CN107057351A - Composite polyethylene imines modification graphene oxide composite material and its preparation method and application - Google Patents
Composite polyethylene imines modification graphene oxide composite material and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of composite polyethylene imines modification graphene oxide composite material and its preparation method and application, composite includes graphene oxide, low molecular polyethylene imines, polyethyleneimine polymer and crosslinking agent, and low molecular polyethylene imines and polyethyleneimine polymer are modified in surface of graphene oxide by crosslinking agent；Polyethyleneimine polymer is the polyethyleneimine of relative molecular mass >=20000, and low molecular polyethylene imines is the polyethyleneimine that relative molecular weight is less than 5000.Its preparation method is：Graphene oxide is dispersed in water, crosslinking agent is added and obtains suspension；Polyethyleneimine polymer and low molecular polyethylene imines are mixed into addition crosslinking agent and obtain mixed solution；Mixed solution is instilled to stirring in suspension dropwise, dialysed with bag filter.Composite polyethylene imines modification graphene oxide composite material transfection efficiency is high, toxicity is low, gene transfection and drug carrier is can be applied to, with the advantage such as efficient, quick, safe.
Description
Technical field
The present invention relates to biomedical and technical field of nano material, more particularly to a kind of composite polyethylene imines modification oxygen
Graphite alkene composite and its preparation method and application.
Background technology
With the arriving in functional genome's epoch, building various functions gene mutation body turns into laboratory research and industrial circle
Essential technology and measure.Simultaneously in biomedicine field, functional gene is imported into the focus containing gene delection/mutation
Cell is simultaneously being expressed, so as to change the genetic property of cell, obtains more preferable therapeutic effect.
Successful gene transfection and gene therapy be unable to do without effective composite.Continue to develop safe and effective and hypotoxicity
Gene composite be Gene transfer techniques research in urgent problem.Current gene transfection agent generally can be divided into two
Class：Viral composite and non-viral composite.Viral composite has high transfection efficiency, but due to immunogenicity
In the presence of, cause body produce repulsion so that gene delivery safety is not high.Traditional non-viral composite has liposome, gathered
Compound and gold nano grain etc., but it is relatively low to there is transfection efficiency, and cytotoxicity is high, lacks the defects such as targeting.In chemotherapeutic
Thing field there is also it is similar the problem of, as Doxorubicin class fat-soluble medicine is easily lost in during oral and drug administration by injection,
It has been metabolized before focus cell is reached, has caused drug utilization low, normal tissue cell has stronger toxic side effect.Cause
This, seeks to build efficient and safe transfection reagent and drug composite, make target gene in cell effective expression and
Efficient transportation of the medicine in histocyte, is the focus and emphasis of current biological field of medicaments concern.
The content of the invention
The technical problem to be solved in the present invention is to overcome the deficiencies in the prior art there is provided a kind of transfection efficiency is high, toxicity is low
Composite polyethylene imines modification graphene oxide composite material and preparation method thereof, can be applied to gene transfection and medicine fortune
Carry, with the advantage such as efficient, quick, safe.
In order to solve the above technical problems, modifying graphene oxide composite material, bag there is provided a kind of composite polyethylene imines
Include graphene oxide, low molecular polyethylene imines, polyethyleneimine polymer and crosslinking agent, the low molecular polyethylene imines and
Polyethyleneimine polymer is modified in the surface of graphene oxide by crosslinking agent；The polyethyleneimine polymer is relative
The polyethyleneimine of molecular mass >=20000, the low molecular polyethylene imines is the polyethylene that relative molecular weight is less than 5000
Imines.
Above-mentioned composite polyethylene imines modification graphene oxide composite material, it is preferred that the high-molecular polythene is sub-
Amine is the polyethyleneimine that relative molecular mass is 20000~25000, and the low molecular polyethylene imines is that relative molecular weight is
1500~3000 polyethyleneimine.
Above-mentioned composite polyethylene imines modification graphene oxide composite material, it is preferred that the high-molecular polythene is sub-
The mass ratio of amine, low molecular polyethylene imines and the graphene oxide is 5~10: 5~10: 1.
Above-mentioned composite polyethylene imines modification graphene oxide composite material, it is preferred that the crosslinking agent be 1- ethyls-
One or more in 3-3- dimethylaminopropyls, N-hydroxy-succinamide ester and triethanolamine.
As a total technical concept, present invention also offers a kind of above-mentioned composite polyethylene imines modification oxidation stone
The preparation method of black alkene composite, comprises the following steps：
S1, graphene oxide is dispersed in water, adds crosslinking agent and obtain suspension；By polyethyleneimine polymer and low
The mixing of molecular polyethylene imines adds crosslinking agent and obtains mixed solution；
S2, the mixed solution is instilled into suspension stirring dropwise make fully contact；
S3, with bag filter dialysis obtain composite polyethylene imines modification graphene oxide composite material.
Above-mentioned preparation method, it is preferred that the crosslinking agent added in the graphene oxide is 1- ethyl -3-3- dimethyl
Aminopropyl (EDC) and N-hydroxy-succinamide ester (NHS)；The polyethyleneimine polymer and the low molecular polyethylene
The crosslinking agent added in the mixture of imines is triethanolamine (TEA).EDC and NHS be in order that the carboxyl of graphene oxide with
The amino of polyethyleneimine preferably reacts, and generates amido link.TEA is for preferably polyethylene dissolving imines.
Above-mentioned preparation method, it is preferred that the temperature stirred described in the step S2 is room temperature, and the time of stirring is
24h；In the step S3, the bag filter is 100KDa bag filter, and the condition of the dialysis is：At room temperature with deionization
Water penetration 3d.Room temperature in the present invention is 25 DEG C.
As a total technical concept, present invention also offers a kind of above-mentioned composite polyethylene imines modification oxidation stone
The composite polyethylene imines modification graphene oxide composite material that black alkene composite or above-mentioned preparation method are prepared exists
Application in microorganism or the transfection of animal and plant cells gene.It is preferred that, microbial cell is Escherichia coli, saccharomycete or unwrapping wire
Bacterium, zooblast is the stable cell lines or primary cultured cell of laboratory cultures, and plant cell is the cell line set up or plant
Thing callus.
As a total technical concept, present invention also offers a kind of above-mentioned composite polyethylene imines modification oxidation stone
The composite polyethylene imines modification graphene oxide composite material that black alkene composite or above-mentioned preparation method are prepared exists
Application in drug carrier.It is preferred that, the medicine is antineoplastic.
Above-mentioned application, it is preferred that the medicine is aromatic series medicine.Due to unique carbon atom of surface of graphene oxide
Structure can occur π-π stackings with the phenyl ring or heterocycle of aroma substance and act on, therefore this composite can also load it
His aromatic herbs, such as adriamycin, taxol, resveratrol, Indomethacin, Irinotecan.
Compared with prior art, the advantage of the invention is that：
(1) graphene oxide composite material, including graphite oxide are modified the invention provides a kind of composite polyethylene imines
Alkene, low molecular polyethylene imines and polyethyleneimine polymer.Wherein graphene oxide is a kind of New Two Dimensional carbon nanomaterial,
Possess good biocompatibility, high-specific surface area, easy modified, be started applied to drug delivery.Graphene oxide leads to
The effect of π-π stackings is crossed, can be by aromatic series medicine such as Doxorubicin high-efficient carrier in surface.And it is multiple in surface of graphene oxide modification
Closing polyethyleneimine can promote composite efficiently to enter cell.Polyethyleneimine is a kind of electropositive polymer, is recognized
For be transfection gold composite.Contain a large amount of amino in branched polyethylene imine, be conducive to the Protonation effect of itself, promote
Enter endosome escape, improve transfection efficiency.Although high molecular polyethyleneimine has compared with high transfection efficiency, toxicity is larger,
Although low molecular polyethylene imines cytotoxicity is smaller, transfection efficiency is relatively low.The present invention gives full play to both excellent spies
Property, surface of graphene oxide is modified using both mixed type solution, transfection has been obtained higher than the poly- second of low molecule amount
Alkene imines, and cytotoxicity is less than novel polyethylene imines-stannic oxide/graphene nano composite of high molecular weight polyethyleneimine.
(2) the invention provides the preparation method that a kind of composite polyethylene imines modifies graphene oxide composite material,
During graphene oxide reacts with the polyethyleneimine mixed, crosslinking agent EDC and NHS are in order that the carboxyl of graphene oxide is with gathering
The amino of aziridine preferably reacts, and generates amido link.The TEA added in polyethyleneimine mixed solution is in order to more
Polyethylene dissolving imines well.The effect of dialysis is that, in order to remove unnecessary polyethyleneimine, acquisition composite polyethylene imines is repaiied
Adorn graphene oxide.
(3) transfected and medicine in gene the invention provides a kind of composite polyethylene imines modification graphene oxide composite material
Thing reprint in application, when its be applied to gene transfection, in human liver cancer cells Hep G2 transfection efficiency up to more than 80%,
When concentration is 18 μ g/mL, cell survival rate is maintained at more than 85%, and whole transfection process can be entered under containing serum condition
OK.Drug composite experiment will be applied to, antineoplastic Doxorubicin is loaded, load factor reaches 25%, and tumour cell is known
Not rate is up to more than 85%.With safely, quickly, effectively etc. advantage.
Brief description of the drawings
To make the purpose, technical scheme and advantage of the embodiment of the present invention clearer, below in conjunction with the embodiment of the present invention
In accompanying drawing, clear, complete description is carried out to the technical scheme in the embodiment of the present invention.
Fig. 1 modifies the atomic force microscopy diagram of graphene oxide composite material for the composite polyethylene imines in embodiment 1.
Fig. 2 modifies the infrared figure of graphene oxide composite material for the composite polyethylene imines in embodiment 1.
Fig. 3 modifies the transfection efficiency fluorogram of graphene oxide composite material for the composite polyethylene imines in embodiment 2.
Fig. 4 is that the composite polyethylene imines modification graphene oxide composite material in embodiment 2 turns with other transfection reagents
Contaminate efficiency comparative's figure.
Fig. 5 be embodiment 3 in composite polyethylene imines modify graphene oxide composite material to DOX under different pH
Release profiles.
Fig. 6 is that the composite polyethylene imines in embodiment 3 modifies the cell in vitro that graphene oxide composite material loads DOX
Take in fluorogram.
Fig. 7 is that the composite polyethylene imines modification graphene oxide composite material in embodiment 3 loads DOX with other materials
Cytotoxicity analysis.
Embodiment
Below in conjunction with Figure of description and specific preferred embodiment, the invention will be further described, but not therefore and
Limit the scope of the invention.
Embodiment
Material and instrument employed in following examples are commercially available.
Embodiment 1：
A kind of composite polyethylene imines modification graphene oxide composite material (1.8K-PEI-25K-PEI- of the invention
GO), including graphene oxide, low molecular polyethylene imines, polyethyleneimine polymer and crosslinking agent, low molecular polyethylene imines
Modified with polyethyleneimine polymer by crosslinking agent in surface of graphene oxide；The relative molecular weight of polyethyleneimine polymer
For 25000, the relative molecular weight of low molecular polyethylene imines is 1800.Polyethyleneimine polymer, low molecular polyethylene imines
Mass ratio with graphene oxide is 5: 5: 1.
A kind of composite polyethylene imines of the present embodiment modifies the preparation method of graphene oxide composite material, including following
Step：
(1) 1mg graphene oxides are taken in deionized water, ultrasonic disperse 30min；Graphene oxide is set to be fully dispersed in water
Graphene oxide dispersion is obtained in body.
(2) in graphene oxide dispersion add 54.3mg 1- ethyl -3-3- dimethylaminopropyls (EDC) with
50.6mg N-hydroxy-succinamide ester (NHS), is mixed to get suspension.
(3) it is 1: 1 progress according to mass ratio by 5mg polyethyleneimine polymer and 5mg low molecular polyethylene imines
Mixing, the triethanolamine (TEA) for then adding 100 μ L obtains mixed solution.
(4) mixed solution of step (3) is added drop-wise in the suspension of step (2) dropwise, after ultrasonic 30min mixing, room
Temperature stirring 24h, the osmotic bag for finally loading 100KDa is dialysed 3 days at 25 DEG C, obtains composite polyethylene imines modification oxidation stone
Black alkene composite.
AFM (AFM) is carried out to the composite polyethylene imines modification graphene oxide composite material of embodiment 1
Analysis, referring to Fig. 1：Composite polyethylene imines modification graphene oxide composite material highly be 2~4nm, size for 20nm~
80nm。
Infrared analysis is carried out to the composite polyethylene imines modification graphene oxide composite material of embodiment 1, referring to Fig. 2：
Polyethyleneimine is effectively combined on graphene oxide.
Comparative example 1
A kind of polyethyleneimine polymer modification graphene oxide composite material (25K-PEI-GO), the area with embodiment 1
It is not that step (3) is：10mg polyethyleneimine polymer is added and obtains mixing molten in 100 μ L triethanolamine (TEA)
Liquid.
Comparative example 2
A kind of low molecular polyethylene imines modification graphene oxide composite material (1.8K-PEI-GO), the area with embodiment 1
It is not that step (3) is：10mg low molecular polyethylene imines is added and obtains mixing molten in 100 μ L triethanolamine (TEA)
Liquid.
Embodiment 2
A kind of application of the composite polyethylene imines modification graphene oxide composite material of embodiment 1 in gene transfection.
By Hepatocellular carcinoma cell line with every hole 104Density be inoculated in 6 orifice plates, and cultivate 24 hours to treat SMMC-
7721 cell attachments.Then, original culture medium is carefully inhaled and abandoned, then toward DMEMs of the addition 0.1mL containing 10%FBS in each hole
Culture medium.
It is respectively that 1.8K-PEI-25K-PEI-GO (embodiment 1), 25K-PEI-GO (comparative example 1), 1.8K-PEI-GO is (right
Ratio 2) and graphene oxide (GO) and egfp expression plasmid according to mass ratio be mix at 10: 1, add DMEM and train
Support base mixing and obtain within 30 minutes compound 1,2,3,4.Above-mentioned compound 1~4 is taken respectively in hole, is continued in 37 DEG C of incubators
It is incubated after 4h, then by original culture medium, carefully suction is abandoned, and adds fresh DMEM culture mediums, continuation is incubated in cell culture incubator
Educate.The expression quantity that green fluorescent protein is observed under fluorescence inverted phase contrast microscope is used after 48h.It is incubated 24h and not up to optimal
Transfection, 48h has reached optimal transfection, and 72h dead cells are more more than 24h and 48h.
Fig. 3 be embodiment 2 in 1.8K-PEI-25K-PEI-GO transfection efficiency fluorogram.Left figure is transfection fluorogram,
In the case where 488nm light is excited, cell sends green fluorescence, represents green fluorescent protein successful expression in cell.Right figure is represented
The figure of bright-field figure, i.e. cell under light microscopic.
Respectively after 24h, 48h and 72h, the culture medium in each hole is inhaled and abandons clean, adds PBS twice, uses cell
Lysate (cell pyrolysis liquid includes Tri-HCl, NaCl, PMSF, EDTA) cell lysis is simultaneously tried using luciferase assays
Agent box (purchase spontaneous work bioengineering limited company) detects the uciferase activity in lysate.Egg in lysate
Bai Hanliang is then detected by BCA protein detection kits (the spontaneous work bioengineering limited company of purchase).Luciferase
Activity is represented with Relative fluorescence units/milligram albumen (RLU/mg), so as to determine transfection efficiency.Measurement result is referring to table 1.
Table 1：Each group transfection efficiency is contrasted
Fig. 4 is composite polyethylene imines modification graphene oxide composite material group, polyethyleneimine polymer group, low molecule
Polyethyleneimine group and graphene oxide group transfection efficiency comparison diagram.
It was found from table 1 and Fig. 4 result：The transfection efficiency of composite polyethylene imines modification graphene oxide composite material is bright
It is aobvious to be better than low molecular polyethylene imines group and graphene oxide group or even higher than polyethyleneimine polymer group.
Embodiment 3
A kind of application of the composite polyethylene imines modification graphene oxide composite material of embodiment 1 in medicine reprinting.
Experimental group：The 1.8K-PEI-25K-PEI-GO of 100 μM of adriamycin and 0.2mg/ml embodiment 1 is added into PH
In=7.4 phosphate buffer solution, mixture is obtained within 24 hours in the stirring of black dark at room temperature.Mixture is centrifuged with 12000rpm
5min, and be washed with deionized, repeated centrifugation operation, until supernatant becomes colorless.Last sediment is load Ah mould
The composite polyethylene imines modification graphene oxide composite material (1.8K-PEI-25K-PEI-GO-DOX) of element, is freeze-dried standby
With.
Compare 1 group：Replace 1.8K-PEI-25K-PEI-GO to carry out same operation the 25K-PEI-GO of comparative example 1 to obtain
It is loaded with the polyethyleneimine polymer (25K-PEI-GO-DOX) of adriamycin.
Compare 2 groups：The 1.8K-PEI-GO of comparative example 2 is equally operated instead of 1.8K-PEI-25K-PEI-GO, born
It is loaded with the molecular polyethylene imines (1.8K-PEI-GO-DOX) of adriamycin
Compare 3 groups：Replace 1.8K-PEI-25K-PEI-GO to carry out same operation graphene oxide (GO) to be loaded with
The graphene oxide (GO-DOX) of adriamycin.
Medicine efficiency of loading is determined：
From Lambert-Beer's law, the absorbance of material is directly proportional to concentration.DOX is configured into concentration is respectively
0.025mg/mL, 0.05mg/mL, 0.1mg/mL, 0.2mg/mL, 0.4mg/mL solution, determine above-mentioned difference dense at 480nm
Spend the absorbance of DOX solution.Linear equation is obtained according to concentration and absorbance.
Above-mentioned 1.8K-PEI-25K-PEI-GO-DOX is dialysed in deionized water, it is many in dialyzate by finally determining
Remaining DOX absorbances at 480nm, further according to gained linear equation before, can obtain its concentration.It is thus determined that not loading
DOX mass, it is counter to push away the DOX mass being supported on carrier.Calculating obtains medicine efficiency of loading (DLE) and encapsulation efficiency (EE).
Efficiency of loading (Drug Loading Efficiency), DLE=(WFirst drug-WUnloaded medicine)/WCarrier。
Envelop rate (Entrapment Efficiency), EE=(WFirst drug-WUnloaded medicine)/WFirst drug。WFirst drugIt is anti-to add
Answer the drug quality of system, WUnloaded medicineFor the drug quality after dialysis in dialyzate, WCarrierRepaiied for composite polyethylene imines
Adorn graphene oxide composite material.
It is respectively 29.35% and 52.87% that calculating, which obtains medicine efficiency of loading (DLE) with encapsulation efficiency (EE),.
The release of external medicine is determined：
Insoluble drug release is analyzed by dialysis, is concretely comprised the following steps：1.8K-PEI-25K-PEI-GO-DOX is divided to for two parts,
In the PBS (2mL) for being dissolved in the pH=5.3 (2mL) and physiological environment PH=7.4 of simulation cancer cell microenvironment respectively, it is placed in
In 100KDa bag filters, permeated in 50mL PBS, and the stirring at low speed at 37 DEG C.Under different time intervals (2,4,6,
8th, 10,12 and 24hr), the outer PBS (2mL) of bag filter is taken, its UV absorbance in 480nm is measured, while adding the PBS of equivalent
Solution.The concentration for being released into DOX in PBS is obtained by DOX standard concentration curve, and this method is equally applicable to GO-DOX
(control is used as using GO).In triplicate, accumulation calculates DOX burst size for all experiments.
Referring to Fig. 5：1.8K-PEI-25K-PEI-GO has obvious slow release effect to medicine after carrying medicine, in pH=5,24h
Drug release rate is 30%, far above the 24% of pH=7.4, illustrates that the load medicine is adapted in meta-acid Environment release.
Cell in vitro intake experiment：
By Hepatocellular carcinoma cell line with every hole 104The density of individual cell (being counted with cell counting count board) is inoculated in containing 2mL
The 6 of culture medium (containing the DEME culture mediums of 10% hyclone, DMEM is a kind of culture medium containing various amino acid and glucose)
Well culture plate culture.It is respectively 0.5 μ g/mL, 1 μ g/mL, 2 μ g/mL, 5 μ g/mL DOX, GO-DOX, 1.8K- by concentration containing DOX
PEI-25K-PEI-GO-DOX adds each hole, be incubated 0.5 respectively in 37 DEG C of incubators, 1, give up nutrient solution after 2,4hr, use pH
Fluoroscopic image is collected under=7.4 PBS cell 2 times, fluorescence microscope.
The wherein 1.8K-PEI-25K-PEI-GO-DOX for the 5 μ g/mL that DOX concentration is 2hr fluorescence imaging result most
Good, imaging results are referring to Fig. 6, and left figure is that adriamycin sends red fluorescence under the fluorescence visual field, illustrates that adriamycin has from carrier
Effect is discharged, and can cause its fluorescent quenching or faint because adriamycin is combined with carrier, confirms above in-vitro simulated swollen herein
Under knurl faintly acid, the release of medicine.Right figure represents the figure of bright-field figure, i.e. cell under light microscopic.As can be known from Fig. 6：Carry medicine
Obvious red fluorescence is presented in 80% cell of group, illustrates that medicine has been successfully entered cell.
Carry medicine effect analysis：
SMMC-7721 liver cancer cells are inoculated in 96 orifice plates with the density of 6000 cell per wells, cultivate 24hr to treat
Cell attachment and long to exponential phase.Then careful inhale abandons original culture medium, adds 120 μ L into each hole respectively containing 1.8K-
PEI-25K-PEI-GO-DOX DMEM culture mediums (wherein 1.8K-PEI-25K-PEI-GO-DOX mass-volume concentration difference
For 2 μ g/mL, 6 μ g/mL, 10 μ g/mL, 14 μ g/mL, 18 μ g/mL), DMEM culture medium (its containing 1.8K-PEI-25K-PEI-GO
Middle 1.8K-PEI-25K-PEI-GO mass-volume concentration is respectively 2 μ g/mL, 6 μ g/mL, 10 μ g/mL, 14 μ g/mL, 18 μ g/
ML (wherein 1.8K-PEI-GO-DOX mass-volume concentration is respectively 2 μ g/ to DMEM culture mediums), containing 1.8K-PEI-GO-DOX
ML, 6 μ g/mL, 10 μ g/mL, 14 μ g/mL, 18 μ g/mL), DMEM culture mediums (the wherein GO-DOX quality volume containing GO-DOX
Concentration is respectively 2 μ g/mL, 6 μ g/mL, 10 μ g/mL, 14 μ g/mL, 18 μ g/mL).Original culture is carefully drawn after being incubated 24hr
Base, adds the DMEM culture mediums that 100 μ L contain 10 μ L MTT solution in each hole, continues to be incubated 4hr.150 μ are added then to every hole
L DMSO, is incubated 5min, and taking-up is placed in low speed on shaking table and shakes up 10min.Finally, detect that each hole exists by using ELIASA
Absorbance at 570nm calculates cytoactive.Testing result is referring to Fig. 7.
Fig. 7 shows：1.8K-PEI-25K-PEI-GO group cytoactives are maintained at more than 85%, and obvious cell toxicant is not presented
Property.And 1.8K-PEI-25K-PEI-GO-DOX groups occur obvious inhibition to cell, its inhibition is substantially better than 1.8K-
PEI-GO-DOX and GO-DOX groups.It increased with the rise toxicity of GO-DOX concentration.This is due to that GO-DOX passes through endocytosis
Effect enters cell, causes insoluble drug release in tumour weakly acidic condition.And 1.8K-PEI-25K-PEI-GO-DOX and 1.8K-PEI-
GO-DOX enters cell by electrostatic interaction and encytosis, causes medicine under tumour weakly acidic condition and endosome escape effect
Release.Therefore, from Fig. 6 and Fig. 7,1.8K-PEI-25K-PEI-GO into cell and can promote insoluble drug release very well, and
Cytotoxicity is small, and 1.8K-PEI-25K-PEI-GO-DOX cytotoxicities are strong.In 1.8K-PEI-25K-PEI-GO-DOX concentration
During for 18 μ g/mL, cytoactive is already below 60%.
18 μ g/mL 1.8K-PEI-25K-PEI-GO-DOX, 18 μ g/mL GO-DOX drug loading efficiency is investigated simultaneously
And cell survival rate, result is investigated referring to table 2.
Table 2：Investigate drug loading efficiency and cell survival rate
| DOX (%) | GO-DOX (%) | 1.8K-PEI-25K-PEI-GO-DOX (%) | |
| Drug loading efficiency (%) | 15.55±3.02 | 29.35±4.41 | |
| Cell survival rate (%) | 65.59±3.62 | 67.54±5.65 | 55.62% ± 7.38 |
As can be known from Table 2：The drug loading efficiency and cell survival rate of 1.8K-PEI-25K-PEI-GO-DOX groups are superior to
GO-DOX groups.
The above described is only a preferred embodiment of the present invention, not making any formal limitation to the present invention.Though
So the present invention is disclosed as above with preferred embodiment, but is not limited to the present invention.It is any to be familiar with those skilled in the art
Member, in the case where not departing from the Spirit Essence and technical scheme of the present invention, all using in the methods and techniques of the disclosure above
Appearance makes many possible variations and modification to technical solution of the present invention, or is revised as the equivalent embodiment of equivalent variations.Therefore,
Every content without departing from technical solution of the present invention, the technical spirit according to the present invention is to made for any of the above embodiments any simple
Modification, equivalent substitution, equivalence changes and modification, still fall within technical solution of the present invention protection in the range of.
Claims (10)
1. a kind of composite polyethylene imines modifies graphene oxide composite material, it is characterised in that including graphene oxide, low point
Sub- polyethyleneimine, polyethyleneimine polymer and crosslinking agent, the low molecular polyethylene imines and polyethyleneimine polymer
Modified by crosslinking agent in the surface of graphene oxide；The polyethyleneimine polymer is relative molecular mass >=20000
Polyethyleneimine, the low molecular polyethylene imines be relative molecular weight be less than 5000 polyethyleneimine.
2. composite polyethylene imines according to claim 1 modifies graphene oxide composite material, it is characterised in that described
Polyethyleneimine polymer is the polyethyleneimine that relative molecular mass is 20000~25000, the low molecular polyethylene imines
The polyethyleneimine for being 1500~3000 for relative molecular weight.
3. composite polyethylene imines according to claim 1 or 2 modifies graphene oxide composite material, it is characterised in that
The mass ratio of the polyethyleneimine polymer, low molecular polyethylene imines and the graphene oxide is 5~10: 5~10: 1.
4. composite polyethylene imines according to claim 1 or 2 modifies graphene oxide composite material, it is characterised in that
The crosslinking agent be 1- ethyl -3-3- dimethylaminopropyls, N-hydroxy-succinamide ester and one kind in triethanolamine or
It is a variety of.
5. the composite polyethylene imines any one of a kind of Claims 1-4 modifies the system of graphene oxide composite material
Preparation Method, it is characterised in that comprise the following steps：
S1, graphene oxide is dispersed in water, adds crosslinking agent and obtain suspension；By polyethyleneimine polymer and low molecule
Polyethyleneimine mixing adds crosslinking agent and obtains mixed solution；
S2, the mixed solution is instilled into suspension stirring dropwise make fully contact；
S3, with bag filter dialysis obtain composite polyethylene imines modification graphene oxide composite material.
6. preparation method according to claim 5, it is characterised in that the crosslinking agent added in the graphene oxide is 1-
Ethyl -3-3- dimethylaminopropyls and N-hydroxy-succinamide ester；The polyethyleneimine polymer and the low molecule
The crosslinking agent added in the mixture of polyethyleneimine is triethanolamine.
7. preparation method according to claim 5, it is characterised in that the temperature stirred described in the step S2 is room
Temperature, the time of stirring is 24h；In the step S3, the bag filter is 100KDa bag filter, and the condition of the dialysis is：
3d is permeated with deionized water at room temperature.
8. composite polyethylene imines modification graphene oxide composite material or power any one of a kind of Claims 1-4
Profit requires the composite polyethylene imines modification graphene oxide composite wood that the preparation method any one of 5 to 7 is prepared
Expect the application in microorganism or the transfection of animal and plant cells gene.
9. composite polyethylene imines modification graphene oxide composite material or power any one of a kind of Claims 1-4
Profit requires the composite polyethylene imines modification graphene oxide composite wood that the preparation method any one of 5 to 7 is prepared
Expect the application in drug carrier.
10. application according to claim 9, it is characterised in that the medicine is aromatic series medicine.
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| CN109054377B (en) * | 2018-07-09 | 2020-12-29 | 珠海中科先进技术研究院有限公司 | Dendritic polyamidoamine-graphene oxide combined composite material, and preparation method and application thereof |
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| CN110193357A (en) * | 2019-06-27 | 2019-09-03 | 中素新科技有限公司 | Composite sponge and its preparation method and application containing modified graphene |
| CN112844308A (en) * | 2019-11-28 | 2021-05-28 | 中国科学院大连化学物理研究所 | Magnetic composite material modified by aptamer and preparation and application thereof |
| CN112844308B (en) * | 2019-11-28 | 2022-05-06 | 中国科学院大连化学物理研究所 | Magnetic composite material modified by aptamer and preparation and application thereof |
| CN111154808A (en) * | 2020-01-10 | 2020-05-15 | 阜阳师范大学 | Gene transporter and preparation method and application thereof |
| CN111154808B (en) * | 2020-01-10 | 2022-03-22 | 阜阳师范大学 | Gene transporter and preparation method and application thereof |
| CN112294788A (en) * | 2020-09-29 | 2021-02-02 | 四川大学华西医院 | Multifunctional nano graphene oxide compound for treating osteoarthritis |
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