CN109646677A - A kind of magnetic nano-particle and its method for preparing intraocular hypertension animal model - Google Patents
A kind of magnetic nano-particle and its method for preparing intraocular hypertension animal model Download PDFInfo
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Abstract
The present invention provides a kind of magnetic nano-particle and its methods for preparing intraocular hypertension animal model.A kind of magnetic nano-particle of the invention based on size adjustable, the magnetic nano-particle have multistage package assembly, can be prepared by emulsion polymerization；When preparing intraocular hypertension animal model, it is imported by injected into anterior chambers intraocular, approach can be recycled according to aqueous humor --- the specific heterogeneous microstructure of trabecular network/schlemm's canal adjusts self-crosslinking or solution is crosslinked, form stable aqueous humor barrier layer, comprehensive multi-level closure aqueous humor runs channel, aqueous humor drains body circulation is limited, increases ah outflow resistance, persistently maintains intraocular hypertension.The preparation method of intraocular hypertension animal model of the present invention, it is simply, conveniently, safe, long-acting.
Description
Technical field
The present invention relates to a kind of magnetic nano-particle and its methods for preparing intraocular hypertension animal model, belong to nano material skill
Art field.
Background technique
At present glaucoma intraocular hypertension animal model preparation method include ball week vein ligation method, dexamethasone method, Qian Fangning
Glue injection method and anterior chamber's magnetic bead injection method, although these methods can prepare intraocular hypertension animal model, each have their own problem.
Total to be characterized in that intraocular pressure stability is poor, success rate of operation is low.Ball week, vein ligation method can stimulate blood vessel to compensate hyperplasia, and bypass ligatures
Blood vessel realizes venous return, modeling is caused to fail；Dexamethasone method can cause intraocular many tissue chronobiological toxicity, increase
Problem complexity is unfavorable for the accurate investigation of Single-issue；Gel method maintains the intraocular hypertension time shorter, for long-term intraocular hypertension disease
The deep investigation effect of disease is limited；Magnetic bead partial size selected by anterior chamber's magnetic bead injection method, can only be in room angle trabecular network position at 3 μm or so
Enrichment cannot be deep into aqueous humor operation channel interior tissue, and the fundamental property of magnetic bead used does not have controllability, cannot be fine
Ground adapts to specific heterogeneous microstructure, and caused intraocular hypertension stability and persistence are all undesirable, intraocular hypertension animal model at
Power is only less than one third.Therefore, preparation can maintain the intraocular hypertension animal model of intraocular hypertension steadily in the long term, it has also become high eye
Press the task of top priority of pathogenesis and drug screening correlative study.
Summary of the invention
The present invention is in view of the above-mentioned problems, providing a kind of magnetic nano-particle and its preparing the side of intraocular hypertension animal model
Method.The present invention is based on a kind of magnetic nano-particle of size adjustable, there is multistage package assembly, by by a kind of magnetic Nano
Particle dispersion is imported in eye with anterior chamber's injection system, and under magnet guidance, magnetic nano-particle is deeply recycled to aqueous humor
Aqueous humor runs channel organizations at different levels, and according to aqueous humor circulation channel --- the specific microstructure knot of trabecular network/schlemm's canal
Structure adjusts itself partial size, and comprehensive multi-level closure aqueous humor circulation channel increases aqueous humor resistance to outflow, persistently maintains intraocular hypertension.
The particle size adjustment of the magnetic nano-particle can realize that the nanoparticle of dissociation is transported by being crosslinked reconciliation crosslinking between particle
Row arrives new position, is crosslinked again with the generation of other particles, and crosslinking reconciliation cross-linking process dynamic occurs between particle, finally tends to flat
Weighing apparatus.
Present invention firstly provides a kind of magnetic nano-particle, the magnetic nano-particle particle size range be 100~
1200nm is crosslinked by the assembling of small-size magnetic nanoparticle；
The small-size magnetic nano particle diameter range is 10~100nm, has core-shell structure, and core is four oxidations three
Ferromagnetism nanometer particle, shell are mesoporous silicon dioxide nano layer, and mesoporous silicon dioxide nano layer is coated on Fe 3 O 4 magnetic
Nanoparticle surface protects ferroferric oxide magnetic nano-particles, and provides better Biocompatibility；Four oxygen
Changing between three ferromagnetism nanometer particles and mesoporous silicon dioxide nano layer also has among the mesoporous silicon oxide containing fluorescence probe
Layer, in order to it is subsequent can pass through nuclear magnetic resonance or fluorescence microscope and track magnetic nano-particle in running body and tissue
Distribution；Magnetic nano-particle has multi-modal tracking and detection feature, is conducive to the system of the system evaluation intraocular hypertension animal model
Preparation Method is also beneficial to the other correlative studys carried out based on intraocular hypertension animal model work；
The mesoporous silicon dioxide nano layer surface also has polymer molecule decorative layer, passes through between polymer molecule
Small-size magnetic nanoparticle is assembled into magnetic nano-particle by cross-linking reaction；The polymer molecule, one end include silicon
Oxyalkyl group, the other end include mercapto groups；Polymer molecule is grafted to mesoporous silicon dioxide nano layer by siloxane group
Surface, then intermolecular cross-linking is realized by the mercapto groups of the other end, and then realizes small size by being crosslinked between polymer molecule
The assembling of magnetic nano-particle is crosslinked.With the mercapto groups of polymer oxidation reaction can occur for small molecule mercapto reagent, be formed
Disulfide bond can hinder to crosslink reaction with disulfide bond mode between nanoparticle.
Preferably, the magnetic nano-particle particle size range is 300~1000nm, the small-size magnetic nanoparticle
Sub- particle size range is 20~80nm.It is highly preferred that the magnetic nano-particle particle size range is 500~800nm, described is small
Sized magnetic nano particle diameter range is 40~60nm.
The polymer molecule has good biological safety, including non-degradable type polymer and degradable polymerization
Object, non-degradable type polymer include polyethylene, polypropylene, polyacrylate, aromatic polyester, polysiloxanes, appointing in polyformaldehyde
Any combined polymerization of one kind or any combination thereof or monomer or segment；Degradable polymer includes collagen, gelatin, fibroin
Albumen, chitosan, chitin, cellulose, polyaminoacid, polyethylene glycol, polylactic acid, polyglycolic acid, poly lactic-co-glycolic acid
Any combined polymerization of any one of copolymer, polycaprolactone or any combination thereof or monomer or segment；Preferably, it polymerize
Object is polysiloxanes, polyethylene glycol, polyaminoacid, polylactic acid, polyglycolic acid, poly lactide-glycolide acid, gathers in oneself
Any combined polymerization of any one of ester or any combination thereof or monomer or segment；Polymer molecular weight between 2000~
Between 20000.
The present invention also provides a kind of method for preparing intraocular hypertension animal model based on above-mentioned magnetic nano-particle, this method
Include the following steps:
S1: the preparation of small-size magnetic nanoparticle
S11: from Fe3O4Nanoparticle is to Fe3O4@SiO2(FITC)
Magnetic iron powder-n-hexane dispersion liquid is diluted in the cyclohexane solution of n-octyl alcohol and Triton X-100,
Ammonium hydroxide is added and forms stable reverse micro emulsion, is added with stirring silester (TEOS) in continuous machinery and is reacted, with shape
At the magnetic nano-particle dispersion liquid of silicon dioxide coating, i.e. Fe3O4@SiO2Sample；In above-mentioned Fe3O4@SiO2Disperse liquor
Middle addition fluorescein isothiocynate (FITC) fluorescence probe mother liquor, and being stirred under the conditions of being protected from light, product purifies through magnetic separation,
It is dried in vacuo after ethanol washing, obtains Fe3O4@SiO2(FITC) magnetic nano-particle；
S12: from Fe3O4@SiO2(FITC) Fe is arrived3O4@SiO2(FITC)@mSiO2
Fe3O4@SiO2(FITC) magnetic nano-particle is scattered in cetyl trimethylammonium bromide (CTAB), deionized water
In the mixed solution of n-hexane, ultrasonic treatment is uniformly dispersed；With continuous stirring, ammonia spirit and TEOS are added dropwise dropwise
It into above-mentioned mixed solution, is stirred under the conditions of being protected from light, and product is collected by magnetic separation and is washed repeatedly with ethyl alcohol；It obtains
Sample it is flowed back in ethanol solution, to remove CTAB template reagent, further use ethanol washing, magnetic separation obtains
Fe3O4@SiO2(FITC)@mSiO2Magnetic nano-particle；
S13: from Fe3O4@SiO2(FITC)@mSiO2To Fe3O4@SiO2(FITC)@mSiO2-
(P)-SH
Sulfydryl-polymer (P)-propyl trimethoxy silicane is added to Fe3O4@SiO2(FITC)@mSiO2Magnetic Nano
It is reacted, will be separated through the nanoparticle of-(P)-SH modification using Magnet Treatment, and use second in the alcohol dispersion liquid of particle
Alcohol washs purifying repeatedly, obtains Fe3O4@SiO2(FITC)@mSiO2-(P)-SH；
S2: the assembling of small-size magnetic nanoparticle is cross-linked into magnetic nano-particle
S21:Fe3O4@SiO2(FITC)@mSiO2A large amount of existing-SH the groups in (P)-SH magnetic nano-particle surface, lead to
Peroxidating formation-S-S-, with realize be chemically crosslinked between polymer and nanoparticle between be chemically crosslinked, form magnetic nano particle
Son；
S22: the cross-linking reaction of step S21 is placed under the constrained environment of emulsion polymerization and is carried out, the magnetism being crosslinked is received
Rice corpuscles can have uniform size, form stable magnetic nano-particle dispersion liquid；
S3: magnetic nano-particle prepares intraocular hypertension animal model
The magnetic nano-particle is injected into camera oculi anterior with aqueous dispersion, magnetic nano particle is guided with annular magnet
Son is gathered in room angle, as aqueous humor circulation is gradually scattered in trabecular network, schlemm's canal and venous capillary, runs in aqueous humor
It is enriched in the organizations at different levels of channel, hinders aqueous humor drains to body circulation, to cause intraocular hypertension.
Further, in the method that above-mentioned magnetic nano-particle prepares intraocular hypertension animal model, small molecule sulfydryl can be passed through
Reagent adjusts the sulfydryl density of the small-size magnetic nanoparticle surface polymer molecule end of the chain, to adjust small-size magnetic nanoparticle
Sub- cross-linking reaction ability makes small-size magnetic nanoparticle in the crosslinked action of surface polymer molecule and magnetic mutually exclusive work
Under double control, the particle size and particle diameter distribution of accuracy controlling magnetic nano-particle, and the stabilization in dispersion liquid
Dispersion.The magnetic nano-particle that uniform particle sizes are prepared by emulsion polymerization terminates cross-linking reaction by small molecule mercapto reagent,
Centrifuge separation, disperses again, forms stable magnetic nano-particle dispersion liquid.
Further, in the method that above-mentioned magnetic nano-particle prepares intraocular hypertension animal model, the magnetic nano particle
After trabecular network enrichment, magnetic nano-particle local concentration increases son, between particle can further occurrence cross-linking reaction, formed bigger
The magnetic nano-particle of partial size or complete plugged zone；Part magnetic nano-particle can solve crosslinking, release a large amount of small size magnetic
Property nanoparticle is enriched in place with aqueous humor circulation to schlemm's canal or venous capillary, the small size of enrichment
Magnetic nano-particle can occur to be crosslinked between particle again since local density increases, and formation is comprehensive, multi-level, structure is opposite
Stable nanoparticle barrier film, persistently maintains intraocular hypertension.
Further, in the method that above-mentioned magnetic nano-particle prepares intraocular hypertension animal model, the magnetic nano particle
The dispersion liquid of son can be individually used for preparing intraocular hypertension animal model；It can also be used as carrier material, further load can promote
The raised drug of intraocular pressure, to prepare the intraocular hypertension animal model with more intraocular hypertension.
The preparation method of intraocular hypertension animal model of the present invention has many advantages, such as simply, conveniently, safe, long-acting:
(1) preparing intraocular hypertension animal model by injected into anterior chambers method is the general side for preparing intraocular hypertension animal model at present
Method, this method adjustable grain diameter characteristic since nanoparticle has, manufactures ah outflow resistance lasting stability, one injection
It realizes target, does not need multiple injection；
(2) this method is based on the adjustable magnetic nano-particle of partial size and prepares intraocular hypertension animal model, can be according to specific room
Water running environment adjusts self-crosslinking degree and particle size, can be in the multistage tissue such as trabecular network/schlemm's canal/vein blood vessel
Realize aggregation, it is complete to block mechanism system for comprehensive, multi-level closure aqueous humor circulation channel；
(3) the intraocular hypertension animal model of this method preparation, magnetic nano-particle block the resistance positions of aqueous humor not preceding
Within the scope of room, will not influence the institutional framework in the visual field and anterior chamber, will not influence it is subsequent based on intraocular hypertension animal model and open
Every experiment of exhibition；
(4) magnetic nano-particle used by this method can rapidly enter predetermined position under annular magnet guidance, build
It renders meritorious service energy；Can also be under the effect and guidance of reducing agent and externally-applied magnetic field, quickly solution crosslinking, removing function.
Detailed description of the invention
Fig. 1: the preparation of small-size magnetic nanoparticle and surface modification schematic diagram (polymer molecule is polyethylene glycol)
Fig. 2: SEM the and TEM photo (a) and hysteresis loop (b) of small-size magnetic nanoparticle
Fig. 3: small-size magnetic nanoparticle aggregation is cross-linked into magnetic nano-particle schematic diagram
Fig. 4: magnetic nano-particle multi-layer (trabecular network, schlemm's canal and venous capillary) under magnet guidance hinders
It fills in aqueous humor and runs access diagram
Fig. 5: distribution of the magnetic nano-particle in the tissue such as trabecular network, schlemm's canal and venous capillary
It abbreviation in figure and writes a Chinese character in simplified form:
TEOS:Tetraethyl orthosilicate: ethyl orthosilicate
APTS:3-Aminopropyltriethoxysilane:3- aminopropyl triethoxysilane
FITC:Fluorescein Isothiocyante: fluorescein isothiocynate
Octanol: octanol
Trixton X-100: Triton X-100
NH3H2O: ammonium hydroxide
CTAB:Cetyltriethylammnonium Bromide: cetyl trimethylammonium bromide
N-hexane: n-hexane
Remove CTAB: cetyl trimethylammonium bromide is removed
Fe3O4Magnetite nanoparticles: ferroferric oxide magnetic nano-particles
Fe3O4@SiO2(FITC): ferroso-ferric oxide@silica (FITC fluorescence probe) nanoparticle
Fe3O4@SiO2(FITC)@mSiO2: ferroso-ferric oxide@silica (FITC fluorescence probe)@mesoporous silicon oxide
Nanoparticle
Fe3O4@SiO2(FITC)@mSiO2(P)-SH: ferroso-ferric oxide@silica (FITC fluorescence probe)@mesoporous two
Silica-polymer (P)-sulfydryl nanoparticle
Fe3O4@SiO2(FITC)@mSiO2- PEG-SH: ferroso-ferric oxide@silica (FITC fluorescence probe)@mesoporous two
Silica-polyethylene glycol-sulfydryl nanoparticle
TM:Trabecular Meshwork: trabecular network
SC:Schlemm ' s Canal: schlemm's canal
CB:Ciliary Body: ciliary body
Specific embodiment
The present invention is described in detail combined with specific embodiments below.Following embodiment will be helpful to the technology of this field
Personnel further understand the present invention, but the invention is not limited in any way.It should be pointed out that the ordinary skill of this field
For personnel, without departing from the inventive concept of the premise, it can also make certain adjustments and improvements.These belong to the present invention
Protection scope.
Embodiment 1
The preparation of magnetic nano-particle, using following steps:
S1: the preparation of small-size magnetic nanoparticle
5mL magnetic iron powder-n-hexane dispersion liquid (20mg/mL) is diluted in 10mL n-octyl alcohol and 10mL polyethylene glycol octyl benzene
In the cyclohexane solution of base ether (trixton X-100).0.5mL ammonium hydroxide (28wt%) is added in the above solution, is formed and is stablized
Reverse micro emulsion.Then, it is added with stirring the silester (TEOS) of 0.04mL in continuous machinery, so that reaction is carried out 24 small
When, to form the magnetic nano-particle dispersion liquid of silicon dioxide coating, i.e. Fe3O4@SiO2Sample.Then, in above-mentioned Fe3O4@
SiO2Disperse that fluorescence probe mother liquor (0.02g 3- aminopropyl triethoxysilane (APTS), the different sulphur cyanogen of 0.01g are added in liquor
Sour fluorescein (FITC), 0.02mL TEOS, 5mL ethyl alcohol).After continuing stirring under the conditions of being protected from light 12 hours, product is through magnetic force point
It from purifying, three times with ethanol washing, and is dried in vacuum overnight at 30 DEG C, obtains Fe3O4@SiO2(FITC) magnetic nano-particle；
S12: from Fe3O4@SiO2(FITC) Fe is arrived3O4@SiO2(FITC)@mSiO2
Fe3O4@SiO2(FITC) magnetic nano-particle be scattered in cetyl trimethylammonium bromide (CTAB, 0.2g), go from
In the mixed solution of sub- water (40mL) and n-hexane (3mL), it is uniformly dispersed by ultrasonic treatment.With continuous stirring, by ammonium hydroxide
Solution (0.4mL, 28wt%) and 0.2mL TEOS are added drop-wise to dropwise in above-mentioned mixed solution, continue stirring 12 under the conditions of being protected from light
Hour, product is collected by magnetic separation and is washed repeatedly with ethyl alcohol.The sample of acquisition returns it in 80 DEG C of ethanol solution
Stream 12 hours further uses ethanol washing to remove CTAB template reagent, and magnetic separation obtains Fe3O4@SiO2(FITC)@
mSiO2Magnetic nano-particle；
S13: from Fe3O4@SiO2(FITC)@mSiO2To Fe3O4@SiO2(FITC)@mSiO2-(P)-SH
Sulfydryl-polyethylene glycol (P)-propyl trimethoxy silicane of 0.05g is added to Fe3O4@SiO2(FITC)@mSiO2
In the alcohol dispersion liquid of magnetic nano-particle (20mL, 0.6wt%).Reaction carry out 12 hours after, using Magnet Treatment will through-
(P) the nanoparticle separation of-SH modification, and wash purifying repeatedly with ethyl alcohol, obtain Fe3O4@SiO2(FITC)@mSiO2-(P)-
SH；Polymer is polyethylene glycol (PEG), molecular weight 5000 herein.
S2: the assembling crosslinking of small-size magnetic nanoparticle
S21:Fe3O4@SiO2(FITC)@mSiO2A large amount of existing-SH the groups in (P)-SH magnetic nano-particle surface, lead to
Peroxidating formation-S-S-, with realize be chemically crosslinked between polymer and nanoparticle between be chemically crosslinked, form magnetic nano particle
Son；
S22: the cross-linking reaction of step S21 is placed under the constrained environment of emulsion polymerization and is carried out, the magnetism being crosslinked is received
Rice corpuscles can have uniform size, form stable magnetic nano-particle dispersion liquid；
S3: magnetic nano-particle prepares intraocular hypertension animal model
The magnetic nano-particle is injected into camera oculi anterior with aqueous dispersion, magnetic nano particle is guided with annular magnet
Son is gathered in room angle, as aqueous humor circulation is gradually scattered in trabecular network, schlemm's canal and venous capillary, runs in aqueous humor
It is enriched in the organizations at different levels of channel, hinders aqueous humor drains to body circulation, to cause intraocular hypertension.
Using the resulting magnetic nano-particle of the above method, partial size 800nm is received by the small-size magnetic that partial size is 30nm
Rice corpuscles assembling is crosslinked；Small-size magnetic nanoparticle has core-shell structure, and core is ferroferric oxide magnetic nano-particles,
Shell is mesoporous silicon dioxide nano layer, and mesoporous silicon dioxide nano layer is coated on ferroferric oxide magnetic nano-particles surface；Institute
It is also mesoporous with the fluorescence probe containing FITC between the ferroferric oxide magnetic nano-particles stated and mesoporous silicon dioxide nano layer
Silicon dioxide intermediate layer；Mesoporous silicon dioxide nano layer surface also has polymer molecule decorative layer, and the polymer molecule is
Polyethylene glycol；Its molecular weight is 5000, and one end includes siloxane group, and the other end includes mercapto groups；Polymer molecule is logical
It crosses siloxane group and is grafted to mesoporous silicon dioxide nano layer surface, then realize intermolecular friendship by the mercapto groups of the other end
Connection, and then realize that the assembling of small-size magnetic nanoparticle is crosslinked by being crosslinked between polymer molecule.
Intraocular hypertension animal model is prepared based on above-mentioned magnetic nano-particle, step is by the magnetic nano-particle with water
Dispersion is injected into camera oculi anterior, and dosage is 100 μ L.It is gathered in room angle with annular magnet guidance magnetic nano-particle, with
Aqueous humor circulation is gradually scattered in trabecular network, schlemm's canal and venous capillary, rich in aqueous humor operation channel organizations at different levels
Collection.Intraocular hypertension is persistently maintained, intraocular pressure can be increased to 3 times of normal intraocular tension, and be stably maintained at some intraocular pressure data field
Between, continue 10 weeks.
Embodiment 2
The present embodiment and embodiment 1 are substantially similar, difference are as follows: polymer molecule is polyaminoacid, and molecular weight is
10000；Small-size magnetic nano particle diameter is 50nm, is cross-linked to form by small size nanoparticle by emulsion polymerization assembling
Magnetic nano-particle partial size be 900nm；The magnetic nano-particle is injected into camera oculi anterior with aqueous dispersion, dosage is
80μL.With annular magnet guidance magnetic nano-particle be gathered in room angle, with aqueous humor circulation, be gradually scattered in trabecular network, apply it is tired
Mu Shi pipe and venous capillary are enriched in aqueous humor operation channel organizations at different levels.Intraocular hypertension is persistently maintained, intraocular pressure can increase
To 4 times of normal intraocular tension, and it is stably maintained at some intraocular pressure data interval, continues 9 weeks.
Embodiment 3
The present embodiment and embodiment 1 are substantially similar, difference are as follows: polymer molecule is polylactic acid, molecular weight 8000；It is small
Sized magnetic nano particle diameter is 70nm, is received by small size nanoparticle by the magnetism that emulsion polymerization assembling is cross-linked to form
Rice corpuscles partial size is 600nm；The magnetic nano-particle is injected into camera oculi anterior with aqueous dispersion, dosage is 60 μ L.With
Annular magnet guidance magnetic nano-particle be gathered in room angle, with aqueous humor circulation, be gradually scattered in trabecular network, schlemm's canal and
Venous capillary is enriched in aqueous humor operation channel organizations at different levels.Intraocular hypertension is persistently maintained, intraocular pressure can be increased to normal eyes
3 times of pressure, and it is stably maintained at some intraocular pressure data interval, continue 8 weeks.
Embodiment 4
The present embodiment and embodiment 1 are substantially similar, difference are as follows: and polymer molecule is poly lactide-glycolide acid,
Molecular weight is 15000；Small-size magnetic nano particle diameter is 80nm, is assembled by small size nanoparticle by emulsion polymerization
The magnetic nano-particle partial size being cross-linked to form is 1000nm；The magnetic nano-particle is injected at the moment with aqueous dispersion
Room, dosage are 100 μ L.It is gathered in room angle with annular magnet guidance magnetic nano-particle, with aqueous humor circulation, is gradually scattered in small
Weir, schlemm's canal and venous capillary are enriched in aqueous humor operation channel organizations at different levels.Persistently maintain intraocular hypertension, eye
Pressure can be increased to 5 times of normal intraocular tension, and be stably maintained at some intraocular pressure data interval, continue 7 weeks.
Embodiment 5
The present embodiment and embodiment 1 are substantially similar, difference are as follows: polymer molecule is polysiloxanes, molecular weight 6000；
Small-size magnetic nano particle diameter is 10nm, assembles the magnetism being cross-linked to form by emulsion polymerization by small size nanoparticle
Nano particle diameter is 100nm；The magnetic nano-particle is injected into camera oculi anterior with aqueous dispersion, dosage is 80 μ L.
It is gathered in room angle with annular magnet guidance magnetic nano-particle, with aqueous humor circulation, is gradually scattered in trabecular network, schlemm's canal
And venous capillary, it is enriched in aqueous humor operation channel organizations at different levels.Intraocular hypertension is persistently maintained, intraocular pressure can be increased to normally
4 times of intraocular pressure, and it is stably maintained at some intraocular pressure data interval, it is for 4 weeks.
Embodiment 6
The present embodiment and embodiment 1 are substantially similar, difference are as follows: polymer molecule is polycaprolactone, and molecular weight is
12000；Small-size magnetic nano particle diameter is 20nm, is cross-linked to form by small size nanoparticle by emulsion polymerization assembling
Magnetic nano-particle partial size be 300nm；The magnetic nano-particle is injected into camera oculi anterior with aqueous dispersion, dosage is
60μL.With annular magnet guidance magnetic nano-particle be gathered in room angle, with aqueous humor circulation, be gradually scattered in trabecular network, apply it is tired
Mu Shi pipe and venous capillary are enriched in aqueous humor operation channel organizations at different levels.Intraocular hypertension is persistently maintained, intraocular pressure can increase
To 2 times of normal intraocular tension, and it is stably maintained at some intraocular pressure data interval, continues 5 weeks.
Embodiment 7
The present embodiment and embodiment 1 are substantially similar, difference are as follows: polymer molecule is polyethylene glycol, molecular weight 2000；
Small-size magnetic nano particle diameter is 40nm, assembles the magnetism being cross-linked to form by emulsion polymerization by small size nanoparticle
Nano particle diameter is 500nm；The magnetic nano-particle is injected into camera oculi anterior with aqueous dispersion, dosage is 100 μ L.
It is gathered in room angle with annular magnet guidance magnetic nano-particle, with aqueous humor circulation, is gradually scattered in trabecular network, schlemm's canal
And venous capillary, it is enriched in aqueous humor operation channel organizations at different levels.Intraocular hypertension is persistently maintained, intraocular pressure can be increased to normally
5 times of intraocular pressure, and it is stably maintained at some intraocular pressure data interval, continue 6 weeks.
Embodiment 8
The present embodiment and embodiment 1 are substantially similar, difference are as follows: polymer molecule is polysiloxanes, and molecular weight is
15000；Small-size magnetic nano particle diameter is 60nm, is cross-linked to form by small size nanoparticle by emulsion polymerization assembling
Magnetic nano-particle partial size be 750nm；The magnetic nano-particle is injected into camera oculi anterior with aqueous dispersion, dosage is
80μL.With annular magnet guidance magnetic nano-particle be gathered in room angle, with aqueous humor circulation, be gradually scattered in trabecular network, apply it is tired
Mu Shi pipe and venous capillary are enriched in aqueous humor operation channel organizations at different levels.Intraocular hypertension is persistently maintained, intraocular pressure can increase
To 4 times of normal intraocular tension, and it is stably maintained at some intraocular pressure data interval, continues 9 weeks.
Embodiment 9
The present embodiment and embodiment 1 are substantially similar, difference are as follows: polymer molecule is polyaminoacid, and molecular weight is
20000；Small-size magnetic nano particle diameter is 100nm, is crosslinked shape by emulsion polymerization assembling by small size nanoparticle
At magnetic nano-particle partial size be 1200nm；The magnetic nano-particle is injected into camera oculi anterior, agent with aqueous dispersion
Amount is 60 μ L.With annular magnet guidance magnetic nano-particle be gathered in room angle, with aqueous humor circulation, be gradually scattered in trabecular network,
Schlemm's canal and venous capillary are enriched in aqueous humor operation channel organizations at different levels.Intraocular hypertension is persistently maintained, intraocular pressure can be with
4 times of normal intraocular tension are increased to, and is stably maintained at some intraocular pressure data interval, continues 8 weeks.
Claims (10)
1. a kind of magnetic nano-particle, which is characterized in that the magnetic nano-particle particle size range is 100~1200nm, by
The assembling of small-size magnetic nanoparticle is crosslinked；
The small-size magnetic nano particle diameter range is 10~100nm, has core-shell structure, and core is ferroso-ferric oxide magnetic
Property nanoparticle, shell is mesoporous silicon dioxide nano layer, and mesoporous silicon dioxide nano layer is coated on ferroferric oxide magnetic nano
Particle surface；Also have between the ferroferric oxide magnetic nano-particles and mesoporous silicon dioxide nano layer and contains fluorescence probe
Mesoporous silicon oxide middle layer, by nuclear magnetic resonance or fluorescence microscope and magnetic nano particle can be tracked in order to subsequent
Son in running body and Tissue distribution；
The mesoporous silicon dioxide nano layer surface also has a polymer molecule decorative layer, by being crosslinked between polymer molecule
Reaction, is assembled into magnetic nano-particle for small-size magnetic nanoparticle；The polymer molecule, one end include siloxanes
Group, the other end include mercapto groups；Polymer molecule is grafted to mesoporous silicon dioxide nano layer surface by siloxane group,
Intermolecular cross-linking is realized by the mercapto groups of the other end again, and then realizes that small-size magnetic is received by being crosslinked between polymer molecule
The assembling of rice corpuscles is crosslinked.
2. magnetic nano-particle as described in claim 1, which is characterized in that the magnetic nano-particle particle size range is
300~1000nm, the small-size magnetic nano particle diameter range are 20~80nm.
3. magnetic nano-particle as claimed in claim 1 or 2, which is characterized in that the magnetic nano-particle particle size range
For 500~800nm, the small-size magnetic nano particle diameter range is 40~60nm.
4. magnetic nano-particle as described in claim 1, which is characterized in that the polymer molecule has good biology
Safety, including non-degradable type polymer and degradable polymer, non-degradable type polymer include polyethylene, polypropylene, gather
Any one of acrylate, aromatic polyester, polysiloxanes, polyformaldehyde or any combination thereof or monomer or segment it is any
Combined polymerization；Degradable polymer includes collagen, gelatin, fibroin albumen, chitosan, chitin, cellulose, polyaminoacid, gathers
Any one of ethylene glycol, polylactic acid, polyglycolic acid, poly lactide-glycolide acid, polycaprolactone or its any group
Conjunction or any combined polymerization of monomer or segment.
5. magnetic nano-particle as described in claim 1 or 4, which is characterized in that the polymer molecule be polysiloxanes,
Any one of polyethylene glycol, polyaminoacid, polylactic acid, polyglycolic acid, poly lactide-glycolide acid, polycaprolactone
Or any combination thereof or monomer or segment any combined polymerization.
6. magnetic nano-particle as described in claim 1, which is characterized in that the molecular weight of the polymer molecule between
Between 2000~20000.
7. a kind of method for preparing intraocular hypertension animal model based on magnetic nano-particle described in claim 1, which is characterized in that
This method comprises the following steps:
S1: the preparation of small-size magnetic nanoparticle
S11: from Fe3O4To Fe3O4@SiO2(FITC)
Magnetic iron powder-n-hexane dispersion liquid is diluted in the cyclohexane solution of n-octyl alcohol and Triton X-100, is added
Ammonium hydroxide forms stable reverse micro emulsion, is added with stirring silester (TEOS) in continuous machinery and is reacted, to form two
The magnetic nano-particle dispersion liquid of silica-coated, i.e. Fe3O4@SiO2Sample；In above-mentioned Fe3O4@SiO2Disperse in liquor to add
Enter fluorescein isothiocynate (FITC) fluorescence probe mother liquor, and stirred under the conditions of being protected from light, product is through magnetic separation purifying, ethyl alcohol
It is dried in vacuo after washing, obtains Fe3O4@SiO2(FITC) magnetic nano-particle；
S12: from Fe3O4@SiO2(FITC) Fe is arrived3O4@SiO2(FITC)@mSiO2
Fe3O4@SiO2(FITC) magnetic nano-particle is scattered in cetyl trimethylammonium bromide (CTAB), deionized water and just
In the mixed solution of hexane, ultrasonic treatment is uniformly dispersed；With continuous stirring, ammonia spirit and TEOS are added drop-wise to above-mentioned mixing
It in solution, is stirred under the conditions of being protected from light, product is collected by magnetic separation, is washed repeatedly with ethyl alcohol；The sample of acquisition by its
It flows back in ethanol solution, to remove CTAB template reagent, further uses ethanol washing, magnetic separation obtains Fe3O4@SiO2
(FITC)@mSiO2Magnetic nano-particle；
S13: from Fe3O4@SiO2(FITC)@mSiO2To Fe3O4@SiO2(FITC)@mSiO2-(P)-SH
Sulfydryl-polymer (P)-propyl trimethoxy silicane is added to Fe3O4@SiO2(FITC)@mSiO2Magnetic nano-particle
It is reacted, will be separated through the nanoparticle of-(P)-SH modification using magnetic separation method, and repeatedly with ethyl alcohol in alcohol dispersion liquid
Washing purifying, obtains Fe3O4@SiO2(FITC)@mSiO2-(P)-SH；
S2: the assembling of small-size magnetic nanoparticle is cross-linked into magnetic nano-particle
S21:Fe3O4@SiO2(FITC)@mSiO2A large amount of existing-SH the groups in (P)-SH magnetic nano-particle surface, pass through oxidation
Formation-S-S-, with realize be chemically crosslinked between polymer and nanoparticle between be chemically crosslinked, form magnetic nano-particle；
S22: the cross-linking reaction of step S21 being placed under the constrained environment of emulsion polymerization and is carried out, the magnetic nano particle being crosslinked
Son can have uniform size, form stable magnetic nano-particle dispersion liquid；
S3: magnetic nano-particle prepares intraocular hypertension animal model
The magnetic nano-particle is injected into camera oculi anterior with aqueous dispersion, it is poly- with annular magnet guidance magnetic nano-particle
Fang Jiao is combined in, as aqueous humor circulation is gradually scattered in trabecular network, schlemm's canal and venous capillary, runs channel in aqueous humor
It is enriched in organizations at different levels, hinders aqueous humor drains to body circulation, to cause intraocular hypertension.
8. the method that magnetic nano-particle as claimed in claim 7 prepares intraocular hypertension animal model, which is characterized in that can pass through
Small molecule mercapto reagent adjusts the sulfydryl density of the small-size magnetic nanoparticle surface polymer molecule end of the chain to adjust small size
Magnetic nano-particle cross-linking reaction ability makes small-size magnetic nanoparticle in the crosslinked action and magnetism of surface polymer molecule
Under the double control of repulsive interaction, the particle size and particle diameter distribution of accuracy controlling magnetic nano-particle, and dispersing
Stable dispersion in liquid.
9. the method that magnetic nano-particle as claimed in claim 7 prepares intraocular hypertension animal model, which is characterized in that described
After trabecular network enrichment, magnetic nano-particle local concentration increases magnetic nano-particle, between particle can further occurrence crosslinking it is anti-
It answers, forms more large-sized magnetic nano-particle or complete plugged zone；Part magnetic nano-particle can solve crosslinking, release big
Amount small-size magnetic nanoparticle is enriched in place with aqueous humor circulation to schlemm's canal or venous capillary, rich
The small-size magnetic nanoparticle of collection can occur to be crosslinked between particle again since local density increases, and form comprehensive, multilayer
Secondary, the metastable nanoparticle barrier film of structure, persistently maintains intraocular hypertension.
10. the method that magnetic nano-particle as claimed in claim 7 prepares intraocular hypertension animal model, which is characterized in that described
The dispersion liquid of magnetic nano-particle can be individually used for preparing intraocular hypertension animal model；It can also be used as carrier material, into one
Step load can promote the raised drug of intraocular pressure, to prepare the intraocular hypertension animal model with more intraocular hypertension.
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