022-10-11 | 2023-04-04 | 苏州大学 | Microfluidic electrostatically spun fibrous membrane microsphere, preparation method thereof and application thereof in vascular repair |
| CN115887762B (en) * | 2022-10-11 | 2024-03-15 | 苏州大学 | Microfluidic electrospun fiber membrane microsphere, preparation method thereof and application thereof in vascular repair |
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