CN1846645A - Photoacoustic blood vessel imaging method and equipment for monitoring photodynamic tumor treating effect - Google Patents
Photoacoustic blood vessel imaging method and equipment for monitoring photodynamic tumor treating effect Download PDFInfo
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- CN1846645A CN1846645A CNA2006100340136A CN200610034013A CN1846645A CN 1846645 A CN1846645 A CN 1846645A CN A2006100340136 A CNA2006100340136 A CN A2006100340136A CN 200610034013 A CN200610034013 A CN 200610034013A CN 1846645 A CN1846645 A CN 1846645A
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Abstract
The present invention relates to real-time monitoring of photodynamic tumor treating effect, and the photoacoustic blood vessel imaging method is that during laser irradiation of tumor tissue for photodynamic treatment, the photoacoustic chromatographic images of the tumor tissue before and after photodynamic action are compared and the blood vessel diameter change is reflected with the positive and negative peak-to-peak width of the photoacoustic blood vessel signals. The equipment for the real-time monitoring of photodynamic tumor treating effect includes laser generator assembly, acoustic signal acquiring assembly and computer successively connected electrically, as well as rotating scan mechanism connected electrically to the computer, sample fixing assembly and connected acoustic assembly. The present invention is sensitive and quick in real-time monitoring of photodynamic tumor treating effect.
Description
Technical field
The present invention relates to medical science and medical apparatus and instruments.A kind of in more detail method of optoacoustic blood vessel imaging monitoring photodynamic tumor therapeutic effect.
The invention still further relates to and realize the used device of said method.
Background technology
Photodynamic therapy is the minimum early stage treating malignant tumor method of damage, also is a kind of palliative treatment method of late malignant tumour.Optical dynamic therapy can optionally be eliminated local superficial tumor and not jeopardize normal structure, and can carry out with chemotherapy and radiation is collaborative.Because the main tumor peripheral vessels that relies on of the existence of tumor cell and growth provides nutritional labeling, so the blood vessel injury effect is considered to one of important mechanisms of optical dynamic treatment of tumor.Therefore, the monitoring of carrying out photodynamic tumor treatment medium vessels damage effect not only has the important in theory meaning with research in the life sciences basic research, and for real-time detection tumor treatment situation, screening have optimum therapeuticing effect photosensitizer, determine suitable pdt agent amount, the clinical application that promotes optical dynamic therapy also has realistic meaning.
The mechanism that optical dynamic therapy destroys the tumor peripheral vessels and suppresses new vessels receives publicity always, but is not studied clearly fully, and this is subject to choosing of animal model and imaging mode to a certain extent.Studies show that, the caliber of blood vessel injury degree and blood vessel can be changed associating.Traditional optical microscopy can be observed the blood vessel around the tumor, but needs tissue slice, can not realize that carrier detects the degree of impairment of blood vessel in real time; Utilize technique of laser imaging can study hemodynamics variation such as the velocity flow profile of blood, filling rate.But because tissue is a turbid medium, the strong scattering of light will cause the muting sensitivity and the low resolution of pure optical imagery.
Summary of the invention
The objective of the invention is to shortcoming and defect at the prior art existence, a kind of method of optoacoustic blood vessel imaging monitoring photodynamic tumor therapeutic effect is provided, utilize photoacoustic technique can produce the characteristics of the high-resolution tissue image of high-contrast, by the optoacoustic tomographic map before and after the optical dynamic therapy is compared the damage effect of assessing blood vessel, utilize the change of change monitoring blood vessels caliber of peak-to-peak value width of the positive-negative polarity of photoacoustic signal simultaneously, its operating characteristics sensitivity, quick can accurate in real time monitoring photodynamic tumor therapeutic effect.
The present invention just is used for the monitoring of tumor tissues and studies its therapeutic process; And the present invention is not limited only to this, and the present invention also can be used for observing biological evolution growth course etc.
Another object of the present invention is to provide a kind of device of realizing said method.
The method of the method for optoacoustic blood vessel imaging monitoring photodynamic tumor therapeutic effect of the present invention comprises: carry out in the process of optical dynamic therapy at the laser irradiation tumor tissues, the optoacoustic tomographic map of contrast tumor tissues before and after photodynamics, the change of the peak-to-peak width of the positive-negative polarity by the blood vessel photoacoustic signal reflects the change of blood vessels caliber, thus monitoring photodynamic tumor therapeutic effect in real time.
More specifically, method of the present invention comprises the steps:
(1) before optical dynamic therapy, utilize scanning to obtain the optoacoustic tomographic map of optical dynamic therapy pre-neoplastic blood vessel;
(2) in the optical dynamic therapy process, the blood vessel photoacoustic signal of continuous acquisition tumor tissues;
(3) after optical dynamic therapy finished, rotation sweep obtained the tumor vascular optoacoustic tomographic map behind the optical dynamic therapy;
(4) the blood vessel photoacoustic signal of gathering is carried out date processing, analyze the degree of impairment of tumor tissues blood vessel in therapeutic process, contrast the blood vessel optoacoustic tomographic map of optical dynamic therapy front and back simultaneously, assess the effect of optical dynamic therapy.
In the described step (1), the optical dynamic treatment of tumor model makes up and obtains by tumor cell transplantation being cultivated tumor on toy such as Mus or the Embryo Gallus domesticus.
In the described step (2), optical dynamic therapy adopts photosensitizer commonly used, as, Protoporphyrin IX.
In the described step (2), when carrying out optical dynamic therapy, select pulse laser for use, this laser is the light source of optical dynamic therapy, also is the light source of optoacoustic monitoring.Optical maser wavelength is corresponding with the absworption peak of photosensitizer, generally at 350-700nm.
In the described step (3), rotation sweep refers to that under the control of LABIEW capture program, motor drives the detector of sound rotation and gathers photoacoustic signal for 360 °.
In the described step (4), date processing refers to the photoacoustic signal of gathering is analyzed and handled.
A kind of device of realizing that said method adopts comprises laser generating assembly, rotation sweep assembly, acoustical signal acquisition component, computer module, sample fixation kit, acoustical coupling assembly; Laser generating assembly, acoustical signal acquisition component, computer are electrically connected successively; Rotary scanning mechanism and computer are electrically connected; The sample fixation kit is connected with acoustical coupling assembly preface successively.
Described laser generating assembly is used for that laser instrument is produced pulse laser and shines sample to be monitored through the light path adjustment system.
Described rotary scanning mechanism is scanned by the step motor control circular scanning support that band drives.
Described acoustical signal acquisition component is electrically connected successively by the computer of detector of sound, signal amplifier, oscillograph, band GPIB card and constitutes.
Acquisition controlling software and data processing software are housed in the described computer.
Described sample fixation kit is three-dimensional adjustable platform and sample fixture.
Be full of acoustical coupling liquid in the tank of described acoustical coupling assembly, the built-in attemperating unit of coupling liquid can be controlled temperature about 37 ℃.
Operation principle of the present invention is: because the existence of tumor cell and the main tumor peripheral vessels that relies on of growing provide nutritional labeling, receive publicity so optical dynamic therapy destroys the mechanism of tumor peripheral vessels and inhibition new vessels always.Studies show that blood vessel injury causes vasoconstriction and platelet aggregation to cause blood flow retardance and local tissue hypoxia, is to cause tumor tissues damage and even downright bad main cause.The present inventor discovers, the caliber of blood vessel injury degree and blood vessel can be changed associating.When with pulsed laser irradiation around tumor tissues, owing to color development group absorbing light energy such as hemoglobin in the blood vessel cause temperature rise, temperature rise causes thermal expansion and produces pressure wave, and optoacoustic effect that Here it is, optoacoustic effect have provided the relation of distribution of sample light absorption light and photoacoustic signal.The light absorption that can be finally inversed by tissue by the measuring light acoustical signal distributes, particularly can reconstruct the diameter of blood vessel, the vascularity imaging that this not only can be used to organize can also be assessed OPK therapeutic effect by the change of monitoring blood vessel diameter size.Owing to the variation that just can directly reflect blood vessel diameter by detection signal, the method is convenient and swift, can monitor the curative effect of optical dynamic therapy in real time.
The present invention compared with prior art has following advantage and effect:
(1) the inventive method combines and ultrasonic body is had stronger penetration capacity and optical imagery has high-resolution advantage, compares the advantage with high-resolution high-contrast with the traditional medicine image method.
(2) the present invention utilizes the degree of impairment of photoacoustic technique monitoring tumor therapeutic procedure medium vessels, and comparing with traditional light microscopic microtechnique does not need Histological section, is a kind of undamaged monitoring technology, and detection in vivo can be provided.
(3) the inventive method can directly be utilized photoacoustic signal reflection blood vessels caliber size, so can accomplish the degree of impairment of accurately real-time monitoring optical dynamic therapy process medium vessels.
(4) pulse laser of selecting for use among the present invention be optical dynamic therapy also be the light source of photo-acoustic detection simultaneously, effectively with tumor treatment and monitoring integration, for clinical research provides a kind of method more easily.
(5) cost of each assembly of device of the present invention is lower, so the cost of single unit system is also relatively low, is easy to application.
Description of drawings
Fig. 1 is the structural representation of apparatus of the present invention.
Fig. 2 is the tumor vessel optoacoustic tomographic map that utilizes the optical dynamic therapy front and back of device realization shown in Figure 1.
Fig. 3 (a) utilizes 0,5,10,15 of device collection shown in Figure 1, the optoacoustic time-domain signal of 20 minutes tumor vessel calibers.
Fig. 3 (b) is the continuous real-time change curve of the tumor vessel caliber in the optical dynamic therapy process that utilizes device shown in Figure 1 to realize.
Fig. 4 (a) utilizes the caliber of device realization shown in Figure 1 at the damage curve of the tumor vessel about 90 microns under different light dosage and photosensitizer concentration.
Fig. 4 (b) utilizes the caliber of device realization shown in Figure 1 at the damage curve of the tumor vessel about 280 microns under different light dosage and photosensitizer concentration.
The specific embodiment
Embodiment 1
As seen from Figure 1, apparatus of the present invention by laser generating assembly 1, rotation sweep assembly 2, acoustical signal acquisition component 3, computer form 4, sample fixation kit 5, acoustical coupling assembly 6 form.Wherein laser assembly 1 is connected and composed by laser instrument 1-1, metallic-membrane plating reflector 1-2, concavees lens 1-3, clouded glass 1-4 preface successively; The rotation sweep assembly is rotated by digital I/O card and relay controller 2-2 control step motor 2-1 driven rotary support 2-3 by computer 4.The acoustical signal acquisition component is amplified through signal amplifier 3-2 by detector of sound 3-1, and the average back of oscillograph 3-3 is finished collection by the computer 4 of band GPIB card by control software.Wherein: laser instrument 1-1 selects the TII of Byelorussia LOTIS company for use, and can send wavelength is 350-500nm, the continuously adjustable and 532nm of 690nm-1000nm and the pulse laser of 1064nm; Detector of sound adopt acicular sulfonation gather difluoroethylene (Polyvinylidene Fluoride, the PVDF) hydrophone of film (Precision acoustic Ltd system), receiving area diameter 1mm, detectivity is 850nv/pa, the receiving area diameter is 1mm.Homemade power amplifier gain amplifier 0～20dB, bandwidth 10kHz～12MHz; Adopt U.S. Tyke (Tektronix) TDS3032 type digital oscilloscope.Motor is 1.8 ° of Japanese east 2 phase step motor VEXIA step-lengths.The acquisition controlling program realizes that with LABVIEW software digital graphics signal is handled with matlab software and realized; Computer 4 can be selected the P4 microcomputer for use, internal memory 256M; Sample cell 6-1 is the cuboid box made from synthetic resin, is full of homemade acoustical coupling liquid 6-2 in the sample cell and is beneficial to velocity of sound coupling, and the temperature of utilizing temperature control system 6-3 maintenance coupling liquid in test can be downright bad to guarantee Embryo Gallus domesticus about 37 ℃.
Said apparatus is applied to an Embryo Gallus domesticus of having cultivated tumor, utilizes three-dimensional adjustable sample stage to fix the position of Embryo Gallus domesticus, the embryo position harmony position of detector that keeps desire observation is in same horizontal plane.Select the 532nm pulse laser in the test for use, pulsewidth 6ns, repetition rate is 10Hz, the output pulse energy is 4mJ/cm
2, the light source that is optical dynamic therapy also is the light source of optoacoustic monitoring.Send into digital oscilloscope behind photoacoustic signal preposition amplification of process in the photodynamics process that detector of sound receives and the main amplifier.Oscillographic sample rate is 250MHz in the experiment, detector is each station acquisition 16 times, after oscillograph is average, also stored by computer acquisition by the GPIB card, computer is by the motor rotation of digital I/O card control rotation platform, step-length is 1.8 degree, and detector is gathered the photoacoustic signal of 200 positions altogether.Adopt filtered back projection to handle the signal of gathering by the MATLAB program and obtain photodynamics optoacoustic tomographic map in the past.Then in the Embryo Gallus domesticus chorioallantoic membrane injection 100 μ L concentration be 40umol/L photosensitizer Protoporphyrin IX (relative molecular mass: 562.66, chemical molecular formula: C
34H
34N
4O
4), use phosphate buffer (PBS) solution washing 5 times behind the 0.5h, do not soak into tumor or endovascular Protoporphyrin IX to remove.Detector of sound is fixed on the upright position of monitoring blood vessel, and per second is gathered a signal, the photoacoustic signal of continuous acquisition blood vessel in the optical dynamic therapy process.After photodynamics is finished, the rotation sweep detector of sound, using the same method to obtain the later optoacoustic tomographic map of photodynamics, as shown in Figure 2.Both compare the damage effect that can observe blood vessel behind the photodynamics intuitively.Shown in Fig. 2 (b), can find out significantly that the blood vessel diameter after the treatment dwindles, blood capillary reduces.Test shows, most blood capillaries in the pdt agent amount excessive and photosensitizer concentration excessive in, break very soon.
Take out the blood vessel optoacoustic time-domain signal of different light time (surveying after 0,5,10,15 minutes) in the photodynamics process, as Fig. 3 (a).Can see significantly that along with the prolongation of light application time, the peak-to-peak value width of photoacoustic signal constantly reduces, illustrate with light application time to increase that under the effect of photosensitizer, the blood vessels caliber around the tumor is constantly dwindling.
Gather a photoacoustic signal as Fig. 3 (b) for per second, the variation of blood vessels caliber and the change of photoacoustic signal amplitude under the dynamically recording continuously.From diagram as can be seen, slight expansion all appears in light dynamic experiment incipient stage blood vessel, the phenomenon that caliber increases slightly, flow velocity increases, and blood vessel shrinks gradually then, and caliber diminishes.
The result shows that the vasodilation of preceding 2min is that photo-thermal effect is occupied an leading position because compare photosensitivity reaction, cause that local temperature raises, and the diameter of blood vessel increases gradually with the rising of temperature, has caused the temporary transient expansion of blood vessel.
The device that present embodiment adopts is with embodiment 1.Said apparatus is applied to the research of light power to the concentration damage effect of different blood vessel diameter, illumination dose and the photosensitizer of blood vessel.In the test respectively to diameter 80
μ mAbout and diameter 284
μ mBlood vessel light power after data analyze, the destruction that blood vessel is subjected to is seen Fig. 4 (a) and (b) respectively.Wherein, when Protoporphyrin IX concentration be 40
μ mol/L, light dosage is 36J/cm
2, 72J/cm
2The time diameter 80
μ mAbout blood vessel almost destroyed fully.Diameter 284
μ mBlood vessel since its tube wall thicker, be subjected to low dose (20
μ mol/LProtoporphyrin IX influence little, all at more than 50% of initial value, and more recovery behind 24h, heavy dose (40
μ mol/L) Protoporphyrin IX blood vessel is had damage, but the damage of the less blood vessel of diameter group is still much smaller, for initial value 30%～50% between.Can find out significantly that the blood vessel diameter after the treatment dwindles, blood capillary reduces.Most blood capillaries in the pdt agent amount excessive and photosensitizer concentration excessive in, break very soon.
The foregoing description is a preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (4)
1, a kind of method of optoacoustic blood vessel imaging monitoring photodynamic tumor therapeutic effect, it is characterized in that carrying out in the process of optical dynamic therapy at the laser irradiation tumor tissues, the optoacoustic tomographic map of contrast tumor tissues before and after photodynamics, the change of the peak-to-peak width of the positive-negative polarity by the blood vessel photoacoustic signal reflects the change of blood vessels caliber, thus monitoring photodynamic tumor therapeutic effect in real time.
2, method according to claim 1 is characterized in that comprising the steps:
(1) before optical dynamic therapy, utilize scanning to obtain the optoacoustic tomographic map of optical dynamic therapy pre-neoplastic blood vessel;
(2) in the optical dynamic therapy process, the blood vessel photoacoustic signal of continuous acquisition tumor tissues;
(3) after optical dynamic therapy finished, rotation sweep obtained the tumor vascular optoacoustic tomographic map behind the optical dynamic therapy;
(4) the blood vessel photoacoustic signal of gathering is carried out date processing, analyze the degree of impairment of tumor tissues blood vessel in therapeutic process, contrast the blood vessel optoacoustic tomographic map of optical dynamic therapy front and back simultaneously, assess the effect of optical dynamic therapy.
3, method according to claim 1 and 2 is characterized in that selecting pulse laser for use in the step (2), and this laser is the light source of optical dynamic therapy, also is the light source of optoacoustic monitoring; Optical maser wavelength is corresponding with the absworption peak of photosensitizer, at 350-700nm.
4, a kind of device of realizing one of claim 1-3 described method is characterized in that comprising laser generating assembly, rotation sweep assembly, acoustical signal acquisition component, computer module, sample fixation kit, acoustical coupling assembly; Laser generating assembly, acoustical signal acquisition component, computer are electrically connected successively; Rotary scanning mechanism and computer are electrically connected; The sample fixation kit is connected with acoustical coupling assembly preface successively;
Described laser generating assembly is used for that laser instrument is produced pulse laser and shines sample to be monitored through the light path adjustment system;
Described rotary scanning mechanism is scanned by the step motor control circular scanning support that band drives;
Described acoustical signal acquisition component is electrically connected successively by the computer of detector of sound, signal amplifier, oscillograph, band GPIB card and constitutes;
Described sample fixation kit is three-dimensional adjustable platform and sample fixture;
Be full of acoustical coupling liquid, the built-in attemperating unit of coupling liquid in the tank of described acoustical coupling assembly.
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