CN101829326A - Use of microbubbles with high-intensity cavitation generated by combining ultrasonic in preparation of anti-tumor neovascularization medicaments - Google Patents
Use of microbubbles with high-intensity cavitation generated by combining ultrasonic in preparation of anti-tumor neovascularization medicaments Download PDFInfo
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Abstract
The invention relates to use of microbubbles with high-intensity cavitation generated by combining ultrasonic in preparation of anti-tumor neovascularization medicaments. The microbubbles are high-intensity cavitation effect microbubbles generating mechanical damage under the action of pulse-type high-peak negative pressure ultrasonic energy; and the ultrasonic energy has higher peak sound pressure and lower transmitting duty cycle. The generated high-intensity cavitation effect can generate obvious tumor microcirculation blocking effect on the tumor neovascularization, repeatedly perform deprivation therapy for further generating tumor growth inhibition, and reduce the transfer rate of malignant tumors. The novel treatment use of the microbubbles is applied to the treatment of various solid malignant tumors and angiogenesis pathological changes and is brand new use of the microbubble with application prospect.
Description
Technical field
The present invention relates to the new purposes of the antineoplaston of microvesicle under ultrasonic action, can be applied to the treatment of malignant tumor, belong to the technical field of atraumatic ultrasonic therapeutic.
Technical background
Malignant tumor is the disease of serious threat human life health, and its treatment is the difficult point and the focus of medical circle always.At present, although the basic skills of treatment cancer has therapies such as operation, radiotherapy, chemotherapy and biological immune, but these treatment meanss often can not satisfy patient's treatment requirement, and operation, radiotherapy, chemotherapy are traumatic big, easily produce serious adverse or drug resistance, not only risk is big, and has reduced patient's life quality.Tumors such as primary hepatocarcinoma are all insensitive to radiation and chemotherapy, and existing naturopathy has necessarily traumatic again, should not repeatedly use repeatedly, so it is very necessary to seek a kind of tumor therapeuticing method of atraumatic.
The generation of entity tumor, development, invasion and attack and transfer rely on tumor-blood-vessel growth (Tumorangiogenesis)
[1]So, be that the tumor " dormancy therapy " of therapeutic purposes is in recent years continue a kind of new oncotherapy pattern of performing the operation, growing up behind the chemicotherapy with inhibition, thromboembolism or blocking-up tumor neogenetic blood vessels (mainly referring to blood capillary)
[2-5]At present, get involved the tumor tremulous pulse that the blood vessel embolism therapy can only the big caliber of thromboembolism, powerless to the collateral circulation of extensive generation; Use chemistry or biological preparation antineoplastic vascular to generate treatment and can directly act on tumor neovasculature target spot, but specificity is not high, curative effect is undesirable.Tumor naturopathy method comprises that various interventions are hot in naturely melted, freezing, electrochemistry etc., and all by puncture or operation direct killing tumor tissues, but side effect is also bigger.
Utilize ultrasonic energy non-invasive treatment tumor is the target that industry is pursued always.Although with the heat effect ablated tumor is high intensity focused ultrasound (the High intensity focused ultrasound of representative, HIFU) development at home and abroad rapidly, part hepatocarcinoma, osteosarcoma and hysteromyoma treatment have been successfully applied to, but the huge costliness of HIFU equipment, complicated operation, stopping at the rib intestinal, conduction of heat is inhomogeneous etc. under the situation may burned skin, rib and around vitals, produce serious adverse, therapeutic effect is unsatisfactory.With regard to its reason, mainly be because it utilizes the heat effect ablated tumor, need assemble up to 5000～20000W/cm in the target area
2Ultrasonic energy, produce 65～100 ℃ of high temperature in the several seconds, be damaged to normal structure in the window unavoidably.
Ultrasonic cavitation effect (cavitation, annotate: cavitation effect is meant that microbubble produces a series of dynamic processes such as concussion, expansion, contraction and implosion in the liquid under ultrasonication, follows multiple energy release behaviors such as transient high temperature, high pressure, shock wave, discharge and microjet)
[6]It is another kind of ultransonic Main physical effect outside the heat effect.When launching with pulsed, the cavitation heat production can not be accumulated when ultrasonic, and its heat effect is small, discharges and mainly show as mechanical energy (shock wave, microjet etc.).
In recent years, the basic research of the enhanced ultrasonic cavitation effect of microcapsular ultrasound contrast agent in atraumatic ultrasonic therapeutic field is very active, but most of correlational studyes relate generally to drug release and gene transfection, and be few to its cavitation mechanical injuries effect study.The generation and the intensity effect factor of biological intravital sound cavitation effect are more, but most important factor is ultrasonic peak negative pressure intensity, generally significant cavitation effect will take place in peak negative pressure during greater than 0.6MPa
[7]American-European only have a few seminars such as Hwang, Cain, Miller and Liu Zheng to carry out the discussion in this field
[8-14], their research only relates to the cavitation physical damnification of normal blood vessels wall and the ultrasonic cavitation of in vitro tissue specimen melts, and the Therapeutic Method of employing also is not quite similar.Hwang etc.
[8]Employing damages the damage that has realized rabbit ear edge vein blood vessel endothelium through intravenous injection microvesicle associating focused ultrasonic cavitation, but needs the low dose of thrombin of injection in the local tube chamber, just may cause blood vessel embolism; Cain
[9,10]Cavitation research in do not adopt the intravenous injection microvesicle, but utilize in the ultrasonic excitation organism self potential microvesicle generation cavitation to carry out ablation of tissue, employing be to exsomatize tissue such as normal myocardium as object of study; The inductive ultrasonic cavitation of main research microvesicle such as Miller does not all adopt tumor neogenetic blood vessels as melting the destruction object for normal myocardium or the microvascular cavitation damage side effect of kidney and other organs in their the cavitation research.That domestic Wu Wei etc. [11] adopt is low frequency (20-50KHz) low-power (1-100W/cm
2) the ultrasonic in combination microvesicle carries out the blood capillary thromboembolism, not only 300KHz-3.0MHz, the peak negative pressure with this project demand is different fully greater than the pulsating ultrasound of 2.0MPa, and experimental result in should research document, through repeating not have thromboembolism blocking-up blood capillary effect repeatedly.Though the diagnostic ultrasound peak negative pressure also can reach more than the 2MPa, its pulse width is extremely lacked (being about 1 cycle), can not induce high-intensity ultrasonic cavitation effect.
Summary of the invention
The purpose of this invention is to provide microvesicle and be used for the purposes of combining ultrasonic generation high-intensity cavitation as anti-tumor neovascularization medicaments in preparation.
The microvesicle that the present invention relates generally to is the microvesicle that produces the high-intensity cavitation effect of mechanical destruction under the ultrasonic energy effect of a kind of pulsed, peak value negative pressure; Described ultrasonic energy has higher peak sound pressure and lower emission dutycycle.The described high-intensity cavitation effect that produces can produce significant tumor microcirculation blocking effect to tumor neogenetic blood vessels, and blocking treatment further produces the tumor growth inhibition repeatedly, and reduces the rate of transform of malignant tumor.
Tumor refers to various malignant entity tumors here, for example hepatocarcinoma, breast carcinoma etc., and (perhaps claim angiogenesis, Angiogenesis) be the common trait of malignant solid tumor to tumor neogenetic blood vessels.Described tumor neogenetic blood vessels is meant that the new vessels of various malignant entity tumors (perhaps claims angiogenesis, Angiogenesis), comprises small tremulous pulse, small vein, blood capillary and mimicry blood vessel.
That microvesicle is meant conventional extensive stockization or be about to commercial acoustic contrast agent microvesicle, the ultrasonic contrast potentiation of microvesicle goes through in clinical diagnosis.Comprise Novi (
),
, Optison, Imagent, perfluoropropane human albumin microsphere and perfluor show; Microvesicle adopts the administering mode through peripheral intravenous injection, microvesicle injection using dosage but be not limited to 0.01～1.0 milliliter of scope of per kilogram of body weight.
Under the ultrasonic energy effect of the present invention's design, high-intensity sound cavitation effect can take place in the microvesicle in the blood circulation, at first can cause significant tumor neogenetic blood capillary mechanical destruction, hemorrhage and hematoma formation, and then generation tumor circulation (blood perfusion) blocking effect, on this basis, blocking-up further produces tumor growth inhibition and rate of transform decline repeatedly.Ultrasonic energy of the present invention is meant the ultrasonic pulse form of energy (Fig. 1) of a kind of pulsed or intermittent pulse type, high peaks acoustic pressure, lower emission dutycycle.The frequency range of its emission is at 300KHz-3.0MHz, and each pulsating ultrasound emitting treatment time, ultrasound emission can be continuous impulse formula or intermittent pulse type more than 30 seconds, and intermittently (time-out) time is not waited from 2-20 second; Ultrasonic peak negative pressure is in the 2-15MPa scope; Each pulse width of ultrasound emission is at 20-2000 periodic regime; Changeableization of ultrasound emission dutycycle is between 0.01-10%.Thereby average sound intensity (I
SPTA) be lower than 3W/cm more
2, heat effect is little.For example: the parameters,acoustic that this research is used is ultrasound emission frequency 1.2MHz, peak sound pressure 4.6MPa, 500 cycles of pulse width, pulse recurrence frequency 20Hz, intermittent pulse type ultrasound emission (launching 6 seconds/intermittently 6 seconds), actual duty cycle 0.5%, average sound intensity is (I only
SPTA) 0.89W/cm
2
The generation and the intensity effect factor of biological intravital sound cavitation effect are more, but most important factor is ultrasonic peak negative pressure intensity and microbubble concentration.Known significant cavitation effect will take place in peak negative pressure during greater than 0.6MPa
[7], but excite microvesicle can produce more intensive cavitation effect with the above ultrasonic pulse of 2.0MPa, never see the research report that the ultrasonic energy excitation acoustic contrast agent microvesicle that uses above parameters,acoustic is used to destroy the treatment of blocking-up tumor neogenetic blood vessels in the document.
Description of drawings
Fig. 1. intermittent pulse type, the peak value negative pressure ultrasound form of energy ideograph of the present invention's design.
Fig. 2. rabbit VX
2Subcutaneous transplantation tumor ultrasonic contrast time-intensity shows that tumor blood is for abundant unusually.
Fig. 3. the subcutaneous VX2 tumor of New Zealand white rabbit, the ultransonic therapeutic effect of employing 1.2MHz, 3.2MPa.A: the ultrasonic contrast tumor vessel strengthens significantly before the treatment; B: the tumor blood flow perfusion of treatment back disappears, and is negativity and develops; C, D: the azovan blue perfusion shows tangent plane substantially after the treatment, and peripheral muscle indigo plant is dyed (white arrow), and tumor is unstained (deceiving arrow).
Fig. 4. ultrasonic cavitation treatment blocking-up rat Walker256 Subcutaneous tumor rheography, a treatment frequency 300KHz, peak negative pressure 3.2MPa.A: tumor two-dimensional ultrasound image; B: tumor colorful blood image; C: the blood perfusion (white arrow) that treatment pre-neoplastic district is abundant; D: (post0) tumor area bloodstream blocking at once after the treatment, filling defect (white arrow) fully.
Fig. 5. ultrasonic cavitation blocking-up rat Walker 256 Subcutaneous tumor rheographies, a treatment frequency 1.2MHz, peak negative pressure 1.6MPa level.A: blood perfusion is abundant before the oncotherapy; B: still have a certain amount of blood flow to enter after the treatment, barrier effect is not thorough.
Fig. 6. peak negative pressure 2.0MPa level, VX2 tumor blood flow perfusion different time points ultrasonic contrast figure under the ultrasonic in combination microvesicle cavitation blocking-up rabbit.A: treatment pre-neoplastic blood perfusion abundant (arrow); B, C: treatment back 0-30min, tumor area negativity development (arrow); D: a small amount of recover (arrow) of treatment back 60min tumor perfusion beginning.
Fig. 7. in ultrasonic therapeutic head frequency 1.2MHz, peak negative pressure 4.6MPa level, cavitation treatment blocking-up rat Walker256 Subcutaneous tumor rheography, its barrier effect is good, can continue more than 24 hours.A: tumor two-dimensional ultrasound and color blood-stream; B: ultrasonic contrast is seen tumor blood flow perfusion abundant (white arrow) before the treatment; C: treatment back 30min radiography, tumor area bloodstream blocking, filling defect fully; D: treat back 24 hours radiographies, the tumor area blood flow is still blocked filling defect fully.
Fig. 8. the pathological observation after the tumor neogenetic blood vessels ultrasonic cavitation treatment, visible blood capillary is broken, hematoma thrombosis (black arrow), phenomenons such as blood capillary blocking-up.
Each experimental group tumor growth vary in diameter curve chart of Fig. 9, ultrasonic microbubble group diameter of tumor are starkly lower than two matched groups in addition.
Each experimental group neoplasm metastasis scoring statistic histogram of Figure 10, microcapsular ultrasound treatment group (US+MB) rate of transform significantly is lower than simple ultrasonic group (US) and the false group (SHAM) of shining.
The specific embodiment
(1) acoustic measurement
Its parameters,acoustic of ultrasonic therapeutic instrument comprises supersonic frequency, peak negative pressure, pulse width and dutycycle etc., is as the criterion with hydrophone method measurement result without exception.After the sonicator signal sent, received signal was received by the needle-like hydrophone (NTR1000, the U.S.) at distance transmitting transducer 3cm place, amplified the back by digital oscilloscope (Agilent 55310, the U.S.) computer sampling analysis through preamplifier (NTR, the U.S.).
(2) the treatment experiment conventional method summary of tumor blood capillary blocking-up
At first, obtain body surface locating information, sound window information and the tumor neogenetic blood vessels blood perfusion information of tumor, the locating information that provides for microcapsular ultrasound cavitation blocking-up tumor neogenetic blood vessels by diagnostic imaging modes such as ultrasonic, CT or MR.Then, treating through intravenous injection under the situation of described dosage microvesicle, the pulsating ultrasound irradiation treatment is carried out in the tumor-localizing target area, destroy the high concentration microvesicle that is in the tumor neogenetic blood vessels, mechanical energy forms such as ultrasonic cavitation release of shock wave and microjet in microcapsular ultrasound cavitation mode.The batch (-type) ultrasound emission is that microvesicle pours into and wins TPER, when making ultrasonic therapeutic, is in high concentration microvesicle perfusion state (Fig. 2) in the tumor vessel all the time.After treatment was finished, again by ultrasonic contrast, perhaps CE such as CT, MR was judged tumor microcirculation barrier effect.
(3) ultrasonic cavitation treatment actual parameter screening
In this research, adopted 4 of the sonicators of the different parameters,acoustics of development voluntarily and " the fat fluorine shows " acoustic contrast agent microvesicle through intravenous injection." the fat fluorine shows ", acoustic contrast agent was a kind of lipid ultrasonic contrast agent of standard, and core gas is perfluoropropane, mean diameter 2 μ m, and wherein 98% less than 8 μ m, and concentration is about 6
-9 * 10
10/ ml, the injected dose per kilogram of body weight is 0.1 milliliter in this research.
Aspect the ultrasonic therapeutic head tranmitting frequency, successively adopt subcutaneous VX2 tumor of New Zealand white rabbit and rat Walker256 Subcutaneous tumor as the tumor experiment model, tested 300KHz, 0.8MHz, 1.2MHz and four ultrasonic therapeutic head frequencies of 3.0MHz, as a result four kinds of frequencies at peak sound pressure greater than 2.0MPa, pulse width is greater than under 20 the situation, all produced significant tumor neogenetic blood vessels blocking effect (so application 300KHz-3.0MHz frequency band claim) with upper frequency, its barrier effect all confirms (Fig. 3-4) by azovan blue circumfusion damaged (not blue dying), but barrier effect preferably the supersonic frequency section be 0.8MHz, 1.2MHz (Fig. 3).
Aspect peak negative pressure, successively adopt subcutaneous VX2 tumor of New Zealand white rabbit and rat Walker256 Subcutaneous tumor as the tumor experiment model, under tranmitting frequency 1.2MHz situation, peak negative pressure levels such as 1.0MPa, 1.6MPa, 2.0MPa, 3.2MPa, 4.6MPa and 15MPa have been tested, barrier effect below 2.0MPa is remarkable (Fig. 5) not, and the above ultrasonic cavitation of 2.0MPa has all produced tangible tumor neogenetic blood vessels blocking effect in testing.Although the cavitation bloodstream blocking effect of peak negative pressure 2.0MPa level is better at once, blocking-up back restoration of blood flow is very fast, promptly slowly begins after 1 hour (Fig. 6).The therapeutic effect of peak negative pressure 4.6-15MPa level is better, and its barrier effect can continue (Fig. 7) more than 24 hours.So therapeutic effect is more remarkable with the rising of peak negative pressure, the persistent period is longer.
Aspect ultrasonic pulse width (Pulse length), same priority adopts the setting of each ultrasonic pulse 10,15,20,50,100,500,1000 and 2000, pulse width does not have obvious barrier effect in the time of 10 and 15 as a result, occurs tangible barrier effect more than 20.This may be because pulse width at 20 below cycle of oscillation, microvesicle does not also enter effective inertia cavitation resonant condition.
Ultrasound emission can be continuous impulse formula or intermittent pulse type, and intermittently the infusion time again after being destroyed by cavitation on the microvesicle in the tumor vessel is decided (time-out) time, does not generally wait second from 2-20.Each ultrasonic therapeutic total time should be more than 30 seconds, are lower than this time can cause the blocking-up of tumor blood capillary not exclusively.
(4) New Zealand white rabbit VX2 tumor blood capillary blocking experiment
The small-sized pulsed focused ultrasound therapy instrument that experiment adopts this project to develop voluntarily, a treatment frequency 1.0MHz, peak negative pressure 4.6MPa, pulse width 500, pulse recurrence frequency 20Hz, 6 seconds per treatment times, intermittently 6 seconds, actual duty cycle 0.5%, average sound intensity (I
SPAT) 0.89W/cm
2Microvesicle adopts " the fat fluorine shows " lipid microbubble, and core gas is perfluoropropane, the outward appearance curdy that is creamy white, and mean diameter 2 μ m, wherein 98% less than 8 μ m, and microbubble concentration is about 6
-9 * 10
10/ ml.
The blood perfusion that adopts ultrasonic contrast to estimate tumor changes.
Concrete experimentation is as follows: New Zealand white rabbit lotus VX2 Subcutaneous tumor rabbit is divided into simple microvesicle matched group, simple ultrasonic matched group and microcapsular ultrasound experimental group at random.The vertical irradiation tumor area 5min of treatment head is adopted in ultrasonic therapeutic.Each group is carried out conventional ultrasonic examination and ultrasonic contrast inspection to tumor respectively before and after treatment.In addition, the capable ultrasonic contrast of microcapsular ultrasound experimental group 30min, 60min behind irradiation is checked.VX2 tumor ultrasonic contrast blood is for abundant before and after simple microvesicle group and the simple ultrasonic irradiation group treatment, radiography GTG value difference different not significantly (table 1, P＞0.05); Before the treatment of microcapsular ultrasound experimental group, VX2 tumor imaging blood is for enriching average GTG value (Grayscale value, GSV) 67.8 ± 13.3; At once ultrasonic contrast after the treatment, tumor area vision blood perfusion disappears, and is negativity and develops, and GSV reduces to 29.7 ± 20.1, and being does not have the basic GTG value of enhancing, and ultrasonic in combination microvesicle treatment group tumor imaging GSV significantly is lower than matched group (P＜0.01); After 30-60 minute, GSV is 34.3 ± 20.7
-31.3 ± 19.9, in each group, tumor periphery normal structure blood perfusion is not found significant change.
The ultrasonic contrast average GTG value of peak strength (GSV) contrast before and after each group treatment of table 1
Annotate: relatively preceding with treatment,
*P＞0.05;
#P＜0.01
Because tumor neogenetic blood vessels has congenital developmental defects such as tube wall weakness, permeability height, shortage elastic fibers layer, originally discover, the mechanism of ultrasonic cavitation therapeutic modality blocking-up tumor tiny blood vessels mainly be the tumor blood capillary by mechanical destruction, tube wall break, hemorrhage and hematoma thrombosis etc. (Fig. 8).Normal blood capillary or diameter 2mm above than trunk, tube wall physically well develops, vertebration, thickness of pipe wall is and tough and tensile, generally this therapeutic modality is small to its influence, tests to find no remarkable blocking-up normal blood vessels phenomenon.In addition, the ultrasound emission sound intensity of this pulsating ultrasound therapeutic modality is the 1/2000-5000 of high intensity focused ultrasound, how at 0.5-3W/cm
2In, ultrasonic cavitation mainly is limited in the lumen of vessels again, thus small for the biological effect of trunk and the generation of other internal organs, thus avoided the side effect generation.
Acoustic contrast agent is a kind of microbubble suspension, and microvesicle on average between particle diameter 2-4 micron, significantly less than the erythrocyte particle diameter, can not see through capillary wall and enter interstice, is a kind of blood pond developing agent.Thereby, the nonvisualized zone of contrast agent, blood formed elements such as erythrocyte more can't enter.
(5) tumor growth suppresses and the metastasis inhibition experiment specific embodiment
The level diagnosis program, at first by microcapsular ultrasound contrast agent (as: sound Novi etc.) through the intravenous injection diagnostic dose, tumor is carried out the ultrasonic contrast diagnosis, obtain body surface locating information, sound window information and the tumor neogenetic blood vessels blood perfusion information of tumor, for ultrasonic cavitation blocking-up tumor neogenetic blood vessels provides locating information.
The locating therapy program then, under the situation of the described therapeutic dose microvesicle of intravenous injection, is carried out the pulsating ultrasound irradiation treatment to institute's locating area.The small-sized pulsed focused ultrasound therapy instrument that experiment adopts this project to develop voluntarily, a treatment frequency 1.0MHz, peak negative pressure 4.6MPa, pulse width 500, pulse recurrence frequency 20Hz, 6 seconds per treatment times, intermittently 6 seconds, actual duty cycle 0.5%, average sound intensity (I
SPTA) 0.89W/cm
2With the high concentration microvesicle generation cavitation in the described special ultrasonic pulse excitation tumor neogenetic blood vessels, main release of shock wave of pulsating ultrasound cavitation and microjet homenergic form produce the physical damage effect.The batch (-type) ultrasound emission is that microvesicle pours into and wins TPER, when making ultrasonic therapeutic, is in high concentration microvesicle perfusion state in the tumor vessel all the time.The treatment observation cycle is 30 days.
The imaging evaluation program, the maximum diameter of tumor of each time point two dimension tangent plane of ultrasonic measurement, ultrasonic contrast video picture etc.Wherein, measuring diameter is the method for the most directly perceived and easy row.
Concrete experimentation is as follows: the new zealand white rabbit of subcutaneous transplantation VX2 tumor is divided into ultrasonic microbubble treatment group (US+MB), simple ultrasonic group (US) and false according to organizing (SHAM+SALINE) research that experimentizes at random.In the time of the intravenous injection lipid microbubble, the direct irradiation tumor region of pulsed focus supersonic.Simple ultrasonic group and false according to group respectively with 5ml normal saline and ultrasonic false according to substituting, each 10 minutes, every treatment in 72 hours once, gather each time point tumor two-dimensional ultrasonic image and ultrasonic contrast image, measure tumor tangent plane maximum gauge.Each experimental group was at 30 days time points, inject excessive anesthetis in the ultrasonic contrast posterior vein and put to death lotus tumor rabbit and dissection, observation lung, liver, kidney, pelvic lymph node have or not metastasis, include laboratory observation scope position number of tumors purpose in and how much carry out classification: 0 grade-do not find that lump, organ are seen substantially and be the normal structure structure; 1-2 lump appears in 1 grade-look-out station, and turns out to be the VX2 metastatic carcinoma through conventional pathological section; 3-10 lump appears in 2 grades-look-out station, and turns out to be the VX2 metastatic carcinoma through conventional pathological section; 3 grades-look-out station occurs surpassing 10 lumps, and turns out to be the VX2 metastatic carcinoma through conventional pathological section.
As a result, microcapsular ultrasound treatment group, simple ultrasonic group and false be respectively 1.1 ± 0.1cm, 1.2 ± 0.1cm, 1.2 ± 0.1cm, three groups of tumor average diameter difference not statistically significants (P＞0.05) according to the tumor average diameter of group before the cavitation treatment.After finishing 30 days experimental period, microcapsular ultrasound treatment group, simple ultrasonic group and false tumor average diameter according to group grow to 2.1 ± 0.5cm, 3.0 ± 0.9cm, 3.4 ± 0.7cm respectively, ultrasonic microbubble group tumor average diameter is significantly less than simple ultrasonic group, difference has statistical significance (table 2, P＜0.05); Ultrasonic microbubble group diameter of tumor is significantly less than ultrasonic false according to group, and difference has statistical significance (P＜0.01); Big or small and the false group of shining of simple ultrasonic group of diameter of tumor compares not statistically significant (P＞0.05) (Fig. 9).Rank test is carried out in scoring to the neoplasm metastasis rate, and the microcapsular ultrasound treatment group rate of transform is starkly lower than simple ultrasonic group and the false group (Figure 10, P＜0.01) of shining.
Each experimental group different time points diameter of tumor of table 2. (cm, x ± s)
With simple ultrasonic group of comparison, * P＜0.05; With ultrasonic false according to organizing relatively * * P＜0.01
The growth of entity tumor, transfer rely on tumor neogenetic blood vessels and generate, in case new vessels is blocked, produce blood supply insufficiency, and tumor growth will be suppressed, and tumor also will be controlled through the blood loop jump.Owing to observe tumor vessel blocking-up back 3-4 days, tumor blood is for the part recovery situation is arranged, block microcirculatory purpose in order to reach persistence, (per 72 hours once), the method for repeatability treatment have repeatedly been adopted periodically in this experiment, experimental result, ultrasonic cavitation treatment have significantly suppressed tumor growth and have controlled the generation of neoplasm metastasis.
By damage tumor blood capillary, cause hematoma, thrombosis, microcirculation blocking-up, ultrasonic cavitation has produced the bloodstream blocking effect.Because tumor neogenetic blood vessels density height, quantity is big, blood is for abundant unusually, this body structure has the characteristics of vulnerability and defective, for the mechanical injuries of ultrasonic cavitation effect provide more advantageous conditions, the preliminary explanation of experiment, produce factor that tumor growth suppresses effect and be ultrasonic with microvesicle collaborative due to, Neither of the two can be dispensed.
Oncotherapy effect pathological evaluation
Tumor is seen substantially to protruding in the nodal-like enclosed mass of body surface, with the surrounding tissue boundary clear, has complete peplos to form, and quality is harder; Treatment group and the about 2.0cm-3.8cm of matched group tumor maximum diameter are with the maximum tangent plane diameter of the two-dimensional ultrasonic image tumor of gathering basically identical; Each organizes tumor center all necrosis in different degree, and incomplete tumor is flesh of fish shape on every side.Conventional H .E. stained light microscopic is observed down, and tumor center mostly is downright bad cancerous cell, incomplete tumor rich blood vessel on every side, and tumor cell arranges that to be reality streak, has cancer nests to form, the big engrain of nucleus, atypia is obvious.
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Claims (6)
1. microvesicle is used for the purposes of combining ultrasonic generation high-intensity cavitation as anti-tumor neovascularization medicaments in preparation, it is characterized in that described microvesicle is the microvesicle that produces the high-intensity cavitation effect of mechanical destruction under the ultrasonic energy effect of a kind of pulsed, peak value negative pressure; Described ultrasonic energy has higher peak sound pressure and lower emission dutycycle.
2. microvesicle according to claim 1 is used for combining ultrasonic in preparation and produces the purposes of high-intensity cavitation as anti-tumor neovascularization medicaments, it is characterized in that microvesicle is acoustic contrast agent microvesicle or ultrasonic therapeutic type microvesicle, is selected from Novi
Optison, Imagent, perfluoropropane human albumin microsphere and perfluor show.
3. microvesicle according to claim 1 is used for combining ultrasonic in preparation and produces the purposes of high-intensity cavitation as anti-tumor neovascularization medicaments, it is characterized in that microvesicle adopts the administering mode through peripheral intravenous injection, microvesicle injection using dosage is 0.01～1.0 milliliter of a per kilogram of body weight.
4. microvesicle according to claim 1 is used for combining ultrasonic in preparation and produces the purposes of high-intensity cavitation as anti-tumor neovascularization medicaments, it is characterized in that: described tumor neogenetic blood vessels is meant the new vessels of various malignant entity tumors, comprises small tremulous pulse, small vein, blood capillary and mimicry blood vessel.
5. microvesicle according to claim 1 is used for combining ultrasonic in preparation and produces the purposes of high-intensity cavitation as anti-tumor neovascularization medicaments, and it is characterized in that: the form of described ultrasonic energy: the frequency range of emission is 300 KHz (KHz)～3.0 megahertzes (MHz); Ultrasonic peak negative pressure is 2～15 MPas (MPa); Each ultrasonic pulse width is at 20-2000 periodic regime; Its each pulsating ultrasound launch time is more than 30 seconds; Pulsating ultrasound occurs as continous way or is batch (-type), and the intermittent time is 2～20 seconds; The ultrasound emission change in duty cycle is 0.01-10%; Described high sound intensity cavitation is meant that microvesicle excites the strong sound cavitation effect that is taken place under the situation in described ultrasonic energy form irradiation.
6. microvesicle according to claim 5 is used for combining ultrasonic in preparation and produces the purposes of high-intensity cavitation as anti-tumor neovascularization medicaments, it is characterized in that: the apparatus for ultrasonic therapeutic treatment of described ultrasonic energy emissions is the Ultrasound Instrument of plane formula emission or focusing emission.
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