CN109172828A - A kind of novel rare-earth nanometer bimodal imaging agent and its preparation method and application - Google Patents

A kind of novel rare-earth nanometer bimodal imaging agent and its preparation method and application Download PDF

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CN109172828A
CN109172828A CN201811091427.1A CN201811091427A CN109172828A CN 109172828 A CN109172828 A CN 109172828A CN 201811091427 A CN201811091427 A CN 201811091427A CN 109172828 A CN109172828 A CN 109172828A
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汪建华
任志豪
王思齐
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Abstract

The invention discloses a kind of novel rare-earth nanometer bimodal imaging agent and its preparation method and application, which is NaHoF4@CeO2Nanoparticle, the preparation of the imaging agent is the following steps are included: first is that NaHoF4The preparation of nanoparticle, second is that CeO2The preparation of nanoparticle, third is that in NaHoF4Middle dopen Nano CeO2And obtain NaHoF4@CeO2Nanoparticle, the imaging agent is used in the imaging of CT and MRI bimodal, physicochemical characterization and toxicity test show that the biological safety of the nanoparticle is preferable, and experiment in vitro shows that the nanoparticle has good CT and MRI bimodal imaging effect, while having radio therapy sensitization effect to pancreatic cancer cell.

Description

A kind of novel rare-earth nanometer bimodal imaging agent and its preparation method and application
Technical field
The present invention relates to medical imaging technical field, specifically a kind of novel rare-earth nanometer bimodal imaging agent and its preparation Methods and applications.
Background technique
In recent years, the exploitation of multi-modality imaging and diagnosis and treatment integration imaging agent has become molecular imaging area research Hot spot.CT (CT scan) and MRI (magnetic resonance imaging) has complementary advantages, and is clinical most common tumor imaging Detection methods.The recall rate and diagnostic accuracy to disease can be improved in CT, MRI bimodal imaging contrast, is conducive to more acurrate Tumor boundaries are sketched the contours, provide accurate positioning for further radiotherapy.Currently, clinically used CT and MRI contrast agent still cannot be very Meet bimodal imaging demand well.
Some scholars carry out the research of CT, MRI bimodal imaging contrast, mainly include Fe-Bi, Fe-Pt, Gd-Au, Au Or TaOxModify Fe3O4, these nanoparticles MRI imaging is mainly using the oxide of iron.But the magnetization of ferriferous oxide is in low magnetic Field (< 1T) has been satisfied, and its r2Relaxation not the raising with magnetic field from 0.5T to 9.4T and increase.Due to high spatial resolution The needs of the magnetic resonance imaging of shorter imaging acquisition, and High magnetic field MRI (> 3T) has been applied, and high performance, Gao Ci is developed Field MRI, CT bimodal contrast agent are particularly important.In all lanthanide ions, paramagnetic holmium ion (Ho3+) have Highest Effect magnetic moment, it may be possible to the most promising contrast agent candidate of superelevation magnetic resonance imaging.
The contrast images of MRI be based between different tissues proton density difference and different longitudinal direction (T1) and laterally (T2) relaxation time.CT imaging is mainly based upon the different densities substance damping capacity different to X-ray.Holmium ion (Ho3+) can As T2Contrast medium is since it is with shorter the electron relaxation time (10-13s) and efficient magnetic moment (about 10.6 μ B).Bu etc. takes the lead in Report doping Ho3+NaYbF4Nanoparticle can be used for 3T MRI imaging, while realize up-conversion luminescence and CT imaging.2016 Year, Ni etc. reports the NaHoF of DSPE-PEG5k coating for the first time4Nano particle can be used as High magnetic field MRI and CT bimodal comparison Agent.
Cancer of pancreas is clinically needed using complex treatment, and particularly with Locally Advanced patient, radiotherapy is even more must, can Few treatment means, nevertheless, still remaining recurrence and DISTANT METASTASES IN after treatment of pancreatic cancer, main cause is in radiotherapy In the process, tumour cell is to the ray damage of radioactive ray tolerance and surrounding vital tissue internal organs, and wherein oxygen radical plays key Effect, studies have shown that CeO2It is the attemperator of internal oxygen radical, can reduce the damage by X-ray to organism normal cell, increases By force to the lethal effect of tumour cell.
CeO2With with the similar antioxidant activity such as anti-oxidant albumen superoxide dismutase, catalase, can Scavenging activated oxygen ROS.CeO2This characteristic and different valence state Ce3+And Ce4+It can be with machine during mutually converting Intracorporal oxygen radical combines related.Colon etc. is research shows that CeO2Nanoparticle can protect colon epithelial cell, avoid in spoke Caused injury in treatment is penetrated, and reduces ROS, has raised Sudismase, nano Ce O2It is normally thin to reduce mouse Colon The apoptosis of born of the same parents.For one of Wason and Baker studies have shown that doing control group with tailored radiation (RT), control group injects physiology salt Water, nano Ce O2+ RT group is experimental group, carries out mouse experiment in vivo, the results showed that, suffer from the mouse of cancer of pancreas in radiotherapy mistake CeO is used in journey2It is more significant to handle radiotherapeutic effect.
With lanthanide series (Ln3+) based on paramagnetic nanoparticle have and very big be likely to become magnetic resonance imaging (MRI) Imaging agent, wherein Ho is the strongest substance of paramagnetism in all lanthanide series.In addition, the atomic number of Ho is 67, it is higher than iodine Atomic number 53, have higher x-ray damping capacity.CeO2It is a kind of oxygen that redox reaction is carried out according to environment pH value On the one hand the attemperator of free radical can reduce the damage by X-ray to organism normal cell (partial neutral environment), on the other hand, There is oxidation in the acidic environment of tumour, enhance the killing to tumour cell, there is Apoptosis.
CT and MRI is that most common two kinds of Imaging Methods, the two are complementary to one another in cancer of pancreas clinical diagnosis.But often at present CT or MRI contrast agent generally can not carry out bimodal imaging, also not have therapeutic effect.As can develop a kind of while having The imaging agent of CT, MRI bimodal imaging and therapeutic effect, is undoubtedly of great significance to the diagnosis and treatment of cancer of pancreas.
Summary of the invention
The purpose of the present invention is to provide a kind of novel rare-earth nanometer bimodal imaging agent and its preparation method and application, with Solve the problems of the prior art.
To achieve the above object, the invention provides the following technical scheme:
A kind of novel rare-earth nanometer bimodal imaging agent, the imaging agent are NaHoF4@CeO2Nanoparticle.
A kind of preparation method of novel rare-earth nanometer bimodal imaging agent, the preparation method the following steps are included:
(1)NaHoF4The preparation of nanoparticle: 2mmol HoCl is added into flask3·6H2O, 8ml oleic acid and 30ml ten Eight carbenes, are stirred at room temperature 1h, are then slowly heated to 120 DEG C, are heated to 160 DEG C, and 1h is maintained at 160 DEG C, cooling To room temperature, 10ml is added into flask and is dissolved with NaOH and NH4Three hours, Zhi Houhuan is stirred at room temperature in the methanol solution of F Slowly 230 DEG C are heated to, and maintain 15min at 230 DEG C, be cooled to room temperature, the mixed liquor after reaction is centrifuged, received Collecting supernatant is NaHoF4Solution, by NaHoF4Solution is added in the chloroformic solution of 5ml DSPE-PEG5k, stirs 15min, Rotary evaporation is carried out at 60 DEG C later, then adds 5ml deionized water, ultrasonic 5min obtains PEG-NaHoF4；
(2)CeO2The preparation of nanoparticle: suitable Ce (NO is weighed3)3·6H2O, being configured to concentration is 0.15molL-1 Ce (NO3)3Aqueous solution, first toward above-mentioned Ce (NO3)3Aqueous solution in be added dropwise appropriate 3% H2O2, then drip under magnetic stirring Adding concentration is 2molL-1NH3·H20, until solution pH value be greater than 9 until, be then centrifuged, to precipitate into Row ultrasonic vibration is washed 3 times, is collected precipitating, then will be deposited in freeze-day with constant temperature 12h at 120 DEG C, and obtain light yellow CeO2Nanometer Particle；
(3)NaHoF4@CeO2The preparation of nanoparticle: CeO obtained by a certain amount of step (2) is weighed2Nanoparticle is put into In deionized water, ultrasonic disperse 30min obtains the finely dispersed hydrosol, is vigorously stirred on magnetic stirring apparatus, adjusts water-soluble The pH value of glue is 6.3, the NaHoF obtained by a dropping step (1) in the above-mentioned hydrosol4Solution continues to stir 1h, makes NaHoF4 Sufficiently it is adsorbed onto CeO2Then reaction solution is heated to 70 DEG C by nanoparticle surface, react 2h at 70 DEG C, obtain NaHoF4@ CeO2Nanoparticle (NPs).
Further, 5mmolNaOH and 8mmolNH is dissolved in step (1) in the methanol solution of 10ml4F。
Further, the concentration of the chloroformic solution of DSPE-PEG5k is 20mg/ml in step (1).
Further, NaHoF in step (3)4With CeO2Molar ratio be 1:15.
A kind of application of novel rare-earth nanometer bimodal imaging agent, the imaging agent answering in the imaging of CT and MRI bimodal With.
Compared with prior art, the beneficial effects of the present invention are:
First is that NaHoF prepared by the present invention4@CeO2Holmium (Ho) and cerium (Ce) in nanoparticle all have higher atom The atomic number of ordinal number, holmium (Ho) and cerium (Ce) is respectively 67 and 58, and being above the atomic number of iodine (I), (atomic number of I is 53), higher than I, (under the conditions of 100KeV, the attenuation coefficient of Ho is 3.49cm to the attenuation coefficient of Ho2.g-1, the attenuation coefficient of I is 1.94cm2.g-1), which can be used as CT imaging contrast；The size of the nanoparticle is small (1-100nm), the nanoparticle Son can be easier to enter in the capillary that internal diameter is micron-scale, due to the high-permeability and retention effect of cancerous issue, Nano particle can be more deposited in cancerous issue, more preferably to cancerous region imaging effect；
Second is that NaHoF prepared by the present invention4@CeO2Ho in nanoparticle has very strong paramagnetism effect, can be obvious Shorten T2Relaxation time；On the other hand, the effect strong to X ray attenuation ability by Ho and Ce high atomic number, can be used for CT Imaging, imaging effect are better than clinical iodine contrast medium, therefore, NaHoF4@CeO2Nanoparticle has aobvious as MRI, CT bimodal As the value of agent, the present invention further passes through experiment in vitro research shows that ray can be remarkably reinforced to tumour cell in the nanoparticle Lethal effect, have extracorporeal six field radiation sensitization；
Third is that NaHoF prepared by the present invention4@CeO2Nanoparticle does not influence CeO2Itself intrinsic structure, by external MTT experiment and cell clone proliferation experiment, it was demonstrated that it has good radio therapy sensitization effect, pancreatic cancer cell used in the present invention Interior acidic environment can make Ce3+To Ce4+The oxygen radical ROS that ray generates is removed in conversion, improves tumour cell to the quick of ray Perception, and when normal cell is by ray damage, CeO2Start reduction reaction, to protect normal tissue, mitigates radiation damage.
Detailed description of the invention
Fig. 1 is a kind of novel rare-earth nanometer bimodal imaging agent NaHoF of the present invention4@CeO2The transmitted electron of nanoparticle is aobvious Micro mirror figure；
Fig. 2 is HAEC, Panc-1 cell survival rate situation map under various concentration NPs in MTT experiment of the present invention；
Fig. 3 is the NPs nude mice main organs tissue of tail vein injection saline of the present invention (control) and various concentration HE stained slice figure (200 ×)；
A figure in Fig. 4 is the CT figure of various concentration Iopamidol and NPs；B figure is the statistics to A figure；
A figure and B figure in Fig. 5 are respectively relational graph between the concentration and relaxation rate r1, r2 of NPs；
A figure in Fig. 6 is that MRI T2WI schemes under various concentration NPs；B figure is the statistics to A figure；
A figure in Fig. 7 be various concentration in MTT experiment NPs culture after radiotherapy to the influence diagram of cells survival rate；B Figure is cell clonal formation experimental evaluation NPs to cell radio therapy sensitization effect picture.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other Embodiment shall fall within the protection scope of the present invention.
Embodiment 1:
A kind of novel rare-earth nanometer bimodal imaging agent, the imaging agent are NaHoF4@CeO2Nanoparticle.
A kind of preparation method of novel rare-earth nanometer bimodal imaging agent, main laboratory apparatus and reality in the preparation method Testing material includes:
Six chloride hydrate holmium (HoCl3·6H2O, > 99.99%), octadecene (OED, 90%), ammonium fluoride (NH4F, > 99.99%), six water nitrous acid cerium (Ce (NO3)3·6H2O, > 99.99%) and methoxyl group polyethanol (DSPE-mPEG5000, > 99%), from Pu Mai biotech firm (Shanghai, China) purchase；
Human aorta endothelial cell (HAEC), human pancreatic cancer cell (Panc-1), by Chinese Academy of Sciences's Shanghai tumor research It is provided；
256 layers of iCT image instrument, are produced by Dutch Philips company；
NIUMAG 0.5T MRI small animal imaging system is produced by Chinese Shanghai Niu Mai company；
Varian 23EX linear accelerator, is produced by VARIAN Oncology Systems.
The preparation method the following steps are included:
(1)NaHoF4The preparation of nanoparticle: 2mmol HoCl is added into flask3·6H2O, 8ml oleic acid and 30ml ten Eight carbenes, are stirred at room temperature 1h, are then slowly heated to 120 DEG C, are heated to 160 DEG C, and 1h is maintained at 160 DEG C, cooling To room temperature, 10ml is added into flask and is dissolved with 5mmolNaOH and 8mmolNH4The methanol solution of F is stirred at room temperature three Hour, it is heated slowly to 230 DEG C later, and maintain 15min at 230 DEG C, is cooled to room temperature, the mixed liquor after reaction is carried out Centrifuge separation, collecting supernatant is NaHoF4Solution, by NaHoF4Solution is added to the chloroformic solution of 5ml DSPE-PEG5k In, 15min is stirred, carries out rotary evaporation at 60 DEG C later, then adds 5ml deionized water, ultrasonic 5min obtains PEG- NaHoF4；
(2)Ce02The preparation of nanoparticle: suitable Ce (NO is weighed3)3·6H2O, being configured to concentration is 0.15molL-1 Ce (NO3)3Aqueous solution, first toward above-mentioned Ce (NO3)3Aqueous solution in be added dropwise appropriate 3% H2O2, then drip under magnetic stirring Adding concentration is 2molL-1NH3·H20, until solution pH value be greater than 9 until, be then centrifuged, to precipitate into Row ultrasonic vibration is washed 3 times, is collected precipitating, then will be deposited in freeze-day with constant temperature 12h at 120 DEG C, and obtain light yellow CeO2Nanometer Particle；
(3)NaHoF4@CeO2The preparation of nanoparticle: CeO obtained by a certain amount of step (2) is weighed2Nanoparticle is put into In deionized water, ultrasonic disperse 30min obtains the finely dispersed hydrosol, is vigorously stirred on magnetic stirring apparatus, adjusts water-soluble The pH value of glue is 6.3, the NaHoF obtained by a dropping step (1) in the above-mentioned hydrosol4Solution, NaHoF4With CeO2Molar ratio For 1:15, continues to stir 1h, make NaHoF4Sufficiently it is adsorbed onto CeO2Then reaction solution is heated to 70 DEG C by nanoparticle surface, 2h is reacted at 70 DEG C, obtains NaHoF4@CeO2Nanoparticle.
A kind of application of novel rare-earth nanometer bimodal imaging agent, the imaging agent answering in the imaging of CT and MRI bimodal With.
Effect example:
(1) experimental subjects: NaHoF obtained by the embodiment of the present invention 14@CeO2Nanoparticle (NPs).
(2) experimental method:
(A)NaHoF4@CeO2Nanoparticle physicochemical property and characterization measurement
Transmission electron microscope (Transmission electron microscopy, TEM) intuitively shows the shape of NPs Shape, dispersion degree, the uniformity and reunion situation；With the eletrokinetic potential and average grain of dynamic light scattering measurement NPs in aqueous solution The distribution situation of diameter；Inductively coupled plasma bulk optics transmitter measures the content of Ho, Ce element in NPs, calculates molal weight；
(B) external, inner cell toxicity assessment
Its cytotoxicity is verified by MTT experiment using HAEC, Panc-1 cell, various concentration (0,50,100,200, 400 and 800 μ g/l) influence of the NPs to cells survival rate, 5 multiple holes of each concentration multiple cropping, experiment in triplicate, chooses life Long good male nude mouse, records weight, is randomly divided into experimental group and control group, experimental group sets three groups of different pharmaceutical concentration groups, often 3, the Ho particle NPs 15ml of tail vein injection various concentration (0,5,10,20mg/kg) are organized, is raised under the same terms, every other day Nude mice weight is recorded, after 15 days, obtains nude mice blood and tissue samples, blood sample row laboratory checks, detects blood routine: red Cell count (Red Blood Cells, RBC), white blood cell count(WBC) (White Blood Cell, WBC), platelet count (Blood Platelet, PLT), liver function: glutamic-pyruvic transaminase (Alanine Aminotransferase, ALT), millet straw turn ammonia Enzyme (Aspartate transaminase, AST), renal function: serum creatinine (Creatinine, Cre), urea nitrogen (Blood Urea Nitrogen, BUN), main organs (lung, heart, kidney, spleen, liver, pancreas) carry out tissue HE dyeing.
(C) external CT, MRI imaging effect evaluation
Use Philips 256iCT.The condition of scanning: 120kv, 130mA, thickness, layer are away from being 1mm, sweep time 2s. Various concentration (0,1,2,4g/l) Iopamidol and NPs are measured to various concentration respectively and carry out cross-section position CT examination, by what is obtained Image transmitting to picture archiving and communication system (Picture Archiving and Com munication Systems, PACS), choose on pacs systems and obtain image maximum section survey CT value, record data, every group of experiment is repeated 3 times, and asks flat Mean value.
Using NIUMAG 0.5T MRI imaging system, using dedicated point of Software contrast agent of NMR-CA Analyzing Software is analysed, various concentration (0,0.1,0.2,0.4,0.6,0.8mM) NPs contrast agent T is measured1、T2Relaxation time and relaxivity r1、r2Test, every group of experiment are repeated 3 times, average.It is obtained using NMR Imaging Software imaging software different dense The two dimensional image for spending NPs evaluates imaging results, contrast images signal strength row statistical analysis.
(D) extracorporeal six field radiation enhanced sensitivity is studied
(a) mtt assay detects NPs to Panc-1 cell radio therapy sensitization effect
Experimental group: 1. control group, 2. nanometer group (NPs), 3. irradiation group (RI), 4. joint group (NPs+RI).It takes and is in The Panc-1 cell of logarithmic growth phase, is inoculated in 96 orifice plates, 5 multiple holes of multiple cropping, after cell adherent growth, is added different dense Spend the NPs culture solution of gradient (0nM, 5nM, 10nM, 20nM).After 24 hours, 3. group and 4. group carry out roentgen radiation x, and dosage is 10Gy (6MV X-ray, S SD100cm, add 2cm equivalent), culture detect the survival of each hole cell with mtt assay after 24 hours Rate.
(b) cell clonal formation experiment further evaluates NPs to Panc-1 cell radio therapy sensitization effect
Experimental group: 1. control group, 2. nanometer group (NPs), 3. irradiation group (RI), 4. joint group (NPs+RI), wherein 3., 4. being divided into different subgroups by different exposure doses (0Cy, 2Cy, 4Cy, 6Cy, 8Cy, 10Cy).By the cell of logarithmic growth phase, Live cell fraction > 90%, for cell inoculation in 6 orifice plates, old culture solution is abandoned after cell adherent growth in every group of 3 hole of multiple cropping, 2., 4. the NPs culture solution that concentration is 20nM is added in group.3., 4. group culture solution is abandoned after 24 hours, fresh culture solution is added in each group, In each subgroup be irradiated by various dose gradient, cell is placed in incubator by after treatment to be continued, and is occurred in straight hole plate Macroscopic clone's inch is cultivated eventually.Finally, fixed, dyeing, microscopically observation count clone's number of > 50 cells, count CNN surviving fraction.
(E) statistical method
It analyzes, the experiment of single treatment group is examined using double tail t, for containing two using 20.0 statistical software of SPSS The experiment of processing group is examined using Two-ANOVA, is that difference is statistically significant with P < 0.05.
(3) experimental result
(A) NaHoF is successfully made4@CeO2Nanoparticle (NPs)
NaHoF4@CeO2Nanoparticle is light light yellow, tasteless powdery solid granules.TEM the result shows that synthesis NaHoF4@CeO2Nanoparticle shape is similar round, forms good monodispersity, has no obvious agglomeration, and NPs's is averaged Diameter is about 16nm (as shown in Figure 1)；Dynamic light scattering NaHoF as the result is shown4@CeO2Nanoparticle average potential is about -6mV, Average particle size is in normal distribution, and distribution 13-20nm, median is 17 ± 0.8nm；Inductively coupled plasma body Optical emitting instrument measures NaHoF4@CeO2The content of Ho and Ce is respectively 2715.3mg/l and 3976.1mg/l in nanoparticle.
(B) assessment of the inside and outside cytotoxicity of body
(a) vitro cytotoxicity MTT experiment
MTT experiment is the result shows that various concentration (0,50,100,200,400 and 800 μ g/l) NPs handles HAEC and Panc-1 After cell, HAEC cells survival rate is respectively 92.51%, 91.45%, 93.25%, 92.43%, 89.52%；Panc-1 cell Survival rate is respectively 96.54%, 93.28%, 90.85%, 92.56%, 89.57%.The experimental results showed that in various concentration Under, the survival rate of HAEC and Panc-1 cell is compared with the control group without significant difference (p > 0.05；Double tail t are examined), such as Fig. 2 institute Show.
(b) in Histological assessment's body
Various concentration (0,5,10,20mg/kg) Ho3+After tail vein injection nude mice 15 days, the inspection pair of blood sample laboratory According to a group blood routine result are as follows: RBC is counted: 10.5 × 106/mm3, WBC counting: 7.91 × 106/mm3, PLT counting: 770.5 × 106/mm3；Maximum concentration (20mg/kg) group blood routine result is tested as RBC counting: 11.0 × 106/mm3, WBC counting: 8.11 ×106/mm3, PLT counting: 690.5 × 106/mm3.Control group hepatic and renal function main indicator result are as follows: ALT:180IU/L, AST: 241IU/L；Cre:28.7mol/L, BUN:7.81mmol/L test maximum concentration group ALT:201IU/L, AST:257IU/L, Cre:27.9mol/L, BUN:8.21mmol/L.The experimental results showed that the blood count of experimental group nude mice, hepatic and renal function result with Control group is compared to no significant difference, statistically significant (p > 0.05 of difference；Double tail t are examined).Nude mice Main Tissues organ (lung, Liver,spleen,kidney is dirty, the heart and pancreas) H&E staining analysis shows (as shown in Figure 3) that NPs has no apparent damage to nude mice main organs Wound.
(C) external imaging moiety
(a) external CT imaging effect
The CT value that Philips iCT measures various concentration Iopamidol is respectively 1HU, 15HU, 27HU, 45HU；NPs's CT value is respectively 2HU, 28HU, 74H U, 234HU.It is control with Iopamidol 0g/L, statistical result showed is (such as Fig. 4 institute Show), with the increase of two kinds of drug concentrations, CT value gradually rises (*, p < 0.05；*, p < 0.01；* *, p < 0.001；Double tails T is examined)；Iopamidol group is compared with NPs group, and under same concentrations, the CT value of NPs group ratio Iopamidol group is higher, is had significant Difference (*, p < 0.05；*, p < 0.01；* *, p < 0.001；Two-ANOVA is examined).The experimental results showed that NPs can be used as CT at As contrast agent, and under same concentrations, the CT contrasting effects of NPs are better than Iopamidol.
(b) MRI imaging effect is evaluated
The T of NMR-CA Analyzing Software contrast agent dedicated analysis software measurement various concentration NPs1Value difference For 14285.7ms, 7692.3ms, 2380.5ms, 1785.7ms, 1282.5ms；T2Value respectively 217.3ms, 92.5ms, 48.9ms,40.3ms,24.4ms.Each sample relaxation rate is calculated according to r=1/T, data are counted using double tail t inspections Handle (as shown in Figure 5).The result shows that with the increase of sample concentration, T2Value is obviously shortened, relaxation rate r2Increase, NPs tool There is apparent short T2Effect makes it that can become T2Contrast agent.
NMR Imaging Software imaging software obtains various concentration NPs2MRI T2WI image (as shown in Figure 6A), As sample concentration increases, MRI T2The gradually blackening of WI image；Statistical result (as shown in Figure 6B) with sample concentration increase, MRI image signal strength is gradually reduced (*, p < 0.05；*, p < 0.01；* *, p < 0.001；Double tail t are examined).
(D) extracorporeal six field radiation enhanced sensitivity
(a) extracorporeal six field radiation MTT experiment
The experimental results showed that (as shown in Figure 7 A), cell growth has no obvious inhibition after simple 10nM NPs culture, survives Rate is about 93.58%；Cell survival rate reduces after simple irradiation, grows obvious inhibit；The growth inhibition of cell after joint irradiation Relatively simple irradiation becomes apparent, and inhibiting rate is increased with the increase of concentration, is positively correlated with concentration, group of cells survival rate Between there were significant differences (* *, p < 0.01；* *, p < 0.001；Two-ANOVA is examined).
(b) cell clonal formation is tested
The experimental results showed that (as shown in Figure 7 B), after various dose radiation exposure, plastidogenetic clone's number is significantly reduced, CNN surviving fraction reduces, and reduces with the increase of roentgen dose X, and there are statistical significance (* * *, p < 0.001 for difference；Two- ANOV A is examined).And between simple NPs group and blank group between cell survival amount and no difference of science of statistics (P > 0.05, Two- ANOVA is examined).
It is obvious to a person skilled in the art that invention is not limited to the details of the above exemplary embodiments, Er Qie In the case where without departing substantially from spirit or essential attributes of the invention, the present invention can be realized in other specific forms.Therefore, no matter From the point of view of which point, the present embodiments are to be considered as illustrative and not restrictive, and the scope of the present invention is by appended power Benefit requires rather than above description limits, it is intended that all by what is fallen within the meaning and scope of the equivalent elements of the claims Variation is included within the present invention.Any reference signs in the claims should not be construed as limiting the involved claims.

Claims (6)

1. a kind of novel rare-earth nanometer bimodal imaging agent, it is characterised in that: the imaging agent is NaHoF4@CeO2Nanoparticle.
2. a kind of preparation method of novel rare-earth nanometer bimodal imaging agent, which is characterized in that the preparation method includes following step It is rapid:
(1)NaHoF4The preparation of nanoparticle: 2mmol HoCl is added into flask3·6H2O, 18 carbon of 8ml oleic acid and 30ml 1h is stirred at room temperature in alkene, is then slowly heated to 120 DEG C, is heated to 160 DEG C, maintains 1h at 160 DEG C, is cooled to room Temperature is added 10ml into flask and is dissolved with NaOH and NH4Three hours are stirred at room temperature in the methanol solution of F, slowly add later Heat maintains 15min to 230 DEG C, and at 230 DEG C, is cooled to room temperature, and the mixed liquor after reaction is centrifuged, in collection Clear liquid is NaHoF4Solution, by NaHoF4Solution is added in the chloroformic solution of 5ml DSPE-PEG5k, stirs 15min, later Rotary evaporation is carried out at 60 DEG C, then adds 5ml deionized water, and ultrasonic 5min obtains PEG-NaHoF4；
(2)CeO2The preparation of nanoparticle: suitable Ce (NO is weighed3)3·6H2O, being configured to concentration is 0.15molL-1Ce (NO3)3Aqueous solution, first toward above-mentioned Ce (NO3)3Aqueous solution in be added dropwise appropriate 3% H2O2, then be added dropwise under magnetic stirring dense Degree is 2molL-1NH3·H20, until the pH value of solution is greater than 9, then it is centrifuged, precipitating is surpassed Washing 3 times is swung in acoustic shock, is collected precipitating, then will be deposited in freeze-day with constant temperature 12h at 120 DEG C, and obtain light yellow CeO2Nanoparticle；
(3)NaHoF4@CeO2The preparation of nanoparticle: CeO obtained by a certain amount of step (2) is weighed2Nanoparticle be put into from In sub- water, ultrasonic disperse 30min obtains the finely dispersed hydrosol, is vigorously stirred on magnetic stirring apparatus, adjusts the hydrosol PH value is 6.3, the NaHoF obtained by a dropping step (1) in the above-mentioned hydrosol4Solution continues to stir 1h, makes NaHoF4Sufficiently It is adsorbed onto CeO2Then reaction solution is heated to 70 DEG C by nanoparticle surface, react 2h at 70 DEG C, obtain NaHoF4@CeO2It receives Rice corpuscles.
3. a kind of preparation method of novel rare-earth nanometer bimodal imaging agent according to claim 2, it is characterised in that: institute It states and dissolves 5mmolNaOH and 8mmolNH in step (1) in the methanol solution of 10ml4F。
4. a kind of preparation method of novel rare-earth nanometer bimodal imaging agent according to claim 3, it is characterised in that: institute The concentration for stating the chloroformic solution of DSPE-PEG5k in step (1) is 20mg/ml.
5. a kind of preparation method of novel rare-earth nanometer bimodal imaging agent according to claim 4, it is characterised in that: institute State NaHoF in step (3)4With CeO2Molar ratio be 1:15.
6. a kind of application of novel rare-earth nanometer bimodal imaging agent, it is characterised in that: the imaging agent is aobvious in CT and MRI bimodal Application as in.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110935021A (en) * 2019-12-05 2020-03-31 南京大学深圳研究院 Spindle type CuS @ CeO2Nano composite material and preparation method thereof
CN112724978A (en) * 2021-01-21 2021-04-30 广州大学 Core-shell structure up-conversion nano material and preparation method thereof
CN112898977A (en) * 2021-01-21 2021-06-04 广州大学 Terbium ion doped up-conversion nano material and preparation method thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102175735A (en) * 2010-12-31 2011-09-07 甘肃省科学院传感技术研究所 Au NPs-CeO2@PANI (polyaniline) nanocomposite, preparation method thereof and glucose biological sensor manufactured by utilizing nanocomposite
CN104548144A (en) * 2014-12-18 2015-04-29 中国科学院上海硅酸盐研究所 Medicinal ultrahigh-field nuclear-magnetism contrast agent and preparation method thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102175735A (en) * 2010-12-31 2011-09-07 甘肃省科学院传感技术研究所 Au NPs-CeO2@PANI (polyaniline) nanocomposite, preparation method thereof and glucose biological sensor manufactured by utilizing nanocomposite
CN104548144A (en) * 2014-12-18 2015-04-29 中国科学院上海硅酸盐研究所 Medicinal ultrahigh-field nuclear-magnetism contrast agent and preparation method thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DALONG NI等: "PEGylated NaHoF4 nanoparticles as contrast agents for both X-ray computed tomography and ultra-high field magnetic resonance imaging", 《BIOMATERIALS》 *
JIMMIE COLON等: "Cerium oxide nanoparticles protect gastrointestinal epithelium from radiation-induced damage by reduction of reactive oxygen species and upregulation of superoxide dismutase 2", 《NANOMEDICINE: NANOTECHNOLOGY, BIOLOGY, AND MEDICINE》 *
沈岳年等: "纳米CeO2在生物医药上的应用", 《中国稀土学报》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110935021A (en) * 2019-12-05 2020-03-31 南京大学深圳研究院 Spindle type CuS @ CeO2Nano composite material and preparation method thereof
CN110935021B (en) * 2019-12-05 2022-02-22 南京大学深圳研究院 Spindle type CuS @ CeO2Nano composite material and preparation method thereof
CN112724978A (en) * 2021-01-21 2021-04-30 广州大学 Core-shell structure up-conversion nano material and preparation method thereof
CN112898977A (en) * 2021-01-21 2021-06-04 广州大学 Terbium ion doped up-conversion nano material and preparation method thereof

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