CN108042802A - A kind of artificial red cells motor and preparation method thereof - Google Patents
A kind of artificial red cells motor and preparation method thereof Download PDFInfo
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Abstract
The present invention is with hemoglobin,Natural fine after birth,Magnetic micro-nano granules are material,Utilize chemical coprecipitation and cell membrane fusion technology,By the main constituents hemoglobin of red blood cell,Magnetic micro-nano granules and the cell membrane with cell biological performance combine together,Prepare the artificial red cells motor with red blood cell concave-concave disc structure,Compared with the prior art,The beneficial effects of the present invention are,Repeatability of the invention is strong,Preparation process is simple,And the artificial motor prepared has the biological property of class cell,Stability is good,It can be under the driving of external source ultrasonic field,It is quickly moved along acoustic pressure gradient,Simultaneously,Artificial red cells motor can also pass through the cells such as the natural fine after birth specific recognition cancer cell of surface modification,Or by applying external source magnetic field,Realize the targeting campaign in artificial red cells motor movement direction,It targets and transports in oxygen,Drug specificity carries,Toxin is removed,The biomedical sectors such as oncotherapy are with a wide range of applications.
Description
Technical field
The present invention relates to medical biotechnology field of material technology, and in particular to a kind of artificial red cells motor and its preparation side
Method.
Background technology
Early in Hollywood science fiction movies in 1966《Unusual travelling》In, people just imagine macro object is narrowed down to it is small
Scale, and the object that these are reduced is injected in body, and it is every multiple to perform health examination, disease treatment, organ reparation etc.
Miscellaneous task.Since half a century, with the continuous development of micro-nano science, these imaginations have been increasingly becoming reality.It is micro-nano
Motor is to refer to the energy such as chemical energy, acoustic energy, luminous energy being converted into mechanical movement simultaneously complete the micro-nano system of complex task
System.In recent years, micro-nano motor has become the research hotspot of micro-nano science and its association area.Wherein, the Nuo Bei of 2016
Your chemistry prize is even more three scientists for authorizing and having outstanding contributions in molecular motor research field.At this stage, micro-nano motor
Type of drive mainly include bubble driving, optical drive, field drives and ultrasound-driven.Wherein, bubble drive motor mostly with
Hydrogen peroxide is fuel.Since hydrogen peroxide is relatively low in the in vivo content of people and has certain toxic side effect to human body,
The biological utility of bubble drive motor is poor.Though in addition, at present most of micro-nano motors with certain locomitivity but
Its targets identification ability is poor, is unfavorable for realizing the efficient targeting transport of privileged site.
In view of drawbacks described above, creator of the present invention proposes the present invention by prolonged research and practice.
The content of the invention
To solve above-mentioned technological deficiency, the technical solution adopted by the present invention is, provides a kind of artificial red cells motor,
Red blood cell shape skeleton, the magnetic nanoparticle and surface modification positioned at the skeletal internal formed including natural hemoglobin
Cell membrane.
Preferably, the magnetism micro-nano granules are ferriferrous oxide particles, and the grain size of the magnetic micro-nano granules
For 5nm-500nm.
Preferably, the natural hemoglobin is included in pig source hemoglobin, horse source hemoglobin or ox source hemoglobin
One kind.
Preferably, the cell membrane includes one kind in erythrocyte membrane, leucocyte film or platelet membrane.
Preferably, preparing a kind of method of artificial red cells motor, comprise the following steps：
Step 1: prepare the magnetic micro-nano granules；
Step 2: by chemical coprecipitation, contain the magnetic micro-nano granules described in step 1 and natural blood in synthesis
The red blood cell shape kernel of Lactoferrin；
Step 3: using the red blood cell shape kernel described in step 2 as template, using cell membrane fusion technology by n cell
It is film modified on the surface of the template, obtain artificial red cells motor skeleton；
Step 4: the kernel in the artificial red cells motor skeleton obtained using solvent dissolution method removal step three,
Finally obtain artificial red cells motor.
Preferably, kernel described in step 2 is calcium hydroxide particle, and the grain size of the kernel is 2 μm -100 μm.
Preferably, it is phosphate-buffered that 0.01-0.1mol/L, pH are 5.0-7.0 that solvent described in step 4, which is concentration,
Liquid, the ethylenediamine tetrem that the dilute hydrochloric acid solution or pH value that concentration is 0.005-0.05mol/L are 7.0, concentration is 0.1-1mol/L
One kind in acid disodium salt solution.
Preferably, a diameter of 2 μm -100 μm of red blood cell motor described in step 4, thickness is 500nm-5 μm.
Preferably, preparing a kind of method of artificial red cells motor, comprise the following steps：
Step 1: prepare the magnetic micro-nano granules：A, by the frerrous chloride and 0.004- of 0.002-0.02mol
The iron chloride of 0.04mol presses 1:2 weight ratio is dissolved into the deionized water of 100-1000mL, and 70 are heated under nitrogen protection
DEG C, 40mL ammonium hydroxide is added in while stirring；B, the reaction was continued 1 it is small when after stop heating, the flask equipped with above-mentioned reaction liquid puts
In on magnet, 30min is stood, after ferriferrous oxide particles to be formed deposit to the bottom of the flask completely, removes upper strata
Liquid；C, after cleaning ferriferrous oxide particles 3-4 time using acetone, into the flask, 25mL is not oxygenous goes for addition
Ionized water obtains ferriferrous oxide particles suspension；
Step 2: synthetic red blood cells shape calcium hydroxide particle：A, by the natural hemoglobin of 100-1000mg, 0.5-5g
Dextran sulfate is distributed to the CaCl that 100mL concentration is 0.05-0.5mol/L2In solution, mixed solution A is obtained；B, to 20mL
The magnetic ferroferric oxide particle suspension liquid described in step 1 is added in the sodium hydroxide solution that concentration is 0.5-5mol/L, is obtained
Mixed solution B；C, the mixed solution A is agreed into ultrasound 10 seconds after the mixed solution B is mixed rapidly；D, after standing 20min
It is collected by centrifugation to get to the red blood cell shape calcium hydroxide containing the magnetic ferroferric oxide particle and the natural hemoglobin
Particle；
Step 3: prepare artificial hemoglobin motor skeleton：A, the spy that different cell densities are different in blood is made full use of
Property, using Ficoll density-gradient centrifugation methods, blood of human body is centrifuged with the speed of 4500rpm/min, cell is collected in separation, profit
With Hypotonic treatment cell 24 at 4 DEG C of hypotonic buffer liquid it is small when, fully to remove intracellular content；B, using mini extruding
Device squeezes cell through the polyurethane film 21 times that aperture is 200nm, prepares the cell membrane nanocapsule that grain size is 200nm repeatedly
Bubble；C, with the red blood cell shape calcium hydroxide particle containing magnetic ferroferric oxide particle and natural hemoglobin described in step 2
For kernel, kernel is sufficiently mixed with the cell membrane nano vesicle described in b, based on cell membrane fusion technology, is existed using Ultrasound Instrument
Ultrasound fusion 30min under 59kHz frequencies, in the core surface fused cell film, completes the artificial hemoglobin motor bone
The preparation of frame.
Step 4: dissolving kernel：Artificial hemoglobin motor skeleton described in step 3 is added to concentration as 0.01-
0.1mol/L, pH scope are the phosphate buffer of 5.0-7.0, and concentration is the dilute hydrochloric acid solution or pH of 0.005-0.05mol/L
It is worth in the disodium EDTA solution for being 0.1-1mol/L for 7.0, concentration, is centrifuged after 30min, supernatant discarding
Liquid, gained precipitation are cleaned three times with the phosphate buffer of pH 7.4, finally obtain the artificial red cells motor.
Compared with the prior art, the beneficial effects of the present invention are：Repeatability of the invention is strong, and preparation process is simple, and
Artificial motor to be modified using the cell membrane surface of n cell, makes artificial motor that there is the biological property of class cell, stability is good,
And the concave-concave disc structure with class red blood cell can quickly move under the driving of external source ultrasonic field along acoustic pressure gradient.Meanwhile people
Making red blood cell motor can also be by cells such as the natural fine after birth specific recognition cancer cells of surface modification or outer by applying
The targeting campaign in artificial red cells motor movement direction is realized in source magnetic field, and in oxygen targeting transport, drug specificity carries, is malicious
The biomedical sectors such as plain removing, oncotherapy are with a wide range of applications.
Description of the drawings
It is required in being described below to embodiment in order to illustrate more clearly of the technical solution in various embodiments of the present invention
The attached drawing used is briefly described.
Fig. 1 is the preparation process 3 of artificial red cells motor in the embodiment of the present invention 2 and the schematic diagram of step 4；
Fig. 2 is the schematic diagram that artificial red cells motor moves about in the embodiment of the present invention 2；
Fig. 3 is the laser confocal microscope photo of artificial hemoglobin motor prepared by the embodiment of the present invention 3；
Fig. 4 is the light microscope moved under the ultrasound-driven of artificial hemoglobin motor prepared by the embodiment of the present invention 3
Photo；
Fig. 5 is to target fortune under the ultrasound of artificial hemoglobin motor prepared by the embodiment of the present invention 3 and magnetic field collective effect
Dynamic optical microscope photograph.
Specific embodiment
Below in conjunction with attached drawing, the forgoing and additional technical features and advantages are described in more detail.
Embodiment 1
A kind of artificial red cells motor is present embodiments provided, a diameter of 2 μm -100 μm, thickness is 500nm-5 μm, bag
Include natural hemoglobin formation red blood cell shape skeleton, positioned at the skeletal internal magnetic nanoparticle and surface modification it is thin
After birth, wherein the magnetism micro-nano granules are the ferriferrous oxide particles that grain size is 5nm-500nm, the natural hemoglobin
Including one kind in pig source hemoglobin, horse source hemoglobin or ox source hemoglobin, the cell membrane includes erythrocyte membrane, white
One kind in cell membrane or platelet membrane.
Wherein hemoglobin there is oxygen to carry, releasability, the still main composition of red blood cell, also not extensive
Applied in artificial oxygen transportation carrier, red blood cell has blood long circulating ability, and leucocyte has immune clearance ability, and blood is small
Plate have thrombus and bleeding part recognition capability, and cell can by memebrane protein, polysaccharide of cell membrane surface etc. form into
Divide and be mutually distinguishable, modify artificial motor using the cell membrane surface of n cell, make artificial motor that there is the biological of class cell
Can, stability is good, can quickly be moved along acoustic pressure gradient under the driving of external source ultrasonic field.Meanwhile artificial red cells motor may be used also
By cells such as the natural fine after birth specific recognition cancer cells of surface modification or by applying external source magnetic field, to realize artificial
The targeting campaign in red blood cell motor movement direction, in oxygen targeting transport, drug specificity carries, toxin is removed, oncotherapy
Biomedical sectors is waited to be with a wide range of applications.
Embodiment 2
Fig. 1 and Fig. 2 are referred to,
Fig. 1 is the schematic diagram of the preparation process 3 and step 4 of originally applying artificial red cells motor in example；
Fig. 2 is the schematic diagram that artificial red cells motor moves about in the present embodiment.
The present embodiment using hemoglobin, natural fine after birth, magnetic micro-nano granules as material, using chemical coprecipitation and
Cell membrane fusion technology, by the main constituents hemoglobin of red blood cell, magnetic micro-nano granules and with cell biological
The cell membrane of performance combines together, prepares the artificial red cells motor with red blood cell concave-concave disc structure.Specifically include with
Lower step：
Step 1: prepare the magnetic micro-nano granules：
A, the iron chloride of the frerrous chloride of 0.002-0.02mol and 0.004-0.04mol are pressed 1:2 weight ratio dissolving
Into the deionized water of 100-1000mL, 70 DEG C are heated under nitrogen protection, stirring while adds in 40mL ammonium hydroxide；B, continue anti-
Answer 1 it is small when after stop heating, the flask equipped with above-mentioned reaction liquid is placed on magnet, stands 30min, to be formed four oxidations three
After iron particle deposits to the bottom of the flask completely, supernatant liquid is removed；C, the ferroso-ferric oxide is cleaned using acetone
After grain 3-4 times, the not oxygenous deionized waters of 25mL are added in into the flask, obtain four oxidations that grain size is 5nm-500nm
Three-iron particle suspension liquid；
Step 2: synthetic red blood cells shape calcium hydroxide particle：
A, the dextran sulfate of the natural hemoglobin of 100-1000mg, 0.5-5g is distributed to 100mL concentration as 0.05-
The CaCl of 0.5mol/L2In solution, mixed solution A is obtained；B, into the sodium hydroxide solution that 20mL concentration is 0.5-5mol/L
The magnetic ferroferric oxide particle suspension liquid described in step 1 is added, obtains mixed solution B；C, the mixed solution A is agreed into institute
State ultrasound 10 seconds after mixed solution B is mixed rapidly；D, it is collected by centrifugation after standing 20min to get to containing magnetic four oxygen
The grain size for changing three iron particles and the natural hemoglobin is 2 μm of -100 μm of red blood cell shape calcium hydroxide particles；
Step 3: prepare artificial hemoglobin motor skeleton：
A, the characteristic for making full use of in blood different cell densities different, using Ficoll density-gradient centrifugation methods, with
The speed centrifugation blood of human body of 4500rpm/min, separation collect cell, utilize Hypotonic treatment cell 24 at 4 DEG C of hypotonic buffer liquid
Hour, fully to remove intracellular content；B, using mini squeezer, cell is squeezed repeatedly through aperture as 200nm's
Polyurethane film 21 times prepares the cell membrane nano vesicle that grain size is 200nm；C, with described in step 2 containing being magnetic four oxidations
The red blood cell shape calcium hydroxide particle of three iron particles and natural hemoglobin is kernel, kernel and the cell membrane nanometer described in b
Vesica is sufficiently mixed, and based on cell membrane fusion technology, using Ultrasound Instrument, ultrasound merges 30min under 59kHz frequencies, described interior
Core surface fused cell film completes the preparation of the artificial hemoglobin motor skeleton.
Step 4: dissolving kernel：Artificial hemoglobin motor skeleton described in step 3 is added to concentration as 0.01-
0.1mol/L, pH scope are the phosphate buffer of 5.0-7.0, and concentration is the dilute hydrochloric acid solution or pH of 0.005-0.051mol/L
It is worth in the disodium EDTA solution for being 0.1-1mol/L for 7.0, concentration, is centrifuged after 30min, supernatant discarding
Liquid, gained precipitation are cleaned three times with the phosphate buffer of pH 7.4, finally obtain the artificial red cells motor.
It can be clearly seen that the step 3 agrees the forming process of artificial red cells motor in step 4 by Fig. 1,
InThe calcium hydroxide particle containing be magnetic micro-nano granules and hemoglobin is represented,Represent Cell membrane vesicles,Magnetic micro-nano granules are represented,Represent artificial red cells motor；
The principle that artificial red cells motor moves about under the driving of external source ultrasonic field can be clearly seen that by Fig. 2,
InUltrasonic field is represented, straight line represents its travelling mode.
Drug used is commercial product in the present embodiment, and the natural hemoglobin in the present embodiment step 2 can be by different sulphur
Cyanic acid fluorescein (FITC) marks, and the cell membrane in step 3 can use cell membrane dyestuff DiD to mark, so as to obtain the people with fluorescence
Make red blood cell motor.Artificial red cells motor manufactured in the present embodiment is quickly transported under the driving of external source ultrasonic field along acoustic pressure gradient
Dynamic, ultrasound condition is：Voltage is 0.5~20V, frequency 2kHz.Meanwhile artificial red cells motor passes through the natural of surface modification
The cells such as cell membrane specific recognition cancer cell realize the targeting campaign in artificial red cells motor movement direction.This method is simple
It is easy, can mass production.
Embodiment 3
Difference lies in the targeting mode of artificial red cells motor is the spy of external source introduction by magnetic field with embodiment 2 for this implementation
Opposite sex targeting.
Embodiment 4
It refers to shown in 3, Fig. 4, Fig. 5,
Fig. 3 is the laser confocal microscope photo of artificial hemoglobin motor manufactured in the present embodiment；
Fig. 4 is the optical microscope photograph moved under the ultrasound-driven of artificial hemoglobin motor manufactured in the present embodiment；
Fig. 5 is the light that movement is targeted under the ultrasound of artificial hemoglobin motor manufactured in the present embodiment and magnetic field collective effect
Learn microphotograph.
The present embodiment using hemoglobin, natural fine after birth, magnetic micro-nano granules as material, using chemical coprecipitation and
Cell membrane fusion technology, by the main constituents hemoglobin of red blood cell, magnetic micro-nano granules and with cell biological
The cell membrane of performance combines together, prepares artificial red cells motor.Specifically include following steps：
Step 1: prepare the magnetic micro-nano granules：
A, the iron chloride of the frerrous chloride of 0.02mol and 0.04mol is dissolved into the deionized water of 100mL, nitrogen is protected
It is heated to 70 DEG C under shield, stirring while adds in 40mL ammonium hydroxide；B, the reaction was continued 1 it is small when after stop heating, equipped with above-mentioned reaction
The flask of liquid is placed on magnet, stands 30min, and ferriferrous oxide particles to be formed deposit to the bottom of the flask completely
Afterwards, supernatant liquid is removed；C, after using the acetone cleaning ferriferrous oxide particles 4 times, 25mL is added in not into the flask
Oxygenous deionized water obtains the ferriferrous oxide particles suspension that grain size is 500nm；
Step 2: synthetic red blood cells shape calcium hydroxide particle：
A, it is 0.05mol/L's the dextran sulfate of the natural hemoglobin of 100mg, 0.5g to be distributed to 100mL concentration
CaCl2In solution, mixed solution A is obtained；B, it is addition step 1 institute in the sodium hydroxide solution of 0.5mol/L to 20mL concentration
The magnetic ferroferric oxide particle suspension liquid stated, obtains mixed solution B；C, that the mixed solution A is agreed the mixed solution B is fast
Ultrasound 10 seconds after speed mixing；D, it is collected by centrifugation after standing 20min to get to containing the magnetic ferroferric oxide particle and institute
The grain size for stating natural hemoglobin is 2 μm of red blood cell shape calcium hydroxide particle；
Step 3: prepare artificial hemoglobin motor skeleton：
A, the characteristic for making full use of in blood different cell densities different, using Ficoll density-gradient centrifugation methods, with
The speed centrifugation blood of human body of 4500rpm/min, separation collect cell, utilize Hypotonic treatment cell 24 at 4 DEG C of hypotonic buffer liquid
Hour, fully to remove intracellular content；B, using mini squeezer, cell is squeezed repeatedly through aperture as 200nm's
Polyurethane film 21 times prepares the cell membrane nano vesicle that grain size is 200nm；C, with described in step 2 containing being magnetic four oxidations
The red blood cell shape calcium hydroxide particle of three iron particles and natural hemoglobin is kernel, kernel and the cell membrane nanometer described in b
Vesica is sufficiently mixed, and based on cell membrane fusion technology, using Ultrasound Instrument, ultrasound merges 30min under 59kHz frequencies, described interior
Core surface fused cell film completes the preparation of the artificial hemoglobin motor skeleton.
Step 4: dissolving kernel：Artificial hemoglobin motor skeleton described in step 3 is added to concentration is
In 0.01mol/L, the phosphate buffer that pH value is 6.0, centrifuged after 30min, abandoning supernatant, gained precipitation pH
7.4 phosphate buffer cleaning three times, finally obtains the artificial red cells motor.
Drug used is commercial product in the present embodiment, and the natural hemoglobin in the present embodiment step 2 can be by different sulphur
Cyanic acid fluorescein (FITC) marks, and the cell membrane in step 3 can use cell membrane dyestuff DiD to mark, so as to obtain the people with fluorescence
Make red blood cell motor.Artificial red cells motor manufactured in the present embodiment is quickly transported under the driving of external source ultrasonic field along acoustic pressure gradient
Dynamic, ultrasound condition is：Voltage is 0.5~20V, frequency 2kHz.Meanwhile artificial red cells motor passes through the natural of surface modification
The cells such as cell membrane specific recognition cancer cell realize the targeting campaign in artificial red cells motor movement direction.This method is simple
It is easy, can mass production.
From Fig. 3 it can be found that the artificial hemoglobin motor that the present embodiment obtains is in the concave-concave disc structure of class red blood cell,
A diameter of 2 μm.And the hemoglobin of FITC marks and the erythrocyte membrane of DiD marks are merged, and illustrate the people being prepared
It is with good stability to make red blood cell motor.
Embodiment 5
The present embodiment using hemoglobin, natural fine after birth, magnetic micro-nano granules as material, using chemical coprecipitation and
Cell membrane fusion technology, by the main constituents hemoglobin of red blood cell, magnetic micro-nano granules and with cell biological
The cell membrane of performance combines together, prepares artificial red cells motor.Specifically include following steps：
Step 1: prepare the magnetic micro-nano granules：
A, the iron chloride of the frerrous chloride of 0.002mol and 0.004mol is dissolved into the deionized water of 1000mL, nitrogen
It is heated to 70 DEG C under protection, stirring while adds in 40mL ammonium hydroxide；B, the reaction was continued 1 it is small when after stop heating, equipped with above-mentioned anti-
The flask of liquid is answered to be placed on magnet, stands 30min, ferriferrous oxide particles to be formed deposit to the bottom of the flask completely
Behind portion, supernatant liquid is removed；C, after using the acetone cleaning ferriferrous oxide particles 4 times, 25mL is added in into the flask
Not oxygenous deionized water obtains the ferriferrous oxide particles suspension that grain size is 5nm；
Step 2: synthetic red blood cells shape calcium hydroxide particle：
A, it is 0.05mol/L's the dextran sulfate of the natural hemoglobin of 100mg, 0.5g to be distributed to 100mL concentration
CaCl2In solution, mixed solution A is obtained；B, it is addition step 1 institute in the sodium hydroxide solution of 0.5mol/L to 20mL concentration
The magnetic ferroferric oxide particle suspension liquid stated, obtains mixed solution B；C, that the mixed solution A is agreed the mixed solution B is fast
Ultrasound 10 seconds after speed mixing；D, it is collected by centrifugation after standing 20min to get to containing the magnetic ferroferric oxide particle and institute
The grain size for stating natural hemoglobin is 2 μm of red blood cell shape calcium hydroxide particle；
Step 3: prepare artificial hemoglobin motor skeleton：
A, the characteristic for making full use of in blood different cell densities different, using Ficoll density-gradient centrifugation methods, with
The speed centrifugation blood of human body of 4500rpm/min, separation collect cell, utilize Hypotonic treatment cell 24 at 4 DEG C of hypotonic buffer liquid
Hour, fully to remove intracellular content；B, using mini squeezer, cell is squeezed repeatedly through aperture as 200nm's
Polyurethane film 21 times prepares the cell membrane nano vesicle that grain size is 200nm；C, with described in step 2 containing being magnetic four oxidations
The red blood cell shape calcium hydroxide particle of three iron particles and natural hemoglobin is kernel, kernel and the cell membrane nanometer described in b
Vesica is sufficiently mixed, and based on cell membrane fusion technology, using Ultrasound Instrument, ultrasound merges 30min under 59kHz frequencies, described interior
Core surface fused cell film completes the preparation of the artificial hemoglobin motor skeleton.
Step 4: dissolving kernel：Artificial hemoglobin motor skeleton described in step 3 is added to concentration is
In 0.01mol/L, the phosphate buffer that pH value is 6.0, centrifuged after 30min, abandoning supernatant, gained precipitation pH
7.4 phosphate buffer cleaning three times, finally obtains the artificial red cells motor.
Drug used is commercial product in the present embodiment, and the natural hemoglobin in the present embodiment step 2 can be by different sulphur
Cyanic acid fluorescein (FITC) marks, and the cell membrane in step 3 can use cell membrane dyestuff DiD to mark, so as to obtain the people with fluorescence
Make red blood cell motor.Artificial red cells motor manufactured in the present embodiment is quickly transported under the driving of external source ultrasonic field along acoustic pressure gradient
Dynamic, ultrasound condition is：Voltage is 0.5~20V, frequency 2kHz.Meanwhile artificial red cells motor passes through the natural of surface modification
The cells such as cell membrane specific recognition cancer cell realize the targeting campaign in artificial red cells motor movement direction.This method is simple
It is easy, can mass production.
Embodiment 6
The present embodiment in place of the difference of embodiment 5 with being, by the frerrous chloride and 0.02mol of 0.01mol in step 1
Iron chloride be dissolved into the deionized water of 100mL, obtain grain size be 150nm ferriferrous oxide particles suspension, other with
Embodiment 5 is identical.
Embodiment 7
The present embodiment in place of the difference of embodiment 5 with being, the phosphorus that concentration is 0.01mol/L in step 4, pH value is 6.0
Phthalate buffer replaces with the dilute hydrochloric acid solution that concentration is 0.005mol/L.
Embodiment 8
The present embodiment in place of the difference of embodiment 5 with being, the phosphorus that concentration is 0.01mol/L in step 4, pH value is 6.0
Phthalate buffer replaces with the disodium EDTA solution that pH value is 7.0, concentration is 0.1mol/L.
Embodiment 9
With being in place of the difference of embodiment 5, step 2 becomes the present embodiment：
Synthetic red blood cells shape calcium hydroxide particle：A, the dextran sulfate of the natural hemoglobin of 1000mg, 5g are disperseed
To the CaCl that 100mL concentration is 0.5mol/L2In solution, mixed solution A is obtained；B, to the hydrogen-oxygen that 20mL concentration is 0.5mol/L
Change the magnetic ferroferric oxide particle suspension liquid added in sodium solution described in step 1, obtain mixed solution B；C, by the mixing
Solution A agrees ultrasound 10 seconds after the mixed solution B is mixed rapidly；D, it is collected by centrifugation after standing 20min to get to containing described
The grain size of magnetic ferroferric oxide particle and the natural hemoglobin be 100 μm of red blood cell shape calcium hydroxide particles, other with
Embodiment 5 is identical.
Embodiment 10
With being in place of the difference of embodiment 5, step 2 becomes the present embodiment：
Synthetic red blood cells shape calcium hydroxide particle：A, the dextran sulfate of the natural hemoglobin of 500mg, 2.5g are disperseed
To the CaCl that 100mL concentration is 0.2mol/L2In solution, mixed solution A is obtained；B, to the hydrogen-oxygen that 20mL concentration is 2.5mol/L
Change the magnetic ferroferric oxide particle suspension liquid added in sodium solution described in step 1, obtain mixed solution B；C, by the mixing
Solution A agrees ultrasound 10 seconds after the mixed solution B is mixed rapidly；D, it is collected by centrifugation after standing 20min to get to containing described
The grain size of magnetic ferroferric oxide particle and the natural hemoglobin is 30 μm of red blood cell shape calcium hydroxide particles, other and reality
It is identical to apply example 5.
The foregoing is merely presently preferred embodiments of the present invention, is merely illustrative for the purpose of the present invention, and not restrictive
's.Those skilled in the art understands, many changes can be carried out to it in the spirit and scope limited in the claims in the present invention,
It changes or even equivalent, but falls in protection scope of the present invention.
Claims (9)
1. a kind of artificial red cells motor, which is characterized in that it includes the red blood cell shape skeleton of natural hemoglobin formation, is located at
The magnetic nanoparticle of the skeletal internal and the cell membrane of surface modification.
2. artificial red cells motor according to claim 1, which is characterized in that the magnetism micro-nano granules are four oxidations
Three iron particles, and the grain size of the ferriferrous oxide particles is 5nm-500nm.
3. artificial red cells motor according to claim 1, which is characterized in that the natural hemoglobin includes pig source blood
One kind in Lactoferrin, horse source hemoglobin or ox source hemoglobin.
4. the artificial red cells motor according to belonging to claim 1, feature exist, the cell membrane includes erythrocyte membrane, white thin
One kind in after birth or platelet membrane.
5. prepare the method such as claim 1-4 any one of them artificial red cells motors, which is characterized in that it includes following
Step：
Step 1: prepare the magnetic micro-nano granules；
Step 2: by chemical coprecipitation, synthesis is interior to contain the magnetic micro-nano granules described in step 1 and natural blood red egg
White red blood cell shape kernel；
Step 3: using the red blood cell shape kernel described in step 2 as template, natural fine after birth is repaiied using cell membrane fusion technology
It adorns on the surface of the template, obtains artificial red cells motor skeleton；
Step 4: the kernel in the artificial red cells motor skeleton obtained using solvent dissolution method removal step three, finally
Obtain artificial red cells motor.
6. the preparation method of artificial red cells motor according to claim 5, which is characterized in that kernel described in step 2
For calcium hydroxide particle, and the grain size of the kernel is 2 μm -100 μm.
7. the preparation method of artificial red cells motor according to claim 5, which is characterized in that solvent described in step 4
It is the phosphate buffer that 0.01-0.1mol/L, pH are 5.0-7.0 for concentration, concentration is the dilute hydrochloric acid of 0.005-0.05mol/L
One kind in the disodium EDTA solution that solution or pH value are 7.0, concentration is 0.1-1mol/L.
8. the preparation method of artificial red cells motor according to claim 5, which is characterized in that red thin described in step 4
A diameter of 2 μm -100 μm of born of the same parents' motor, thickness are 500nm-5 μm.
9. the method according to claim 5 for preparing artificial red cells motor, which is characterized in that it comprises the following steps：
Step 1: prepare the magnetic micro-nano granules：A, by the frerrous chloride and 0.004-0.04mol of 0.002-0.02mol
Iron chloride press 1:2 weight ratio is dissolved into the deionized water of 100-1000mL, 70 DEG C is heated under nitrogen protection, stirring
While add in 40mL ammonium hydroxide；B, the reaction was continued 1 it is small when after stop heating, the flask equipped with above-mentioned reaction liquid is placed in magnet
On, 30min is stood, after ferriferrous oxide particles to be formed deposit to the bottom of the flask completely, removes supernatant liquid；c、
After acetone cleaning ferriferrous oxide particles 3-4 times, the not oxygenous deionized waters of 25mL are added in into the flask,
Obtain ferriferrous oxide particles suspension；
Step 2: synthetic red blood cells shape calcium hydroxide particle：A, by the natural hemoglobin of 100-1000mg, the sulfuric acid of 0.5-5g
Glucan is distributed to the CaCl that 100mL concentration is 0.05-0.5mol/L2In solution, mixed solution A is obtained；B, to 20mL concentration
To add the magnetic ferroferric oxide particle suspension liquid described in step 1 in the sodium hydroxide solution of 0.5-5mol/L, mixed
Solution B；C, the mixed solution A is agreed into ultrasound 10 seconds after the mixed solution B is mixed rapidly；D, centrifuged after standing 20min
It collects to get to the red blood cell shape calcium hydroxide containing the magnetic ferroferric oxide particle and the natural hemoglobin
Grain；
Step 3: prepare artificial hemoglobin motor skeleton：A, the characteristic that different cell densities are different in blood is made full use of, is made
With Ficoll density-gradient centrifugation methods, blood of human body is centrifuged with the speed of 4500rpm/min, cell is collected in separation, and utilization is hypotonic
When Hypotonic treatment cell 24 is small at 4 DEG C of buffer solution, fully to remove intracellular content；B, using mini squeezer, repeatedly
Cell is squeezed through the polyurethane film 21 times that aperture is 200nm, prepares the cell membrane nano vesicle that grain size is 200nm；C, with
Red blood cell shape calcium hydroxide particle containing magnetic ferroferric oxide particle and natural hemoglobin described in step 2 is kernel,
Kernel is sufficiently mixed with the cell membrane nano vesicle described in b, based on cell membrane fusion technology, using Ultrasound Instrument in 59kHz frequencies
Ultrasound fusion 30min under rate, in the core surface fused cell film, the system of the completion artificial hemoglobin motor skeleton
It is standby.
Step 4: dissolving kernel：Artificial hemoglobin motor skeleton described in step 3 is added to concentration as 0.01-
0.1mol/L, pH scope are the phosphate buffer of 5.0-7.0, and concentration is the dilute hydrochloric acid solution or pH of 0.005-0.05mol/L
It is worth in the disodium EDTA solution for being 0.1-1mol/L for 7.0, concentration, is centrifuged after 30min, supernatant discarding
Liquid, gained precipitation are cleaned three times with the phosphate buffer of pH7.4, finally obtain the artificial red cells motor.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111632038A (en) * | 2020-06-05 | 2020-09-08 | 南京师范大学 | Platelet drug-loaded micro-nano motor and preparation method and application thereof |
| CN114366811A (en) * | 2021-06-07 | 2022-04-19 | 核工业四一六医院 | Bionic nano motor for targeted drug delivery and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1410529A (en) * | 2002-09-18 | 2003-04-16 | 重庆飞翔生物技术有限公司 | In vitro reconstituted human erythrocyte, its preparation and application in blood substitute material |
| WO2008013952A2 (en) * | 2006-07-27 | 2008-01-31 | The University Of North Carolina At Chapel Hill | Nanoparticle fabrication methods, systems, and materials for fabricating artificial red blood cells |
| US20170143830A1 (en) * | 2015-11-20 | 2017-05-25 | The Regents Of The University Of California | Cellular Micromotors and Uses Thereof |
-
2017
- 2017-12-11 CN CN201711306509.9A patent/CN108042802A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1410529A (en) * | 2002-09-18 | 2003-04-16 | 重庆飞翔生物技术有限公司 | In vitro reconstituted human erythrocyte, its preparation and application in blood substitute material |
| WO2008013952A2 (en) * | 2006-07-27 | 2008-01-31 | The University Of North Carolina At Chapel Hill | Nanoparticle fabrication methods, systems, and materials for fabricating artificial red blood cells |
| US20170143830A1 (en) * | 2015-11-20 | 2017-05-25 | The Regents Of The University Of California | Cellular Micromotors and Uses Thereof |
Non-Patent Citations (6)
| Title |
|---|
| CHUNMEI YU等: "Construction of biconcave hemoglobin-based microcapsules and electrochemical evaluation for its ability of oxygen carry", 《SENSORS AND ACTUATORS B: CHEMICAL》 * |
| ZHIGUANG WU等: "RBC micromotors carrying multiple cargos towards potential theranostic applications", 《NANOSCALE》 * |
| 刘聪慧等: "微纳米马达的运动控制及其在精准医疗中的应用", 《中国科学：化学》 * |
| 吴志光: "自驱动合成微纳米马达的仿生设计及其生物医学应用", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 贺强等: "超声驱动红细胞类血红蛋白微马达", 《中国化学会2017全国高分子学术论文报告会摘要集-主题J：高分子组装与超分子体系》 * |
| 顾晓彤等: "细胞膜修饰的纳米载药系统的研究进展", 《中国医药工业杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111632038A (en) * | 2020-06-05 | 2020-09-08 | 南京师范大学 | Platelet drug-loaded micro-nano motor and preparation method and application thereof |
| CN111632038B (en) * | 2020-06-05 | 2022-02-11 | 南京师范大学 | Platelet drug-loaded micro-nano motor and preparation method and application thereof |
| CN114366811A (en) * | 2021-06-07 | 2022-04-19 | 核工业四一六医院 | Bionic nano motor for targeted drug delivery and preparation method and application thereof |
| CN114366811B (en) * | 2021-06-07 | 2022-08-23 | 核工业四一六医院 | Bionic nano motor for targeted drug delivery and preparation method and application thereof |
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