CN106031795A - Ultrasound-microbubble transferring device and method thereof - Google Patents
Ultrasound-microbubble transferring device and method thereof Download PDFInfo
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Abstract
The invention discloses an ultrasound-microbubble transferring device and a method thereof. The device comprises an ultrasonic generator, an energy amplifier, an impedance matching circuit, a sound wave focusing probe, a probe supporting bracket, a depression cone, an ultrasound permeable membrane, an ultrasonic coupling agent layer, a glass dish cover and a culturing dish. Parts are reasonably combined to construct an ultrasonic platform, microbubbles with medicine or treating genes are transfected into cultured cells, the microbubbles are smashed with ultrasonic waves with the certain energy on the cultured cells, and the medicine or the treating genes which is/are brought by the microbubbles is/are released to the cells to achieve the treating effect. The ultrasound-microbubble transferring device and the method thereof have the advantages that the targeting performance and the transfection efficiency are high, and operation is safe, easy and convenient.
Description
Technical field
The present invention relates to microvesicle as medicine or the vehicle of therapeutic gene, particularly relate to a kind of ultrasonic micro-
Bubble rehandling facilities and method thereof.
Background technology
The medical therapy of present stage is to be administered, and medicine imports common method oral administration and quiet
Arteries and veins drug administration by injection, oral administration differs because of medicine taste, and patient compliance is the most different, and can not be special
The opposite sex plays a role affected organ, takes effect slow, is easily absorbed by other organ-tissues and has side effects, vein
It is poor that injection equally exists targeting, there is the situation of potential safety hazard and side effect.And gene transfection is to pass through structure
Build carrier by the process of gene into cells.The method of gene transfection at present is divided into viral vector transfection and non-disease
Poison transfection, including: viral vector such as adenovirus, slow virus, retrovirus and DNA direct injection,
Partickle bombardment technology, electroporation, magnetic robin and the non-viral transfection method such as liposome-mediated.And viral vector
There is a specificity low, there is the drawback such as security risk and immune response, non-viral transfection approach is also
There is the problems such as restricted application, invasive, transfection efficiency be low.So shortage one is both safe and effective,
Existing specificity has again the gene transfection system of targeting.The microbubble technique of ultrasonic mediation is the most increasingly being subject to
To paying attention to.Current research shows, the microbubble technique of ultrasonic mediation also shows that huge potentiality in the treatment,
Can be as a kind of efficient gene or drug delivery vehicles.By microvesicle particular combination medicine or therapeutic gene,
Enter cell or peripheral blood vessel to be expelled in blood, take the microvesicle of gene with ultrasonic smashing, carry out specific group
The location release knitted, it is provided that new administration and gene rotaring transfecting mode.
The action principle of ultrasound-microbubble system is to be noted by the vein of this gene of microvesicle with the carrier tool of medicine
It is mapped to internal, with blood flow to destination organization, under ultrasonic mediation, by improving ultrasound intensity, reduces threshold value,
Produce cavitation effect, microvesicle can be broken, and be that vascular endothelial cell permeability increase makes to take in microvesicle
Gene or the drug release of band play a role to surrounding tissue.
Microvesicle mostly is what lipid film parcel noble gas was formed, and its adventitia mostly is unilamellar lipid film, belongs to mantle,
There is good elasticity.In it, the gas of parcel is to be a kind of colourless, tasteless, nontoxic, nonflammable under normality
Gas, chemical composition stability.It is slightly soluble in water and there is preferable crushing resistance.The microvesicle particle diameter of 90% is less than
8um, without preservative, have good harmonic performance, internal stability and safety, it is to avoid gas damage
Lose, spread, dissolve and microvesicle gathering.
Microvesicle has good acoustic properties and stability so that it is also easily produce significantly under relatively low acoustic pressure
Microvesicle resonance, i.e. produces nonlinear second-harmonic generation in the case of microvesicle does not ruptures, and makes micro-
Bubble maintains the long period in blood.
Ultrasound microbubble mediated treatment has many advantages, such as reduced immunogenicity, hypotoxicity, low aggressive, nothing
Wound, organ-tissue specificity, repeatable application etc., it has also become have potential in various disease treatment means
New technique.There is no the invention of the fixing device of ultrasound-microbubble system at present, also without assembling this internal system structure
Further elements.
Summary of the invention
It is an object of the invention to overcome the deficiencies in the prior art, it is provided that a kind of ultrasonic microbubble rehandling facilities and
Its method.
Technical scheme is as follows:
Ultrasonic microbubble rehandling facilities includes that ultrasonic generator, energy amplifier, impedance matching circuit, sound wave are poly-
Burnt probe, probe support bracket, cone of depression, ultrasonic permeable membrane, ultrasonic coupling oxidant layer, glass dish lid and
Culture dish.
Described ultrasonic generator, energy amplifier, impedance matching circuit, sound wave focusing probe is sequentially connected,
Described sound wave focusing probe vertical is arranged on downwards on probe support bracket, sound wave focusing probe and landing leakage
One end of bucket is connected, and the other end of cone of depression is provided with ultrasonic permeable membrane, and described glass dish lid is coated with
Ultrasonic coupling oxidant layer, described ultrasonic permeable membrane is in close contact with ultrasonic coupling oxidant layer, described glass dish lid
It is arranged on culture dish.
Civil power is converted into the high-frequency ac signal of telecommunication by ultrasonic generator, ultrasonic is converted into high frequency electrical signal to drive
Running, this instrument master is providing sonic energy source and is carrying out the conversion of high-quality form of energy；Energy amplifier,
The most ultrasonic amplifier so that sound wave obtains energy from electronics there, ultrasound wave is exaggerated rear to have enough
Electric energy transmission circuit later is transmitted；Impedance matching circuit makes energy in transmitting procedure, bears
Carry impedance consistent with the internal driving of transmission, reduce the loss of energy in transmitting procedure, it is ensured that all energy are all
It is supported absorption, obtains the duty of maximum power output, so that there being enough energy to gather
On probe；The electric energy transmitted in impedance matching circuit is converted to acoustic energy by piezoelectric effect by sound wave focusing probe,
And by point focusing or the form of line focus, after being gathered together by acoustic energy, send ultrasound wave, come
Act the effect smashing microvesicle release active substance afterwards.
The Tissue Culture Dish lid of general merchandise is as culture dish body, for plastic material.The present invention is by polymer
Ware lid is replaced with glass dish lid, and ultrasound wave is propagated in glass material and faster absorbed less, arrives in cell
Also the most more, it is simple to package unit plays a role.
As preferably, described ultrasonic microbubble rehandling facilities also includes culture dish seat and flexible connecting rod；Described
Culture dish be on culture dish seat, described culture dish seat is by vertically arranged flexible connecting rod and probe
Support bracket connects, and described culture dish seat is horizontally moveable, and can be according to the shape of culture dish and big
Little (such as 10cm diameter circular culture dish, 6cm diameter circular culture dish, rectangle 6,24 orifice plates etc.)；
The mode that culture dish seat moves horizontally is varied, as can be to arrange horizontal direction at flexible connecting rod end
Slide rail, culture dish seat is arranged at the pulley that slide rail matches, can realize culture dish seat level move
Dynamic, additionally can also be arranged at chute at flexible connecting rod end join by offering chute on culture dish seat
The slide bar closed, it is also possible to use tenon connected mode to realize the movement of horizontal direction, for mechanical connection manner
Moving resistance can be reduced by lubricating oil.
As preferably, described cone of depression is pyramidal structure, and shell is plastic material, inner hollow,
Can be added water entrance, and cross-sectional area suitable for reading is relatively big, and probe stretches in saliva, and end opening cross-sectional area is less,
It is connected with ultrasonic permeable membrane, mainly plays focusing ultrasound wave and in water, be more beneficial for ultrasound wave conduction etc.
Effect.
As preferably, described sound wave focusing probe is Voice segment formula probe, and power is 200-500kHz.
The microvesicle used in this value appropriate means, can smash the microvesicle carrying medicine or therapeutic gene, release
Active substance.
It is polyurethane film as the most described ultrasonic permeable membrane, stable components, waterproof, several to ultrasound wave
Do not absorb, in the case of without ware lid, it is also possible to avoid ware inner cell to pollute, be primarily used to infiltration super
Sound is to conduct.
Ultrasonic coupling agent as the most described ultrasonic coupling oxidant layer is water soluble polymer colloid.Water-soluble
In property polymer colloid, each constituent proportion count by weight percentage is: hydroxyethyl cellulose 1%～
3%, benzalkonium chloride antibacterial 0.3%～1.5%, propylene glycol 6%～11%, glycerol 5%～8%, remaining is for going
Ionized water.Ultrasonic coupling agent is colourless or light-coloured transparent gel, nontoxic, and toughness is suitable, passes through
Slight void between filling contact surface, does not make micro-air between space affect ultrasonic penetrating and reduce ultrasonic
Energy is in the reflection loss at this interface；Meanwhile, there is lubrication to reduce friction, it is easy to launch；Meanwhile,
Ultrasonic coupling agent, pH is neutral, fanout free region corrosion-free to ultrasonic probe；Thus effectively in ultrasonic effectively
In incoming measured object.
The ultrasonic microbubble that the invention also discloses described device reprints method: culture dish is arranged on culture dish seat,
In culture dish, cultivation has transfected the cell of microvesicle, covers on culture dish by glass dish lid, at glass dish lid
Upper coating ultrasonic coupling agent so that it is launch on glass dish lid, eliminates body surface air, forms ultrasonic coupling
Mixture layer, the length of the flexible connecting rod of regulation, make the ultrasonic permeable membrane of cone of depression lower end be close to ultrasonic coupling
Oxidant layer, power-on, sound wave focusing probe release ultrasound wave smashes the active substance in microvesicle release microvesicle.
Preferably, described microvesicle outer layer is lipid film, and inside is perfluoroalkanes gas, preferably perfluor
Pentane gas, microvesicle particle diameter is less than 25 μm, and 90% microvesicle particle diameter is less than 8 μm.
Compared with prior art, after different medicines or therapeutic gene are combined by the present invention with microvesicle, subtract
Lack these therapeutic components and arrive the loss before cell or organ-tissue and destruction.With the tool building out
There is the focused ultrasonic transducer of appropriate energy and beneficially ultrasound wave is lossless, the connected structure of high efficiency conduction
Ultrasonic platform, smashes microvesicle specifically treating in cell or organ-tissue, produce cavitation and acoustic aperture
Effect so that while discharging active substance after microbubble ruptures, microvascular permeability increases, thus enhancing office
The transfection in portion and expression, thus play therapeutical effect.The method has many advantages, as reduced immunogenicity,
Hypotoxicity, low aggressive, noinvasive, organ-tissue specificity, repeatable application etc., it has also become various diseases
Treatment means has potential new technique.
Accompanying drawing explanation
Fig. 1 is assembly of the invention structural representation；
Fig. 2 is the connection figure of the present invention ultrasonic generation system；
Fig. 3 is that band of the present invention stretches the structure drawing of device of connecting rod.
Detailed description of the invention
Below in conjunction with Figure of description and embodiment, the present invention will be further described.
As it is shown in figure 1, in one embodiment of the invention, ultrasonic microbubble rehandling facilities includes ultrasonic generation
Device, energy amplifier, impedance matching circuit, sound wave focusing probe, probe support bracket, cone of depression,
Ultrasonic permeable membrane, ultrasonic coupling oxidant layer, glass dish lid and culture dish.
Described ultrasonic generator, energy amplifier, impedance matching circuit, sound wave focusing probe is sequentially connected,
Described sound wave focusing probe vertical is arranged on downwards on probe support bracket, sound wave focusing probe and landing leakage
One end of bucket is connected, and the other end of cone of depression is provided with ultrasonic permeable membrane, and described glass dish lid is coated with
Ultrasonic coupling oxidant layer, described ultrasonic permeable membrane is in close contact with ultrasonic coupling oxidant layer, described glass dish lid
It is arranged on culture dish.
As in figure 2 it is shown, civil power is converted into the high-frequency ac signal of telecommunication by ultrasonic generator, to drive ultrasonic conversion
For the running of high frequency electrical signal, this instrument master is providing sonic energy source and is carrying out the conversion of high-quality form of energy；
Energy amplifier, the most ultrasonic amplifier so that sound wave obtains energy from electronics there, after ultrasound wave is exaggerated
It is transmitted to have in enough electric energy transmission circuit later；Impedance matching circuit makes energy pass
During defeated, load impedance is consistent with the internal driving of transmission, reduces the loss of energy in transmitting procedure, protects
Demonstrate,prove all energy and be all supported absorption, obtain the duty of maximum power output, so that having enough
Energy can gather on probe；The electricity that sound wave focusing probe will be transmitted in impedance matching circuit by piezoelectric effect
Acoustic energy can be converted to, and by point focusing or the form of line focus, after acoustic energy is gathered together, send out
Go out ultrasound wave, finally act the effect smashing microvesicle release active substance.
As it is shown on figure 3, in another embodiment of the present invention, described ultrasonic microbubble rehandling facilities also includes
Culture dish seat and flexible connecting rod；Described culture dish is on culture dish seat, and described culture dish seat passes through
Vertically arranged flexible connecting rod is connected with probe support bracket, and described culture dish seat is horizontally moveable.
The workflow of this device is: culture dish is arranged on culture dish seat, and in culture dish, cultivation has transfected
The cell of microvesicle, covers on culture dish by glass dish lid, coats ultrasonic coupling agent on the glass dish lid, makes
It launches on glass dish lid, eliminates body surface air, forms ultrasonic coupling oxidant layer, and regulation is flexible to be connected
The length of bar, makes the ultrasonic permeable membrane of cone of depression lower end be close to ultrasonic coupling oxidant layer, power-on, sound wave
Focusing probe release ultrasound wave smashes the active substance in microvesicle release microvesicle.
The microvesicle used in device be outer layer be lipid film, there is good elasticity；Inside is stable fluorine carbon
It is a kind of gas colourless, tasteless, nontoxic, nonflammable under material such as perflenapent gas, normality, chemistry
Stable components.It is slightly soluble in water and there is preferable crushing resistance.The microvesicle particle diameter of 90% is less than 8um, without anticorrosion
Agent, have good harmonic performance, internal stability and safety, it is to avoid gas loss, diffusion, molten
Solve and microvesicle is assembled.Microvesicle has good acoustic properties and stability so that it is the easiest under relatively low acoustic pressure
Produce significant microvesicle resonance, in the case of microvesicle does not ruptures, i.e. produce nonlinear second harmonic effect
Should, and make microvesicle maintain the long period in blood.
Microvesicle involved in the present invention is the commercialized product that can directly buy from other places, as Taiwan produces
Ulsphere, Sonovue and Optison produced in USA etc..
Can use two kinds of microvesicle kinds, one is bottled medicine-carrying microvesicle, directly carries medicine or treatment inside film
Gene；Two is bottled unloaded microvesicle, is injected in microvesicle bottle by the plasmid carrying medicine or therapeutic gene,
Shake, produce milky turbidity liquid, imported in cell or organ-tissue by syringe.This
Bright recommend the first, more convenient, and there is not microvesicle and medicine or therapeutic gene fusion process
The loss of middle appearance.
Claims (9)
1. a ultrasonic microbubble cell rehandling facilities, it is characterised in that include ultrasonic generator, energy amplifier, resistance
Anti-match circuit, sound wave focusing probe, probe support bracket, cone of depression, ultrasonic permeable membrane, ultrasonic coupling
Mixture layer, glass dish lid and culture dish seat；
Described ultrasonic generator, energy amplifier, impedance matching circuit, sound wave focusing probe is sequentially connected,
Described sound wave focusing probe vertical is arranged on downwards on probe support bracket, sound wave focusing probe and landing leakage
One end of bucket is connected, and the other end of cone of depression is provided with ultrasonic permeable membrane, and described glass dish lid is coated with
Ultrasonic coupling oxidant layer, described ultrasonic permeable membrane is in close contact with ultrasonic coupling oxidant layer, described glass dish lid
It is arranged on culture dish.
Ultrasonic microbubble rehandling facilities the most according to claim 1, it is characterised in that described device also includes training
Support ware seat and flexible connecting rod；Described culture dish is positioned on culture dish seat, and described culture dish seat is by hanging down
The straight flexible connecting rod installed is connected with probe support bracket, and described culture dish seat is horizontally moveable.
Ultrasonic microbubble rehandling facilities the most according to claim 1 and 2, it is characterised in that described cone of depression
For plastic conical hollow structure, cross-sectional area suitable for reading is more than end opening, is full of water in it, and sound wave focusing probe is stretched
Entering suitable for reading and submerge in water, end opening is connected with ultrasonic permeable membrane.
Ultrasonic microbubble rehandling facilities the most according to claim 1 and 2, it is characterised in that described sound wave focusing
Probe is Voice segment formula probe, and power is 200-500kHz.
Ultrasonic microbubble rehandling facilities the most according to claim 1 and 2, it is characterised in that described ultrasonic infiltration
Film is polyurethane film.
Ultrasonic microbubble rehandling facilities the most according to claim 1, it is characterised in that described ultrasonic coupling oxidant layer
Ultrasonic coupling agent be water soluble polymer colloid.
7. the ultrasonic microbubble of a device as claimed in claim 1 reprints method, it is characterised in that: culture dish is arranged
On culture dish seat, in culture dish, cultivation has transfected the cell of microvesicle, covers glass dish lid at culture dish
On, glass dish lid coats ultrasonic coupling agent so that it is launch on glass dish lid, eliminate superficial air shape
Become ultrasonic coupling oxidant layer, the length of the flexible connecting rod of regulation, make the ultrasonic permeable membrane of cone of depression lower end be close to
Ultrasonic coupling oxidant layer, power-on, ultrasonic generator produces ultrasonic, after amplified device carries out energy amplification,
Being converted into electric energy to be conducted by impedance matching circuit, it is ultrasonic through probe release that arrival probe is converted into acoustic energy
Ripple, ultrasound wave propagates through the water of cone of depression and assembles, then through ultrasonic permeable membrane and ultrasonic coupling
The sound wave infiltration of mixture and conduction, carry out the effect of ultrasound wave through glass dish lid to cell, so that micro-
The raw cavitation effect of steep raising also crushes, finally the active substance in release microvesicle.
Method the most according to claim 7, it is characterised in that described microvesicle outer layer is lipid film, and inside is
Perfluoroalkanes gas, microvesicle particle diameter is less than 25 μm, and 90% microvesicle particle diameter is less than 8 μm.
Method the most according to claim 8, it is characterised in that described perfluoroalkanes gas is perflenapent.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106620734A (en) * | 2016-12-20 | 2017-05-10 | 江西宜信堂医疗科技有限公司 | Sterile medical ultrasonic coupling patch and preparation method thereof |
| CN106635793A (en) * | 2017-02-07 | 2017-05-10 | 重庆科技学院 | Culture dish for ultrasonic experiment and use method |
| WO2019113930A1 (en) * | 2017-12-15 | 2019-06-20 | 深圳先进技术研究院 | Ultrasonic microbubble cell stimulation device and operation method thereof |
| CN109929756A (en) * | 2017-12-15 | 2019-06-25 | 深圳先进技术研究院 | Ultrasonic microbubble stimulates cell device and its operating method |
| WO2022021734A1 (en) * | 2020-07-27 | 2022-02-03 | 中国科学院深圳先进技术研究院 | Ultrasonic sound field conformal gene transfection device |
| WO2022120732A1 (en) * | 2020-12-10 | 2022-06-16 | 深圳先进技术研究院 | Ultrasonic drug delivery experimental device and ultrasonic drug delivery experimental method |
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| CN106620734A (en) * | 2016-12-20 | 2017-05-10 | 江西宜信堂医疗科技有限公司 | Sterile medical ultrasonic coupling patch and preparation method thereof |
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| WO2019113930A1 (en) * | 2017-12-15 | 2019-06-20 | 深圳先进技术研究院 | Ultrasonic microbubble cell stimulation device and operation method thereof |
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| WO2022021734A1 (en) * | 2020-07-27 | 2022-02-03 | 中国科学院深圳先进技术研究院 | Ultrasonic sound field conformal gene transfection device |
| WO2022120732A1 (en) * | 2020-12-10 | 2022-06-16 | 深圳先进技术研究院 | Ultrasonic drug delivery experimental device and ultrasonic drug delivery experimental method |
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