CN106390120A - Magnetic nanometer material used for imaging and photothermal therapy and preparation method and application of magnetic nanometer material - Google Patents
Magnetic nanometer material used for imaging and photothermal therapy and preparation method and application of magnetic nanometer material Download PDFInfo
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- 238000003384 imaging method Methods 0.000 title claims abstract description 35
- 238000007626 photothermal therapy Methods 0.000 title claims abstract description 29
- 239000000463 material Substances 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 claims abstract description 52
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 60
- 206010028980 Neoplasm Diseases 0.000 claims description 41
- 239000002086 nanomaterial Substances 0.000 claims description 15
- 230000008859 change Effects 0.000 claims description 13
- 229910052742 iron Inorganic materials 0.000 claims description 12
- 239000002105 nanoparticle Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- 235000013339 cereals Nutrition 0.000 claims description 6
- 238000005481 NMR spectroscopy Methods 0.000 claims description 5
- 229910021577 Iron(II) chloride Inorganic materials 0.000 claims description 4
- 229910021578 Iron(III) chloride Inorganic materials 0.000 claims description 4
- 240000007594 Oryza sativa Species 0.000 claims description 4
- 235000007164 Oryza sativa Nutrition 0.000 claims description 4
- NMCUIPGRVMDVDB-UHFFFAOYSA-L iron dichloride Chemical compound Cl[Fe]Cl NMCUIPGRVMDVDB-UHFFFAOYSA-L 0.000 claims description 4
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 4
- 235000009566 rice Nutrition 0.000 claims description 4
- 238000013019 agitation Methods 0.000 claims description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
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- 238000003756 stirring Methods 0.000 claims description 3
- 238000007920 subcutaneous administration Methods 0.000 claims description 2
- 239000000155 melt Substances 0.000 claims 1
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 abstract description 88
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N Iron oxide Chemical compound [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 abstract description 14
- 239000002245 particle Substances 0.000 abstract description 14
- 238000000034 method Methods 0.000 abstract description 7
- 230000008901 benefit Effects 0.000 abstract description 3
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- 241000699670 Mus sp. Species 0.000 description 15
- NDLPOXTZKUMGOV-UHFFFAOYSA-N oxo(oxoferriooxy)iron hydrate Chemical compound O.O=[Fe]O[Fe]=O NDLPOXTZKUMGOV-UHFFFAOYSA-N 0.000 description 12
- 238000003786 synthesis reaction Methods 0.000 description 11
- 238000002347 injection Methods 0.000 description 8
- 239000007924 injection Substances 0.000 description 8
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- DCYOBGZUOMKFPA-UHFFFAOYSA-N iron(2+);iron(3+);octadecacyanide Chemical compound [Fe+2].[Fe+2].[Fe+2].[Fe+3].[Fe+3].[Fe+3].[Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] DCYOBGZUOMKFPA-UHFFFAOYSA-N 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
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- 239000013225 prussian blue Substances 0.000 description 4
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- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 3
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 description 3
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- WAEMQWOKJMHJLA-UHFFFAOYSA-N Manganese(2+) Chemical compound [Mn+2] WAEMQWOKJMHJLA-UHFFFAOYSA-N 0.000 description 1
- ZOKXTWBITQBERF-UHFFFAOYSA-N Molybdenum Chemical compound [Mo] ZOKXTWBITQBERF-UHFFFAOYSA-N 0.000 description 1
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- 206010035664 Pneumonia Diseases 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
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- DEGAKNSWVGKMLS-UHFFFAOYSA-N calcein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(O)=O)CC(O)=O)=C(O)C=C1OC1=C2C=C(CN(CC(O)=O)CC(=O)O)C(O)=C1 DEGAKNSWVGKMLS-UHFFFAOYSA-N 0.000 description 1
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- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- OMZSGWSJDCOLKM-UHFFFAOYSA-N copper(II) sulfide Chemical compound [S-2].[Cu+2] OMZSGWSJDCOLKM-UHFFFAOYSA-N 0.000 description 1
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- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
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- 229910021389 graphene Inorganic materials 0.000 description 1
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- 239000012216 imaging agent Substances 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
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- 230000010354 integration Effects 0.000 description 1
- ICIWUVCWSCSTAQ-UHFFFAOYSA-M iodate Chemical compound [O-]I(=O)=O ICIWUVCWSCSTAQ-UHFFFAOYSA-M 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 231100001252 long-term toxicity Toxicity 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000000696 magnetic material Substances 0.000 description 1
- 239000002122 magnetic nanoparticle Substances 0.000 description 1
- 229910001437 manganese ion Inorganic materials 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910052750 molybdenum Inorganic materials 0.000 description 1
- 239000011733 molybdenum Substances 0.000 description 1
- 229920005615 natural polymer Polymers 0.000 description 1
- 229910000510 noble metal Inorganic materials 0.000 description 1
- 229960002378 oftasceine Drugs 0.000 description 1
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- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 229920001690 polydopamine Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920000128 polypyrrole Polymers 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
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- ITRNXVSDJBHYNJ-UHFFFAOYSA-N tungsten disulfide Chemical compound S=[W]=S ITRNXVSDJBHYNJ-UHFFFAOYSA-N 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
- A61K49/1818—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
- A61K49/1821—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
- A61K49/1824—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
- A61K49/1827—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
- A61K49/1833—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with a small organic molecule
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
- A61K49/222—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by a special physical form, e.g. emulsions, liposomes
- A61K49/225—Microparticles, microcapsules
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- Health & Medical Sciences (AREA)
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- Nanotechnology (AREA)
- Public Health (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Radiology & Medical Imaging (AREA)
- Acoustics & Sound (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a magnetic nanometer material used for imaging and photothermal therapy and a preparation method and application of the magnetic nanometer material. The magnetic nanometer material used for imaging and photothermal therapy refers to nanometer iron oxide particles coated with melanin. The magnetic nanometer material is non-toxic, high in biocompatibility and capable of imaging. According to the preparation method, a one-step coprecipitation method is adopted to composite water-soluble Fe3O4@mel NCs. The magnetic nanometer material has the advantages of mildness and simplicity.
Description
Technical field
The present invention relates to pharmaceutical technology field, particularly to a kind of magnetic Nano material for imaging and photo-thermal therapy and
Preparation method and application.
Background technology
With the fast development of biotechnology and nano science, the photo-thermal therapy of near-infrared laser mediation is gradually as one
Plant the alternative medicine with bright prospects for oncotherapy.Photo-thermal therapy is a kind of aggressive less local treatment mode.
When tumor region is exposed to near-infrared laser irradiation range, luminous energy can be turned by the photothermal deformation reagent being enriched in tumor region
It is changed to heat energy, effective killing tumor cell.As compared to traditional remedies (such as excision, chemotherapy, radiotherapy etc.), photo-thermal therapy has
Non-Invasive, high selectivity, high therapeutic efficiency, damage the advantage such as little to surrounding health tissue.The more photo-thermal of research turns at present
Changing reagent mainly has noble metal nanometer material, carbon nanomaterial, copper chalcogen compound, organic polymer etc..Although achieving one
Fixed be in progress, most photothermal deformation reagent have and are difficult to metabolism, Nondegradable, may cause potential long term toxicity (as oxygen
Change stress, pneumonia etc.).And, they lack clinical diagnosises and the ability of real-time dynamic monitoring deep tissue, and clinical practice is subject to
Limit.
Nuclear magnetic resonance, as a kind of multi-faceted multiparameter image mode with high spatial resolution, is examined in clinical tumor
Disconnected aspect has great value.By excellent magnetic performance, ferric oxide nanometer particle has been widely used in magnetic resonance pair
Ratio enhancing, medicine delivery, magnetic thermotherapy etc. is studied.At present, many document reports are had to receive based on the heterozygosis of ferric oxide nanometer particle
Rice material is used for mri guided lower photo-thermal therapy, as by gold, copper sulfide, molybdenum bisuphide, Graphene, tungsten disulfide etc.
Photo-thermal module is coated on ferric oxide nanometer particle surface.However, these materials are required to loaded down with trivial details synthesis step by difference in functionality
Component is integrated into same nano platform, therefore synthesizes loaded down with trivial details, high cost.
Find that the magnetic material of one pack system also has certain photothermal deformation ability recently, such as ferriferrous oxide nano
Grain, ferriferrous oxide nano cluster etc..Also there is document that the polymer near infrared region absorption is coated on ferroso-ferric oxide to receive
Rice material surface to improve its photo-thermal conversion efficiency, such as polypyrrole, poly-dopamine etc..However, adopting molten these nano-particle more
Agent full-boiled process or the synthesis of heat sink area method, need longer response time, harsh reaction condition, loaded down with trivial details ligand exchange, complicated
Chemical modification.
Melanin is that one kind is widely present in each in-house natural polymer of organism.It has good bio-compatible
Property, biodegradability and stronger near infrared region absorb, and are a kind of preferable photothermal deformation reagent.There is document utilization melanin
Excellent metal ion chelating capacity, adsorbs iron ion, manganese ion, copper 64 ion in the melanin nano grain surface of synthesis
Deng, carry out multi-modality imaging instruct under photo-thermal therapy, achieve preferable effect.However, because metal ion is by chela
Cooperation is adsorbed in nano grain surface, has the risk coming off.
Therefore, to prepare with good biocompatibility and good light in the urgent need to developing a kind of simply gentle method
The novel magnetic nano-particle of hot-cast socket ability.
Content of the invention
It is an object of the invention to provide a kind of magnetic Nano material for imaging and photo-thermal therapy, this material has non-poison
Property and good biocompatibility, and there is imaging capability.
The technical scheme is that：
For the magnetic Nano material of imaging and photo-thermal therapy, this material is the coated ferric oxide nanometer particle of melanin.
This material is the coated ferric oxide nanometer particle of water miscible melanin.
Every 0.26mg melanin is coated 1mg ferrum.
Second object of the present invention is to provide a kind of preparation side for imaging and the magnetic Nano material of photo-thermal therapy
Method, the method adopts the water miscible Fe of coprecipitation one-step synthesis method3O4@mel NCs, has gentle, simple advantage.
Preparation method for imaging and the magnetic Nano material of photo-thermal therapy of the present invention, has following steps：
1) by FeCl2·4H2O and FeCl3·6H2O is dissolved in 5mL 1M HCl solution, is heated to 90 under the conditions of inflated with nitrogen
℃；
2), under intense agitation, rapidly join the NH that 0.5mL concentration is 28vol.%4OH solution, reacts 20 seconds.With
Afterwards, add 5mL melanin solution in 1 minute, stir 30 minutes at 90 DEG C；
3) solution is cooled to room temperature, and Magneto separate is until solution ph is neutrality.Collect precipitation, the melanin obtaining is coated
Ferric oxide nanometer particle be dispersed in water.Its preparation method is as follows：
Described FeCl2·4H2O：FeCl3·6H2O weight is than for 1:2.72.
Step 2) described melanin solution concentration be 1mg/mL, pH=9.0.
The 3rd purpose of the present invention is to provide the magnetic Nano material for imaging and photo-thermal therapy preparing for magnetic altogether
The application shaken in imaging agent；
And, the magnetic Nano material for imaging and photo-thermal therapy is used for the application in photoacoustic imaging preparation in preparation.
And, for being imaged magnetic Nano material the answering in preparation treatment tumor photo-thermal ablation preparation with photo-thermal therapy
With.
Described tumor is subcutaneous tumors.
Applicant adopts Fe of the present invention3O4@mel NCs, carries out magnetic resonance, the imaging of optoacoustic bimodal and tumor light
Thermal therapeutical.Test result indicate that：The Fe of uv-visible absorption spectroscopy display synthesis3O4Light at 808nm for the@mel NCs is inhaled
There is good linear relationship in receipts value and concentration；To Fe3O4The magnetic resonance of@mel NCs, photoacoustic imaging and photothermal deformation performance are entered
Row characterizes, and photo-thermal therapy instrument thermograph result is shown under the irradiation of 808nm laser, Fe3O4@mel NCs solution temperature raises
Concentration and laser power dependency；Compared with multiple business-like ferric oxide nanometer particles, Fe of the present invention3O4@mel
NCs near infrared region absorbs highest, and light thermal property is best, photoacoustce signal intensity highest under 700nm laser excitation, and R2 relaxation
Rate is higher；Cytotoxicity experiment result shows the Fe synthesizing3O4@mel NCs cytotoxicity is very low；Fe3O4@mel NCs injects lotus
After tumor Mus tumor locus, tumor locus photoacoustic signal and magnetic resonance signal change at once, and compare more at 1 hour about
Dissipate；Accept Fe3O4The mice with tumor tumor of@mel NCs+ laser irradiation group is obviously reduced, decreased growth；Each main organs cell shape
State no significant difference.
The Fe of present invention synthesis3O4@mel NCs, is a kind of new multi-functional nanometer material for tumor diagnosis and treatment integration
Material, this material will provide new approaches for developing and building multifunctional nano diagnosis and treatment platform, have certain in biomedical sector
Application prospect.
Brief description
Fig. 1 is one-step synthesis method Fe3O4@mel NCs and its schematic diagram of diagnosis and treatment application；
Fig. 2 is Fe3O4The synthesis of@mel NCs and sign；Wherein, Fig. 2 a is not add melanic original ferroso-ferric oxide
Nano-particle (left) and Fe3O4@mel NCs aqueous solution (right)；Fig. 2 b is Fe3O4The images of transmissive electron microscope of@mel NCs；Fig. 2 c is
Magnetization curve；Fig. 2 d is Fourier transform infrared spectroscopy；
Fig. 3 is melanin solution and iron content standard absorption curve；Wherein, Fig. 3 a, 3b represent melanin solution in 500nm
The standard absorption curve at place；Fig. 3 c, 3d represent that o-phenanthroline detects iron content at 510nm；
Fig. 4 is Fe3O4The absorption spectrum of@mel NCs and size of hydration；Wherein, Fig. 4 a represents variable concentrations Fe3O4@mel
The uv-visible absorption spectroscopy of NCs；Fig. 4 b represents Fe3O4Absorption value-concentration of iron at 808nm for the@mel NCs aqueous solution is bent
Line；Fig. 4 c represents Fe3O4The size of hydration of@mel NCs；Fig. 4 d is surface potential.
Fig. 5 is Fe3O4The magnetic of@mel NCs, light, hot property characterize；Wherein, Fig. 5 a represents Fe3O4The R2 of@mel NCs relaxes
Henan rate；Fig. 5 b represents variable concentrations Fe3O4The photoacoustce signal intensity of@mel NCs；Fig. 5 c represents variable concentrations Fe3O4@mel NCs
In 2W/cm2Thermograph under laser irradiation；Fig. 5 d represents Fe3O4@mel NCs (concentration of iron is 10mM) is in different capacity laser
Thermograph under irradiation；
Fig. 6 is Fe3O4The photothermal deformation performance map of@mel NCs；Wherein, Fig. 6 a, Fig. 6 b are under different laser powers
Fe3O4The temperature change of@mel NCs；Fig. 6 c, Fig. 6 d are variable concentrations Fe3O4Temperature under same laser power for the@mel NCs
Change.
Fig. 7 is Fe3O4(concentration of iron is under comparable sodium for@mel NCs and multiple business-like ferric oxide nanometer particles
Performance comparision 10mM)；Wherein, Fig. 7 a is uv-visible absorption spectroscopy；Fig. 7 b is photothermal deformation performance；Fig. 7 c is 700nm
Photoacoustce signal intensity under laser excitation；Fig. 7 d is R2 relaxation rate；
Fig. 8 is Fe3O4The impact to U87MG cytoactive for the@mel NCs；Wherein, Fig. 8 a is U87MG cell through Fe3O4@
Mel NCs process 24 hours after cytoactive；Fig. 8 b adds laser irradiated cells row calcein and iodate for different disposal
The fluoroscopic image that third pyridine contaminates altogether；Fig. 8 c adds laser irradiated cells Activity determination result for different disposal；
Fig. 9 is Fe3O4@mel NCs is by the situation of cell effectively picked-up；Wherein, Fig. 9 a is U87MG cell and Fe3O4@mel
NCs be incubated 6 hours altogether after cell nuclear magnetic resonance；Fig. 9 b is imaged for cell opto-acoustic；Fig. 9 c is cell prussian blue staining；
Figure 10 is internal optoacoustic, the imaging of magnetic resonance bimodal；Wherein, Figure 10 a is intratumor injection Fe3O4@mel NCs(25μ
L, 2mg Fe/mL) before and after different time points tumor locus photoacoustic image；Figure 10 b is intratumor injection Fe3O4@mel NCs (25 μ L,
2mg Fe/mL) before and after different time points tumor locus magnetic resonance image (MRI)；Figure 10 c is the photoacoustic imaging signal intensity of tumor region
Change；Figure 10 d is the magnetic resonance signal intensity change of tumor region；
Figure 11 is Fe3O4@mel NCs treats the situation of mouse tumor；Wherein, Figure 11 a is U87MG mice with tumor through noting in tumor
Penetrate 25 μ L Fe3O4The thermograph of@mel NCs row photo-thermal therapy after 1 hour；Figure 11 b be different grouping mice before the treatment after
The gross tumor volume of different time；Figure 11 c be different grouping mice before the treatment after different time Mouse Weight；Figure 11 d is not
With packet mice before the treatment and treatment after the 15th day picture；
Figure 12 is the 15th day row hematoxylin-eosin staining picture after the treatment of different grouping mice.
Specific embodiment
The reagent that the present invention adopts：
Melanin is purchased from Sigma-Aldrich；
Business-like ferric oxide nanometer particle respectively be purchased from ocean nanosecond science and technology company limited of the U.S., Guerbet SA and
Chinese Wan De High Seience Technology Co., Ltd.；
Remaining reagent is purchased from Chemical Reagent Co., Ltd., Sinopharm Group.
Embodiment 1 Co deposited synthesis Fe3O4@mel NCs
By 100umol, the FeCl of 19.881mg2·4H2The FeCl of O and 200umol, 54.058mg3·6H2O is dissolved in 5mL
In 1M HCl solution, it is heated to 90 DEG C under the conditions of inflated with nitrogen, under intense agitation, rapidly join the 28vol.% of 0.5mL
NH4OH solution, reacts 20 seconds.Subsequently, add 5mL melanin solution (1mg/mL, pH=9.0) in 1 minute, stir at 90 DEG C
30 minutes, it is cooled to room temperature, Magneto separate is until solution ph reaches neutrality.Collect precipitation, the Fe that will obtain3O4@mel NCs divides
Dissipate in water, referring to Fig. 1.
Embodiment 2Fe3O4The synthesis of@mel NCs and sign
Transmission electron microscope observing pattern：Pipette 50uL Fe3O4@mel NCs solution Deca is online in surface carbon coated copper, with filter
Paper siphons away unnecessary solution, natural air drying under room temperature condition.Observed under 200kV voltage conditions using transmission electron microscope and receive
The pattern of rice grain, particle diameter and deployment conditions.
Vibrating specimen magnetometer detects magnetic performance：Weigh dried powder sample about 10mg, be placed in test special micro-
In amount test tube, stopper compacting with absorbent cotton.Normal Temperature is carried out using vibrating specimen magnetometer：Power work frequency
86Hz, input power 5W, X-axis recording magnetic field change on computers, Y-axis record magnetization change.
Fourier transform infrared spectrometer detects chemical group：Vacuum drying potassium bromide powder agate mortar is ground
Mill, pipettes the testing sample after appropriate lyophilization, by sample and potassium bromide according to about 1：100 mass ratio in mortar again
Mix, grind.Irradiate lower sheeting sample preparation in infrared baking lamp, sample is placed on Fourier infrared spectrograph and is detected.
Transmission electron microscope results show successfully to prepare the nano-particle of size uniformity, and its result is shown in Fig. 2.Fig. 2 a shows and does not add
Melanic reactant liquor can occur coagulation after room temperature stands overnight, and after adding melanin, can significantly improve nano-particle
Water dispersible.Magnetic Measurement as shown by data Fe3O4The saturated magnetization rate of@mel NCs is 47.8emu/g, and has excellent magnetic
Response performance.Fourier transform infrared spectroscopy shows that melanin is successfully coated on nano grain surface.
Embodiment 3 detects Fe3O4Melanin and iron content in@mel NCs
Prepare the melanin solution of variable concentrations, its optical absorption situation is measured using all-wave length microplate reader.Take at 500nm
Absorbance value, makees absorbance-melanin solution concentration curve, the Fe of measurement synthesis3O4Melanin content in@mel NCs.
With (NH4)2Fe(SO4)2·6H2O prepares ferrum standard bathing solution, makees concentration of iron standard using standard o-phenanthroline and inhale at 510nm
Receive curve, the Fe of measurement synthesis3O4Iron content in@mel NCs.Calculate Fe3O4The content ratio of melanin and ferrum in@mel NCs
Example.
Make to absorb standard curve, detection respectively at 500nm and at 510nm using all-wave length microplate reader and o-phenanthroline
Melanin and iron content in product.Record every 1mg Fe and correspond to 0.26mg melanin, referring to Fig. 3.
Embodiment 4 uv-visible absorption spectroscopy and size of hydration detection
Prepare the Fe of variable concentrations3O4@mel NCs, measures its optical absorption situation using all-wave length microplate reader.Take 808nm
Place's absorbance value, makees absorbance-Fe3O4@mel NCs solution concentration curve.
Dynamic light scattering detection size of hydration and surface potential：Take 1mL Fe3O4@mel NCs solution to sample cell,
It is placed in Analyzer testing result cavity.Data acquisition conditions are 173 ° of the angle of diffraction, 25 DEG C of test temperature.
Uv-visible absorption spectroscopy shows Fe3O4Absorbance value at 808nm for the@mel NCs and concentration exist good
Linear relationship, dynamic light scattering records Fe3O4The size of hydration of@mel NCs is about 91nm, surface potential about -26.1mV,
Referring to Fig. 4.
Embodiment 5Fe3O4The magnetic resonance of@mel NCs, photoacoustic imaging and photothermal deformation performance characterization
Nuclear magnetic resonance：Prepare the Fe of different ferrum molar concentrations3O4@mel NCs solution.At ambient temperature, sample is put
Tested in German Brooker 7.0T toy magnetic resonance imager.Test parameter sets：TR=4000ms, TE=45ms,
The visual field is 25 × 25mm.
Photoacoustic imaging：Prepare the Fe of different ferrum molar concentrations3O4@mel NCs solution.At ambient temperature, sample is placed in
Endra128 toy photoacoustic imaging instrument is tested.Imaging parameters：Energy, 2.8 MJs；Wavelength, 700nm；Angle, 60 °.
Photothermal deformation performance detection：(1) prepare the Fe that ferrum molar concentration is 10mM3O4@mel NCs solution, solution is moved to
In microplate reader orifice plate, the 808nm laser instrument being respectively adopted different capacity is irradiated to solution 5 minutes, is monitored with infrared temperature
The temperature variations of solution when instrument recording laser irradiates.It is depicted as temperature-time curve, analyze Fe3O4@mel NCs solution temperature
Degree change and the relation of laser power.(2) prepare the Fe of different ferrum molar concentrations3O4@mel NCs solution, solution is moved to enzyme mark
In instrument orifice plate, it is 2W/cm using power2808nm laser instrument solution is irradiated 5 minutes, monitored with infrared temperature simultaneously
Instrument recording materials carry out temperature variations when laser irradiates.It is depicted as temperature-time curve, analyze Fe3O4@mel NCs is molten
Liquid temp change and the relation of solution concentration.
To Fe3O4The magnetic resonance of@mel NCs, photoacoustic imaging and photothermal deformation performance are characterized.7.0T toy magnetic is altogether
Vibration Meter records Fe3O4The T2 signal intensity of@mel NCs is gradually lowered with the rising of solution concentration, and its R2 relaxation rate is about
246mM-1S-1, Endra128 photoacoustic imaging result display Fe3O4The photoacoustce signal intensity of@mel NCs has concentration dependent；Light
Thermal cure instrument thermograph result is shown under the irradiation of 808nm laser, Fe3O4@mel NCs solution temperature raises in concentration and swashs
Luminous power dependency.Referring to Fig. 5, Fig. 6.
Embodiment 6Fe3O4@mel NCs and the comparison of commercialization ferric oxide nanometer particle performance
Fe by synthesis3O4@mel NCs and three kinds of business-like ferric oxide nanometer particles carry out Performance comparision：Prepare ferrum to rub
Your concentration is four kinds of nanoparticles solution of 10mM, is respectively adopted all-wave length microplate reader and measures its absorption spectrum；Using power it is
2W/cm2808nm laser instrument solution is irradiated 5 minutes；Carry out detecting molten using Endra128 toy photoacoustic imaging instrument
Photoacoustic signal at 700nm wavelength for the liquid；Measure the R2 relaxation rate of solution using Brooker 7.0T toy magnetic resonance imager.
Due to there being melanic parcel, under comparable sodium, Fe of the present invention3O4@mel NCs inhales near infrared region
Receive highest, therefore light thermal property preferably, and photoacoustce signal intensity highest under 700nm laser excitation.Find the present invention simultaneously
Described Fe3O4@mel NCs R2 relaxation rate is apparently higher than business-like ferric oxide nanometer particle.Referring to Fig. 7.
Embodiment 7 cytotoxicity experiment and cell photo-thermal therapy
Cytotoxicity experiment：5000 U87MG cells are seeded in 96 porocyte culture plates, after cell attachment, plus
Enter the Fe of variable concentrations gradient3O4@mel NCs, is incubated 24 hours altogether with cell.MTT experiment detects Fe3O4@mel NCs is to thin
The impact of cytoactive.Cell photo-thermal therapy：5000 U87MG cells are seeded in 96 well culture plates, after cell attachment, plus
Enter the Fe of variable concentrations gradient3O4@mel NCs, is incubated 6 hours altogether with cell.Cleaned with phosphate buffer three times, add 100
Microlitre blood-free medium, adopts 808nm laser instrument with 1W/cm2Power illumination cell 5 minutes.Matched group include untreated fish group,
Only NCs group and only laser group.After the completion of irradiation, cell continues incubation 4 hours, and MTT experiment detection photo-thermal therapy is to cytoactive
Impact.
Cytotoxicity experiment result shows the Fe synthesizing3O4@mel NCs cytotoxicity is very low.Human glioma cells U87MG
With Fe3O4After@mel NCs is incubated 6 hours altogether, carry out photo-thermal therapy, find Fe3O4@mel NCs adds the cell of laser irradiation group
Activity is significantly lower than other matched groups.Show Fe3O4@mel NCs can pass through the effective killing tumor cell of photo-thermal effect.Referring to figure
8.
Embodiment 8 cellular uptake is tested
Cell imaging is tested：U87MG cell is inoculated in 10cm Tissue Culture Dish, when cell fusion is to 80% about,
Add the Fe of variable concentrations3O4@mel NCs, is incubated 6 hours altogether with cell.The cell being not handled by is as comparison.Digestion, from
The heart, collection cell.By cell with 1% agarose disperse, be respectively compared different grouping cell magnetic resonance T2 signal or
Photoacoustic signal at 700nm wavelength.Cell prussian blue staining is tested：U87MG cell is inoculated in 12 porocyte culture plates, treats thin
When born of the same parents are fused to 80% about, add the Fe of variable concentrations3O4@mel NCs, is incubated 6 hours altogether with cell.It is not handled by
Cell is as comparison.Row prussian blue staining, inverted microscope carries out observing, takes pictures.
With Fe3O4After@mel NCs is incubated 6 hours altogether, cell magnetic resonance T2 signal significantly reduces, and photoacoustic signal substantially increases
By force.Prussian blue staining result shows Fe3O4@mel NCs effectively can be absorbed by cell.Referring to Fig. 9.
Embodiment 9 tumor optoacoustic and the imaging of magnetic resonance bimodal
By Fe3O4After@mel NCs (25 μ L, 2mg Fe/mL) adopts Flos Mume injection injection mice with tumor tumor locus, respectively
Before the injection, inject latter 5 minutes and inject latter 1 hour to mouse tumor position row photoacoustic imaging and nuclear magnetic resonance.Than less
With time point signal intensity situation.
By Fe3O4@mel NCs (25 μ L, 2mg Fe/mL) injection mice with tumor tumor locus after, tumor locus photoacoustic signal and
Magnetic resonance signal changes at once, and compares disperse at 1 hour about.Referring to Figure 10.
Embodiment 10 tumor thermal therapy and curative effect monitoring
By Fe3O4After@mel NCs (25 μ L, 2mg Fe/mL) injection mice with tumor tumor locus, row light after injecting 1 hour
Thermal therapeutical.Matched group is divided into untreated fish group, only NCs group and only laser group.With 2W/cm2Laser power irradiate 8 points of tumor locus
Clock, using infrared temperature monitor dynamically recording tumor locus temperature change.Curative effect monitoring：Before and after the treatment of different grouping mice
Different time points, every other day weigh a Mouse Weight, and use vernier caliper measurement gross tumor volume.And respectively before the treatment and
Mice was substantially taken pictures in the 15th day after treatment.
Row photo-thermal therapy after injecting 1 hour, Fe3O4@mel NCs group tumor locus temperature can be increased to 51.3 DEG C, and not
Injection group is only 39.4 DEG C.The dynamic monitoring different grouping mice gross tumor volume of rear different time points and Mice Body before the treatment
Weight, compared with matched group, accepts Fe3O4The mice with tumor tumor of@mel NCs+ laser irradiation group is obviously reduced, decreased growth.Each group
Mouse Weight changes no significant difference.Referring to Figure 11.
Embodiment 11 tissue slice Yihong-brazilwood extract dyeing
The 15th day after the treatment, 10% chloral hydrate anesthesia mice.50mL normal saline is first used to rinse after blood through heart,
50mL 4% paraformaldehyde is used to irrigate again.It is rapidly separated and obtains the main organs such as the heart, liver, spleen, lung, kidney, be soaked in 10% good fortune
In your Malin.Carry out paraffin embedding after 24 hours.Cut into slices with 5 microns of thickness, dewaxing.Tissue slice immersion hematoxylin dye
10 minutes in liquid, eosin stains 5 minutes, inverted microscope is observed, is taken pictures.
Each main organs morphocytology no significant difference, shows that nano-particle has preferable biocompatibility.Its knot
Fruit is referring to Figure 12.
Claims (10)
1. a kind of for imaging and photo-thermal therapy magnetic Nano material it is characterised in that：This material is the coated oxygen of melanin
Change iron nano-particle.
2. material according to claim 1 it is characterised in that：This material is received for the coated ferrum oxide of water miscible melanin
Rice grain.
3. material according to claim 1 and 2 it is characterised in that：Every 0.26mg melanin is coated 1mg Fe.
4. the preparation method of magnetic Nano material being used for imaging and photo-thermal therapy is it is characterised in that there are following steps：
1) by FeCl2·4H2O and FeCl3·6H2O is dissolved in 5mL 1M HCl solution, is heated to 90 DEG C under the conditions of inflated with nitrogen；
2), under intense agitation, rapidly join the NH that 0.5mL concentration is 28vol.%4OH solution, reacts 20 seconds.Subsequently, 1
Add 5mL melanin solution in minute, stir 30 minutes at 90 DEG C；
3) solution is cooled to room temperature, and Magneto separate, until solution ph is neutrality, collects precipitation, by coated for the melanin obtaining oxygen
Change iron nano-particle to be dispersed in water.
5. preparation method according to claim 4 it is characterised in that：Step 1) described FeCl2·4H2O and FeCl3·6H2O
Weight than for 1:2.72.
6. preparation method according to claim 4 it is characterised in that：Step 2) described melanin solution concentration be 1mg/
ML, pH=9.0.
7. it is used for imaging and the magnetic Nano material of photo-thermal therapy is used for the application in nuclear magnetic resonance preparation in preparation.
8. it is used for imaging and the magnetic Nano material of photo-thermal therapy is used for the application in photoacoustic imaging preparation in preparation.
9. it is used for imaging and the magnetic Nano material of photo-thermal therapy melts the application in preparation in preparation treatment tumor photo-thermal.
10. according to claim 9 application it is characterised in that：Described tumor is subcutaneous tumors.
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