CN106618729B - A kind of device for tumor microenvironment variation in the RF therapy of dynamic observation in real time - Google Patents
A kind of device for tumor microenvironment variation in the RF therapy of dynamic observation in real time Download PDFInfo
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Abstract
The invention discloses a kind of devices for tumor microenvironment variation in the RF therapy of dynamic observation in real time.The device includes: the ridge skin wing form for the tumor microenvironment variation of dynamic observation in real time；For carrying out the radio-frequency coil of RF therapy；Attachment device for the ridge skin wing form and the radio-frequency coil to be detachably connected；For the ridge skin wing form and the radio-frequency coil to be separated, the non-conducting material that the radio-frequency coil and the attachment device separate.The present invention is by ridge skin wing form and the ingenious combination of RF therapy coil, guarantee the effective radio-frequency ablation in treatment of drug, observed during RF therapy simultaneously it is intuitive, easy to operate, can timely and accurately judge to study in therapeutic process tumor vessel and the case where normal blood vessels, to observe and studying radio frequency drug to tumor vascular blocking process and mechanism, effective and reliable means are provided to evaluate tumor vessel blocking drugs under antitumor especially action of radio.
Description
Technical field
The invention belongs to oncobiology fields, and in particular to one kind is micro- for tumour in the RF therapy of dynamic observation in real time
The device of environmental change.
Background technique
So far, cancer is still the public health problem of most serious in the world.The cancer in China or even world wide
Disease incidence and the death rate still constantly soaring, malignant tumour, which has become, leads to one of principal disease of human death.
Tumour is complicated multifactor, a multistage process, is genes of individuals unstability and gives birth to including the mankind
The coefficient results such as the environmental factor including mode living.Folkman in 1971 etc. propose solid tumor growth and transfer according to
Rely in tumor angiogenesis, further proposes to pass through angiogenesis inhibitor using Tumor Angiongesis as target spot
Inhibit or destroy Tumor Angiongesis, cuts off the new treatment of tumors of nutrients source and the cancer of " hungry to death " tumour.
There are two general orientation: anti-angiogenic agent (angiogenesis for tumor neovasculature research at present
Inhibitors, AIs) and vascular disrupting agents (vascular disrupting agents, VDAs).VDAs refers to that one kind can be selected
Selecting property destroys malignant tumour new vessels, to reach the drug for the treatment of cancer purpose.Its mechanism of action is to utilize malignant tumour
The difference of new vessels and normal blood vessels on pathologic, physiologic, selectively combines with malignant tumour new vessels, directly or
Make tumor neogenetic blood vessels that occlusion or damage occur indirectly, cause the ischemic necrosis of tumour cell, so that it is pernicious swollen to play treatment
The effect of tumor.The clinical test of some small molecule VDAs report have cardiovascular adverse effects for example hypertension, tachycardia, slowly
Property arrhythmia cordis, auricular fibrillation, myocardial infarction etc., these adverse reactions limit its being widely used clinically.Invention one
Kind novel effectively intuitive diagnosis detects the technological means of influence of this technology to tumor microcirculation, research and development for new drug and faces
Bed application is of great significance.For treating the RF therapy system of tumour, for swelling in the RF therapy of dynamic observation in real time
The device of tumor micro-environmental variation and using the device carry out RF therapy system do not have been reported that.
Summary of the invention
The device that tumor microenvironment changes in RF therapy is observed for dynamic in real time the object of the present invention is to provide a kind of,
It can be during the drugs combination RF therapy such as fullerene derivate and nano material to tumor locus side radio frequency using the device
Side observation is treated, easy to operate, visual observation window is stablized in the operation offer of lesser window and rigorous aseptic, straight in real time
Variation caused by the microcirculation over the course for the treatment of in ground observation tumour and normal tissue is seen, guarantees the auxiliary function for the treatment of radio frequency
It can stablize effectively, to need RF energy come the blood vessel blockings drugs such as the fullerene that assists in the treatment of and such new treatment method
The research of therapeutic mechanism provides strong and reliable means and foundation.
Provided by the present invention for the device of tumor microenvironment variation in the RF therapy of dynamic observation in real time, it includes:
The ridge skin wing form of tumor microenvironment variation for dynamic observation in real time；
For carrying out the radio-frequency coil of RF therapy；
Attachment device for the ridge skin wing form and the radio-frequency coil to be detachably connected；
For the ridge skin wing form and the radio-frequency coil to be separated, the radio-frequency coil and the attachment device every
The non-conducting material opened.
In above-mentioned device, conventional ridge skin wing form is can be used in the ridge skin wing form, at of the invention one
Or in multiple specific embodiments, the ridge skin wing form includes:
For fixing the form fixinig plate I and form fixinig plate II of the ridge skin wing；The form fixinig plate I and form are fixed
Piece II is to be detachably connected；At least one in the form fixinig plate I and the form fixinig plate II offers window；
Form slide for being fixed on the window.
In above-mentioned device, the interval between the ridge skin wing form and the radio-frequency coil can be 1~10 centimetre, tool
Body can be spaced described in the thickness control of the non-conducting material by being arranged.
In above-mentioned device, the ridge skin wing form is applicable in animal range concretely all types of rats and small
Mouse.The diameter of window can be 10~50 millimeters in the ridge skin wing form, concretely 27 millimeters, 35 millimeters or 46 millimeters.
The ridge skin wing form can be used for a variety of Observations Means such as optical microscopy, fluorescence microscope and magnetic resonance imaging.The back
The material of ridge skin wing form can be titanium alloy and nylon.
In above-mentioned device, described be detachably connected can be to be threadedly coupled, specifically can be by the ridge skin wing form
Screw and nut are attached, and radio-frequency coil accordingly punches.
In above-mentioned device, the non-conducting material concretely plastics are such as regarded the ridge skin wing using plastic sheet
Window and the coil separate, using plastic bushing by form screw and coil separate.The non-conducting material setting can be prevented
The only radiofrequency signal that interference radio-frequency coil is issued.
Invention further provides a kind of systems for carrying out RF therapy using above-mentioned apparatus, including radio frequency to fill
It sets, above-mentioned device and nano material for tumor microenvironment variation in the RF therapy of dynamic observation in real time；The radio frequency hair
The radiofrequency signal of generating apparatus transmitting is applied to tumor locus by the radio-frequency coil in described device, makes to be enriched in tumor vessel
Nano material occur movement destroy tumor vessel, treat tumour.
In above-mentioned system, the radio frequency generator that the radio frequency generator is known to the skilled person is adjustable
Save rf frequency, the radio-frequency power etc. of the radio frequency generator.The radio frequency generator is specific can include: one for emitting
The radio-frequency signal generator of radiofrequency signal out；One radiofrequency signal for frequency needed for amplifying the radiofrequency signal and integrating output
Signal amplifier.The system may also include the frequency spectrograph of the power density for detecting the radio-frequency coil.The frequency spectrum
Instrument can provide reference for oncotherapy condition.
In above-mentioned system, the nano material is to have the nano material of following property: (1) surface is hydrophily
And/or surface has electronegativity；(2) material has rigidity, so that it is not easily passed through tumor vessel cinclides hole and is stuck in described
Hole；(3) extraneous radiation energy can be absorbed and transform into thermal energy in inside, while the thermal energy being accumulated, so that described
Acute variation occurs for the morphosis of nano material.
The nano material can be water-soluble elemental metals, water-soluble nonmetal nano material or Water Soluble Compound nanometer
Material, concretely water-soluble fullerene nano particle, water-soluble metal fullerene nano particle, water-soluble precious metal nanometer
Grain, such as gold nano grain, Pt nanoparticle etc., water soluble oxidized iron nano-particle, water-soluble manganous ferrite nano particle are water-soluble
Ferrous acid cobalt nano-particle etc..In one or more specific embodiments of the invention, the nano material can be water solubility
Gd metal fullerene nano particle (such as metal fullerene hydroxylation derivative Gd@C82 (OH) n).
It is stuck in described hole in order to make the material not easily pass through tumor vessel cinclides hole, the grain of the nano material
Diameter can be 50~250nm, such as 100~200nm.
The nano material can exist in form of an aqueous solutions, the concentration of the aqueous solution of the nano material can for 0.1~
10mM, such as 5mM.
In above-mentioned system, the radio-frequency coil matches from different therapeutic agents；The transmitting of the radio-frequency coil is penetrated
Frequency parameter can be adjusted in following ranges: transmitting rf frequency is 10~300 megahertzs；Power is 0~100 milliwatt.It is described
Concretely 10 megahertzs, 100 megahertzs, 200 megahertzs or 300 megahertzs of rf frequency of transmitting.The radio-frequency coil hair
Penetrate radio-frequency power concretely 1 milliwatt, 10 milliwatts or 15 milliwatts.
In above-mentioned system, the system also includes imaging system, concretely optical microscopy or fluorescence microscope.It is real
When dynamic observation any means record of photo or video acquisition can be used；The photo or video acquisition information can be by any
Imaging system obtain the blood vessel microenvironment of normal tissue can be used when observation tumor microenvironment and be observed as control；It is described
The variation of the blood vessel microenvironment of tumour and normal tissue includes the change of tumor vessel number of modalities, blood circulation and tumor tissues
Change, concretely angiorrhoxis, bleeding, Oligemia or tissue color variation.
The invention has the following beneficial effects:
The present invention by animal ridge skin wing form model and the ingenious combination of RF therapy coil, design different therapeutic frequencies and
The radio-frequency coil to match with the ridge skin wing form of unlike material guarantees that the drugs such as fullerene and specific nano material have
The radio-frequency ablation in treatment of effect, at the same observe during RF therapy it is intuitive, easy to operate, can timely and accurately judge to study
Tumor vessel and the case where normal blood vessels in therapeutic process, for observe and study radio frequency drug to tumor vascular blocking process and
Mechanism provides effective and reliable means to evaluate tumor vessel blocking drugs under antitumor especially action of radio.
Detailed description of the invention
Fig. 1 is mouse ridge skin wing form illustraton of model in embodiment.
Fig. 2 be embodiment in mouse ridge skin wing form model in conjunction with radio-frequency coil after device figure.It is each to mark in figure
It is as follows: 1 ridge skin wing form, 2 radio-frequency coils, 3 screws.
Fig. 3 is to observe tumor microenvironment device figure in mouse ridge skin wing form in embodiment under optical microscopy.
Fig. 4 is mouse tumor blood vessel microenvironment image under treatment times different in embodiment.
Fig. 5 is mouse Normal tissue vascular microenvironment image under treatment times different in embodiment.
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Embodiment 1, the device changed for tumor microenvironment in the RF therapy of dynamic observation in real time
A kind of device (Fig. 2 institute for tumor microenvironment variation in the RF therapy of dynamic observation in real time provided by the invention
Show), it includes the ridge skin wing form (shown in Fig. 1) for the tumor microenvironment variation of dynamic observation in real time, ridge skin wing view
Window includes the form fixinig plate I and form fixinig plate II for fixing the ridge skin wing；In form fixinig plate I and form fixinig plate II
Between offer window；And the form slide for being fixed on window；
For carrying out the radio-frequency coil of RF therapy；
Screw and nut for form fixinig plate I, form fixinig plate II and radio-frequency coil to be threadedly coupled；
For ridge skin wing form and radio-frequency coil to be separated, the plastic spacer and plastic sheath that radio-frequency coil and nut separate
Pipe.The thickness of plastic spacer makes radio-frequency coil and ridge skin wing form model window be separated by 1~10 centimetre.
Apparatus of the present invention when in use,
Firstly, pull-up skin of back, ridge skin wing form is fixed on the skin of tumor locus, concrete operations are as follows:
Form fixinig plate I is sewn on skin window fixed area (tumor locus), the skin of window portion is cut, is put into form glass
Piece.Form fixinig plate II is placed on above, is aligned with the fixinig plate I that the back side fixes, is sutured with operation sewing needle.
Secondly, making the form back side upward, ridge skin wing form nut is unscrewed, by plastic sheath pipe sleeve on the screw of form,
By radio-frequency coil on the screw at the form back side, plastic spacer, tightening nuts are clamped.
Embodiment 2, a kind of system of tumor treatment that can dynamically observe tumor microenvironment variation in RF therapy in real time
A kind of oncotherapy system for tumor microenvironment variation in the RF therapy of dynamic observation in real time provided by the invention
System, it includes radio frequency generator, the dress for tumor microenvironment variation in the RF therapy of dynamic observation in real time in embodiment 1
It sets and the hydroxylation derivative of Gd@C82 (Gd@C82 (OH) n)；Radio frequency generator includes: for launching radiofrequency signal
Radio-frequency signal generator；The signal amplifier of radiofrequency signal for frequency needed for amplifying the radiofrequency signal and integrating output.
The system may also include the frequency spectrograph of the power density for detecting radio-frequency coil and for observing tumor microenvironment variation
Optical microscopy or fluorescence microscope.The radiofrequency signal that radio frequency generator is launched is applied to by the radio-frequency coil in device
Tumor locus, the hydroxylation derivative (Gd@C82 (OH) n) for making to be enriched in the Gd@C82 in tumor vessel occur movement destruction and swell
Tumor blood vessel treats tumour.
The hydroxylation derivative (Gd@C82 (OH) n) of drug Gd@C82 for RF therapy tumour, specifically can be according to text
Offer Mingming Zhen, Chunying Shu, Jie Li, et al, Science China materials.2015,58,799
Disclosed in method be prepared, partial size 100-200nm.
Embodiment 3 carries out tumour in RF therapy and the RF therapy of dynamic observation in real time using apparatus of the present invention and system
Micro-environmental variation
One, ridge skin wing form is established
(1) pre-operative preparation
Prepare outfit and reagent: titanium alloy ridge skin wing form, operating scissors, ophthalmic tweezers, suture, haemostatic clamp,
Coverslip, cotton swab, sterile saline, antibiotic, 75% alcohol and Iodophor etc., preoperative ultraviolet-sterilization disappears together with super-clean bench
Poison.
(2) transplanting ridge skin wing form operation
Experiment mice is injected intraperitoneally in yellow Jackets, is anaesthetized.It has been anaesthetized substantially to mouse, has dipped iodine with cotton rod
Volt smears disinfection back baring skin.Pull-up back of mice skin, pull-up back of mice skin, alignment light selection have abundant
Vascular distribution as skin window fixed area.The form fixinig plate for clamping mouse skin is sutured with swaged needle.Allow mouse side
It lies, upward, can pad instrument below keeps window area holding horizontal for the side for making not fixinig plate, is carried on the back with the careful pull-up of tweezers
Portion's skin starts to cut skin, according to the big light maintenance skin openings of the form slide of loading, openings of sizes is made to be slightly less than slide.In skin
There is bubble when form slide to prevent placing in sterile saline containing antibiotic in skin opening drop, and form slide is small
The heart is embedded in skin openings, is sure not bubble occur.Another form fixinig plate is placed on above, the fixinig plate pair fixed with the back side
Together, it is sutured with operation sewing needle.Postoperative mouse can identity it is weak and be easy infection, guarantee the whole of its diet and living environment
It is clean.Help it warming with fritter gauze package mouse at the first time after operation.In terms of diet, guarantee small with egg yolk feeding daily
Mouse, drinking water is the sterile water containing antibiotic, to prevent its wound inflammation.Padding is also required to replacement in every two days once, hard daily
Hold observation mouse.
(3) ridge skin wing form model inoculated tumour
Normal mouse H22 ascites is taken, carries out cell count after blowing and beating uniformly.After being computed, cell density is every milliliter 1
×107-7.5×107A cell draws 10 microlitres with micro-sampling pin, mouse is lain on one's side (the non-slide of form lateral on) place
In left hand, the right hand, which grasps micro-sampling pin and is close to skin outside window area, is slowly pushed into syringe needle, allow syringe needle enter skin and
Gap between form slide, it should be noted that not wear out skin.Cell is slowly injected to the center of observation area, while being revolved gradually
Rotating the needle head is pulled out from gap to be come, and injection is completed.Usual H22 tumour time of occurrence is about postoperative 4-5 days, grows pale pink
It can be observed after tumour, as shown in Figure 1.After H22 tumour is obviously grown, with 2% isoflurane oxygen mixed gas pair
Mouse is anaesthetized.Form slide is cleaned with wine clear cotton swab.
Two, ridge skin wing form and radio-frequency coil are connected
The narcosis for keeping experiment mice stable with 2% isoflurane oxygen mixed gas.Mouse is lain on one's side, form back side court
On, gently unscrew the ridge skin wing three nuts of form, with tweezers by plastic sheath pipe sleeve on the screw of form, gently will be corresponding
200 megahertzs of radio-frequency coils clamp three plastic spacers with tweezers on three screws at the form back side respectively, keep form
It is parallel to desktop, distinguishes gently tightening nuts, as shown in Figure 2.
Three, tumor microenvironment when dynamic observes RF therapy in real time under RF therapy
(1) fixation is installed on optical microscopy dressing table
The narcosis for keeping experiment mice stable with 2% isoflurane oxygen mixed gas.Upward by mouse form window
It lies on one's side, keeps parallel with desktop, the radio-frequency coil having connected is fixed with board device with full dog point socket set, is put in after tightening
Optical microscopy sample stage simultaneously clamps board device with fixing clamp, guarantees that form window plane is parallel with sample stage at this time, such as Fig. 3
It is shown.
(2) apply RF therapy
Radio-frequency coil interface is connect with radio frequency generator, radio frequency generator parameter is set, rf frequency is 200,000,000
Hertz, whether radio-frequency power is 10 milliwatts, and be consistent with actual radio frequency power at frequency spectrograph detection form window.
Hydroxylation derivative (Gd@C82 (OH) n) (partial size 100- of the Gd@C82 of 5mM is administered in mouse mainline
Aqueous solution 200nm), dosage 15mg/kg after 15 minutes, apply RF therapy 1 hour.
(3) acquisition picture is observed in real time
The narcosis for keeping experiment mice stable with 2% isoflurane oxygen mixed gas, it is fixed in parallel micro-
Mirror sample stage carries out observation of taking pictures to tumor vessel in window and surrounding normal blood vessel with 4 times of camera lenses of optical microscopy.When taking pictures
Between for treatment before, treatment 30 minutes and 1 hour.
(4) analysis and arrangement data
The brightness and contrast for adjusting the photo of different time points different parts shooting, compares same position different time
Variation.
As shown in figure 4, treatment before, treatment 30 minutes and 60 minutes when the tumor vascular change procedure of H22.Before treatment, swell
Tumor rich blood vessel complete display；When treating 30 minutes, there is blutpunkte around tumor vessel；When treating 60 minutes, tumor vessel
Surrounding blutpunkte area becomes larger, and blood vessel is broken.Illustrate Gd C82 amino acid derivatization drug under this radio frequency conditions,
Play the role of blocking H22 tumor vascular, works when treating 30 minutes and destroy H22 tumor microenvironment, to reach brokenly
The bad growth for inhibiting tumour, demonstrates this drug and blocks tumor vascular cure mechanism at radio frequencies.
As shown in figure 5, treatment before, treatment 30 minutes and 60 minutes when Normal tissue vascular change procedure.Before the treatment
The orderly complete display of tumour surrounding normal blood vessel, with the progress for the treatment of, in tumor vessel by drugs block therapeutic effect
Meanwhile the blood vessel of normal tissue does not generate variation, is not destroyed or blocks.Illustrate that this blocks tumor vascular treatment pair
The blood vessel of normal tissue does not generate destruction, so that normal tissue does not generate toxicity or destruction, in treatment neoplastic process
Normal tissue organ has no toxic side effect, and demonstrating this drug, targeting therapy on tumor acts at radio frequencies.
Claims (9)
1. a kind of device for tumor microenvironment variation in the RF therapy of dynamic observation in real time, it is characterised in that: it includes: use
In the ridge skin wing form of real-time dynamic observation tumor microenvironment variation；
The ridge skin wing form includes: the form fixinig plate I and form fixinig plate II for fixing the ridge skin wing；The form
Fixinig plate I and form fixinig plate II are to be detachably connected；At least one in the form fixinig plate I and the form fixinig plate II
Offer window；Form slide for being fixed on the window；
For carrying out the radio-frequency coil of RF therapy；
Attachment device for the ridge skin wing form and the radio-frequency coil to be detachably connected；
1~10 centimetre is divided between the ridge skin wing form and the radio-frequency coil；
For separating the ridge skin wing form and the radio-frequency coil, the radio-frequency coil and the attachment device separate
Non-conducting material.
2. the apparatus according to claim 1, it is characterised in that: the diameter of window is 10~50 in the ridge skin wing form
Millimeter.
3. device according to claim 1 or 2, it is characterised in that: the non-conducting material is plastics.
4. a kind of system for carrying out RF therapy using the device of any of claims 1-3, it includes radio frequency
Device, the device of any of claims 1-3 for tumor microenvironment variation in the RF therapy of dynamic observation in real time
And nano material；The radiofrequency signal of the radio frequency generator transmitting is applied to tumour by the radio-frequency coil in described device
Position makes the nano material being enriched in tumor vessel that movement occur and destroys tumor vessel, treats tumour.
5. system according to claim 4, it is characterised in that: the nano material be water-soluble precious metal nano particle,
Water-soluble fullerene nano particle, water-soluble metal fullerene nano particle, water soluble oxidized iron nano-particle, water-soluble ferrous acid
Any one of manganese nano particle and water-soluble ferrous acid cobalt nano-particle.
6. system according to claim 4 or 5, it is characterised in that: the partial size of the nano material is 50~250nm.
7. system according to claim 4 or 5, it is characterised in that: the radio frequency generator includes: for emitting outgoing
The radio-frequency signal generator of frequency signal；The signal of radiofrequency signal for frequency needed for amplifying the radiofrequency signal and integrating output
Amplifier.
8. system according to claim 4 or 5, it is characterised in that: the transmitting radio frequency parameter of the radio-frequency coil can be
Adjust in following ranges: transmitting rf frequency is 10~300 megahertzs；Power is 0~100 milliwatt.
9. system according to claim 4 or 5, it is characterised in that: the system also includes imaging systems, swollen for observing
Tumor micro-environmental variation.
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| CN108714046A (en) * | 2018-05-03 | 2018-10-30 | 上海理工大学 | A kind of lumen organization's welding system and control method based on RF energy |
| CN109157280A (en) * | 2018-08-10 | 2019-01-08 | 重庆大学 | Irreversible electroporated tissue ablation effect dynamic realtime assessment equipment |
| CN112656535A (en) * | 2021-01-04 | 2021-04-16 | 中国人民解放军空军军医大学 | Optical fiber connection photobiological treatment window for rats |
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| CN102119042A (en) * | 2008-08-14 | 2011-07-06 | 盎可撒姆股份有限公司 | Portable radiofrequency hyperthermia device with flexible treatment electrode for electric field capacitive coupled energy transfer |
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