CN103773366A - Application of fluorescent carbon nanoparticles in myocardial imaging - Google Patents
Application of fluorescent carbon nanoparticles in myocardial imaging Download PDFInfo
- Publication number
- CN103773366A CN103773366A CN201310710701.XA CN201310710701A CN103773366A CN 103773366 A CN103773366 A CN 103773366A CN 201310710701 A CN201310710701 A CN 201310710701A CN 103773366 A CN103773366 A CN 103773366A
- Authority
- CN
- China
- Prior art keywords
- carbon nanoparticle
- nanoparticle
- fluorescent
- carbon
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention belongs to the field of biomaterial and organ imaging, and designs application of fluorescent carbon nanoparticles in myocardial imaging. A mulberry silk is processed by a hydrothermal process, so as to obtain the fluorescent carbon nanoparticles. The nanoparticles have good targeting property on the myocardial cell, and have good safety. Therefore, the fluorescent carbon nanoparticles can be applied to cardiac imaging.
Description
Technical field
The present invention relates to materialogy and organ imaging field, the particularly application of fluorescent carbon nanoparticle in myocardial imaging.This carbon nanoparticle is the nanoparticle at 10-150nm by a series of particle diameters that are prepared from by " hydrothermal method " containing the organic materials of " carbon, hydrogen, oxygen, nitrogen ", because the character of itself can be used in myocardial imaging.
Background technology
Nanoparticle, as a kind of conventional and comparatively novel pharmaceutical carrier, is the focus of research in recent years always, and key is the existence form that it constantly changes, preparation method, physico-chemical property.Nanoparticle generally refers to by natural or synthetic polymer carrier and makes, it is a kind of solid colloid, particle diameter is at 1～1000nm, similarly dispersed system also has nanometer ball, nanocapsule, nano-micelle, nanometer liposome, nano-emulsion (nanoemulsion, NE) and nanogel (nanogel) etc., nanoparticle more commonly wherein.
Quantum dot (quantum dot) is the nano material of accurate zero dimension (quasi-zero-dimensional), is made up of a limited number of atom, and three dimension are all in nanometer scale.Quantum dot is generally spherical or class is spherical, by semiconductor material (conventionally elementary composition by II B～VI B or IIIB～VB) make, stable diameter is at the nanoparticle of 2～20 nil2.Due to the elementary composition and structure of CdS quantum dots self uniqueness, make it have photoluminescence fluorescent effect, and there is stable fluorescence quantum yield, therefore can be used as biological fluorescent labeling for the diagnosis in body, there is certain application prospect at life science neighborhood.
Fluorescent carbon nanoparticle is the one of quantum dot, and its main raw is carbon, compares conventional transition metal quantum point, has the advantages such as security is good, source is abundant, therefore causes extensive concern.But current research mainly concentrates on the method for preparing carbon nanoparticle, its potential biological applications is paid close attention in less research.
At present, the preparation method of fluorescent carbon nanoparticle is mainly divided into physical method and chemical process, is chemically main, and chemical process has two kinds, and a kind of is to adopt the method for colloid chemistry synthetic in organic system, and another kind is synthetic in the aqueous solution.Water direct synthesis technique: the method for directly synthetic quantum dot has easy and simple to handle in water, reproducible, cost is low, surface charge and surface properties are controlled, biocompatibility is better, without significance toxicity, and can introduce the advantage of functional group, its good performance makes it the biological fluorescent labeling as a kind of novelty, is subject in recent years the favor of more and more neighborhoods.
Summary of the invention
Based on above background, one of the object of the invention is to provide a kind of hypotoxicity, and the CdS quantum dots of good biocompatibility, by the character of its photoluminescence fluorescent effect, as a kind of biological fluorescent labeling, for iconography imaging and diagnosis in organism.
CdS quantum dots of the present invention is that a kind of organic materials of carbon containing is prepared from by hydrothermal method, in alkali lye, boil 1h by mulberry silk, use respectively again ethanol, water washing, after oven dry, add certain water gaging to be placed in reactor, in 200 ° of C reaction 72h, be therefore called carbon ball nanoparticle (carbon nanoparticle).
One of object of the present invention is to provide a kind of easy being easy to get, and cost is low, and the carbon nanoparticle that surface-element and electric charge are controlled, because its raw-material source is natural Inorganic biomatetials, itself is the material without overt toxicity, and only by several simply elementary composition.Cytologic experiment proves that its toxicity is low, biocompatibility is better, carbon nanoparticle fluorescent probe prepared by this Inorganic biomatetials myocardial cell's picked-up with accumulate, to myocardial cell without obvious damage, therefore aspect cardiomyography, there is certain advantage, avoiding fluorescent marker confrontation myocardial cell's prepared by some organic fluorescence mark substances or heavy metal element infringement.
The sign of carbon nanoparticle solution of the present invention is to utilize its optical property, for determining of its maximum excitation wavelength (EX) and emission wavelength (EM), be to adopt spectrophotofluorometer to scanning, and its characteristic ultraviolet absorption is to adopt ultraviolet spectrophotometer to scan carbon nanoparticle dilute solution.
The one that carbon nanoparticle fluorescence quantum yield of the present invention is CdS quantum dots characterizes, the general reference method that adopts is measured, under identical shooting conditions, measure respectively the integration fluorescence intensity (proofreading and correct the included area of fluorescence spectrum) of two kinds of dilute solutions of reference fluorescence standard material of fluorescence sample to be measured and known quantum yield and the absorbancy of the incident light (ultraviolet-visible light) to an identical excitation wavelength.By the substitution specific formulation calculating respectively of these values, obtain the quantum yield of fluorescence sample to be measured again; Yu=Ys Fu/Fs As/Au
Yu, Ys-the be respectively fluorescence quantum yield of test substance and reference material
Fu, Fs-the be respectively integration fluorescence intensity of test substance and reference material
As, Au-be respectively test substance and reference material are in the absorption value (A) of the incident light of this excitation wavelength.
While using this formula, general requirement absorbance As, Au be lower than 0.05, the fluorescent substance that reference standard material is close with test substance excitation wavelength, according to the literature: have the Yu of the fluorescent chemicals that analytical applications is worth generally between 0.1-1.
One of object of the present invention is to provide a kind of carbon nanoparticle with modifiable surface functional group, due to comparatively single elementary composition of himself starting material, prepare in the process of carbon nanoparticle in hydrothermal method, its surface can produce some containing " carbon ", " nitrogen ", the functional group of " oxygen ", X ray field emission electron power spectrum (XPS) is carried out surface-element analysis, can analyze carboxyl, amino, the existence of the functional groups such as hydroxyl, infrared spectrometer analyzes to it existence that also further confirms these groups, therefore carbon nanoparticle is in as biological fluorescent labeling, also can be to its modification and as useful for drug delivery to the carrier in organism.
One of object of the present invention is to provide the carbon nanoparticle biological fluorescent labeling that a kind of polyoxyethylene glycol (PEG) is modified, because self existing certain gathering, carbon nanoparticle cause its particle diameter to increase, stability reduces, and cause its fluorescence quantum yield to reduce, carboxyl to carbon nanoparticle surface is modified, by the activation of 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC.HCL) and N-hydroxy-succinamide (NHS), make carboxyl form the active ester of a kind of NHS, adding one end is amino-terminated polyoxyethylene glycol (NH2-PEG, MW 5000) room temperature lucifuge reaction 12h, make carbon nanoparticle surface superscribe one deck PEG, thereby extend its cycling time in vivo and increase himself stability.
One of object of the present invention is to provide a kind of carbon nanoparticle of having modified PEG and can having increased its stability in blood circulation.Due to the impact of serum protein, the stability of carbon nanoparticle can reduce, and makes it assemble with the combination of serum protein, and therefore, in the time carrying out serum stability experiment, the stability of having modified the carbon nanoparticle of PEG is significantly better than the carbon nanoparticle of unmodified PEG.
One of object of the present invention is to provide a kind of carbon nanoparticle that can passively accumulate to myocardial cell, due to the particle diameter of its nanometer scale, distribute in experiment in vivo, can be observed the picked-up of heart tissue and accumulate more, therefore, utilize this character can be for the radiography of cardiac muscular tissue.Due to the existence of cardiac toxic, the many uses that may serve as contrast medium fluorescent probe are limited, CdS quantum dots of preparing as the fluorescent material of chemosynthesis and heavy metal element etc.Therefore utilize carbon nanoparticle self hypotoxicity, the feature of good biocompatibility, can carry out iconography imaging to heart tissue texture, observes the structure of heart tissue.Meanwhile, for the lesions position existing in heart tissue, carbon nanoparticle biological fluorescent labeling can there are differences with healthy tissues in the time that picked-up enters this position, therefore can be used for the diagnosis of heart tissue lesions position.
beneficial effect: the carbon nanoparticle that the present invention introduces not only can be used as useful for drug delivery to the carrier in body, and surmount the nanoparticulate carriers on conventional meaning, it is a kind of CdS quantum dots, himself also can be used as a kind of biological fluorescent labeling, for the diagnosis in body, existing research has disclosed its diagnosis that can be used for tumor locus, the present invention studies different places from these and is its trial for the diagnosis aspect of cardiomyography and heart tissue lesions position, simultaneously due to himself hypotoxicity and biocompatibility better; Surface has modifiable functional group, for further modifying possibility is provided, therefore has certain using value.
accompanying drawing explanation:
Fig. 1: the size distribution figure of carbon nanoparticle.
Fig. 2: the stability of the carbon nanoparticle that carbon nanoparticle is modified from polyoxyethylene glycol (PEG) under different serum-concentration conditions.Serum-concentration is set to 0%, 10%, 50%, and phosphate buffered saline buffer (PBS) is set in contrast.
Fig. 3: the picked-up of myocardial cell to different concns carbon nanoparticle solution (31 μ g/mL, 125 μ g/mL, 500 μ g/mL).
Fig. 4: carbon nanoparticle is to different tissues organ (heart, liver, spleen, lung, kidney, brain) and " tumour " site distribution and fluorescence imaging design sketch.The fluorescence imaging effect that can find out heart tissue is the most obvious.
embodiment:
In order further to set forth the present invention, provide a series of embodiment below.These embodiment are illustrative completely, and they are only used for the present invention to be specifically described, and not should be understood to limitation of the present invention.
embodiment mono-: the preparation of carbon ball nanoparticle
1. take the about 1g of mulberry silk and add 5% Na
2cO
3heated and boiled in solution, and keep boiling 1h left and right, room temperature is cooling.
2. by the silk boiling deionized water repetitive scrubbing, this process repeats 3 times, use again two deionized water wash 3 times, use afterwards 75% washing with alcohol 3 times, use again absolute ethanol washing 3 times, its object, in order to remove as far as possible impurity wherein, is placed in blast drier by the silk having washed and fully dries (temperature setting is set to 60oC).
3. the silk of oven dry is placed in the polytetrafluoroethyllining lining of pyroreaction still, adds two deionized waters of 20mL, this step will guarantee that silk is submerged by water, in 200 ° of C reaction 72h.
4. take out the solution in reactor, pushed film with the water filter membrane that aperture is 0.22 μ m, obtain mulberry silk carbon nanoparticle solution, measure its particle diameter about 100nm, as shown in Figure 1.
embodiment bis-: the optical property of carbon nanoparticle characterizes
1. determining of the maximum excitation wavelength of carbon nanoparticle and emission wavelength, specific experiment scheme is as follows:
2. determining of carbon nanoparticle characteristic ultraviolet absorption, specific experiment operation is as follows:
By 125 times of original solution dilutions, concentration is approximately 126 μ g/mL, under ultraviolet spectrophotometer, it is scanned, and sweep limit is 200-800nm, its Absorption Characteristics is along with the reduction of wavelength raises, and has comparatively significantly absorption peak at 208nm and 274nm place.
embodiment tri-: the mensuration of the fluorescence quantum yield of carbon nanoparticle
The mensuration of carbon nanoparticle fluorescence quantum yield is carried out take Quinine Sulphate Di HC as reference material, and concrete operations are as follows:
1. parameter setting: the excitation wavelength of spectrophotofluorometer is set to 365nm(and approaches the maximum excitation wavelength of carbon nanoparticle), the incident light of the excitation wavelength of ultraviolet spectrophotometer is set to 365nm equally.
2. the preparation of reference solution Quinine Sulphate Di HC dilute solution: the Quinine Sulphate Di HC that takes 34.2mg is dissolved in the H of 2mL 0.1N
2sO
4in, ultrasonic, jolting forms milky white solution (consideration has on a small quantity not dissolving) after dissolving.By this solution dilution 6 × 10
4doubly, measuring its absorbance is 0.039, and to measure its integration fluorescence intensity (Fs) be 60314.775.
3. the preparation of test substance carbon nanoparticle dilute solution: by the solution dilution 2 × 10 of carbon nanoparticle original concentration
4doubly, measuring its absorbance is 0.088, and to measure its integration fluorescence intensity (Fu) be 60334.412.
4. according to Melhuish law, at 25oC, excitation wavelength is 365nm, and the fluorescence quantum yield of Quinine Sulphate Di HC is 0.546.Therefore the fluorescence quantum yield that above-mentioned data substitution formula is calculated to carbon nanoparticle is 0.342.
embodiment tetra-: the PEGization of carbon nanoparticle
The concentration of getting 1mL is the round-bottomed flask that 20mg/mL carbon nanoparticle solution is placed in 25mL, add PBS(PH7.4) to 5mL, precision takes EDCHCL 40.2mg(0.209mmol), NHS 22.6mg(0.196mmol) be placed in solution, room temperature lucifuge stirs 3h, obtains the NHS active ester of carbon nanoparticle.Add afterwards the NH2-PEG(MW 5000 of 200 μ g), adjust PH to 8.0 left and right, room temperature lucifuge stirs 24h, and the water membrane filtration by completely reacted solution with 0.22 μ m obtains the carbon nanoparticle solution that PEG modifies.
embodiment five: the serum stability experiment of fluorescent carbon nanoparticle
1. the concentration of the carbon nanoparticle solution of carbon nanoparticle initial soln and PEGization is determined
In the time carrying out serum stability experiment, to limit the absorbance of carbon nanoparticle and PEGization carbon nanoparticle solution, two kinds of solution should be in 0.1 left and right at the absorbance at 350nm place, by 100 times of carbon nanoparticle solution dilutions, again by 20 times of the carbon nanoparticle solution dilutions of PEGization, absorption value with two kinds of solution of ultraviolet spectrophotometer mensuration at 350nm place, the absorption value that records carbon nanoparticle initial soln is 0.1024, the absorption value of the carbon nanoparticle solution of PEGization is 0.0826, therefore the concentration of carbon nanoparticle initial soln is approximately 1.25 times of carbon nanoparticle solution of PEGization.Using 1.25 times of the carbon nanoparticle solution redilution of 100 times of dilutions with diluted the carbon nanoparticle solution of PEGization of 20 times as this experimental concentration.
2. the preparation of different serum-concentrations
Get human serum 100 μ L PBS(PH7.4) be diluted to 1ML and be prepared into 10% human serum, separately get human serum 500 μ L PBS(PH7.4) be diluted to 1ML and be prepared into 50% human serum.
With PBS as blank, get respectively the PBS of 100 μ L, 10% human serum, 50% human serum and be placed in 96 orifice plates, then add respectively the carbon nanoparticle solution of 100 μ L and the carbon nanoparticle solution of PEGization.Each point arranges 3 attached holes.On Chemiluminescence Apparatus, measure respectively the ultraviolet absorption value (excitation wavelength of incident light is set to 560nm) of 0h, 1h, 2h, 4h, 8h, 12h, 24h, when 24h, the serum stability of the carbon nanoparticle of PEGization is significantly better than PEGization not, as shown in Figure 2.
the myocardial imaging effect of embodiment six, fluorescent carbon nanoparticle
1, the myocyte that cores is cultured in culture vessel with glass bottom, treats that it grows into certain density, adds fluorescent carbon nanoparticle to hatch, and observes afterwards under Laser Scanning Confocal Microscope.
As shown in Figure 3, myocardial cell can obviously absorb fluorescent carbon nanoparticle and present green fluorescence.
the intracorporeal heart imaging effect of embodiment seven, fluorescent carbon nanoparticle
1. tail vein injection fluorescent carbon nanoparticle is to Mice Body, anesthetized mice after 2h, and cardiac perfusion physiological saline and formalin, get afterwards each internal organs and do freezing microtome section, and under Laser Scanning Confocal Microscope, observe the fluorescence distribution of each tissue.
As shown in Figure 4, carbon nanoparticle has stronger distribution at heart, and is better than the tissue such as liver, spleen, illustrates that this nanoparticle has good myocardium targeting and cardiac imaging effect.
Claims (2)
1. the application of fluorescent carbon nanoparticle in cardiomyography, is characterized in that a kind of quantum dot carbon nanomaterial, can target to heart and as a kind of biological fluorescent labeling.
2. " fluorescent carbon nanoparticle " according to claim 1 quanta point material, is characterized in that being mainly prepared from by natural organic (as mulberry silk), and it has photoluminescent property, and has stable fluorescence quantum yield.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310710701.XA CN103773366A (en) | 2013-12-23 | 2013-12-23 | Application of fluorescent carbon nanoparticles in myocardial imaging |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310710701.XA CN103773366A (en) | 2013-12-23 | 2013-12-23 | Application of fluorescent carbon nanoparticles in myocardial imaging |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103773366A true CN103773366A (en) | 2014-05-07 |
Family
ID=50566141
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310710701.XA Pending CN103773366A (en) | 2013-12-23 | 2013-12-23 | Application of fluorescent carbon nanoparticles in myocardial imaging |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103773366A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111437885A (en) * | 2020-04-10 | 2020-07-24 | 济南大学 | Preparation method of porous magnetic quantum dot doped biological composite photocatalyst |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1884430A (en) * | 2006-06-30 | 2006-12-27 | 上海师范大学 | Fluorescent carbon nanometer tube and its preparation method and application |
-
2013
- 2013-12-23 CN CN201310710701.XA patent/CN103773366A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1884430A (en) * | 2006-06-30 | 2006-12-27 | 上海师范大学 | Fluorescent carbon nanometer tube and its preparation method and application |
Non-Patent Citations (4)
| Title |
|---|
| ASHWIN A BHIRDE ET AL.: "Distribution and clearance of PEG-single-walled carbon nanotube cancer drug delivery vehicles in mice", 《NANOMEDICINE》 * |
| WEI LI ET AL.: "Simple and Green Synthesis of Nitrogen-Doped Photoluminescent Carbonaceous Nanospheres for Bioimaging", 《ANGEWANDTE CHEMIE INTERNATIONAL EDITION》 * |
| ZHONGMIN OU ET AL.: "Functional single-walled carbon nanotubes based on an integrin αυβ3 monoclonal antibody for highly efficient cancer cell targeting", 《NANOTECHNOLOGY》 * |
| ZHU LIAN WU ET AL.: "One-pot hydrothermal synthesis of highly luminescent nitrogen-doped amphoteric carbon dots for bioimaging from Bombyx mori silk-natural proteins", 《JOURNAL OF MATERIALS CHEMISTRY B》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111437885A (en) * | 2020-04-10 | 2020-07-24 | 济南大学 | Preparation method of porous magnetic quantum dot doped biological composite photocatalyst |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | The near-infrared-II fluorophores and advanced microscopy technologies development and application in bioimaging | |
| Lou et al. | Organic dots based on AIEgens for two‐photon fluorescence bioimaging | |
| Ding et al. | Recent advances in near-infrared II fluorophores for multifunctional biomedical imaging | |
| Gao et al. | Hybrid graphene/Au activatable theranostic agent for multimodalities imaging guided enhanced photothermal therapy | |
| Chen et al. | Current advances in lanthanide‐doped upconversion nanostructures for detection and bioapplication | |
| Tang et al. | Croconaine nanoparticles with enhanced tumor accumulation for multimodality cancer theranostics | |
| Park et al. | Theranostic probe based on lanthanide‐doped nanoparticles for simultaneous in vivo dual‐modal imaging and photodynamic therapy | |
| Guan et al. | Multifunctional upconversion–nanoparticles–trismethylpyridylporphyrin–fullerene nanocomposite: a near-infrared light-triggered theranostic platform for imaging-guided photodynamic therapy | |
| Gu et al. | Recent advances in design and fabrication of upconversion nanoparticles and their safe theranostic applications | |
| Rostami et al. | Breakthroughs in medicine and bioimaging with up-conversion nanoparticles | |
| Sun et al. | Targeted and imaging-guided in vivo photodynamic therapy for tumors using dual-function, aggregation-induced emission nanoparticles | |
| An et al. | Small ultra-red fluorescent protein nanoparticles as exogenous probes for noninvasive tumor imaging in vivo | |
| Sreejith et al. | Organic–inorganic nanohybrids for fluorescence, photoacoustic and Raman bioimaging | |
| CN106620725A (en) | Optical and opto-acoustic integration dual-mode molecular image probe as well as preparation method and application thereof | |
| Sengupta et al. | A review on synthesis, toxicity profile and biomedical applications of graphene quantum dots (GQDs) | |
| Cui et al. | A generic self-assembly approach towards phototheranostics for NIR-II fluorescence imaging and phototherapy | |
| CN103784978B (en) | A kind of albumen-dye composition and application thereof | |
| Wu et al. | Cascade targeting tumor mitochondria with CuS nanoparticles for enhanced photothermal therapy in the second near-infrared window | |
| Wang et al. | A class of biocompatible dye–protein complex optical nanoprobes | |
| Sengar et al. | Progress on carbon dots and hydroxyapatite based biocompatible luminescent nanomaterials for cancer theranostics | |
| Du et al. | Encapsulation-Dependent Enhanced Emission of Near-Infrared Nanoparticles Using in Vivo Three-Photon Fluorescence Imaging | |
| Wang et al. | Research spotlight: upconversion nanoparticles for potential cancer theranostics | |
| CN103908681B (en) | The application of fluorescent carbon nanoparticle in brain tumor diagnosis that angiopep-2 modifies | |
| ZHANG et al. | Lanthanide-doped fluorescence probes for NIR-Ⅱ fluorescence imaging | |
| CN103773366A (en) | Application of fluorescent carbon nanoparticles in myocardial imaging |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20140507 |