CN108578698A - Purposes of the Prussian blue-Manganese Ferrite composite nano materials as magnetic heat/photo-thermal combination therapy agent - Google Patents
Purposes of the Prussian blue-Manganese Ferrite composite nano materials as magnetic heat/photo-thermal combination therapy agent Download PDFInfo
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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
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- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Abstract
Purposes the invention discloses Prussian blue Manganese Ferrite composite nano materials as magnetic heat/photo-thermal combination therapy agent.Wherein, the Prussian blue Manganese Ferrite composite nano materials are PB MnFe2O4Nanocomposite.
Description
Technical field
The present invention relates to field of new materials more particularly to magnetic heat/photo-thermal combination therapy agent.
Background technology
With the increasingly exacerbation of the social concerns such as current environmental pollution, food security, the incidence of cancer persistently increases, cancer
Disease is just becoming one of the main fatal disease of current threat human health, and the number that cancer is died of in the whole world every year accounts for total number of persons
12.5%, number about 9,000,000, only in 2010, cancer results in nearly 570,000 people dead and more serious, suffers from cancer
Crowd's increasingly rejuvenation.Why so serious cancer present situation is, is determined by the property of itself.Cancer (cancer) is also referred to as
Malignant tumour it is opposite with it be benign tumour, it is since body cell loses normal regulation, disease caused by hyper-proliferative
Disease.The cell of hyper-proliferative is commonly called as cancer cell, and cancer cell easily invades surrounding tissue, easily transfer, easily breeding, easily diffusion.
Invention content
One of the object of the invention is that provide Prussian blue (the PB)-Manganese Ferrite composite nano materials of one kind controls as magnetic heat
The purposes of agent is treated,
It is another object of the present invention to provide Prussian blue-Manganese Ferrite composite nano materials as photo-thermal therapy agent
Purposes.
It is a further object of the present invention to provide Prussian blue-Manganese Ferrite composite nano materials as magnetic heat, photo-thermal joint
The purposes of therapeutic agent.
In Prussian blue-Manganese Ferrite composite nano materials of the present invention, PB and MnFe2O4Mass ratio can be 1:1-
5。
Preferably, Prussian blue-Manganese Ferrite composite nano materials, PB and MnFe of the present invention2O4Mass ratio is 1:
2-3。
Prussian blue-Manganese Ferrite composite nano materials preparation method of the present invention includes：Take PB and MnFe2O4Sample,
According to mass concentration 1：The ratio of 1-5 mixes, and it is 1-4 then to adjust solution ph, after being stirred overnight, Magnetic Isolation, clear repeatedly
It washes until solution is clarified.
Preferably, Prussian blue-Manganese Ferrite composite nano materials preparation method of the present invention includes：Take PB and
MnFe2O4Sample, according to mass concentration 1：The ratio of 2-3 mixes, and it is 1.5-2.5 then to adjust solution ph, after being stirred overnight,
Magnetic Isolation, cleaning are until solution is clarified repeatedly.
The present invention selects PB nano-particles as substrate, and outside adheres to one layer of manganous ferrite nano-particle, but not exclusively package
PB regulates and controls the combination ratio of the two, forms Prussian blue-Manganese Ferrite composite nano materials.And be found through experiments that, it is compound it
After can play " 1+1>2 " effect, i.e., this existing Prussian blue photo-thermal effect of composite material, and ferrous acid can be shown
The magnetic thermotherapeutic function of manganese, be it is a kind of can realize magnetic heat, photo-thermal combination therapy composite nano materials, and when be thermotherapy magnetic
Heat and photo-thermal can further increase the effect of thermotherapy when using simultaneously, faster, required time is less, and effect becomes apparent from for heating.
Therefore, this Prussian blue-Manganese Ferrite composite nano materials can be used in therapy field.
Description of the drawings
The invention will be further described with reference to the accompanying drawings and examples.
Fig. 1：The photo-thermal curve A of material) various concentration composite material photo-thermal curve；B) same concentration (100 μ g/ml)
The curve of the material of different capacity density；C) the photo-thermal curve of same concentration (100 μ g/ml) different materials；D) the cycle of material
Photo-thermal curve：E) the infrared thermal imaging picture of the material of various concentration；
Fig. 2, the hot heating curve A of magnetic of material) various concentration material；B) different size of current；C) different with concentration
Material；D) the infrared thermal imaging picture of various concentration material；E) SAR value of material and it is 3 minutes corresponding when temperature；
Fig. 3, the photo-thermal of cell test A) the MTT figures of composite material；B) the fluorescent staining figure of cell, b1-b4) it is followed successively by not
Add material not illumination, plus material not illumination, be not added with material add illumination and add material add illumination；
Fig. 4, A) cell magnetic hot MTT figure；B) the hot fluorescent staining figure of the magnetic of cell, b1-b4) no material is followed successively by without magnetic
Heat has material to have magnetic heat without magnetic heat, without material, have material to have magnetic hot；
Fig. 5 is the photo-thermal stream data figure A of cell) negative control group；B) positive controls；C-E) material group, material
Concentration is followed successively by 50,100,200mg/ml；
The hot heating curve of photo-thermal magnetic of Fig. 6 materials in varied situations；A is magnetic heat, 500KHz, 30A；B) it is photo-thermal 1W/
cm2；C) it is magnetic hot (500KHz, 30A) and photo-thermal (1W/cm2) joint heating curve；The concentration range of A-C is 10-100 μ g/
MlD when) to be sample concentration be 200 μ g/ml, magnetic heat (500KHz, 20A), photo-thermal (1W/cm2) and the hot photo-thermal joint heating of magnetic
Curve；
The photo-thermal of Fig. 7 mouse/magnetic heat integration Experiment on therapy:A) the infrared thermal imaging figure of mouse；B) the heating curve of tumour
Figure；C) the mass-change curve of mouse；D) the volume change curve of tumour；E) the photo of pretherapy and post-treatment mouse；F) the H＆E of tumour
Stained slice.
Fig. 8 is pathological examination figure, the H＆E stained slices of each internal organs of mouse after 12 days.
Specific implementation mode
1, the preparation of PB
For the prior art, synthetic method refers to related existing literature, briefly as follows：The PVP (K30) of 1.5g is taken to be added to
It in the water of 20mL, is then stirred by ultrasonic, until being completely dissolved, it is 3 that HCl (6M/L), which is then added dropwise, to pH value, is then added
K3 [the Fe (CN) of 45mg6], it then stirs to being completely dissolved, then again plus the citric acid of 0.192g, ultrasound is to being completely dissolved.Again
It is poured into inner liner of reaction kettle, then moves into baking oven, cleaned after 80 DEG C, 2h, then room temperature cooling.Add acetone, the two is according to 1：1
Ratio mixing after, then high speed centrifugation, rotating speed 12000rpm/min, 12min are centrifuged with absolute ethyl alcohol again, finally again with super
Pure water centrifuges, and then will be preserved after 60 DEG C of vacuum drying 12h of sample again.
2 MnFe2O4Preparation
For the prior art, MnFe2O4Synthetic method refer to existing literature, briefly it is as follows：Take four chloride hydrates of 0.01mM
Manganese pours into the beaker of 50mL, and rear to be added in 25ml ultra-pure waters, ultrasonic vibration makes it completely dissolved, it is determined as then solution A takes
The ferric chloride hexahydrate of 0.02mM pours into the beaker of 50mL, and rear to be added in 25mL ultra-pure waters, ultrasonic vibration makes it completely dissolved,
It is set to solution B.Then the two is imported in the three-necked flask of 100mL, 80 DEG C of mechanical agitation 30min, rotating speed 600rpm/min, so
The sodium hydroxide of 0.08mM is added afterwards, continues to stir 30min, then adds the polyethyleneimine of 0.05g, continue to stir
Room temperature cools down after 30min.Then magnet detaches, and is cleaned with ultra-pure water, until solution is clarified.Last 60 DEG C of vacuum drying
It is preserved after 12h.
3 PB/MnFe2O4Preparation
Take the PB and MnFe of above-mentioned synthesis2O4Sample, according to mass concentration 1：2.3 ratio mixing, is then added dropwise 6M/L's
Hydrochloric acid, it is 2 to make solution ph, and then mechanical agitation is stayed overnight, rotating speed 300rpm/min, and solution is in blackish green, and then magnet divides again
From, cleaned with ultra-pure water, repeatedly Magnetic Isolation cleaning until solution is clarified.It is preserved after last 60 DEG C of vacuum drying 12h.
4, the photo-thermal of cell and the experiment of magnetic heat
In order to which from the photo-thermal and magnetic heating performance of cellular level authentication material, steps are as follows：1) exponential phase is chosen
Cell, with trypsin digestion, PBS buffer solution is cleaned 2 times, then with 5 × 105The concentration of mL-1 is inoculated in 12 orifice plates, is divided into
Two groups, one group is examined with mtt assay, and another group of carry out Calcein AM/PI dyeing processing is 1.5mL per pore volume, then moves into
It is incubated for 24 hours in constant incubator；2), then original fluid is removed, PBS buffer solution is cleaned 2 times, then added dense containing different materials
The culture solution of (400 μ g/mL, 200 μ g/mL, 100 μ g/mL, 50 μ g/mL, 25 μ g/mL, being not added with material group) is spent, then moves into constant temperature
4h is incubated in incubator；Then photo-thermal irradiation experiment, the experiment of magnetic heat and photo-thermal magnetic heat integration Experiment on therapy are carried out respectively, then
Again move into constant incubator in be incubated, MTT groups for 24 hours, dyeing group 4h；Then 20ml MTT are added in MTT groups, are cultivated for
4h, then supernatant is removed, it is dissolved and is precipitated with DMSO, the absorbance at spectrophotometric determination 570nm is then used again, to be not added with material
Group as a control group, calculates cell survival rate.After another group of incubation 4h, the Calcein AM and PI of removal supernatant 5mL, then
It under conditions of being protected from light again, is put on horizontal shaker, 20r/min, 10min, then is incubated 30min.Then it further takes out, removes supernatant
Liquid, PBS buffer solution are gently cleaned 2 times, then the culture solution of 100 μ L is added in each hole, are finally taken fluorescence microscopy under the microscope, are clapped
According to.
5, cell streaming is tested
Handle cell, collect the cell of processing, fixed overnight with 70% cold ethyl alcohol, secondary daily PBS buffer solution clearly twice,
At room temperature, it is protected from light dyeing 30min with Calcein AM and PI dyeing liquors, finally uses flow cytomery apoptosis rate,
In be not added with material group be blank control group.
6, the plantation of tumour
4T1 cell strains are subjected to passage amplification, are prepared into 1 × 108The cell suspending liquid of mL-1.Then mouse is grouped,
It is divided into 8 groups, every group 3, be simple PBS groups, PBS+MFH groups, PBS+PTT groups, PBS+MFH+PTT groups, material group, material respectively
+ MFH groups, material+PTT groups and material+MFH+PTT groups.Then respectively in the wall of the chest 5-6 intercostals of mouse and flank wall notch graft
Then the cell suspending liquid of kind respective concentration observes mouse living and diet and the upgrowth situation of inoculation position tumor mass, such as daily
Knurl time of occurrence, tumor volume (V=ab2/2, a are the major diameter of knurl, and b is minor axis), mouse quality etc., and draw curve.
After the completion of experiment, takes knurl tissue to carry out HE dyeing and carry out pathological observation.
7, treatment of animals
In order in the treatment function of animal level verification material, be primarily referred to as the magnetic heating performance of material, light thermal property and
Performance after the hot photo-thermal superposition of magnetic, selects Blab/C mouse as experimental subjects.First individually carry out mouse magnetic heat test with
And photo-thermal experiment, after first anaesthetizing mouse, then mouse is placed in magnetic heating by 50 μ l of in-situ injection, the material of 200 μ g/ml
Under equipment and laser, the variation of the temperature of mouse tumor region is recorded with infrared thermography.Then the hot photo-thermal superposition of magnetic is carried out
Laser and the joint collocation of magnetic heating equipment are got up in experiment, then mouse anesthesia, in-situ injection material, then mouse is put into and is taken
In the instrument built, with the temperature in infrared thermography record mouse tumour section.It is (simple according to the grouping situation of previous mouse
PBS groups, material+MFH groups, material+PTT groups and material+MFH+PTT groups), it detects successively.It takes pictures daily and detects mouse and swell
The variation of tumor and weight stops treatment further according to the tumor disappearance situation of mouse.
8, HE is dyed
After the completion of mouse treatment, mouse is dissected, detaches internal organ, including the heart, liver, spleen, lung, kidney and tumour, does tissue and cut
Piece carries out pathological observation.Dyeing is that dyestuff is configured to solution, and histotomy is immersed in coloring agent, by certain time,
So that the ingredient of tissue and cell is caught different colors, generate different refractive index, convenient for being seen under an optical microscope
It examines.Dyeing uses haematoxylin eosin staining procedures (HE dyeing).
The toxicity detection of 1 composite material of embodiment
The PB/MnFe2O4 materials of sterilizing are subjected to cytotoxicity test, by detect cell various concentration material
Survival condition (material in (400 μ g/mL, 200 μ g/mL, 100 μ g/mL, 50 μ g/mL, 25 μ g/mL, being not added with material group) co-cultivation
Material is to quantify mixing by the culture medium of+90 μ L of 10 μ L materials to be prepared using concentration), MTT (3- (4,5-dimethyl-2-
Thiazolyl) -2,5-diphenyl-2-H-tetrazolium bromide) method, the cell object chosen here is that HeLa is thin
Born of the same parents and 4T1 cells, specific experiment operation are as follows：
Cell count：It will be placed in superclean bench in exponential phase cell, and remove the former culture medium in culture dish
(being handled with care, cell is avoided to be sucked away since suction is excessive), then 2mL PBS buffer solutions are added to it and clean 2 times, then use
The trypsase (25%) that liquid-transfering gun pipettes 1mL carries out cell dissociation, after 2min, the piping and druming of 1mL culture mediums is added and terminates digestion (this
When microscope under it can be seen that cells float, it is not adherent), then 1.5mL centrifuge tubes is used to collect, rear centrifugal treating, rotating speed
3000r/min, time 5min after centrifugation, remove supernatant, 2mL culture mediums are added, then gently blow and beat cell with liquid-transfering gun
(30~50 times).The cell suspension for taking 10 μ L, takes and is counted into blood counting chamber, passes through being averaged for four count blocks of calculating
Number, so that it is determined that the cell concentration of total cell suspension.
Inoculation：Then it is in 104 cell inoculations to 96 orifice plates by target, a certain amount of culture medium containing serum, which is added, (to be made
The total amount for obtaining cell per well culture medium is 100 μ L), it is then placed in constant incubator, is incubated for 24 hours, keeps cell sufficient
Adherent growth.
Add material：Culture medium is removed, the PBS buffer solution of 100 μ L is added, cleans 2 times, is then respectively adding aimed concn
Material：400 μ g/mL, 200 μ g/mL, 100 μ g/mL, 50 μ g/mL, 25 μ g/mL, it is not added with material group.5 repeating groups of every group of setting,
It is co-cultured for 24 hours in 37 DEG C of constant incubator.After reaching the object time, culture medium is removed, PBS buffer solutions is used in combination to clean 3
Secondary (to remove extra remaining material solution).
MTT is detected：The blood-free medium of the MTT solution and 80 μ L of 20 μ L 5mg/mL is added per hole again, then places into 37
It is protected from light in DEG C constant incubator and is incubated 4h.After 4h, culture solution (avoiding cell from being sucked out to influence experimental result) is carefully removed,
Then 150 μ L DMSO are added per hole, and under conditions of being protected from light, 96 orifice plates are put into horizontal shaker 20r/min, 10min,
Be then placed into microplate reader and carry out ultraviolet detection, OD values are arranged and are drawn by the final OD values obtained per hole, obtain HeLa and
The activity data figure of 4T1 cells.
See Fig. 8, according to the above results, obtaining this Prussian blue material of magnetism has preferable biocompatibility.
The light thermal property of 2 material of embodiment
The sample solution that will be obtained, takes certain density sample in ultraviolet ware, then places it under laser and irradiates
10min, while picture is acquired with infrared thermography, recording materials temperature changes with time situation.It is as follows：
According to laser power density [12] formula：
η=P/ π R2 (1)
η：Laser power density, unit W/cm2；
P：Laser power, unit W；
R：The radius of laser irradiation hot spot on object, unit cm.
The size of laser power is selected, then the probe of fixed laser, it is rear in laser irradiation to ground a light occur
Then spot takes ruler to measure the diameter of hot spot and pop one's head in the distance on ground, according to laser power density adjustable range, then
Ultraviolet cuvette is placed at hot spot, the temperature change of start recording material, draws temperatur-timel curve, at the same with it is infrared it is hot at
As instrument collecting temperature changes picture.It is the different materials of same concentration, H2O, PB, MnFe2O4 respectively from 4 angles
And PB/MnFe2O4；The composite material of different materials concentration under same laser power density, 10 μ g/mL, 20 μ g/mL, 50 μ g/mL
And 100 μ g/mL；The composite material of same concentration under different laser power densities, 0,0.5W/cm2、1W/cm2、2W/cm2；Light
Thermal cycle experiment, the main photo and thermal stability for verifying composite material after material stimulated light irradiates 10min, turn off laser, record material
The variation of material temperature degree opens laser after dropping to room temperature, irradiates 10min, then turn off laser again again, and so on, it is repeated 3 times.
The light thermal property of material is tested
Fig. 1 be material photo-thermal curve A) various concentration composite material photo-thermal curve；B) same concentration (100 μ g/
Ml) the curve of the material of different capacity density；C) the photo-thermal curve of same concentration (100 μ g/ml) different materials；D) material follows
Ring photo-thermal curve：E) the infrared thermal imaging picture of the material of various concentration.
Find that material has very strong absorption peak near infrared region based on ultraviolet spectra test analysis above, to the light of material
Hot property is tested.It first verifies that whether composite material has light thermal property, has chosen 4 material concentrations, such as Figure 1A, successively
For 10 μ g/mL, 20 μ g/mL, 50 μ g/mL and 100 μ g/mL.It was found that when laser power density is 2W/cm2, even if material concentration
Very low, the temperature difference in 10min before and after material remains to reach 10 DEG C, and heating is apparent, and concentration is higher, and heat up more apparent, highest
Thermal gradient energy reaches nearly 30 DEG C, illustrates that material has good light thermal property.
The influence of condition, such as laser power density, are shown in Figure 1B, have selected 4 parameters, respectively 0W/cm2,0.5W/cm2,
1W/cm2,2W/cm2, a concentration of 100 μ g/ml of material.It was found that power density is bigger, heating is more apparent, temperature be followed successively by 0 DEG C,
38 DEG C, 48 DEG C and 57 DEG C.
The influence of Material cladding selects the H of same concentration such as Fig. 1 C2O、PB、MnFe2O4And PB/MnFe2O4Material, laser
Power is 2W/cm2.It was found that H2The front and back variations almost without temperature of O, MnFe2O4There is certain rising, but is not apparent.And
Difference variation is very big before and after PB and PB/MnFe2O4, there is a very strong light thermal property, but after Material cladding temperature it is slightly higher with it is single
PB materials, have certain temperature superimposition effect after the two is compound.
Followed by the solar thermochemical cycle stability test of material, primarily to the photo and thermal stability of test material, as a result
As schemed shown in D.Every laser of 10min switches, but do not stop the detection of material temperature, material temperature is found after 3 cycles
55 DEG C can be risen in degree 10min, shows that material has good photo and thermal stability.
Figure E is the infrared thermal imaging picture of material, mainly uses the temperature picture of infrared thermography capturing material, color
The bright depth directly reflect material temperature height, can therefrom find that the light thermal property of material is preferable.
Cell photo-thermal is tested
It is the MTT figures after material and the irradiation of material stimulated light referring to Fig. 3, Fig. 3 A, it can be seen that the cell toxicant of material
Property is smaller, even material concentration is very high when (400 μ g/mL), the cell survival still more than 80%, and general cell survival
When rate is more than 75%, then show that bio-toxicity is small, biocompatibility is high.And after stimulated light irradiation, cell survival rate is remarkably decreased,
Cell survival rate is less than 40%, this is because after material is by laser emission, temperature rise, and cell death, from cellular level table
The light thermal property that material is illustrated is good.
Fig. 3 B are the uptake ratios of material, it can be seen that material can be and with the extension of time, thin by cellular uptake
The uptake ratio of born of the same parents is higher, can reach 25%.
Fig. 3 C-F are to be not added with material not by the picture after calcein AM and PI dyeing, discovery after cell stimulated light is irradiated
Add the HeLa cells of laser to be all dyed to green fluorescence, shows this cellular control unit normal growth, and simple material does not draw
Photochemical and thermal reaction is played, cell is still green fluorescence.And when only laser light being added to shine, significantly changing does not occur in cell, still all
It is dyed to green fluorescence, shows cell survival.In the case of no light thermit powder, simple 808nm laser irradiations not will produce
Photochemical and thermal reaction, therefore will not be to affecting cells.And under the influence of material and laser exist simultaneously, in fluorescence microscope
Under observation, one jiao there is red fluorescence, although the range at this angle is bigger than the region of laser irradiation, is also accorded with irradiation area
It closes.It is because material has photo-thermal effect that red area is larger, is heat to kill cell by the converting heat of absorption, and by
In the diffusion of heat, adjoining cell activity is also affected, so it is larger, even if but in this way, still having just outside irradiation area
Normal cell.In conclusion this Prussian blue composite material of magnetism can efficiently kill cell by near infrared light fuel factor, therefore
It is of great significance in the photo-thermal therapy of cancer.
Followed by the characterization of cell streaming, the cell after handling well is taken into flow cytomery, the following institute of data
Show.
Fig. 5 be cell photo-thermal stream data figure A) negative control group B) positive controls C-E) material group, material it is dense
Degree is followed successively by 50,100,200mg/ml.
As can be seen from the figure as the figure A of negative control group, Apoptosis is less, and most cells are in living cells shape
State, and as positive controls, due to adding the H2O2 of 20mM, so the cell for having more than 60% is in apoptotic state, and
It is experimental group to scheme C, D, E, has been separately added into various concentration MPB NPs materials, heat has been generated under the irradiation of laser, and then cause
The apoptosis of cell, it can be seen that with the increase of concentration, the accounting of living cells gradually decreases, and the accounting of apoptotic cell is gradual
It increases, illustrates that the photo-thermal effect of material is good, also showing this material has excellent photo-thermal effect.
Treatment of animals
Then the thermotherapy experiment of mouse is carried out, as shown in Figure 7.Wherein Fig. 7 A are the tumor temperature real-time curve in treatment,
It can be seen that the equal nothing of (materials+MFH, materials+PTT, materials+MFH+PTT) temperature in addition to treatment group
Significant change is fluctuated about 36 DEG C.And treatment group change in 2 minutes it is more apparent, wherein magnetic heat reaches 43 DEG C, photo-thermal reaches
To 44 DEG C, and 55 DEG C can be reached within two minutes of combination therapy, it can be seen that material has well in animal level
Effect more can intuitively find out front and back temperature change from Fig. 7 B.Fig. 7 C figures are the mass change of mouse, compared with initially,
Quality almost keeps balancing.Fig. 7 D are the volume change curve of mouse tumour, it can be seen that in addition to treatment group, the body of tumour
Product significantly increases, after 12 days, about initial 8 times of sizes.And magnetic heat is 4 times of initial sizes, photo-thermal is 3 initially
Times size.And the tumour of magnetic heat integration photo-thermal group just disappears for 3 days, directly reflects the photo-thermal magnetic heat integration treatment effect of material
Fruit.And the situation of change of tumour can be directly found out from the photo effect of Fig. 7 E figures, to reflect that material joins with photo-thermal magnetic heat
Close the efficient therapeutic effect for the treatment of.
Pathological analysis
The H＆E stained slices of each internal organs of mouse and tumour after Fig. 8 12 days, scale：100μm
Followed by the tissue and cellular morphology of each organ of H＆E staining analysis, as shown in Figure 8.By experimental group with compare
Group carries out front and back comparison, by comparing it is found that the heart, liver, spleen, lung, the kidney of eight groups of mouse there is no apparent on tissue morphology
Variation illustrates that material has preferable biological safety, does not have apparent toxic side effect to mouse, and during the entire course for the treatment of
Mouse physiological status is good.And the histotomy discovery for observing tumour has larger change, occurs in the tumor tissues of experimental group
The phenomenon that cell shrinkage of large area, injury of blood vessel and tissue necrosis, this shows that tumor tissues have lost in treatment group
Physiological activity no longer has and grows value-added ability.
The magnetic heat of 3 material of embodiment is tested
The present embodiment mainly investigates the magnetic heating performance of material, molten with fluorescence fiber temperature measurement meter recording materials while test
The temperature change of liquid, mainly from 3 angles：1, the concentration of material, 200 μ g/mL, 400 μ g/mL, 800 μ g/mL；2, experiment
Test condition, such as electric current, field strength, frequency, and it is primarily referred to as the size of line of magnetic induction loop current here, when material concentration is constant
When, change size of current, 5A, 10A, 20A, 30A and 40A；3, same experiment condition, concentration of the same race but different materials, H2O,
PB, MnFe2O4 and PB/MnFe2O4.Temperatur-timel curve is drawn according to the temperature measured, and material is calculated according to curve
Heat production rate (Specific Absorption Rate, SAR) [13], that is, the sample of unit mass convert energy to thermal energy
Amount, heat production rate is higher, and magnetic heating performance is better：
In formula：C is the specific heat capacity of material solution, J/g K；
Δ T/ Δs t is the slope of temperature time curve；
mFeThe content of the ferro element contained by material for unit quality.
The magnetic heating performance of material is tested
The hot heating curve A of magnetic of Fig. 2 materials) various concentration material；B) different size of current；C) the material different with concentration
Material；D) the infrared thermal imaging picture of various concentration material；E) SAR value of material and it is 3 minutes corresponding when temperature
It is tested by VSM, it is found that material has higher magnetism, so the next magnetic heating performance of main test material.It is main
Will from 3 angles, be respectively material concentration, the condition of experiment test, magnetic heating SAR value and Material cladding before and after
Influence.
Fig. 2A is that the influence of discussion material concentration has chosen 3 concentration, 200 μ at 500KHz, 12A/H experiment conditions
G/mL, 400 μ g/mL and 800 μ g/mL.Therefrom find that concentration is higher, the heating performance of material is better, also just illustrates the magnetic of material
Hot property is better, even if when concentration very low (200 μ g/mL), temperature remains to reach 42 DEG C.
Followed by the influence of experiment condition, as shown in Figure 2 B.It was found that electric current is bigger, heating is faster, especially works as experiment
When electric current is 40A, electric current rises very rapidly up to 70 DEG C in 3min, it will be apparent that.
Fig. 2 C are to inquire into whether Material cladding has an impact magnetic heating performance.It was found that H2O and PB temperature does not almost change, and
MnFe2O4 and PB/MnFe2O4 heatings are apparent, and 10min interior energies reach 42 DEG C or more, can achieve the effect that treatment.And it is compound
Afterwards, material temperature is slightly higher than single MnFe2O4, is primarily due to material particle size and becomes larger, and magnetic heating performance is by material grain
The influence of diameter, in a certain range, grain size is bigger, and temperature is higher.
Fig. 2 D are the infrared thermal imaging pictures of material, mainly by infrared thermography from macroscopic perspective reaction wood material temperature
The variation of degree, temperature is higher, and color is brighter.It was found that before and after magnetic treatment, changes in material is more apparent.
Fig. 2 E are the SAR value of material, when corresponding 3min, directly reflect the magnetic heating performance of material, and SAR is higher,
Magnetic heating performance is better, can reach nearly 1000W/g, is reported relative to some pertinent literatures, and numerical value is higher.
The magnetothermal effect of cell
From fig. 4, it can be seen that magnetic thermal effect is similar to the effect of Fig. 3 photo-thermal, after magnetic heat treatment, under the activity of cell
Drop, survival rate decline, and illustrate that material has magnetic thermal effect, convert the magnetic energy of absorption to thermal energy, to which cell be killed, and
This point is can be seen that from fluorescent staining effect, and material is after magnetic heats, and the activity of cell reduces even death, therefore cell is
It fluorescence is caught, a small amount of green fluorescence is only seen under fluorescence microscope, largely shows black, the side light magnetic of material
Thermal effect shows that this material can be used for the magnetic heat cure of cancer.
The hot photo-thermal superposition of magnetic of 4 material of embodiment uses
According to the photo-thermal of above-mentioned material and magnetic thermal result, the concentration of choosing 200mg/ml is detected (material at a temperature of this
Material all has temperature rise effect, especially photo-thermal), the hot photo-thermal platform of magnetic is built, then again under photo-thermal and the experiment condition of magnetic heat
The identical time is measured, while temperatur-timel curve is then drawn to photograph to record the variation of temperature with infrared thermal imaging.
Fig. 6 is that the hot photo-thermal of magnetic of material combines curve graph.Wherein the experiment condition of Fig. 6 A is 500KHz, 30A, finds this
Under part, rise 10 DEG C or so in temperature 10min, and under same concentration conditions, in comparison the heating of PTT becomes apparent from, have
16 DEG C or so, as shown in Figure 6B, and after being superimposed, the temperature difference has 20 DEG C or so, and heating is more obvious, illustrates the effect that MFH is superimposed with PTT
Fruit is more preferable, has further showed that feasibility and and effect that superposition uses.Next we select 200 μ g/mL materials to carry out
Magnetic heat adds the trial of photo-thermal, as shown in Figure 6 D, it is found that the temperature difference becomes apparent from, temperature can reach 70 DEG C or more, and the temperature difference is more than
30 DEG C, effect is very good.Finally find both be used in combination after, temperature rise becomes apparent from, the rate of climb faster, temperature higher,
Confirm be thermotherapy magnetic heat and the possibility that is used in combination of photo-thermal, this composite material of side light is in cancer thermotherapy field
Have extremely excellent potential.
The above, only present pre-ferred embodiments, therefore cannot limit the scope of implementation of the present invention according to this, i.e., according to
Equivalent changes and modifications made by the scope of the claims of the present invention and description all should still belong in the range of the present invention covers.
Claims (10)
1. Prussian blue-purposes of the Manganese Ferrite composite nano materials as physiotherapy agent, wherein the Prussian blue-iron
Sour manganese composite nano materials are PB-MnFe2O4。
2. purposes as described in claim 1, which is characterized in that the physiotherapy agent includes that photo-thermal therapy agent or magnetic heat are controlled
Treat agent.
3. purposes as described in claim 1, which is characterized in that the physiotherapy agent is photo-thermal/magnetic heat integration therapeutic agent.
4. the purposes as described in claims 1 or 2 or 3, which is characterized in that the Prussian blue-Manganese Ferrite composite Nano material
In material, PB and MnFe2O4Mass ratio is 1:1-5.
5. the purposes as described in claims 1 or 2 or 3, which is characterized in that the Prussian blue-Manganese Ferrite composite Nano material
In material, PB and MnFe2O4Mass ratio is 1:2-3.
6. the purposes as described in claims 1 or 2 or 3, which is characterized in that the described Prussian blue preparation method of alkalinity includes：
Take PB and MnFe2O4Sample, according to mass concentration 1：The ratio of 1-5 mixes, and it is 1-4 then to adjust solution ph, is stirred overnight
Afterwards, repeatedly Magnetic Isolation, cleaning until solution is clarified.
7. the purposes as described in claims 1 or 2 or 3, which is characterized in that the described Prussian blue preparation method of alkalinity includes：
Take PB and MnFe2O4Sample, according to mass concentration 1：The ratio of 2-3 mixes, and it is 1.5-2.5 then to adjust solution ph, stirred
After night, Magnetic Isolation, cleaning are until solution is clarified repeatedly.
8. a kind of physiotherapy agent contains Prussian blue-Manganese Ferrite composite nano materials.
9. a kind of physiotherapy agent as claimed in claim 8, which is characterized in that the physiotherapy agent includes photo-thermal therapy
Agent or magnetic heat cure agent.
10. a kind of physiotherapy agent as claimed in claim 8, which is characterized in that the physiotherapy agent is photo-thermal/magnetic heat
Combination therapy agent.
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