CN110433143A - Perfluocarbon oxygen carrying microballoon and its preparation method and application - Google Patents
Perfluocarbon oxygen carrying microballoon and its preparation method and application Download PDFInfo
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Abstract
The present invention provides a kind of perfluocarbon oxygen carrying microballoons and its preparation method and application, the step of preparation method are as follows: lotion is made in perfluocarbon, the water-soluble perfluorocarbon emulsion for carrying oxygen is oxygenated and is disinfected, the lotion is wrapped in carboxymethyl chitosan and forms microballoon, and connects folic acid in microsphere surface and realizes target function；Perfluocarbon oxygen carrying microballoon of the invention is applied in treatment tumour and alleviation anoxic are alleviated；The present invention is using perfluocarbon as oxygen carrying material, it is wrapped in carboxymethyl chitosan, and it is grafted folate molecule, form the microballoon of core-shell structure, the slow release effect of oxygen may be implemented in the microballoon, and Shell Materials have good biocompatibility and targeting, so as to alleviate the oxygen deficit problem of tumor locus, it realizes the function in hypoxic sites slow release oxygen, achievees the purpose that treat tumour；The preparation method of perfluocarbon oxygen carrying microballoon of the invention is simple, easy to operate, and the oxygen carrying microballoon prepared can long term storage.
Description
Technical field
The invention belongs to nano materials and biomedical material technology, and in particular to a kind of perfluocarbon oxygen carrying microballoon
And its preparation method and application.
Background technique
In recent years, with the continuous improvement of living standards, people increasingly focus on life security.One system such as tumour, cancer
Column disease seriously threatens the health of people.Anoxic is the common pathophysiological features of most of tumours, this is because oxygen
Gas supply and new blood vessel exception and thrombosis caused by oxygen unbalanced depletion in cancer cell, meanwhile, the anoxic ring of tumour
Border also results in the extraneous adjuvant treatment such as effect of radiotherapy and has a greatly reduced quality.Currently, researcher has discovered that by different sides
Method promotes tumour oxygen content, such as by improving blood flow in tumour, generates oxygen in situ in tumour, and oxygen is transported to
Tumour is treated in tumour with the carrier of oxygen.
Perfluocarbon (PFCs) is a kind of inert chemi-cal substance with good biocompatibility, has been widely used for facing
Bed, such as ultrasonic imaging, organ transplant prevent tissue and the ischemia/reperfusion injury of organ etc..And PFCs possesses high
Oxygen solubility, this becomes a kind of very promising artificial blood substitute.But the time of PFCs release oxygen is very short, and
Excessive oxygen concentration can be damaged to taeniae telarum, it is therefore desirable to be wrapped into microballoon, carry out slow releasing function.
Summary of the invention
Aiming at the shortcomings in the prior art, primary and foremost purpose of the invention is to provide a kind of preparation of perfluocarbon oxygen carrying microballoon
Method, during the preparation process, the selection of microballoon Shell Materials are extremely important, this requires being designed from the structure of material, make
Obtain material not only has pH sensibility simultaneously, but also possesses Targeting Performance, acts on to realize in the oxygen release of tumor locus.
A second object of the present invention is to provide above-mentioned perfluocarbon oxygen carrying microballoons.
Third object of the present invention is to provide the applications of above-mentioned perfluocarbon oxygen carrying microballoon.
In order to achieve the above objectives, solution of the invention is:
A kind of preparation method of perfluocarbon oxygen carrying microballoon comprising following steps:
(1), egg yolk lecithin is added in tyrode's solution, 4-6 times emulsified at 25-30 DEG C, each duration is
5-10s is spaced 30-60s, obtains basic emulsion；
(2), in basic emulsion be added volume fraction >=97% perfluocarbon (PFCs) stoste, emulsify 4-7 times, every time
The time of emulsification is 5-10s, and every minor tick 30-60s obtains perfluocarbon original lotion；
(3), by perfluocarbon original absorbing emulsions oxygen, perfluorocarbon emulsion is obtained；
(4), by perfluorocarbon emulsion filtration sterilization, the sterile lotion of perfluocarbon is obtained；
(5), carboxymethyl chitosan is dissolved into distilled water, it is sub- that 1- (3- dimethylamino-propyl) -3- ethyl carbon two is added
Amine hydrochlorate and N- hydroxysuccinimide, the pH with lye regulation system are neutrality, obtain the first mixed liquor；
(6), folic acid is dispersed in dimethyl sulfoxide-aqueous solution, obtains the second mixed liquor, adjust the second mixed liquor with lye
PH be alkalescent；1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride and N- hydroxysuccinimide are added,
Third mixed liquor is obtained, is 8 with the pH value that lye adjusts third mixed liquor, and is protected from light 4-8h at 20-30 DEG C, obtains the
Four mixed liquors；
(7), the first mixed liquor is added in the 4th mixed liquor, reacts 8-10h at 20-25 DEG C, obtains the 5th mixed liquor,
5th mixed liquor is centrifuged 10-15min, supernatant is removed, obtains solids；
(8), solids are dissolved in phosphate buffer solution, after to be dissolved, obtain polymer solution, by perfluocarbon
Sterile lotion is added drop-wise in polymer solution, is incubated for 18-24h at 37 DEG C, is then centrifuged for 10-15min, is obtained perfluocarbon and is taken
Oxygen microballoon.
Preferably, in step (1), mass percent of the egg yolk lecithin in tyrode's solution is 17-20%；When emulsification, make
The power of ultrasonic emulsifier is 350-450W.
Preferably, in step (2), the volume ratio of basic emulsion and perfluocarbon stoste is 1:1-1.4:1；When emulsification, make
The power of ultrasonic emulsifier is 350-450W.
Preferably, in step (2), perfluocarbon in perfluocarbon stoste is selected from perfluorotributylamine, perfluor cyclopropylamine, complete
One or more of fluorine n-octyl bromide, phenyl-hexafluoride and FtBu.
Preferably, in step (3), the time of perfluocarbon original absorbing emulsions oxygen is 15-20min.
Preferably, in step (4), when filtration sterilization, the diameter of the sterile filters used is 0.65-0.8 μm.
Preferably, in step (5), lye is sodium hydroxide solution, and the pH value of system is 6.5-7.
Preferably, in step (6), lye is sodium hydroxide solution；The pH value of second mixed liquor is 7-8.
Preferably, in step (7), when centrifugation, the revolving speed using centrifuge is 8000-12000r/min.
Preferably, in step (8), when centrifugation, the revolving speed using centrifuge is 10000-12000r/min.
A kind of perfluocarbon oxygen carrying microballoon, is obtained by above-mentioned preparation method.
A kind of above-mentioned perfluocarbon oxygen carrying microballoon is in treatment tumour and alleviates the application in anaerobic environment.
By adopting the above scheme, the beneficial effects of the present invention are:
The first, perfluocarbon (PFCs) of the invention is a kind of change with good biocompatibility, biologically inert in itself
Substance is learned, and as its unique chemical structure (hydrogen atom on molecule is all replaced fluorine atom), keeps PFCs also same
When have high hydrophobicity and hypoergia, this make it have dissolution bulk gas ability.The oxygen course of dissolution of many PFCs
It is a kind of physical action, meltage is nearly 20 times of water, and more taller than hemoglobin dissolved oxygen amount, a large amount of oxygen of carrying are usual
It can be used for being discharged into tissue.In addition, PFCs lotion can be prepared into nanoscale, this small size allows them to convey oxygen
Into the smallest capillary；In addition, and its is from a wealth of sources, there is very extensive application in field of tissue engineering technology.
The second, the Shell Materials of perfluocarbon oxygen carrying microballoon of the present invention are carboxymethyl chitosan and folic acid, carboxymethyl chitosan
Sugar is a kind of derivative of chitosan, chitosan have good biocompatibility, microbic resolvability, height safety with
And special pharmacological activity, and have amino abundant on chitosan, at low ph conditions easily at salt, due to the row of like charges
Reprimand effect, polymer network can be expanded, thus also have excellent pH sensibility.Folic acid can excessive table in tumour cell
It reaches, and is limited expression in normal tissue or organ, the function that this feature can be positioned with active targeting, and leaf
There are also good compatibilities for acid, and folic acid is grafted in microballoon system simultaneously, microballoon can be made to realize active targeting and by moving-target
To combination, alleviate tumour cell near anaerobic environment, therefore, the oxygen carrying microballoon of this core-shell structure can greatly improve needle
To the ability of tumor locus sustained release oxygen.
Third, the preparation method of perfluocarbon oxygen carrying microballoon of the invention are simple, easy to operate, the oxygen carrying microballoon prepared
It can long term storage.
In short, the present invention with perfluocarbon (PFCs) be oxygen carrying material, be wrapped in carboxymethyl chitosan, and be grafted folic acid
Molecule forms the microballoon of core-shell structure, which may be implemented the slow release effect of oxygen, and Shell Materials have good life
Object compatibility and targeting achieve the purpose that treat tumour so as to alleviate the anaerobic condition of tumor locus.
Specific embodiment
The present invention provides a kind of perfluocarbon oxygen carrying microballoons and its preparation method and application.
The present invention is wrapped in carboxymethyl chitosan using PFCs as oxygen carrying lotion, while chemical graft folic acid improves targeting
Property, the perfluocarbon oxygen carrying microballoon of pH sensitivity is obtained, makes it that can discharge oxygen in the acidic environment of tumour, it is swollen to reach killing
The purpose of tumor.
<preparation method of perfluocarbon oxygen carrying microballoon>
The preparation method of perfluocarbon oxygen carrying microballoon of the invention includes the following steps:
(1), the preparation of the sterile lotion of PFCs: egg yolk lecithin is added in tyrode's solution (Tyrode solution), in 25-30
It is emulsified 4-6 times at DEG C using ultrasonic emulsifier, each duration is 5-10s, is spaced 30-60s, obtains basic emulsion；In
The perfluocarbon stoste of volume fraction >=97% is added in basic emulsion, is emulsified 4-7 times using ultrasonic emulsifier, is emulsified every time
Time is 5-10s, every minor tick 30-60s, obtains perfluocarbon original lotion, spare；Perfluocarbon original lotion is put into hyperbaric oxygen
In cabin, oxygen is fully absorbed, obtains perfluorocarbon emulsion；The perfluorocarbon emulsion that oxygenation finishes is crossed using sterile filters and is filtered out
Bacterium obtains the sterile lotion of perfluocarbon, spare；
(2), the preparation of Shell Materials: carboxymethyl chitosan is dissolved into 5mL distilled water, and the 1- (3- bis- of 100mg is added
Methylaminopropyl) -3- ethyl-carbodiimide hydrochloride (EDC) and 100mg N- hydroxysuccinimide (NHS), with lye tune
The pH of section system is neutrality, obtains the first mixed liquor；By 70mg folic acid ultrasonic disperse in 10mL dimethyl sulfoxide-aqueous solution, obtain
To the second mixed liquor, the pH that the second mixed liquor is adjusted with lye is alkalescent；Add the 1- (3- dimethylamino-propyl)-of 66mg
The N- hydroxysuccinimide (NHS) of 3- ethyl-carbodiimide hydrochloride (EDC) and 37mg, obtains third mixed liquor, uses diluted alkaline
The pH value that liquid adjusts third mixed liquor is 8 or so, and is protected from light 4-8h at 20-30 DEG C, obtains the 4th mixed liquor；By first
Mixed liquor is slowly dropped in the 4th mixed liquor, reacts 8-10h at 20-25 DEG C, obtains the 5th mixed liquor, by the 5th mixed liquor
It is centrifuged 10-15min in supercentrifuge, removes supernatant, obtains solids, it is spare；
(3), the preparation of oxygen carrying microballoon: by phosphate (PBS) buffer solution that solids are dissolved in 37 DEG C, pH value is 7.4
In, after to be dissolved, obtain polymer solution；Under magnetic stirring, the sterile lotion of perfluocarbon is added drop-wise in polymer solution,
After mixed with polymers is uniform, it is incubated for 18-24h at 37 DEG C, is centrifuged the polymer solution after incubation using supercentrifuge
10-15min obtains cured nanoparticle, i.e. perfluocarbon oxygen carrying microballoon, by the use for adjusting carboxymethyl chitosan and folic acid
Amount and stirring rate, the different microballoon of available partial size.
Wherein, in step (1), mass percent of the egg yolk lecithin in tyrode's solution can be 17-20%, preferably
17%；When emulsification, the power of the ultrasonic emulsifier used can be 350-450W, preferably 450W.
In step (1), the volume ratio of basic emulsion and perfluocarbon stoste can be 1:1-1.4:1, preferably 1:1；
When emulsification, the power of the ultrasonic emulsifier used can be 350-450W, preferably 450W.
In step (1), it is pungent that the perfluocarbon in perfluocarbon stoste is selected from perfluorotributylamine, perfluor cyclopropylamine, perfluor
One or more of bromide, phenyl-hexafluoride and FtBu.
In step (1), the time that perfluocarbon original lotion is placed in hyperbaric oxygen chamber can be 15-20min, preferably
20min。
In step (1), the diameter of sterile filters can be 0.65-0.8 μm, preferably 0.8 μm.
In step (2), lye is sodium hydroxide solution, and the pH value of system can be 6.5-7, preferably 6.5.
In step (2), the pH value of the second mixed liquor can be 7-8, preferably 8.
In step (2), the revolving speed of supercentrifuge can be 8000-12000r/min, preferably 8000r/min.
In step (3), the revolving speed of supercentrifuge can be 10000-12000r/min, preferably 10000r/min.
<perfluocarbon oxygen carrying microballoon>
Perfluocarbon oxygen carrying microballoon of the invention is obtained by above-mentioned preparation method.
<application of perfluocarbon oxygen carrying microballoon>
Perfluocarbon oxygen carrying microballoon of the invention can be applied in treatment tumour and alleviation anaerobic environment.
The present invention is further illustrated with reference to embodiments.
Embodiment 1:
The preparation method of the perfluorotributylamine oxygen carrying microballoon of the present embodiment includes the following steps:
(1), egg yolk lecithin is added in Tyrode solution, mass percent shared by egg yolk lecithin is 17%, In
It is emulsified 6 times, power 450W at 25 DEG C using ultrasonic emulsifier, each duration is 10s, is spaced 30s, obtains basic cream
Liquid；
(2), the perfluorotributylamine stoste that volume fraction is 97%, three fourth of basic emulsion and perfluor are added in basic emulsion
The volume ratio of amine stoste is 1:1, is emulsified 7 times, power 450W using ultrasonic emulsifier by above-mentioned steps, the time emulsified every time
For 10s, every minor tick 60s, perfluorotributylamine original lotion is obtained, it is spare；
(3), perfluorotributylamine original lotion is put into 20min in hyperbaric oxygen chamber, fully absorbs oxygen, obtains perfluorotributylamine
Lotion；
(4), the perfluorotributylamine lotion filtration sterilization that the sterile filters for the use of diameter being 0.8 μm finish oxygenation, obtains
The sterile lotion of perfluorotributylamine, it is spare；
(5), carboxymethyl chitosan is dissolved into 5mL distilled water, 1- (3- dimethylamino-propyl) -3- of 100mg is added
The N- hydroxysuccinimide (NHS) of ethyl-carbodiimide hydrochloride (EDC) and 100mg, the pH value with NaOH regulation system are
6.5, obtain the first mixed liquor；
(6), by 70mg folic acid ultrasonic disperse in 10mL dimethyl sulfoxide-aqueous solution, the volume ratio of dimethyl sulfoxide and water is
1:1 obtains the second mixed liquor, is 8 with the pH value that NaOH adjusts the second mixed liquor；Add 1- (the 3- dimethylamino third of 66mg
Base) -3- ethyl-carbodiimide hydrochloride (EDC) and 37mg N- hydroxysuccinimide (NHS), obtain third mixed liquor, use
The pH value that dilute NaOH adjusts third mixed liquor is 8, and is protected from light 8h at 25 DEG C, obtains the 4th mixed liquor；
(7), the first mixed liquor is slowly dropped in the 4th mixed liquor, reacts 8h at 25 DEG C, obtain the 5th mixed liquor,
5th mixed liquor is centrifuged 15min, revolving speed 8000r/min in supercentrifuge, supernatant is removed, obtains solids,
It is spare；
(8), by solids be dissolved in 37 DEG C, pH value be 7.4 phosphate (PBS) buffer solution in, after to be dissolved, obtain
The sterile lotion of perfluorotributylamine is added drop-wise in polymer solution by polymer solution under magnetic stirring, when mixed with polymers is equal
After even, it is incubated for 18h at 37 DEG C, the polymer solution after incubation is centrifuged 10min using supercentrifuge, revolving speed is
10000r/min obtains cured nanoparticle, i.e. perfluorotributylamine oxygen carrying microballoon.
Embodiment 2:
The perfluor cyclopropylamine of the present embodiment and the preparation method of perfluorooctyl bromide oxygen carrying microballoon include the following steps:
(1), egg yolk lecithin is added in Tyrode solution, mass percent shared by egg yolk lecithin is 20%, In
It is emulsified 4 times, power 350W at 25 DEG C using ultrasonic emulsifier, each duration is 5s, is spaced 30s, obtains basic cream
Liquid；
(2), perfluor cyclopropylamine and the perfluorooctyl bromide stoste that volume fraction is 98%, basis cream are added in basic emulsion
The volume ratio of liquid and perfluor cyclopropylamine and perfluorooctyl bromide stoste is 1.4:1, using ultrasonic emulsifier by above-mentioned steps emulsification 4
Secondary, power 350W, the time emulsified every time is 5s, every minor tick 30s, obtains perfluor cyclopropylamine and the former cream of perfluorooctyl bromide
Liquid, it is spare；
(3), perfluor cyclopropylamine and perfluorooctyl bromide original lotion are put into 15min in hyperbaric oxygen chamber, fully absorb oxygen, obtains
To perfluor cyclopropylamine and perfluoro capryl bromoiodide emulsion；
(4), perfluor cyclopropylamine and perfluoro capryl bromoiodide emulsion that the sterile filters for the use of diameter being 0.65 μm finish oxygenation
Filtration sterilization obtains perfluor cyclopropylamine and the sterile lotion of perfluorooctyl bromide, spare；
(5), carboxymethyl chitosan is dissolved into 5mL distilled water, 1- (3- dimethylamino-propyl) -3- of 100mg is added
The N- hydroxysuccinimide (NHS) of ethyl-carbodiimide hydrochloride (EDC) and 100mg, the pH value with NaOH regulation system are
7, obtain the first mixed liquor；
(6), by 70mg folic acid ultrasonic disperse in 10mL dimethyl sulfoxide-aqueous solution, the volume ratio of dimethyl sulfoxide and water is
1:1 obtains the second mixed liquor, is 7 with the pH value that NaOH adjusts the second mixed liquor；Add 1- (the 3- dimethylamino third of 66mg
Base) -3- ethyl-carbodiimide hydrochloride (EDC) and 37mg N- hydroxysuccinimide (NHS), obtain third mixed liquor, use
The pH value that dilute NaOH adjusts third mixed liquor is 8, and is protected from light 4h at 30 DEG C, obtains the 4th mixed liquor；
(7), the first mixed liquor is slowly dropped in the 4th mixed liquor, reacts 8h at 25 DEG C, obtain the 5th mixed liquor,
5th mixed liquor is centrifuged 10min, revolving speed 10000r/min in supercentrifuge, supernatant is removed, obtains solids,
It is spare；
(8), by solids be dissolved in 37 DEG C, pH value be 7.4 phosphate (PBS) buffer solution in, after to be dissolved, obtain
Perfluor cyclopropylamine and the sterile lotion of perfluorooctyl bromide are added drop-wise in polymer solution by polymer solution under magnetic stirring, when
It after mixed with polymers is uniform, is incubated at 37 DEG C for 24 hours, the polymer solution after incubation is centrifuged 10min using supercentrifuge,
Revolving speed is 12000r/min, obtains cured nanoparticle, i.e. perfluor cyclopropylamine and perfluorooctyl bromide oxygen carrying microballoon.
Wherein, perfluor cyclopropylamine and the mass ratio of perfluorooctyl bromide are 1:1.
Embodiment 3:
The preparation method of the phenyl-hexafluoride oxygen carrying microballoon of the present embodiment includes the following steps:
(1), egg yolk lecithin is added in Tyrode solution, mass percent shared by egg yolk lecithin is 18%, In
It is emulsified 6 times, power 400W at 25 DEG C using ultrasonic emulsifier, each duration is 10s, is spaced 40s, obtains basic cream
Liquid；
(2), the phenyl-hexafluoride stoste that addition volume fraction is 98% in basic emulsion, basic emulsion and phenyl-hexafluoride stoste
Volume ratio is 1.2:1, is emulsified 6 times, power 400W using ultrasonic emulsifier by above-mentioned steps, and the time emulsified every time is 10s,
Every minor tick 40s obtains phenyl-hexafluoride original lotion, spare；
(3), phenyl-hexafluoride original lotion is put into 18min in hyperbaric oxygen chamber, fully absorbs oxygen, obtain phenyl-hexafluoride lotion；
(4), the phenyl-hexafluoride lotion filtration sterilization that the sterile filters for the use of diameter being 0.7 μm finish oxygenation, obtains hexafluoro
The sterile lotion of benzene, it is spare；
(5), carboxymethyl chitosan is dissolved into 5mL distilled water, 1- (3- dimethylamino-propyl) -3- of 100mg is added
The N- hydroxysuccinimide (NHS) of ethyl-carbodiimide hydrochloride (EDC) and 100mg, the pH value with NaOH regulation system are
6.8, obtain the first mixed liquor；
(6), by 70mg folic acid ultrasonic disperse in 10mL dimethyl sulfoxide-aqueous solution, the volume ratio of dimethyl sulfoxide and water is
1:1 obtains the second mixed liquor, is 7.8 with the pH value that NaOH adjusts the second mixed liquor；Add 1- (the 3- dimethylamino of 66mg
Propyl) -3- ethyl-carbodiimide hydrochloride (EDC) and 37mg N- hydroxysuccinimide (NHS), obtain third mixed liquor,
It is 8 with the pH value that dilute NaOH adjusts third mixed liquor, and is protected from light 4h at 26 DEG C, obtains the 4th mixed liquor；
(7), the first mixed liquor is slowly dropped in the 4th mixed liquor, reacts 8h at 25 DEG C, obtain the 5th mixed liquor,
5th mixed liquor is centrifuged 12min, revolving speed 12000r/min in supercentrifuge, supernatant is removed, obtains solids,
It is spare；
(8), by solids be dissolved in 37 DEG C, pH value be 7.4 phosphate (PBS) buffer solution in, after to be dissolved, obtain
The sterile lotion of phenyl-hexafluoride is added drop-wise in polymer solution by polymer solution under magnetic stirring, when mixed with polymers is uniform
Afterwards, it is incubated for 20h at 37 DEG C, the polymer solution after incubation is centrifuged 10min, revolving speed 12000r/ using supercentrifuge
Min obtains cured nanoparticle, i.e. phenyl-hexafluoride oxygen carrying microballoon.
Embodiment 4:
The preparation method of the FtBu oxygen carrying microballoon of the present embodiment includes the following steps:
(1), egg yolk lecithin is added in Tyrode solution, mass percent shared by egg yolk lecithin is 20%, In
It is emulsified 6 times, power 400W at 25 DEG C using ultrasonic emulsifier, each duration is 10s, is spaced 40s, obtains basic cream
Liquid；
(2), in basic emulsion be added volume fraction be 97% FtBu stoste, basic emulsion and
The volume ratio of FtBu stoste is 1.4:1, is emulsified 6 times using ultrasonic emulsifier by above-mentioned steps, power is
400W, the time emulsified every time are 10s, every minor tick 30s, obtain FtBu original lotion, spare；
(3), FtBu original lotion is put into 18min in hyperbaric oxygen chamber, fully absorbs oxygen, obtained complete
Fluorine (t-butylcyclohexane) lotion；
(4), the FtBu lotion that oxygenation finishes is crossed and is filtered out by the sterile filters for the use of diameter being 0.8 μm
Bacterium obtains the sterile lotion of FtBu, spare；
(5), carboxymethyl chitosan is dissolved into 5mL distilled water, 1- (3- dimethylamino-propyl) -3- of 100mg is added
The N- hydroxysuccinimide (NHS) of ethyl-carbodiimide hydrochloride (EDC) and 100mg, the pH value with NaOH regulation system are
7, obtain the first mixed liquor；
(6), by 70mg folic acid ultrasonic disperse in 10mL dimethyl sulfoxide-aqueous solution, the volume ratio of dimethyl sulfoxide and water is
1:1 obtains the second mixed liquor, is 8 with the pH value that NaOH adjusts the second mixed liquor；Add 1- (the 3- dimethylamino third of 66mg
Base) -3- ethyl-carbodiimide hydrochloride (EDC) and 37mg N- hydroxysuccinimide (NHS), obtain third mixed liquor, use
The pH value that dilute NaOH adjusts third mixed liquor is 8, and is protected from light 6h at 25 DEG C, obtains the 4th mixed liquor；
(7), the first mixed liquor is slowly dropped in the 4th mixed liquor, reacts 8h at 25 DEG C, obtain the 5th mixed liquor,
5th mixed liquor is centrifuged 13min, revolving speed 8000r/min in supercentrifuge, supernatant is removed, obtains solids,
It is spare；
(8), by solids be dissolved in 37 DEG C, pH value be 7.4 phosphate (PBS) buffer solution in, after to be dissolved, obtain
The sterile lotion of FtBu is added drop-wise in polymer solution under magnetic stirring, works as polymerization by polymer solution
Object after mixing, is incubated for 22h at 37 DEG C, and the polymer solution after incubation is centrifuged 10min, revolving speed using supercentrifuge
For 12000r/min, cured nanoparticle, i.e. FtBu oxygen carrying microballoon are obtained.
The above-mentioned description to embodiment is that this hair can be understood and used for the ease of those skilled in the art
It is bright.Those skilled in the art obviously readily can make various modifications to these embodiments, and described herein one
As principle be applied in other embodiments, without having to go through creative labor.Therefore, the present invention is not limited to the above embodiments.
Those skilled in the art's principle according to the present invention, not departing from improvement that scope of the invention is made and modification all should be at this
Within the protection scope of invention.
Claims (10)
1. a kind of preparation method of perfluocarbon oxygen carrying microballoon, it is characterised in that: it includes the following steps:
(1), egg yolk lecithin is added in tyrode's solution, 4-6 times emulsified at 25-30 DEG C, each duration is 5-
10s is spaced 30-60s, obtains basic emulsion；
(2), the perfluocarbon stoste of volume fraction >=97% is added in the basic emulsion, emulsifies 4-7 times, emulsifies every time
Time is 5-10s, and every minor tick 30-60s obtains perfluocarbon original lotion；
(3), by the perfluocarbon original absorbing emulsions oxygen, perfluorocarbon emulsion is obtained；
(4), by the perfluorocarbon emulsion filtration sterilization, the sterile lotion of perfluocarbon is obtained；
(5), carboxymethyl chitosan is dissolved into distilled water, 1- (3- dimethylamino-propyl) -3- ethyl carbodiimide salt is added
Hydrochlorate and N- hydroxysuccinimide, the pH with lye regulation system are neutrality, obtain the first mixed liquor；
(6), folic acid is dispersed in dimethyl sulfoxide-aqueous solution, obtains the second mixed liquor, the pH of the second mixed liquor is adjusted with lye
For alkalescent；1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride and N- hydroxysuccinimide are added, is obtained
Third mixed liquor is 8 with the pH value that lye adjusts third mixed liquor, and is protected from light 4-8h at 20-30 DEG C, and it is mixed to obtain the 4th
Close liquid；
(7), first mixed liquor is added in the 4th mixed liquor, reacts 8-10h at 20-25 DEG C, it is mixed to obtain the 5th
Liquid is closed, the 5th mixed liquor is centrifuged 10-15min, supernatant is removed, obtains solids；
(8), the solids are dissolved in phosphate buffer solution, after to be dissolved, obtain polymer solution；By perfluocarbon
Sterile lotion is added drop-wise in the polymer solution, is incubated for 18-24h at 37 DEG C, is then centrifuged for 10-15min, is obtained perfluorinated
Carbon oxygen carrying microballoon.
2. preparation method according to claim 1, it is characterised in that: in step (1), the egg yolk lecithin is in tyrode's solution
In mass percent be 17-20%；When the emulsification, the power of the ultrasonic emulsifier used is 350-450W.
3. preparation method according to claim 1, it is characterised in that: in step (2), the basic emulsion and perfluocarbon
The volume ratio of stoste is 1:1-1.4:1；When the emulsification, the power of the ultrasonic emulsifier used is 350-450W.
4. preparation method according to claim 1, it is characterised in that: complete in the perfluocarbon stoste in step (2)
Fluorocarbons is selected from one of perfluorotributylamine, perfluor cyclopropylamine, perfluorooctyl bromide, phenyl-hexafluoride and FtBu
More than.
5. preparation method according to claim 1, it is characterised in that: in step (3), the perfluocarbon original absorbing emulsions
The time of oxygen is 15-20min；And/or
In step (4), when the filtration sterilization, the diameter of the sterile filters used is 0.65-0.8 μm.
6. preparation method according to claim 1, it is characterised in that: in step (5), the lye is that sodium hydroxide is molten
Liquid, the pH value of system are 6.5-7.
7. preparation method according to claim 1, it is characterised in that: in step (6), the lye is that sodium hydroxide is molten
Liquid；The pH value of second mixed liquor is 7-8.
8. preparation method according to claim 1, it is characterised in that: in step (7), when the centrifugation, use centrifuge
Revolving speed be 8000-12000r/min；And/or
In step (8), when the centrifugation, the revolving speed using centrifuge is 10000-12000r/min.
9. a kind of perfluocarbon oxygen carrying microballoon, it is characterised in that: it is obtained by the described in any item preparation methods of claim 1-8
It arrives.
10. a kind of application by perfluocarbon oxygen carrying microballoon as claimed in claim 9 in treatment tumour and alleviation anaerobic environment.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112121162A (en) * | 2020-09-07 | 2020-12-25 | 四川康城生物科技有限公司 | Carboxymethyl chitosan microcapsule for carrying chemotherapy medicament and thermotherapy sensitizer for sustained release of novel embolism and preparation method and application thereof |
| CN117483931A (en) * | 2024-01-03 | 2024-02-02 | 四川钛程钛业有限公司 | Explosion welding preparation method of novel marine metal composite board |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140213548A1 (en) * | 2013-01-27 | 2014-07-31 | New Jersey Institute Of Technology | System and method for hemostatic wound dressing |
| CN106075471A (en) * | 2016-07-12 | 2016-11-09 | 上海纳米技术及应用国家工程研究中心有限公司 | A kind of have lympha targeted fluorocarbon emulsion and preparation method thereof |
-
2019
- 2019-07-29 CN CN201910686599.1A patent/CN110433143A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140213548A1 (en) * | 2013-01-27 | 2014-07-31 | New Jersey Institute Of Technology | System and method for hemostatic wound dressing |
| CN106075471A (en) * | 2016-07-12 | 2016-11-09 | 上海纳米技术及应用国家工程研究中心有限公司 | A kind of have lympha targeted fluorocarbon emulsion and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| DOUGLAS ET AL.: "Enrichment of chitosan hydrogels with perfluorodecalin promotes gelation and stem cell vitality", 《MATERIALS LETTERS》 * |
| 张岩等: "载阿霉素的叶酸偶联羧甲基壳聚糖自组装纳米粒的研制及体外释放", 《沈阳大学学报(自然科学版)》 * |
| 朱澍: "新型携氧微球改善外周神经损伤后局部缺氧微环境的实验研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 陈玲等: "构建新型液态氟碳壳聚糖纳米超声造影微粒", 《中国医学影像技术》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112121162A (en) * | 2020-09-07 | 2020-12-25 | 四川康城生物科技有限公司 | Carboxymethyl chitosan microcapsule for carrying chemotherapy medicament and thermotherapy sensitizer for sustained release of novel embolism and preparation method and application thereof |
| CN117483931A (en) * | 2024-01-03 | 2024-02-02 | 四川钛程钛业有限公司 | Explosion welding preparation method of novel marine metal composite board |
| CN117483931B (en) * | 2024-01-03 | 2024-04-23 | 四川钛程钛业有限公司 | Explosion welding preparation method of novel marine metal composite board |
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