CN107739448A - Magnetic control discharges nitric oxide production composite film material and its preparation method and application - Google Patents
Magnetic control discharges nitric oxide production composite film material and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of magnetic control to discharge nitric oxide production composite film material and its preparation method and application.The preparation method of the material comprises the following steps：Chitosan Azide is modified, then glyean dendroid daiamid (CS PAMAM) is synthesized by click-reaction with the daiamid of dendroid containing alkynyl, again in ferroso-ferric oxide bare ball surface modification CS PAMAM, nitric oxide is finally loaded, magnetic control is obtained and discharges nitric oxide production composite film material.Polycaprolactone is coated into above-mentioned magnetic control and discharges nitric oxide production composite film material, finally giving can be in the nitric oxide production composite film material anti-biotic material of control release under alternating magnetic field, resulting anti-biotic material has uniform particle diameter, response is sensitive, and NO loads and storage capacity are big, release time length, biocompatibility is good, fungistatic effect is obvious, it is safe efficient and green the advantages that, can be used for bio-medical engineering material field.
Description
Technical field
The invention belongs to bio-medical engineering material field, more particularly to a kind of magnetic control discharges nitric oxide production compound film material
Material and its preparation method and application.
Background technology
Find that vascular endothelial cell can synthesize, secrete blood vessel endothelium derivative relaxing factor from Furchgott in 1980 etc.
(EDRF) since, the proof EDRF of two people's independences of Furchgott in 1986 and Ignarro is NO, about NO basis and face
Bed research is flourished, and NO turns into one of most noticeable biomolecule in recent years.Substantial amounts of research shows that NO is
Important biological messenger molecule, the processes such as regulation of blood vessels, neurotransmission, inflammation and immune response being participated in, its is widely distributed, throughout
A variety of organs such as brain, blood vessel, immune, lung, reproduction.
Discovered in recent years NO can destroy the cell membrane of bacterium and gene information and prevent bacterium from obtaining energy, have efficient
It is antibacterial and be not likely to produce the characteristics such as drug resistance, receive more and more attention its application in antibacterial field, more and more
Releasable NO anti-biotic material is appeared in the visual field of people.
Such as：Smith etc. proposed to be used for making by nucleophic NO donor N-diazeniumdiolate first in 1996
Standby releasable NO polymeric material (Chemistry 1996,39:1148-1156), but the NO donors of small molecule
Diazeniumdiolates is to be disperseed by way of noncovalent interaction in the polymeric material, so as to cause donor to hold
Easily come off from polymeric matrix, non-degradable, produce carcinogen nitrosamine；(the International such as Jong Oh Kima
Journal of Biological Macromolecules 2015,79:The chitosan for 217-225) being prepared for releasable NO is thin
Film, for the research in terms of antibacterial and its wound healing, research shows that NO can actually effectively suppress the growth of bacterium and numerous
Grow, have obvious effect simultaneously for the healing of wound.(the Advanced Healthcare such as Dongsik Park
Materials 2016,5:Releasable NO poly-dopamine hollow Nano microballoon 2019-2024) is prepared for, for antibiosis
Research, research show that NO can effectively destroy the cell membrane of bacterium, cause bacterial death.However, although NO show it is excellent
Different antibacterial effect and drug resistance is not likely to produce, but NO draws its gas property and half-life short, material load NO content mistake
It is low, the problems such as NO is difficult to long-time storage, so that its application clinically receives great obstruction.
Discharge as the indispensable characteristic in NO delivery systems accordingly, with respect to NO fixed point Partial controll, more turn into
Current biological engineering in medicine field important topic urgently to be resolved hurrily.So far, by ferroso-ferric oxide (Fe3O4) surface with
Chitosan click coupling dendritic interphase (CS-PAMAM) carries out modification and is used as NO donors, finally with polycaprolactone (PCL)
Coated, finally prepared using ferroso-ferric oxide as core, centre is chitosan graft dendritic interphase, and outermost layer is
Polycaprolactone, there is not been reported for the nitric oxide production composite film material of control release and its application under alternating magnetic field.
The content of the invention
The shortcomings that primary and foremost purpose of the present invention is to overcome prior art and deficiency, there is provided a kind of magnetic control discharges nitric oxide
Composite film material preparation method.
The magnetic control being prepared another object of the present invention is to provide methods described discharges nitric oxide production compound film material
Material.
A further object of the present invention is to provide the application that the magnetic control discharges nitric oxide production composite film material.
The purpose of the present invention is achieved through the following technical solutions：A kind of magnetic control discharges the system of nitric oxide production composite film material
Preparation Method, comprise the following steps：
(1) Azide chitosan (CS-N3) synthesis：
By nitrine acetic acid (N3-CH2- COOH) be dissolved in N-N- dimethylformamides (DMF), then sequentially add 1- ethyls-
(3- dimethylaminopropyls) carbodiimide hydrochloride (EDCHCl) and n-hydroxysuccinimide (NHS) are activated, then
Chitosan (CS) solution that addition chitosan (CS) is dissolved in water acquisition is reacted, and is dialysed, and freeze-drying, obtains CS-N3(nitrine
Change chitosan)；
(2) synthesis of chitosan graft dendritic interphase (CS-PAMAM)：
Alkynyl dendritic interphase (PAMAM) will be contained to be dissolved into water, then add in step (1) obtained CS-N3
(Azide chitosan), adds cupric sulfate pentahydrate and sodium ascorbate, is reacted in 40~75 DEG C, dialyses, and freezes, obtains
CS-PAMAM (chitosan graft dendritic interphase)；
(3) ferriferrous oxide composite material (Fe of chitosan graft dendritic interphase (CS-PAMAM) modification3O4@
CS-PAMAM synthesis)：
By Fe3O4It is distributed in absolute ethyl alcohol, under protective atmosphere, heating water bath adds step (2) to 30~50 DEG C
In the obtained CS-PAMAM aqueous solution reacted, wash, drying, obtain Fe3O4@CS-PAMAM (chitosan graft dendroids
The ferriferrous oxide composite material of polyamide-amide modification)；
(4) magnetic control discharges nitric oxide production composite film material (Fe3O4@CS-PAMAM/NONOate) synthesis：
The Fe that will be obtained in step (3)3O4@CS-PAMAM are dissolved in absolute methanol, are then added sodium methoxide ultrasound and are stablized 10
~30min, then be passed through NO gases and reacted, after reaction terminates, wash, dry, it is nitric oxide production compound to obtain magnetic control release
Membrane material (Fe3O4@CS-PAMAM/NONOate)。
The dosage of nitrine acetic acid described in step (1) matches 0.1 by every milliliter of DMF (N-N- dimethylformamides)~
0.5g nitrine acetic acid calculates.
The molecular weight of chitosan described in step (1) is 1000~20000, and deacetylation is 40~85%.
Chitosan, 1- ethyls-(3- dimethylaminopropyls) carbodiimide hydrochloride (EDC described in step (1)
HCl), the mol ratio of n-hydroxysuccinimide (NHS) and nitrine acetic acid is 1:1~10:1~10:1~10；Mol ratio is preferred
For 1:1.5~10:1.5~10:2~10.
The dosage of chitosan described in step (1) matches 0.05~0.5g chitosans by every milliliter of water and calculated.
The time of activation described in step (1) is preferably 0.5~4h.
The temperature of reaction described in step (1) is preferably 5~35 DEG C.
The time of reaction described in step (1) is preferably 12~24h.
Dialysis described in step (1) is to be dialysed in pure water；Dialysed 2~3 days preferably in pure water.
Dendritic interphase containing alkynyl (PAMAM) described in step (2) is according to Application No.
The China of the 201610356643.9th, entitled " one kind loads nitric oxide production cationic polymer and its preparation method and application "
Embodiment 4 in patent application is prepared.
Azide chitosan described in step (2), dendritic interphase containing alkynyl (PAMAM), cupric sulfate pentahydrate and
The mol ratio of sodium ascorbate is 1:1~12:1~6:2.5~18.
The dosage of PAMAM (dendritic interphase containing alkynyl) described in step (2) matches 0.05 by every milliliter of water~
0.15g PAMAM are calculated.
Dialysis described in step (2) is to be dialysed using bag filter；Preferably use molecular cut off for 2000~
20000 bag filter is dialysed；More preferably dialysed 2~4 days with the bag filter that molecular cut off is 2000~20000.
Reaction described in step (2) is to be reacted under inert gas shielding.
Described inert gas is preferably nitrogen.
Reaction time described in step (2) is preferably 24~48h.
Chitosan graft dendritic interphase containing alkynyl (CS-PAMAM) molecular formula described in step (2) is as follows:
Wherein, m:N=5:1.
Fe described in step (3)3O4It is prepared preferably by following method：By FeCl3·6H2O and FeCl2·
4H2O is soluble in water, and ammoniacal liquor (NH is injected under the conditions of 30~100 DEG C of stirring in water bath3·H2O) reacted, after reaction terminates, with nothing
Water-ethanol ultrasound, vibration to solid disperse, and wash, and vacuum drying, obtain Fe3O4。
Described FeCl3·6H2O and FeCl2·4H2O mol ratio is 1:0.1~2.
The dosage of described ammoniacal liquor is calculated by its final concentration of percent by volume 5~15% in the reaction system.
Described reaction is to be reacted under inert gas shielding.
The time of described reaction is preferably 0.5~2h.
The number of described washing is preferably 2~4 times.
Ultrasonic disperse is separated into described in step (3).
The described ultrasonic time is 10~30min.
Described ultrasonic power is 250W.
Absolute ethyl alcohol is preferably prepared via a method which to obtain described in step (3)：Calcium hydride is added in ethanol, stirred
Mix 6~24 hours, then air-distillation, obtain absolute ethyl alcohol；Wherein, the addition of the calcium hydride is to be matched by every liter of ethanol
2~4 grams of calcium hydrides calculate.
Fe described in step (3)3O4Dosage match 0.005~0.02g Fe by every milliliter of absolute ethyl alcohol3O4Calculate
(i.e. Fe3O4The concentration of ethanol solution be mass volume ratio 0.5~2%).
The aqueous solution of CS-PAMAM described in step (3) is water-soluble for CS-PAMAM to be dissolved in the CS-PAMAM of water acquisition
Liquid；(i.e. CS-PAMAM is water-soluble by every milliliter of water 0.01~0.02g of proportioning CS-PAMAM calculating for described CS-PAMAM dosage
The concentration of liquid is mass percent 1%~2%).
Fe described in step (3)3O4Mass ratio with the CS-PAMAM is 0.5~1：1.
The time of reaction described in step (3) is preferably 6~12h.
Washing described in step (3) is to be washed with absolute ethyl alcohol；Preferably washed 1~3 time with absolute ethyl alcohol.
The condition of drying described in step (3) is：Dried 12~48 hours in 50~80 DEG C of vacuum drying chamber.
Fe described in step (4)3O4The mol ratio of@CS-PAMAM and sodium methoxide is 1：400~450.
Fe described in step (4)3O4@CS-PAMAM dosage matches 0.05~0.2g by every milliliter of absolute methanol
Fe3O4@CS-PAMAM are calculated.
The addition of sodium methoxide described in step (4) by its in final concentration of 5~20% (w/w) of the reaction system based on
Calculate.
Reaction described in step (4) is realized preferably by following steps：First it is passed through high pure nitrogen maintenance reaction kettle (10
~20psi) 5~15min reacted again after excluding the air in reactor, after reaction terminates, then it is passed through 10~20psi height
Pure nitrogen gas maintains 10~20min discharges NO.
The time of reaction described in step (4) is preferably 3~7 days.
Washing described in step (4) is to be washed with absolute methanol and ice absolute ether；Preferably first with without water beetle
Alcohol washs 1~3 time, then is washed 1~2 time with ice absolute ether.
Absolute methanol described in step (4) is preferably prepared via a method which to obtain：Calcium hydride is added in methanol,
Stirring 6~24 hours, then air-distillation, obtains absolute methanol；The addition of the calcium hydride matches 2~4 by every liter of methanol
Gram calcium hydride calculates.
A kind of magnetic control discharges nitric oxide production composite film material, is prepared by the method described in any of the above-described.
A kind of magnetic control of polycaprolactone cladding discharges nitric oxide production composite film material, is coated by PCL (polycaprolactone)
Above-mentioned magnetic control discharges nitric oxide production composite film material and obtained.
The magnetic control of described polycaprolactone cladding discharges the preparation method of nitric oxide production composite film material, including following step
Suddenly：
(I) PCL (polycaprolactone) is added in DMF (DMF) and chloroform mixed solution, stirring
It is well mixed, obtain PCL solution；
(II) by above-mentioned Fe3O4@CS-PAMAM/NONOate (magnetic control discharges nitric oxide production composite film material) are distributed to step
Suddenly in the PCL solution obtained in (I), solvent flashing is then stirred for, obtains the magnetic control release nitric oxide of polycaprolactone cladding
Composite film material (Fe3O4@CS-PAMAM/NONOate@PCL)。
The molecular weight of PCL (polycaprolactone) described in step (I) is preferably 5000~50000.
The volume ratio of N,N-dimethylformamide (DMF) and chloroform described in step (I) is 1:1~4.
The time of stirring described in step (I) is 1~2h.
The concentration of PCL solution described in step (I) is mass percent 5~30%.
Fe described in step (II)3O4@CS-PAMAM/NONOate and PCL mass ratio is 0.05~5：1.
It is scattered preferably by the realization of following method described in step (II)：First 150~300W ultrasonic vibrations 30~
After 60min, it is subsequently placed in ice bath, 1~3KW ultrasonic cell disruptor ultrasounds, 5~30min.
The time of stirring described in step (II) is preferably 5~24h.
Described magnetic control discharges nitric oxide production composite film material or the magnetic control of described polycaprolactone cladding discharges an oxygen
Change application of the composite film material of nitrogen in bio-medical engineering material.
Described magnetic control discharges nitric oxide production composite film material or the magnetic control of described polycaprolactone cladding discharges an oxygen
Change application of the composite film material of nitrogen in antibacterials are prepared.
Described antibacterials include suppressing bacterium and the growth of fungi and the medicine of breeding.
Described bacterium is preferably staphylococcus aureus.
Described magnetic control discharges nitric oxide production composite film material or the magnetic control of described polycaprolactone cladding discharges an oxygen
The composite film material for changing nitrogen can be also used for preparing the medicine for promoting wound healing and anti-inflammatory.
The present invention is had the following advantages relative to prior art and effect：
(1) reaction preparation is first clicked in the present invention and synthesizes chitosan coupling dendritic interphase (CS-PAMAM)；
Then prepare ferroso-ferric oxide bare ball, and in its surface modification CS-PAMAM, nitric oxide then loaded in autoclave,
Obtain the ferroferric oxide magnetic nano nitric oxide donors of chitosan coupling dendritic interphase modification：Fe3O4@CS-
PAMAM/NONOate；Then scattered be blended in organic solvent with polycaprolactone (PCL) of NO donors is freezed into compacting film forming, most
Obtaining eventually can be in the nitric oxide production composite film material anti-biotic material of control release under alternating magnetic field.
(2) present invention greatly improves NO load capacity, for bacterium using high algebraically polyamide-amide as NO donors
The influence of biomembrane has obvious inhibition, and can be good at absorption and certain killing effect to electronegative bacterium
Fruit.
(3) present invention uses natural chitosan not only to improve Fe for decorative material3O4Stability, and greatly
The bio-toxicity for reducing cationic polymer polyamide-amide.
(4) dendroid polyester-polyamide-amine and chitosan are coupled by the present invention using click chemistry method, reaction efficiency height,
Structure is easy to accuracy controlling, and molecular weight distribution is single.
(5) present invention is with magnetic Fe3O4For core, temperature can be raised under the conditions of alternating magnetic field, is had to bacterium certain
Fragmentation effect, and surface PCL coatings are quickly dissolved, make the cationic polymer for having loaded NO exposed outside, serve control
System release NO effect.
(6) present invention is under additional magnetic fields, magnetic Fe3O4For core, the cationic polymer polyamide-amide of outer layer
Substantial amounts of bacterium can be effectively adsorbed, so as to play a part of fast purification, material can reclaim, green.
(7) NO donors can be wrapped in inside by the present invention well using PCL as face coat hydrophobic material, protection with
Store substantial amounts of NO；And fusing point is relatively low, can quick response, so as to carry out its material intelligent response effect.
(8) material employed in the present invention is relatively small, and in addition to the cytotoxicity for advantageously reducing product, it is as anti-
Bacterium medicine controlled release carrier, in antibacterials, there is potential application value in the field of transmitting altogether.
(9) the NO donor products obtained in the present invention possess the growth and breeding for effectively suppressing bacterium and fungi, to common
Pathogenic bacteria, dermatophyte, wound infection bacterium etc. there is significant inhibition, and with promoting the work(such as wound healing and anti-inflammatory
Can, provide support in the application for preparing biological medicine engineering material for it.
(10) the invention provides one kind can store a large amount of NO, and control release NO novel antibacterial material according to demand
Material, the nano material uniform particle diameter, response is sensitive, and NO loads and storage capacity are big, and release time length, biocompatibility is good, suppression
Bacterium positive effect, important application prospect is shown in antibiosis.
Brief description of the drawings
Fig. 1 is the molecule for the chitosan graft dendritic interphase containing alkynyl (CS-PAMAM) that the present invention is prepared
The structural representation of formula and composite film material；Wherein, figure A be it is of the invention in CS-PAMAM molecular formula (black circular segment is
NO binding sites)；Scheme the structural representation that B is composite film material.
Fig. 2 is chitosan graft dendritic interphase polymer nuclear-magnetism hydrogen spectrogram.
Fig. 3 is Fe3O4@CS-PAMAM and Fe3O4@CS-PAMAM/NONOate composite uv atlas.
Fig. 4 is Fe3O4@CS-PAMAM/NONOate@PCL are in the drug release profiles after alternating magnetic field processing and space management
Figure.
Fig. 5 is Fe3O4@CS-PAMAM/NONOate@PCL nano material test tube turbidimetries compare different composite material antibacterial
Design sketch；Wherein, 1 is Fe3O4@CS-PAMAM/NONOate@PCL and alternating magnetic field collective effect, 2 be Fe3O4@CS-PAMAM/
NONOate, 3 be blank control.
Embodiment
With reference to embodiment, the present invention is described in further detail, but the implementation of the present invention is not limited to this.
1st, the preparation method of absolute methanol used in following examples is according to following operating procedure：Calcium hydride is added to
In methanol, stir 6~24 hours, then air-distillation, obtain absolute methanol, the addition of the calcium hydride is with every 500mL first
1~2 gram of meter is added in alcohol.
2nd, ethylenediamine used in following examples and methyl acrylate are anhydrous ethylenediamine and anhydrous methyl acrylate, its
Preparation method is according to following operating procedure：
(1) anhydrous potassium hydroxide is added in ethylenediamine, stirs 6~24 hours, be then evaporated under reduced pressure, obtain anhydrous second
Diamines, the addition of the anhydrous potassium hydroxide in every 500mL ethylenediamines to add in terms of 1~2 gram；
(2) anhydrous sodium sulfate is added in methyl acrylate, stirs 6~24 hours, then air-distillation, obtain anhydrous
Methyl acrylate, the addition of the anhydrous sodium sulfate in every 500mL methyl acrylates to add in terms of 1~2 gram.
3rd, propargylamine used in following examples is dry propargylamine, and its preparation method is according to following operating procedure：
Calcium hydride is added in propargylamine, is stirred 6~24 hours, is then evaporated under reduced pressure, obtains dry propargylamine；The calcium hydride
Addition by every 10mL propargylamines add 0.1~0.2 gram in terms of.
4th, the preparation method of absolute ethyl alcohol used in following examples is according to following operating procedure：Calcium hydride is added to
In ethanol, stir 6~24 hours, then air-distillation, obtain absolute ethyl alcohol, the addition of the calcium hydride is with every 500mL second
1~2 gram of meter is added in alcohol.
5th, the Fe dried used in following examples3O4@CS-PAMAM preparation method is according to following operating procedure：Will
Fe3O4@CS-PAMAM are placed in vacuum drying chamber, are dried 12~48 hours under the conditions of 40~80 DEG C.
6th, unless otherwise instructed, the temperature range of the room temperature in the present invention is 5~35 DEG C.
Embodiment 1
(1) chitosan (CS-N of azido group modification3) synthesis：
By nitrine acetic acid (N3- CH2-COOH) it is dissolved in N-N- dimethylformamides (DMF), then sequentially add 1- second
Base-(3- dimethylaminopropyls) carbodiimide hydrochloride (EDCHCl) and n-hydroxysuccinimide (NHS) activation
30min, obtain in nitrine acetic acid mixed solution；By chitosan, (CS molecular weight is 2000, deacetylation after activation terminates
85%) pure water is dissolved in, is then slowly added in above-mentioned nitrine acetic acid mixed solution, reacts 12h at 35 DEG C.Product go from
Dialysed 2 days in sub- water, be freeze-dried, obtain product Azide chitosan；Wherein, described chitosan, 1- ethyls-(3- diformazans
Base aminopropyl) mol ratio of carbodiimide hydrochloride (EDCHCl), n-hydroxysuccinimide (NHS) and nitrine acetic acid is
1:1.5:1.5:2；Described N,N-dimethylformamide in every 10 milliliters to add in terms of 5g hydrazoic acid；Described pure water is with every
5g chitosans are added in 10ml.
(2) synthesis of the polyamide-amide dendrimer (PAMAM) containing alkynyl：
With reference to patent application " one kind loads nitric oxide production cationic polymer and its preparation method and application " (patent Shen
Number PAMAM please be synthesized for the method in 201610356643.9).Wherein, the preparation method of absolute methanol walks according to following operation
Suddenly：Calcium hydride is added in methanol, stirs 6~24 hours, then air-distillation, obtains absolute methanol, the calcium hydride
Addition in every 500mL methanol to add in terms of 1 gram.The methyl acrylate of drying process and the preparation method of ethylenediamine are according to following
Operating procedure：Anhydrous sodium sulfate is added in methyl acrylate and stirred 6 hours, then air-distillation, obtains anhydrous acrylic acid
Methyl esters, the addition of the anhydrous sodium sulfate in every 500mL methyl acrylates to add in terms of 1~2 gram；Anhydrous potassium hydroxide is added
Enter into ethylenediamine stirring 6 hours, be then evaporated under reduced pressure, obtain anhydrous ethylenediamine, the addition of the anhydrous potassium hydroxide with
1 gram of meter is added in per 500mL ethylenediamines.The preparation method of dry propargylamine is according to following operating procedure：Calcium hydride is added
Stirred 6 hours into propargylamine, be then evaporated under reduced pressure, obtain dry propargylamine；The addition of the calcium hydride is with every 10mL
0.1 gram of meter is added in propargylamine.
Embodiment 2
(1) chitosan (CS-N of azido group modification3) synthesis：
By nitrine acetic acid (N3- CH2-COOH) it is dissolved in N-N- dimethylformamides (DMF), then sequentially add 1- second
Base-(3- dimethylaminopropyls) carbodiimide hydrochloride (EDCHCl) and n-hydroxysuccinimide (NHS) activation 4h, are obtained
Into nitrine acetic acid mixed solution；Chitosan (CS molecular weight is 20000, deacetylation 40%) is dissolved in by activation after terminating
Pure water, it is then slowly added in above-mentioned nitrine acetic acid mixed solution, reacts 24h at 25 DEG C of room temperature.Product is in deionized water
Dialysis 3 days, freeze-drying, obtains product Azide chitosan.Wherein, the chitosan, 1- ethyls-(3- dimethylaminos third
Base) carbodiimide hydrochloride (EDCHCl), n-hydroxysuccinimide (NHS) and nitrine acetic acid mol ratio be 1:10:10:
10；Described N,N-dimethylformamide in every 10 milliliters to add in terms of 1g hydrazoic acid；Described pure water in every 10ml to add
0.5g chitosans.
(2) synthesis of the polyamide-amide dendrimer (PAMAM) containing alkynyl：
With reference to patent application " one kind loads nitric oxide production cationic polymer and its preparation method and application " (patent Shen
Number PAMAM please be synthesized for the method in 201610356643.9).Wherein, the system of the methyl acrylate of drying process and ethylenediamine
Preparation Method is according to following operating procedure：Anhydrous sodium sulfate is added in methyl acrylate and stirred 24 hours, then air-distillation,
Anhydrous methyl acrylate is obtained, the addition of the anhydrous sodium sulfate in every 500mL methyl acrylates to add in terms of 2 grams；By nothing
Water potassium hydroxide is added in ethylenediamine and stirred 24 hours, is then evaporated under reduced pressure, and obtains anhydrous ethylenediamine, the anhydrous hydroxide
The addition of potassium in every 500mL ethylenediamines to add in terms of 2 grams.The preparation method of dry propargylamine is according to following operating procedure：
Calcium hydride is added in propargylamine and stirred 24 hours, is then evaporated under reduced pressure, obtains dry propargylamine；The calcium hydride adds
Enter amount to be added in every 10mL propargylamines in terms of 0.2 gram.
Embodiment 3
(1) chitosan (CS-N of azido group modification3) synthesis：
By nitrine acetic acid (N3- CH2-COOH) it is dissolved in N-N- dimethylformamides (DMF), then sequentially add 1- second
Base-(3- dimethylaminopropyls) carbodiimide hydrochloride (EDCHCl) and n-hydroxysuccinimide (NHS) activation 2h, are obtained
Into nitrine acetic acid mixed solution；Chitosan (CS molecular weight is 10000, deacetylation 60%) is dissolved in by activation after terminating
Pure water, it is then slowly added in above-mentioned nitrine acetic acid mixed solution, reacts 16h at 5 DEG C of room temperature.Product dialyses 2 in pure water
My god, freeze-drying, obtain product Azide chitosan.Wherein, the chitosan, 1- ethyls-(3- dimethylaminopropyls) carbon
The mol ratio of diimmonium salt hydrochlorate (EDCHCl), n-hydroxysuccinimide (NHS) and nitrine acetic acid is 1:5:5:5；It is described
N,N-dimethylformamide by every 10 milliliters add 3g hydrazoic acid in terms of；Described pure water is gathered with adding 3g shells in every 10ml
Sugar.
(2) synthesis of the polyamide-amide dendrimer (PAMAM) containing alkynyl：
With reference to patent application " one kind loads nitric oxide production cationic polymer and its preparation method and application " (patent Shen
Number PAMAM please be synthesized for the method in 201610356643.9).Wherein, the system of the methyl acrylate of drying process and ethylenediamine
Preparation Method is according to following operating procedure：Anhydrous sodium sulfate is added in methyl acrylate and stirred 12 hours, then air-distillation,
Anhydrous methyl acrylate is obtained, the addition of the anhydrous sodium sulfate in every 500mL methyl acrylates to add in terms of 2 grams；By nothing
Water potassium hydroxide is added in ethylenediamine and stirred 12 hours, is then evaporated under reduced pressure, and obtains anhydrous ethylenediamine, the anhydrous hydroxide
The addition of potassium in every 500mL ethylenediamines to add in terms of 2 grams.The preparation method of dry propargylamine is according to following operating procedure：
Calcium hydride is added in propargylamine and stirred 12 hours, is then evaporated under reduced pressure, obtains dry propargylamine；The calcium hydride adds
Enter amount to be added in every 10mL propargylamines in terms of 0.15 gram.
The synthesis of the chitosan graft dendritic interphase (CS-PAMAM) of embodiment 4
The dendritic interphase (PAMAM) obtained in the step of embodiment 1 (2) is dissolved into pure water, then added real
The Azide chitosan obtained in the step of example 1 (1) is applied, is passed through nitrogen half an hour.Then cupric sulfate pentahydrate and anti-bad is added
Hematic acid sodium, continues logical nitrogen, reacts 24h in 40 DEG C.Reaction solution carries out dialysis 2 days with bag filter (molecular cut off 2000),
It is lyophilized, obtain product chitosan graft dendritic interphase (CS-PAMAM)；Wherein, described Azide chitosan, containing alkynes
The mol ratio of base dendritic interphase, cupric sulfate pentahydrate and sodium ascorbate is 1:1:1:2.5；Dissolved in described pure water
The usage amount of dendritic interphase is added in terms of 5 grams of dendritic interphases by every 100 milliliters of pure water.
The synthesis of the chitosan graft dendritic interphase (CS-PAMAM) of embodiment 5：
The dendritic interphase (PAMAM) obtained in the step of embodiment 2 (2) is dissolved into pure water, then added real
The Azide chitosan obtained in the step of example 2 (1) is applied, is passed through nitrogen half an hour.Then cupric sulfate pentahydrate and anti-bad is added
Hematic acid sodium, continues logical nitrogen, reacts 48h in 75 DEG C.Reaction solution carries out dialysis 4 days with bag filter (molecular cut off 20000),
It is lyophilized to obtain product chitosan graft dendritic interphase (CS-PAMAM)；Wherein, described Azide chitosan, containing alkynes
The mol ratio of base dendritic interphase, cupric sulfate pentahydrate and sodium ascorbate is 1:12:6:18；Dissolved in described pure water
The usage amount of dendritic interphase is added in terms of 15 grams of dendritic interphases by every 100 milliliters of pure water.
The synthesis of the chitosan graft dendritic interphase (CS-PAMAM) of embodiment 6：
The dendritic interphase (PAMAM) obtained in the step of embodiment 3 (2) is dissolved into pure water, then added real
The Azide chitosan obtained in the step of example 3 (1) is applied, is passed through nitrogen half an hour.Then cupric sulfate pentahydrate and anti-bad is added
Hematic acid sodium, continues logical nitrogen, reacts 36h in 65 DEG C.Reaction solution carries out dialysis 3 days with bag filter (molecular cut off 10000),
It is lyophilized to obtain product chitosan graft dendritic interphase (CS-PAMAM)；Described Azide chitosan, branch containing alkynyl
The mol ratio of shape polyamide-amide, cupric sulfate pentahydrate and sodium ascorbate is 1:6:3:9；Dendroid is dissolved in described pure water to gather
The usage amount of amide-amine is added in terms of 10 grams of dendritic interphases by every 100 milliliters of pure water.
The ferriferrous oxide composite material of the chitosan graft dendritic interphase (CS-PAMAM) of embodiment 7 modification
(Fe3O4@CS-PAMAM) synthesis
(1) FeCl is weighed3·6H2O and FeCl2·4H2O is dissolved in deionized water, is protected in nitrogen, 30 DEG C of stirring in water bath
NH is rapidly injected after 30min3·H2O, under nitrogen protection, continue to stir 30min in 30 DEG C of water-baths, react after terminating with anhydrous
EtOH Sonicate, vibration to solid disperse, and wash 2 times, vacuum drying, obtain Fe3O4；Wherein, described FeCl3·6H2O and
FeCl2·4H2O mol ratio is 1:0.1；Described ammoniacal liquor (NH3·H2O final concentration of percent by volume 5%)；Described
The preparation method of absolute ethyl alcohol is according to following operating procedure：Calcium hydride is added in ethanol, stirred 6 hours, then normal pressure steaming
Evaporate, obtain absolute ethyl alcohol, the addition of the calcium hydride in every 500mL ethanol to add in terms of 1 gram.
(2) Fe obtained in step (1) is weighed3O4Absolute ethyl alcohol is dissolved in, obtains Fe3O4Ethanol solution, ultrasonic (250W)
10min, nitrogen protection, heating water bath is to 30 DEG C, and after temperature stabilization, add the CS-PAMAM aqueous solution (will obtain in embodiment 4
To CS-PAMAM be dissolved in pure water), react 6h, wash 1 time with absolute ethyl alcohol, drying, obtain chitosan graft dendroid and gather
Ferriferrous oxide composite material (the Fe of amide-amine (CS-PAMAM) modification3O4@CS-PAMAM)；Wherein, described Fe3O4Second
The concentration of alcoholic solution is that (every 100 milliliters of absolute ethyl alcohols add 0.5gFe to mass volume ratio 0.5%3O4)；Described CS-PAMAM's
The concentration of the aqueous solution is mass percent 1% (every 100 milliliters of pure water add 1g CS-PAMAM)；Described CS-PAMAM with
Fe3O4Mass ratio is 1:0.5.
The ferriferrous oxide composite material of the chitosan graft dendritic interphase (CS-PAMAM) of embodiment 8 modification
(Fe3O4@CS-PAMAM) synthesis
(1) FeCl is weighed3·6H2O and FeCl2·4H2O is dissolved in deionized water, is protected in nitrogen, 100 DEG C of stirring in water bath
NH is rapidly injected after 2h3·H2O, under nitrogen protection, continue to stir 2h in 100 DEG C of water-baths, reaction is surpassed after terminating with absolute ethyl alcohol
Sound, vibration to solid disperse, and wash 4 times, vacuum drying, obtain Fe3O4；Wherein, described FeCl3·6H2O and FeCl2·
4H2O mol ratio is 1:2；The final concentration of percent by volume 15% of described ammoniacal liquor；The preparation method of described absolute ethyl alcohol
According to following operating procedure：Calcium hydride is added in ethanol, stirs 24 hours, then air-distillation, obtains absolute ethyl alcohol, institute
The addition of calcium hydride is stated to be added in every 500mL ethanol in terms of 2 grams.
(2) Fe obtained in step (1) is weighed3O4Absolute ethyl alcohol is dissolved in, obtains Fe3O4Ethanol solution, ultrasonic (250W)
30min, nitrogen protection, heating water bath is to 50 DEG C, and after temperature stabilization, add the CS-PAMAM aqueous solution (will obtain in embodiment 5
To CS-PAMAM be dissolved in pure water), react 12h, wash 3 times with absolute ethyl alcohol, dry and obtain chitosan graft dendroid and gather
Ferriferrous oxide composite material (the Fe of amide-amine (CS-PAMAM) modification3O4@CS-PAMAM).Wherein, described Fe3O4Second
The concentration of alcoholic solution is that (every 100 milliliters of absolute ethyl alcohols add 2g Fe to mass volume ratio 2%3O4).The CS-PAMAM's is water-soluble
Liquid concentration is mass percent 2% (every 100 milliliters of pure water add 2g CS-PAMAM)；Described CS-PAMAM and Fe3O4Quality
Than for 1:1.
The ferriferrous oxide composite material of the chitosan graft dendritic interphase (CS-PAMAM) of embodiment 9 modification
(Fe3O4@CS-PAMAM) synthesis
(1) FeCl is weighed3·6H2O and FeCl2·4H2O is dissolved in deionized water, is protected in nitrogen, 30 DEG C~100 DEG C water
NH is rapidly injected after bath stirring 1h3·H2O, under nitrogen protection, continue to stir 1h in 60 DEG C of water-baths, react after terminating with anhydrous
EtOH Sonicate, vibration to solid disperse, and wash 3 times, vacuum drying, obtain Fe3O4.Wherein, described FeCl3·6H2O and
FeCl2·4H2O mol ratios are 1:1；The final concentration of percent by volume 10% of described ammoniacal liquor；The preparation side of described absolute ethyl alcohol
Method is according to following operating procedure：Calcium hydride is added in ethanol, stirs 12 hours, then air-distillation, obtains absolute ethyl alcohol,
The addition of the calcium hydride in every 500mL ethanol to add in terms of 1.5 grams.
(2) Fe obtained in step (1) is weighed3O4Absolute ethyl alcohol is dissolved in, obtains Fe3O4Ethanol solution, ultrasonic (250W)
15min, nitrogen protection, heating water bath is to 40 DEG C, after temperature stabilization, adds the CS-PAMAM aqueous solution (by embodiment 6
In obtained CS-PAMAM be dissolved in pure water), react 10h, wash 2 times with absolute ethyl alcohol, dry and obtain chitosan graft branch
The ferriferrous oxide composite material of shape polyamide-amide (CS-PAMAM) modification.(Fe3O4@CS-PAMAM)；Wherein, it is described
Fe3O4The concentration of ethanol solution for mass volume ratio 1.5%, (every 100 milliliters of absolute ethyl alcohols add 1.5g Fe3O4).Described
The concentration of the CS-PAMAM aqueous solution is mass percent 1.5% (every 100 milliliters of pure water add 1.5g CS-PAMAM)；It is described
CS-PAMAM and Fe3O4Mass ratio is 1:0.75.
Synthesis (the Fe of the releasable nitric oxide nanoparticle of embodiment 103O4@CS-PAMAM/NONOate) synthesis
By the Fe of the gained of embodiment 73O4After@CS-PAMAM are dissolved in absolute methanol after drying, it is stable to add sodium methoxide ultrasound
Autoclave is positioned over after 10min to seal and detect air-tightness.High pure nitrogen maintenance reaction kettle (10psi) 5min excludes reaction
Air in kettle, then pass to NO gases (40psi) and react 3 days at room temperature.After reaction terminates, NO is by 10psi high pure nitrogen
Discharge and open reactor after continuing to 10min, take out reaction product, washed 2 times with absolute methanol, then with the anhydrous second of ice
Ether washs 1 time, vacuum drying, obtains final product (Fe3O4@CS-PAMAM/NONOate)；Wherein, described Fe3O4@CS-
The mol ratio of PAMAM and sodium methoxide is 1:400；The concentration of described sodium methoxide is mass percent 5%；Described absolute methanol
With every 10ml dissolving 0.5g Fe3O4@CS-PAMAM are counted；The Fe of the drying3O4@CS-PAMAM preparation method is according to following behaviour
Make step：By Fe3O4@CS-PAMAM are placed in vacuum drying chamber, are dried 12 hours under the conditions of 50 DEG C.
Synthesis (the Fe of the releasable nitric oxide nanoparticle of embodiment 113O4@CS-PAMAM/NONOate) synthesis
By the Fe of the gained of embodiment 83O4After@CS-PAMAM are dissolved in absolute methanol after drying, it is stable to add sodium methoxide ultrasound
Autoclave is positioned over after 30min to seal and detect air-tightness.High pure nitrogen maintenance reaction kettle (20psi) 15min excludes anti-
The air in kettle is answered, NO gases (80psi) is then passed to and reacts 7 days at room temperature.After reaction terminates, NO is by 20psi High Purity Nitrogen
Gas is discharged and opens reactor after continuing to 20min, takes out reaction product.Washed 3 times with absolute methanol, it is then anhydrous with ice
Ether washs 2 times, vacuum drying, obtains final product (Fe3O4@CS-PAMAM/NONOate)；Wherein, it is described
Fe3O4The mol ratio of@CS-PAMAM and sodium methoxide is 1:450；The concentration of the sodium methoxide is mass percent
20%；Described absolute methanol is with every 10ml dissolving 2g Fe3O4@CS-PAMAM are counted；The Fe of the drying3O4@CS-PAMAM's
Preparation method is according to following operating procedure：By Fe3O4@CS-PAMAM are placed in vacuum drying chamber, and it is small that 48 are dried under the conditions of 80 DEG C
When.
Synthesis (the Fe of the releasable nitric oxide nanoparticle of embodiment 123O4@CS-PAMAM/NONOate) synthesis
By the Fe of the gained of embodiment 93O4After@CS-PAMAM are dissolved in absolute methanol after drying, it is stable to add sodium methoxide ultrasound
Autoclave is positioned over after 25min to seal and detect air-tightness.High pure nitrogen maintenance reaction kettle (15psi) 15min excludes anti-
The air in kettle is answered, NO gases (60psi) is then passed to and reacts 6 days at room temperature.After reaction terminates, NO is by 15psi High Purity Nitrogen
Gas is discharged and opens reactor after continuing to 15min, takes out reaction product.Washed 3 times with absolute methanol, it is then anhydrous with ice
Ether washs 2 times, vacuum drying, obtains final product (Fe3O4@CS-PAMAM/NONOate)；Wherein, described Fe3O4@CS-
The mol ratio of PAMAM and sodium methoxide is 1:420；The concentration of the sodium methoxide is mass percent 15%, described absolute methanol
With every 10ml dissolving 1g Fe3O4@CS-PAMAM are counted；The Fe of the drying3O4@CS-PAMAM preparation method is according to following operation
Step：By Fe3O4@CS-PAMAM are placed in vacuum drying chamber, are dried 36 hours under the conditions of 60 DEG C.
The nitric oxide production composite synthesis of controllable release of the polycaprolactone of embodiment 13 cladding
A certain amount of PCL particles (molecular weight 5000) are weighed, are dissolved in volume ratio for 1:1 N,N-dimethylformamide
(DMF) and in chloroform mixed solution, the PCL solution of mass percent 5% is configured to, is sufficiently stirred 1h, it is homogeneous to solution
After stable, the Fe of the gained of embodiment 10 is weighed3O4@CS-PAMAM/NONOate magnetic composites are added to the PCL configured
(Fe in solution3O4The mass ratio of@CS-PAMAM/NONOate and PCL particles is 0.05：1)., will after 150W ultrasonic vibrations 30min
Solution, which is placed in ice bath, uses ultrasonic cell disruptor ultrasound 1KW/5min, magnetic nanoparticle is uniformly divided in PCL solution
Dissipate, then the quick stirring 5h solvent flashings of machinery, obtain the nitric oxide production composite of controllable release of polycaprolactone cladding
(Fe3O4@CS-PAMAM/NONOate@PCL)。
The nitric oxide production composite synthesis of controllable release of the polycaprolactone of embodiment 14 cladding
A certain amount of PCL particles (molecular weight 50000) are weighed, are dissolved in volume ratio for 1:4 N,N-dimethylformamide
(DMF) and in chloroform mixed solution, the PCL solution of mass percent 30% is configured to, is sufficiently stirred 2h, it is homogeneous to solution
After stable, the Fe of the gained of embodiment 11 is weighed3O4@CS-PAMAM/NONOate magnetic composites are added to the PCL configured
(Fe in solution3O4The mass ratio of@CS-PAMAM/NONOate and PCL particles is 5：1)., will be molten after 300W ultrasonic vibrations 60min
Liquid, which is placed in ice bath, uses ultrasonic cell disruptor ultrasound 3KW/5min, makes magnetic nanoparticle dispersed in PCL solution,
Then the quick stirring 24h solvent flashings of machinery, obtain the nitric oxide production composite of controllable release of polycaprolactone cladding
(Fe3O4@CS-PAMAM/NONOate@PCL)。
The nitric oxide production composite synthesis of controllable release of the polycaprolactone of embodiment 15 cladding
A certain amount of PCL particles (molecular weight 20000) are weighed, are dissolved in volume ratio for 1:2 N,N-dimethylformamide
(DMF) and in chloroform mixed solution, the PCL solution of mass percent 20% is configured to, is sufficiently stirred 1.5h, it is equal to solution
After one is stable, the Fe of the gained of embodiment 12 is weighed3O4@CS-PAMAM/NONOate magnetic composites are added to what is configured
(Fe in PCL solution3O4The mass ratio of@CS-PAMAM/NONOate and PCL particles is 5：1)., will after 200W ultrasonic vibrations 40min
Solution, which is placed in ice bath, uses ultrasonic cell disruptor ultrasound 2KW/5min, magnetic nanoparticle is uniformly divided in PCL solution
Dissipate, then the quick stirring 15h solvent flashings of machinery, obtain the nitric oxide production composite of controllable release of polycaprolactone cladding
(Fe3O4@CS-PAMAM/NONOate@PCL)。
Embodiment 16
The chitosan graft dendritic CS-PAMAM of the gained of embodiment 5 is dissolved in heavy water and carries out nucleus magnetic hydrogen spectrum
Characterize.As a result as shown in Fig. 2 proton peak of the chemical shift at 2.0-4.0ppm corresponds to dendroid daiamid and chitosan
Proton hydrogen；Proton peak of the chemical shift at 7.8ppm is corresponding to the dendritic interphase containing alkynyl and containing azido
Chitosan reacts the proton hydrogen for the azacyclo- to be formed by click chemistry.Illustrate dendritic interphase and Azide chitosan
Successfully synthesis is reacted by click chemistry and obtains polymer CS-PAMAM.
Embodiment 19
By the gained Fe of embodiment 103O4@CS-PAMAM/NONOate and the gained of embodiment 8 Fe3O4@CS-PAMAM distinguish
The aqueous solution that concentration is 0.1mg/ml is configured in dissolving distilled water, carries out ultraviolet spectroscopy, as a result Fe as shown in Figure 33O4@
Fes of the CS-PAMAM/NONOate compared to no load NO3O4@CS-PAMAM absorb at 252nm for NONOate characteristic groups
Peak；By Fe3O4@CS-PAMAM/NONOate are dissolved in the aqueous solution that 0.1mg/ml is configured in PBS, and one section is the time
Afterwards, due to Fe3O4@CS-PAMAM/NONOate can be converted into NO after discharging NO2-, occur at 540nm in ultraviolet spectrogram bright
Aobvious azo dyes absworption peak, this is due to NO2-The azo compound generated in dative after this reagent reacting.
Embodiment 20
Weigh the gained Fe of 5mg embodiments 143O4@CS-PAMAM/NONOate@PCL, it is dissolved in 5ml Citrate buffer
Liquid (pH7.4), at 37 DEG C, it is divided into A, two groups of B, A groups is placed under alternating magnetic field, B groups is as blank control, at regular intervals
Respectively take 50 μ L solution to be mixed with 50 μ L DPBS buffer solutions, then add 100 μ L Griess reagents, 15min is kept in dark place,
Its absorbance is determined using ELIASA under 540nm wavelength, draws drug release profiles.As shown in figure 4, the nano material modified through PCL
In the presence of alternating magnetic field can quick release NO, be compared to for blank group, after being modified through PCL, can reach and prolong
Slow releasing effect, it can reach deenergized period 5-6 days or so.
Embodiment 21
The gained Fe of embodiment 14 is weighed respectively3O4@CS-PAMAM/NONOate@PCL and the gained of embodiment 10 Fe3O4@
Each 0.25mg of CS-PAMAM/NONOate are dissolved in 5ml physiological saline and add 50 μ l (absorbance OD thereto590=1) it is golden yellow
In color staphylococcus (ATCC29213) test tube, wherein test tube 1 is Fe3O4@CS-PAMAM/NONOate@PCL+ alternating magnetic fields, examination
Pipe 2 is CS-PAMAM/NON Oate, and test tube 3 continues after cultivating 12h, through wheat for blank control while after alternating magnetic field is handled
Family name's nephelometry compares, and experimental result is as shown in figure 5, the test tube 3 that compares finds that test tube 1,2 is clarified, and bacterial growth is by significant
Suppress, illustrate Fe3O4@CS-PAMAM/NONOate@PCL and Fe3O4@CS-PAMA M/NONOate have obvious antibacterial effect
Fruit, the material are expected to turn into a kind of new antibacterial agent.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (10)
1. a kind of magnetic control discharges the preparation method of nitric oxide production composite film material, it is characterised in that comprises the following steps：
(1) synthesis of Azide chitosan：
Nitrine acetic acid is dissolved in N-N- dimethylformamides, then sequentially adds 1- ethyls-(3- dimethylaminopropyls) carbon
Diimmonium salt hydrochlorate and n-hydroxysuccinimide are activated, and add the chitosan solution progress that chitosan is dissolved in water acquisition
Reaction, dialyse, freeze-drying, obtain CS-N3；
(2) synthesis of chitosan graft dendritic interphase：
Alkynyl dendritic interphase will be contained to be dissolved into water, then add in step (1) obtained CS-N3, add five water
Copper sulphate and sodium ascorbate, reacted, dialysed in 40~75 DEG C, freezed, obtain CS-PAMAM；
(3) synthesis of the ferriferrous oxide composite material of chitosan graft dendritic interphase modification：
By Fe3O4It is distributed in absolute ethyl alcohol, under protective atmosphere, heating water bath is added in step (2) and obtained to 30~50 DEG C
To the CS-PAMAM aqueous solution reacted, wash, drying, obtain Fe3O4@CS-PAMAM；
(4) magnetic control discharges the synthesis of nitric oxide production composite film material：
The Fe that will be obtained in step (3)3O4@CS-PAMAM are dissolved in absolute methanol, then add sodium methoxide ultrasound stablize 10~
30min, then be passed through NO gases and reacted, after reaction terminates, wash, dry, obtain magnetic control and discharge nitric oxide production composite membrane
Material.
2. magnetic control according to claim 1 discharges the preparation method of nitric oxide production composite film material, it is characterised in that：
Chitosan, 1- ethyls-(3- dimethylaminopropyls) carbodiimide hydrochloride, N- hydroxysuccinimidyls described in step (1)
The mol ratio of acid imide and nitrine acetic acid is 1:1~10:1~10:1~10；
CS-N described in step (2)3, dendritic interphase containing alkynyl, the mol ratio of cupric sulfate pentahydrate and sodium ascorbate
For 1:1~12:1~6:2.5~18；
Fe described in step (3)3O4Mass ratio with the CS-PAMAM is 0.5~1：1；
Fe described in step (4)3O4The mol ratio of@CS-PAMAM and sodium methoxide is 1：400~450.
3. magnetic control according to claim 1 discharges the preparation method of nitric oxide production composite film material, it is characterised in that：
The dosage of nitrine acetic acid described in step (1) matches 0.1~0.5g nitrine second by every milliliter of N-N- dimethylformamide
Acid calculates；
The dosage of chitosan described in step (1) matches 0.05~0.5g chitosans by every milliliter of water and calculated.
The dosage of the dendritic interphase containing alkynyl described in step (2) matches 0.05~0.15g by every milliliter of water and contains alkynyl
Dendritic interphase calculates；
Fe described in step (3)3O4Dosage match 0.005~0.02g Fe by every milliliter of absolute ethyl alcohol3O4Calculate；
The aqueous solution of CS-PAMAM described in step (3) is the solution that CS-PAMAM is dissolved in water acquisition；Described CS-
PAMAM dosage matches 0.01~0.02g CS-PAMAM by every milliliter of water and calculated；
Fe described in step (4)3O4@CS-PAMAM dosage matches 0.05~0.2g Fe by every milliliter of absolute methanol3O4@CS-
PAMAM is calculated.
4. magnetic control according to claim 1 discharges the preparation method of nitric oxide production composite film material, it is characterised in that step
Suddenly the CS-PAMAM molecular formula described in (2) are as follows：
Wherein, m:N=5:1.
5. magnetic control according to claim 1 discharges the preparation method of nitric oxide production composite film material, it is characterised in that：
The molecular weight of chitosan described in step (1) is 1000~20000, and deacetylation is 40~85%；
The time for the activation stated in step (1) is 0.5~4h；
The time of reaction described in step (1) is 12~24h；
Reaction described in step (2) is to be reacted under inert gas shielding；
Reaction time described in step (2) is 24~48h；
Dialysis described in step (2) is dialysed for the bag filter for being 2000~20000 with molecular cut off；
Fe described in step (3)3O4It is prepared via a method which to obtain：By FeCl3·6H2O and FeCl2·4H2O is dissolved in water
In, ammoniacal liquor is injected under the conditions of 30~100 DEG C of stirring in water bath and is reacted, after reaction terminates, with absolute ethyl alcohol ultrasound, vibration to admittedly
Body disperses, and washs, and vacuum drying, obtains Fe3O4；Described FeCl3·6H2O and FeCl2·4H2O mol ratio is 1:0.1~
2；The dosage of described ammoniacal liquor is calculated by its final concentration of percent by volume 5~15% in the reaction system；
The time of reaction described in step (3) is 6~12h；
The condition of drying described in step (3) is：Dried 12~48 hours in 50~80 DEG C of vacuum drying chamber；
The time of reaction described in step (4) is 3~7 days.
6. a kind of magnetic control discharges nitric oxide production composite film material, it is characterised in that：By described in any one of Claims 1 to 5
Method be prepared.
7. a kind of magnetic control of polycaprolactone cladding discharges nitric oxide production composite film material, it is characterised in that：Coated and weighed by PCL
Profit requires that the magnetic control described in 6 discharges nitric oxide production composite film material and obtained；Its preparation method comprises the following steps：
(I) PCL is added in DMF and chloroform mixed solution, is uniformly mixed, it is molten to obtain PCL
Liquid；
(II) magnetic control described in claim 6 is discharged into nitric oxide production composite film material and is distributed to what is obtained in step (I)
In PCL solution, solvent flashing is then stirred for, the magnetic control for obtaining polycaprolactone cladding discharges nitric oxide production composite film material.
8. the magnetic control of polycaprolactone cladding according to claim 7 discharges nitric oxide production composite film material, its feature exists
In：
The molecular weight of PCL described in step (I) is 5000~50000；
The volume ratio of N,N-dimethylformamide and chloroform described in step (I) is 1:1~4；
The concentration of PCL solution described in step (I) is mass percent 5~30%；
It is 0.05~5 that magnetic control described in step (II), which discharges nitric oxide production composite film material and the mass ratio of the PCL,：1.
It is 9. poly- described in the nitric oxide production composite film material of magnetic control release or any one of claim 7~8 described in claim 6
The magnetic control of caprolactone cladding discharges application of the nitric oxide production composite film material in bio-medical engineering material.
10. the magnetic control described in claim 6 is discharged described in nitric oxide production composite film material or any one of claim 7~8
The magnetic control of polycaprolactone cladding discharges application of the nitric oxide production composite film material in antibacterials are prepared.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102241790A (en) * | 2011-06-13 | 2011-11-16 | 中山大学 | Amphipathic chitosan derivative and preparation method and application thereof |
| CN106046382A (en) * | 2016-05-25 | 2016-10-26 | 暨南大学 | Nitric oxide loaded cationic polymer, preparation method therefor and application of nitric oxide loaded cationic polymer |
-
2017
- 2017-10-25 CN CN201711008529.8A patent/CN107739448A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102241790A (en) * | 2011-06-13 | 2011-11-16 | 中山大学 | Amphipathic chitosan derivative and preparation method and application thereof |
| CN106046382A (en) * | 2016-05-25 | 2016-10-26 | 暨南大学 | Nitric oxide loaded cationic polymer, preparation method therefor and application of nitric oxide loaded cationic polymer |
Non-Patent Citations (3)
| Title |
|---|
| JONG OH KIM ET AL.: ""Nitric oxide-releasing chitosan film for enhanced antibacterial and in vivo wound-healing efficacy"", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
| RUI LIU ET AL.: ""Core−Shell Fe3O4 Polydopamine Nanoparticles Serve Multipurpose"", 《APPLIED MATERIALS & INTERFACES》 * |
| 聂吉林 等: ""装载壳聚糖修饰氧化铁的聚酰胺-胺类树状分子纳米粒的制备、表征及体外MR显像"", 《中国现代医学杂志》 * |
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| CN108969769B (en) * | 2018-07-11 | 2021-07-27 | 暨南大学 | Polymer simultaneously carrying penicillin and nitric oxide as well as preparation method and application thereof |
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