CN113503907B - In-vitro circulation experiment table and experiment method for medical implant intervention body observation - Google Patents

In-vitro circulation experiment table and experiment method for medical implant intervention body observation Download PDF

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CN113503907B
CN113503907B CN202110921091.2A CN202110921091A CN113503907B CN 113503907 B CN113503907 B CN 113503907B CN 202110921091 A CN202110921091 A CN 202110921091A CN 113503907 B CN113503907 B CN 113503907B
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observation
medical implant
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experimental
fluorescence
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CN113503907A (en
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陈皓生
李永健
蒙奎霖
郑晓冰
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Tsinghua University
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Tsinghua University
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    • G01—MEASURING; TESTING
    • G01D—MEASURING NOT SPECIALLY ADAPTED FOR A SPECIFIC VARIABLE; ARRANGEMENTS FOR MEASURING TWO OR MORE VARIABLES NOT COVERED IN A SINGLE OTHER SUBCLASS; TARIFF METERING APPARATUS; MEASURING OR TESTING NOT OTHERWISE PROVIDED FOR
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    • G—PHYSICS
    • G01—MEASURING; TESTING
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Abstract

The invention discloses an extracorporeal circulation experiment table and an experiment method for medical implant intervention body observation. The experiment table comprises an experiment section module, a circulation loop module, a temperature control module and an observation module, wherein the experiment section module comprises a medical implant intervention body with a power pump, and the circulation loop module and the experiment section module form a circulation loop for the circulation flow of experiment liquid; the observation module has a fluorescence observation function and realizes quantitative observation of the area of the surface thrombus of the medical implant body; the observation module also has a flow field observation function and is used for observing the flow field in the experiment section module. The experiment table can better simulate the environment in a human body, can also observe the growth condition of thrombus on the medical implant intervention body by using a fluorescence observation system, and can effectively guide the design optimization of the medical implant intervention body by using a PIV flow field observation system to observe the flow field condition near the medical implant intervention body.

Description

In-vitro circulation experiment table and experiment method for medical implant intervention body observation
Technical Field
The invention relates to the technical field of experimental test equipment for medical implant insertion bodies, in particular to an in vitro circulation experiment table and an experimental method for medical implant insertion body observation.
Background
Cardiovascular diseases are a serious threat to human health and life. At present, the implanted medical devices are one of the most effective means for treating cardiovascular and cerebrovascular diseases, and the common implanted medical devices in direct contact with blood include artificial hearts, artificial heart valves, artificial stents and the like. The implanted medical devices are required to be subjected to preclinical tests, including in vitro tests of an extracorporeal circulation test platform, standard laboratory tests (such as biocompatibility tests and heat dissipation noise tests of used materials) and animal tests. Because animal experiments are expensive and long-lasting in time, and standard experiments have great difference with actual use working conditions, in-vitro experiments can check whether the design performance meets physiological requirements or not, and guide the design improvement of the implanted medical instrument, and the experimental device has strong repeatability and short time consumption, and is an important experimental research means for the implanted medical instrument.
The existing extracorporeal circulation experiment platform in domestic and foreign researches generally refers to a thrombolysis test device in a standard test method standard ASTM.F1841 provided by the American society for testing and materials, the key point of the research is to simulate the running environment of an implanted medical instrument in a human body, the hydromechanical properties of a blood pump, including flow, pressure, temperature, the pulsation characteristic of the heart and other key factors, are tested through a simulation test, and the experiment result is an important basis for detecting whether the thrombolysis performance of the implanted medical instrument meets the clinical use requirement.
However, for experimental test objects, there is no research observation means, such as the growth of thrombus on the device and the blood flow near the device, and the like, and the design optimization of the implanted medical device cannot be effectively guided. Therefore, there is a need to develop an extracorporeal circulation experiment platform which can better simulate the human body environment and provide abundant observation means.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art. Therefore, an object of the present invention is to provide an extracorporeal circulation experiment table for observing a medical implant intervention body, which can better simulate the human body internal environment, and can also observe the growth condition of thrombus on the medical implant intervention body and the flow field condition near the medical implant intervention body by using an observation module, so as to effectively guide the design optimization of the medical implant intervention body.
An extracorporeal circulation laboratory table for medical implant body observation according to an embodiment of the first aspect of the present invention includes:
the experimental section module comprises a medical implant intervention body with a power pump and a transparent tube with biocompatibility, wherein the medical implant intervention body is arranged in the transparent tube, and the transparent tube is provided with an inlet end and an outlet end;
a circulation loop module, one end of which is connected with the inlet end of the transparent pipe and the other end of which is connected with the outlet end of the transparent pipe, so that the circulation loop module and the experiment section module form a circulation loop for the circulation flow of the experiment liquid; the circulation loop module is used for monitoring and controlling the pressure and the flow of the experimental liquid circulating in the circulation loop;
a temperature control module for controlling the temperature of the test liquid in the circulation loop;
the observation module has a fluorescence observation function and realizes quantitative observation of the area of the thrombus on the surface of the medical implant intervention body; the observation module also has a flow field observation function and is used for observing the flow field in the experiment section module.
According to the extracorporeal circulation experiment table for medical implant intervention body observation in the embodiment of the first aspect of the invention, on one hand, a circulation loop formed by connecting the transparent tube of the experiment section module and the circulation loop module can truly simulate a human body internal circulation loop; the pressure and the flow of the experimental liquid which circularly flows in the circulating loop are monitored and controlled by the circulating loop module, so that the pressure and the flow speed in the circulating loop can be conveniently adjusted by an auxiliary experimenter to simulate the blood flowing condition in a human body, and meanwhile, the temperature of the experimental liquid in the circulating loop is controlled by the temperature control module, so that the temperature of the experimental liquid in the circulating loop can be maintained in the temperature range of 37 +/-1 ℃ of the physiological temperature of the human body, the constant temperature function is realized, and the temperature environment in the human body can be simulated. On the other hand, the observation module has a fluorescence observation function, can capture a fluorescence image of the surface thrombus of the medical implant intervention body, can realize quantitative observation of the thrombus growth condition on the medical implant intervention body by shooting a plurality of fluorescence images at certain intervals in the thrombus growth process, and can also quantitatively observe the area of the surface thrombus of the medical implant intervention body so as to detect whether the thrombolysis performance of the medical implant intervention body meets the clinical use requirement; the observation module has a flow field observation function, so that the observation module can observe the flow field condition of a flow field near the medical implant intervention body in the experiment section module, and can judge the flow condition of blood around the medical implant intervention body when the medical implant intervention body is in a human body according to the result of the flow field observation.
Therefore, the extracorporeal circulation experiment table for observation of the medical implant intervention body in the embodiment of the first aspect of the invention can better simulate the environment in the human body and simulate the growth condition of thrombus in the human body in vitro, and provides a stable and reliable experiment platform for the anticoagulant and thrombus inhibition performance research of the medical implant intervention body such as an artificial heart, an artificial stent, an artificial heart valve and the like; the medical implant intervention body is subjected to fluorescence observation through the observation module, the area of surface thrombus of the medical implant intervention body can be observed quantitatively, the growth process condition of the surface thrombus of the medical implant intervention body can be known visually and quantitatively, the medical implant intervention body is subjected to flow field observation through the observation module, the observation module can observe the flow field condition of a flow field near the medical implant intervention body in the experiment section module, and the flow condition of blood around the medical implant intervention body can be judged when the medical implant intervention body is in a human body according to the result of the flow field observation.
According to an embodiment of the first aspect of the invention, the circulation loop module comprises a biocompatible hose, a first pressure sensor, a first throttle, an ultrasonic flow sensor, a reservoir, a first injection port, a second injection port, and a second pressure sensor; one end of the hose is connected with the inlet end of the transparent pipe, and the other end of the hose is connected with the other end of the transparent pipe; the first pressure sensor, the first throttle valve, the ultrasonic flow sensor, the liquid storage container, the first injection port, the second injection port and the second pressure sensor are sequentially arranged on the hose in the direction from the other end of the hose to one end of the hose;
wherein the first pressure sensor is proximate to the outlet end for monitoring an outlet pressure of the test liquid circulating in the circulation loop after exiting the test section module;
the second pressure sensor is close to the inlet end and used for monitoring the inlet pressure of the experimental liquid which circulates in the circulation loop before entering the experimental section module;
the first throttle valve and the liquid storage container are used for regulating the flow rate and the pressure of the experimental liquid circulating in the circulation loop, and the liquid storage container is also used for discharging gas in the experimental liquid circulating in the circulation loop;
the ultrasonic flow sensor is used for monitoring the flow of the test liquid circulating in the circulating loop;
the first injection port and the second injection port are used for respectively injecting corresponding experimental liquid according to experimental requirements.
According to a further embodiment of the first aspect of the present invention, the hose is a medical PVC hose.
According to a still further embodiment of the first aspect of the present invention, the circulation loop module further comprises a syringe pump and a second throttle valve, the syringe pump is connected to the second injection port through a connecting pipe, and the second throttle valve is disposed on the connecting pipe; the syringe pump is for injecting a pharmaceutical agent.
According to a still further embodiment of the first aspect of the present invention, the temperature control module comprises a heating resistor, a temperature controller and an infrared temperature detector; the heating resistor wraps the liquid storage container and is used for heating the liquid storage container; the temperature controller is used for controlling the temperature of the heating resistor so as to maintain the experiment liquid circulating in the circulating loop at a constant physiological temperature; the infrared temperature detector is used for measuring the temperature of the experiment section module.
According to some embodiments of the first aspect of the present invention, the observation module comprises a fluorescence observation system for implementing the fluorescence observation function, the fluorescence observation system comprising a laser, a first optical filter and a single lens reflex camera; the laser is used for emitting continuous laser to irradiate the experimental section module so as to enable the fluorescent reagent on the thrombus on the surface of the medical implant body to excite fluorescence; the first optical filter is arranged between the experimental section module and the single lens reflex; the single lens reflex is used for shooting a first fluorescence image obtained by filtering fluorescence of a fluorescence reagent on thrombus on the surface of the medical implant body through the first optical filter.
According to some embodiments of the first aspect of the present invention, the observation module further includes a PIV flow field observation system for implementing a flow field observation function, the PIV flow field observation system includes the laser, a CCD camera and a second optical filter, the laser is used for exciting pulsed laser light to irradiate on the experiment section module so as to excite fluorescent particles in the experiment liquid in the experiment section module to emit fluorescent light; the second optical filter is arranged between the experimental section module and the CCD camera; the CCD camera is used for shooting a second fluorescence image obtained by filtering fluorescence of fluorescent particles in the experimental liquid in the experimental section module through the second optical filter, and the shooting frequency of the CCD camera is synchronous with the pulse frequency of the pulse laser.
According to some embodiments of the first aspect of the present invention, the PIV flow field observation system further includes a signal synchronizer electrically connected to the laser and the CCD camera, respectively, to synchronize the pulse frequency of the pulsed laser and the photographing frequency of the CCD camera.
The invention also provides an experimental method of the extracorporeal circulation experiment table for observing the medical implant.
The experiment method of the extracorporeal circulation experiment table for medical implant intervention body observation according to the second aspect of the invention is an extracorporeal circulation experiment table for medical implant intervention body observation according to any one of the first aspect of the invention, the experiment method is a fluorescence observation experiment method, and the experiment method comprises the following steps:
step S1: mixing the first fluorescent agent and the collected blood in proportion, fully and uniformly mixing in a water bath at 37 ℃, injecting into the circulation loop, and discharging the blood after the medical implant intervention body works for a period of time to form thrombus; or a second fluorescent agent is directly coated on the thrombus on the surface of the medical implant intervention body as an observation object and is placed in the transparent tube;
step S2: and quantitatively observing the area of the thrombus on the surface of the medical implant body by utilizing the observation module.
According to the experimental method of the extracorporeal circulation experiment table for medical implant intervention body observation in the embodiment of the second aspect of the invention, the extracorporeal circulation experiment table for medical implant intervention body observation in the embodiment of the first aspect of the invention can truly and well simulate the human intracorporeal circulation environment in the aspects of pressure, flow, temperature and the like, and provides a good experimental basis for a fluorescence observation experimental method. According to the fluorescence observation experiment method provided by the embodiment of the second aspect of the invention, the first fluorescent dye is mixed with blood to form a fluorescence-dyed thrombus on the surface of the medical implant intervention body, or the second fluorescent agent is directly coated on the surface of the medical implant intervention body with the thrombus formed to realize the fluorescence dyeing of the thrombus, the observation module can capture a fluorescence image of the thrombus on the surface of the medical implant intervention body, so that the quantitative observation of the growth condition of the thrombus on the medical implant intervention body and the area of the thrombus on the surface of the medical implant intervention body is realized, whether the blood-dissolving coagulation performance of the medical implant intervention body meets the clinical use requirements is detected, more observation means are provided for the design optimization of the medical implant intervention body, and the design optimization of the medical implant intervention body can be effectively guided.
According to an embodiment of the second aspect of the invention, said first phosphor is a phosphor comprising DiOC6 or/and FITC; the second fluorescent agent is DiOC6 or/and FITC.
According to a further embodiment of the second aspect of the present invention, in the step S2, the observation module includes the fluorescence observation system in an embodiment of the first aspect of the present invention, and the step S2 specifically includes the following sub-steps:
step S201: irradiating the experimental section module with blue laser with the wavelength of 490-515nm provided by the laser to enable the fluorescent reagent on the thrombus on the surface of the medical implant body to excite green fluorescence;
step S202: a green band-pass filter with the wavelength of 550nm-570nm is selected as the first filter and is additionally arranged in front of the single lens reflex;
step S203: and shooting a first fluorescence image of the thrombus on the surface of the medical implant interventional body by using the single-lens reflex camera under the condition of a full dark field, and quantitatively observing the area of the thrombus according to the first fluorescence image.
The third aspect of the invention also provides an experimental method of the extracorporeal circulation experiment table for medical implant body observation.
The experimental method of the extracorporeal circulation test table for medical implant interventional body observation according to the third aspect of the invention is an extracorporeal circulation test table for medical implant interventional body observation according to any one of the embodiments of the first aspect of the invention, and the experimental method is a PIV flow field experimental method, comprising the following steps:
step S6: adding the third fluorescent particles into the transparent liquid to obtain a fluorescent particle solution;
step S7: injecting the fluorescent particle solution into the circulation loop, and fully mixing;
step S8: and the observation module is used for observing the flow field in the experiment section module.
According to the experimental method of the extracorporeal circulation experiment table for medical implant intervention body observation in the embodiment of the third aspect of the invention, the third fluorescent particles are added into the transparent liquid to obtain the fluorescent particle solution, the fluorescent particle solution is injected into the circulation loop, the flow field in the experiment section module is observed by utilizing the flow observation function of the observation module, and the flow condition of peripheral blood when the medical implant intervention body is in the human body can be judged according to the result of flow field observation, so that the design optimization of the medical implant intervention body can be effectively guided, and a new observation means is provided for the research of the medical implant intervention body.
According to an embodiment of the third aspect of the invention, the third fluorescent particle is a Fluo-610 fluorescent particle.
According to a further embodiment of the third aspect of the present invention, the observation module includes a PIV flow field observation system in some embodiments of the first aspect of the present invention, and the step 8 specifically includes the following sub-steps:
step S801: irradiating the experiment section module with pulsed laser provided by the laser to enable third fluorescent particles in the fluorescent particle solution in the experiment section module to excite fluorescence;
step S802: shooting a second fluorescence image of fluorescence excited by a third fluorescence particle in the experimental section module through the second optical filter under a full dark field condition by using the CCD camera, wherein the shooting frequency of the CCD camera is synchronous with the pulse frequency of the pulse laser:
step S803: and processing the obtained second fluorescence image by using special processing software to obtain the flow field condition of the drainage basin near the medical implant.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
fig. 1 is a schematic view of an extracorporeal circulation test stand for medical implant body observation according to an embodiment of the first aspect of the present invention.
Fig. 2 is a schematic view of the operation principle of the PIV flow field observation system in the extracorporeal circulation experiment table for medical implant observation according to the embodiment of the first aspect of the present invention.
Fig. 3 is a flow chart of a fluorescence observation experiment method in the experiment method of the extracorporeal circulation experiment table for medical implant body observation according to the embodiment of the second aspect of the invention.
Fig. 4 is a schematic flow chart of a PIV flow field experimental method in an experimental method of an extracorporeal circulation experimental table for medical implant observation according to an embodiment of the third aspect of the present invention.
Reference numerals:
extracorporeal circulation experiment table 1000 for medical implant intervention body observation
Transparent tube 104 of medical implant body 103 at inlet end 101 and outlet end 102 of experimental segment module 1
Circulation loop module 2
Hose 21 first pressure sensor 22 first throttle 23 ultrasonic flow sensor 24
Reservoir 25 second pressure transducer 26 syringe pump 27 second throttle 28 exhaust port 29
Temperature control module 3
Heating resistor 31 infrared temperature measurer 32 temperature controller 33
Observation module 4 laser 403
Fluorescence observation system 401 first optical filter 4011 single-lens reflex camera 4012
PIV flow field observation system 402 CCD camera 4021 second optical filter 4022 signal synchronizer 4023
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.
An extracorporeal circulation laboratory table 1000 for medical implant intervention body visualization in accordance with an embodiment of the present invention is described below with reference to fig. 1.
As shown in fig. 1, an extracorporeal circulation experiment table 1000 for medical implant body observation according to an embodiment of the first aspect of the present invention includes an experiment section module 1, a circulation loop module 2, a temperature control module 3, and an observation module 4, wherein the experiment section module 1 includes a medical implant body 103 with a power pump and a transparent tube 104 with biocompatibility, the medical implant body 103 is disposed in the transparent tube 104, and the transparent tube 104 has an inlet end 101 and an outlet end 102; one end of the circulation loop module 2 is connected with the inlet end 101 of the transparent pipe 104, and the other end is connected with the outlet end 102 of the transparent pipe 104, so that the circulation loop module 2 and the experiment section module 1 together form a circulation loop for the circulation flow of the experiment liquid; the circulation loop module 2 is used for monitoring and controlling the pressure and the flow of the experimental liquid which circularly flows in the circulation loop; the temperature control module 3 is used for controlling the temperature of the experimental liquid in the circulating loop; the observation module 4 has a fluorescence observation function and realizes quantitative observation of the area of thrombus on the surface of the medical implant; the observation module 4 also has a flow field observation function, and is used for observing the flow field in the experiment section module 1.
Specifically, the experimental segment module 1 comprises a medical implant body 103 with a power pump and a transparent tube 104 with biocompatibility, wherein the medical implant body 103 is arranged in the transparent tube 104, and the transparent tube 104 is provided with an inlet end 101 and an outlet end 102. It can be understood that the medical implant intervention body 103 can be an artificial heart, an artificial stent, an artificial heart valve, or the like, wherein the artificial heart itself has a function of a power pump, and can provide power for the circulation flow of the experimental liquid in the circulation loop module 2, when the medical implant intervention body 103 such as the artificial stent, the artificial heart valve, or the like does not have a power pump, the artificial heart can be placed in the transparent tube 104 as the power pump together with the artificial stent, the artificial heart valve, or the like for testing, so as to provide power for the circulation flow of the experimental liquid in the circulation loop module 2, so that the experimental liquid in the circulation loop flows circularly, the transparent tube 104 is used for accommodating the medical implant intervention body 103, the transparent tube 104 has a transparent characteristic, so that an experimenter can visually observe the growth condition of thrombus on the surface of the medical implant intervention body 103 in the transparent tube 104, and the transparent tube 104 also has biocompatibility, the blood can be prevented from forming thrombus on the inner wall of the transparent tube 104 in the experimental process to influence the experimental result; the inlet end 101 of the transparent tube 104 is the end of the circulating loop where the experimental liquid flows into the transparent tube 104, the outlet end 102 of the transparent tube 104 is the end of the transparent tube 104 where the experimental liquid flows out, the transparent tube 104 is an acrylic tube, and the acrylic tube is the transparent tube 104 made of acrylic material, so that the transparent tube has good biocompatibility and transparency, and is high in light transmittance.
One end of the circulation loop module 2 is connected with the inlet end 101 of the transparent tube 104, and the other end is connected with the outlet end 102 of the transparent tube 104, so that the circulation loop module 2 and the experiment section module 1 together form a circulation loop for experimental liquid circulation flow, and the circulation loop is used for simulating the circulation in the human body; the circulation loop module 2 is used for monitoring and controlling the pressure and flow of the experimental liquid circulating in the circulation loop, that is, the circulation loop module 2 also has the function of detecting and controlling the pressure and flow of the experimental liquid circulating in the circulation loop, so as to assist the experimenter to adjust the pressure and flow rate in the circulation loop, so as to simulate the blood flowing condition in the human body, and truly and better simulate the circulation environment in the human body.
The temperature control module 3 is used for controlling the temperature of the experimental liquid in the circulation loop, is beneficial to maintaining the temperature of the experimental liquid in the circulation loop within the temperature range of 37 +/-1 ℃ of the physiological temperature of the human body, realizes the constant temperature function and can simulate the temperature environment in the blood vessel of the human body.
The observation module 4 has a fluorescence observation function, and realizes quantitative observation of the area of the surface thrombus of the medical implant intervention body 103, that is, the observation module 4 can capture a fluorescence image of the surface thrombus of the medical implant intervention body 103, realize quantitative observation of the thrombus growth condition on the medical implant intervention body 103 and the area of the surface thrombus of the medical implant intervention body 103, detect whether the thrombolysis performance of the medical implant intervention body 103 meets the clinical use requirement, and provide more observation means for the design optimization of the medical implant intervention body 103.
The observation module 4 further has a flow field observation function, and is configured to observe a flow field in the experiment section module 1, wherein the flow field observation is to observe a flow field condition of a flow field near the medical implant intervention body 103 in the experiment section module 1, and a flow condition of peripheral blood when the medical implant intervention body 103 is in a human body can be determined according to a result of the flow field observation, so that design optimization of the medical implant intervention body 103 can be effectively guided, and a new observation means is provided for research on the medical implant intervention body 103.
According to the extracorporeal circulation experiment table 1000 for medical implant intervention body observation in the embodiment of the first aspect of the invention, on one hand, the circulation loop formed by connecting the transparent tube 104 of the experiment section module 1 and the circulation loop module 2 can truly simulate the intracorporeal circulation loop; the pressure and the flow of the experimental liquid which circularly flows in the circulating loop are monitored and controlled by the circulating loop module 2, so that the pressure and the flow speed in the circulating loop can be conveniently adjusted by an auxiliary experimenter to simulate the blood flowing condition in the human body, meanwhile, the temperature of the experimental liquid in the circulating loop is controlled by the temperature control module 3, the temperature of the experimental liquid in the circulating loop can be maintained in the temperature range of 37 +/-1 ℃ of the physiological temperature of the human body, the constant temperature function is realized, and the temperature environment in the human body can be simulated. On the other hand, because the observation module 4 has a fluorescence observation function, the observation module 4 can capture a fluorescence image of the thrombus on the surface of the medical implant intervention body 103, and can realize quantitative observation of the thrombus growth condition on the medical implant intervention body 103 by shooting a plurality of fluorescence images at intervals in the thrombus growth process, and can also quantitatively observe the area of the thrombus on the surface of the medical implant intervention body 103 to detect whether the thrombolysis performance of the medical implant intervention body 103 meets the clinical use requirement; because the observation module 4 has a flow field observation function, the observation module 4 can observe the flow field condition of the flow field near the medical implant intervention body 103 in the experiment section module 1, and the flow condition of blood around the medical implant intervention body 103 can be judged when the medical implant intervention body 103 is in a human body according to the result of the flow field observation.
Therefore, the in vitro circulation experiment table 1000 for observing the medical implant intervention body in the embodiment of the first aspect of the invention can better simulate the environment in the human body and simulate the growth condition of thrombus in the human body in vitro, thereby providing a stable and reliable experiment platform for the anticoagulant and thrombus inhibition performance research of the medical implant intervention body 103 such as an artificial heart, an artificial stent or an artificial heart valve; the area of the thrombus on the surface of the medical implant intervention body 103 can be observed in a quantification manner, the growth process condition of the thrombus on the surface of the medical implant intervention body 103 can be visually and quantificationally known, the flow field of the medical implant intervention body 103 can be observed through the observation module 4, the flow field condition of a drainage basin near the medical implant intervention body 103 in the experiment section module 1 can be observed through the observation module 4, and the flow condition of blood around the medical implant intervention body 103 can be judged when the medical implant intervention body 103 is in a human body according to the observation result of the flow field.
According to one embodiment of the first aspect of the present invention, the circulation loop module 2 comprises a hose 21 having biocompatibility, a first pressure sensor 22, a first throttle 23, an ultrasonic flow sensor 24, a reservoir 25, a first injection port, a second injection port, and a second pressure sensor 26; one end of the hose 21 is connected to the inlet end 101 of the transparent tube 104 and the other end is connected to the other end of the transparent tube 104; the first pressure sensor 22, the first throttle 23, the ultrasonic flow sensor 24, the liquid storage 25, the first inlet, the second inlet, and the second pressure sensor 26 are sequentially provided on the hose 21 in a direction from the other end of the hose 21 to one end of the hose 21. It can be understood that the biocompatible hose 21 is arranged in the circulation loop module 2, so that the aorta vessel of a human body can be simulated, the hose 21 has the characteristic of good biocompatibility, and blood coagulation on the inner surface of the hose 21 can be avoided to form thrombus, so that a normal experiment is not influenced, and therefore, the hose 21 with the biocompatibility is adopted to ensure that the experiment is accurately and efficiently carried out; the first pressure sensor 22, the first throttle valve 23, the ultrasonic flow sensor 24, the liquid storage container 25, the first injection port, the second injection port and the second pressure sensor 26 are sequentially arranged on the hose 21 in a direction from the other end of the hose 21 to one end of the hose 21, and are arranged in this order, so that on one hand, the circulation loop module 2 and the experimental section module 1 form a complete circulation loop for the experimental liquid to circularly flow, and the experimental liquid entering the circulation loop can firstly flow into the experimental section module 1; on the other hand, the pressure and flow of the circulation loop can be further monitored and controlled.
Specifically, the first pressure sensor 22 is close to the outlet end 102 for monitoring the outlet pressure of the experimental liquid circulating in the circulation loop after flowing out of the experimental section module 1, and the second pressure sensor 26 is close to the inlet end 101 for monitoring the inlet pressure of the experimental liquid circulating in the circulation loop before entering the experimental section module 1. Therefore, the pressure change condition of the experimental liquid in the experimental section module 1 can be known through the values monitored by the first pressure sensor 22 and the second pressure sensor 26, and the experimenter can judge the growth condition of the thrombus on the surface of the implant body according to the pressure change condition.
Specifically, the first throttle valve 23 and the reservoir 25 are used to regulate the flow rate and pressure of the test liquid circulating in the circulation circuit, and the reservoir 25 is also used to discharge the gas in the test liquid circulating in the circulation circuit; it will be appreciated that the flow and pressure of the test fluid in the circulation loop are adjusted to bring the flow rate and pressure of the test fluid as close as possible to the blood flow of the human body. Because in concrete experimental process, need add experimental liquid to the circulation circuit, the injection of experimental liquid can inevitably produce the bubble when can occupy the volume in the circulation circuit, shake gently and strike the position that has the bubble, make the bubble along hose 21 get into in the stock solution container 25, the stock solution container 25 can be a soft transparent bag of making by the material that has the biocompatibility, be provided with sealed gas vent 29 on the stock solution container 25, open gas vent 29 on the stock solution container 25, the bubble in the stock solution container 25 of discharging, close gas vent 29 again, thereby avoid the bubble to cause the influence to experimental liquid flow in the circulation circuit and the formation of thrombus.
Specifically, the ultrasonic flow sensor 24 is used to monitor the flow rate of the test liquid circulating in the circulation circuit; it can be understood that, on the one hand, the ultrasonic flow sensor 24 can detect the flow rate of the experimental liquid in the circulation loop, and the thrombus formation can be judged by observing the flow rate value of the experimental liquid, for example, when the flow sensor parameter is reduced to 20% of the initial parameter, it indicates that a large amount of thrombus has been formed on the surface of the tested medical implant insertion body 103, and the experiment is stopped; on the other hand, the ultrasonic flow sensor 24 is a non-contact flow monitor, that is, the ultrasonic flow sensor 24 does not contact with the experimental liquid when detecting the flow of the experimental liquid, thereby avoiding the condition that the experimental liquid forms thrombus on the foreign matter on the surface of the common flow monitor and reducing the influence of external factors on the experimental process.
Specifically, the first injection port and the second injection port are used for respectively injecting corresponding experimental liquid according to experimental requirements. In practice, the first injection port can be used for injecting physiological saline into the circulation loop and withdrawing the physiological saline in the circulation loop from the first injection port, and for injecting test blood for the experimental test into the circulation loop and withdrawing the test blood from the first injection port; the second injection port can be used for adding the experimental reagent, such as CaCl, into the circulation loop at a constant speed during the experiment 2 Solutions, etc. to neutralize anticoagulants or regulate the rate of thrombus formation in the blood, and the test reagents may be selected according to the needs of the test procedure. Physiological saline, test blood, test reagents and the like belong to test liquids.
According to a further embodiment of the first aspect of the invention, the hose 21 is a medical PVC hose. It can be understood that the medical PVC hose has better biocompatibility, and blood can not form thrombus on the medical PVC hose when flowing through the medical PVC hose, thereby being beneficial to the normal operation of the experimental test process of the medical implant intervention body 103; meanwhile, the medical PVC hose has better elasticity and can better simulate blood vessels of human bodies.
According to a still further embodiment of the first aspect of the present invention, the inner diameter of the flexible tube 21 is 3/8 feet, the total length of the circulation loop is 200 + -20 cm, the inner diameter of the flexible tube 21 is 3/8 feet close to the inner diameter of the aorta vessel of the human body, and the total length of the circulation loop is set within 200 + -20 cm, so that the human body internal loop can be better simulated.
According to a still further embodiment of the first aspect of the present invention, the circulation loop module 2 further comprises a syringe pump 27 and a second throttle valve 28, the syringe pump 27 is connected to the second injection port through a connecting pipe, and the second throttle valve 28 is disposed on the connecting pipe; the syringe pump 27 is used to inject the pharmaceutical agent. It should be noted that, when the pharmaceutical agent needs to be injected into the circulation loop, the second throttle 28 is opened, the injection pump 27 can uniformly inject the pharmaceutical agent into the circulation loop at a constant speed, so as to avoid the situation of transient local concentration in the circulation loop being too high, and after the injection is finished, the second throttle 28 is closed, so as to avoid the experimental liquid in the circulation loop from flowing out of the circulation loop, that is, the second throttle 28 can be used for adjusting the injection speed of the pharmaceutical agent and controlling the opening and communication between the circulation loop and the connecting pipe, so as to function as a switch.
According to a still further embodiment of the first aspect of the present invention, the temperature control module 3 comprises a heating resistor 31, a temperature controller 33 and an infrared temperature detector 32; the heating resistor 31 is wrapped on the liquid storage container 25 and used for heating the liquid storage container 25; the temperature controller 33 is used for controlling the temperature of the heating resistor 31 so as to maintain the experiment liquid circulating in the circulating loop at a constant physiological temperature; the infrared temperature detector 32 is used for measuring the temperature of the experimental section module 1. It should be noted that, the heating resistor 31 and the liquid storage container 25 are in a non-contact state, the heating resistor 31 does not cause local high temperature in the liquid storage container 25, and when the heating resistor 31 heats the experimental liquid in the liquid storage container 25, blood cells in the experimental liquid are not damaged, and the heating resistor 31 heats the liquid storage container 25, so that the experimental liquid is at a constant physiological temperature. The temperature controller 33 is used to control the temperature of the heating resistor 31 so as to control the test liquid at a constant physiological temperature. The infrared temperature detector 32 is a non-contact temperature detector, so that the condition that experimental liquid forms thrombus on foreign matters on the surface of a common temperature detector is avoided, and the influence of external factors on the experimental process is reduced.
According to some embodiments of the first aspect of the present invention, the observation module 4 comprises a fluorescence observation system 401 for realizing fluorescence observation function, the fluorescence observation system 401 comprises a laser 403, a first optical filter 4011 and a single-lens reflex camera 4012; the laser 403 is used for emitting continuous laser to irradiate on the experimental section module 1, so that the fluorescent reagent on the thrombus on the surface of the medical implant intervention body 103 excites fluorescence; the first optical filter 4011 is arranged between the experimental section module 1 and the single lens reflex 4012; the single lens reflex 4012 is used for shooting a first fluorescence image, which is a fluorescence image of thrombus components, of fluorescence from a fluorescence reagent on the thrombus on the surface of the medical implant intervention body 103 after being filtered by the first optical filter 4011. It should be noted that, the laser 403 is used to emit laser to excite the fluorescent reagent to generate fluorescence, the incident light direction of the laser 403 is perpendicular to the shooting direction of the single-lens reflex camera 4012, the single-lens reflex camera 4012 is located on one side of the experimental section module 1, the laser emitted by the laser 403 is irradiated on the experimental section module 1, the first optical filter 4011 is arranged to filter stray light from the laser 403 and the environment, and light in a single waveband is obtained, so that the shot picture is clearer and easy to distinguish, wherein the single-lens reflex camera 4012 mainly plays a role in shooting and recording, a common camera with a clear shooting function can also meet experimental requirements, and can quantify parameters such as area of observed thrombus after recording the fluorescence picture of thrombus on the surface of the medical implant 103, thereby providing more experimental bases for the detection of blood coagulation dissolving performance of the medical implant 103.
According to some embodiments of the first aspect of the present invention, the observation module 4 further includes a PIV flow field observation system 402 for implementing a flow field observation function, the PIV flow field observation system 402 includes a laser 403, a CCD camera 4021 and a second optical filter 4022, the laser 403 is used for exciting pulsed laser to irradiate on the experiment section module 1, so that fluorescent particles in the experiment liquid in the experiment section module 1 excite fluorescence; the second optical filter 4022 is arranged between the experimental segment module 1 and the CCD camera 4021; the CCD camera 4021 is configured to capture a second fluorescence image, i.e., a fluorescence image of the fluorescent particles in the flow field, obtained by filtering fluorescence of the fluorescent particles in the experimental liquid in the experimental section module 1 through the second optical filter 4022, and the capturing frequency of the CCD camera 4021 is synchronized with the pulse frequency of the pulse laser. It should be noted that the laser 403 is used for emitting high-intensity pulse laser to excite fluorescent particles to generate fluorescence, the incident light direction of the laser 403 is perpendicular to the shooting direction of the CCD camera 4021, the CCD camera 4021 is located at one side of the experimental segment module 1, the laser emitted by the laser 403 irradiates the experimental segment module 1, the second optical filter 4022 is arranged to filter stray light from the laser 403 and the environment to obtain light in a single band, so that the shot image is clearer and easy to distinguish, wherein the pulse laser emitted by the laser 403 has high intensity and high instantaneous pulse energy, and the laser 403 cannot be in a working state for a long time, so that the shooting frequency of the CCD camera 4021 is synchronous with the pulse frequency of the pulse laser, and the CCD camera 4021 can capture the instant fluorescence emitted by the fluorescent particles in the experimental liquid to continuously record the motion trajectory of the fluorescent particles to obtain the flow field condition around the medical implant 103, thereby simulating the flow field condition of the medical implant 103 in the human body.
According to some embodiments of the first aspect of the present invention, the PIV flow field observation system 402 further comprises a signal synchronizer 4023, and the signal synchronizer 4023 is electrically connected to the laser 403 and the CCD camera 4021, respectively, to synchronize the pulse frequency of the pulsed laser and the photographing frequency of the CCD camera 4021. It can be understood that, as shown in fig. 1, the signal synchronizer 4023 may synchronize the pulse frequency of the pulse laser emitted by the laser 403 and the shooting frequency of the CCD camera 4021, so that the fluorescence image of the flow field condition of the flow field near the medical implant 103 may be recorded when the experimental segment module 1 is excited by the pulse laser to emit fluorescence, thereby obtaining the condition of the flow field in the experimental segment module 1.
The invention also provides an experimental method of the extracorporeal circulation experiment table for observing the medical implant.
Referring to fig. 3, the experimental method of the extracorporeal circulation laboratory table for medical implant interventional body observation according to the embodiment of the second aspect of the invention is an extracorporeal circulation laboratory table 1000 for medical implant interventional body observation according to any one of the embodiments of the first aspect of the invention (see fig. 1 to 2). The experimental method is a fluorescence observation experimental method, and comprises the following steps:
step S1: mixing the first fluorescent agent and the collected blood in proportion, fully and uniformly mixing the mixture in a water bath at 37 ℃, injecting the mixture into a circulation loop, and discharging the blood after the medical implant intervention body 103 works for a period of time to form thrombus; or the second fluorescent agent is directly coated on the thrombus on the surface of the medical implant intervention body 103 as an observation object and is placed in the transparent tube 104.
It is understood that the first fluorescent agent and the second fluorescent agent can fluorescently label insoluble fibrin or platelets in the thrombus, so that the observation module 4 can capture a fluorescence image of the thrombus and can quantitatively observe the thrombus area of the medical implant intervention body 103. In a specific experiment, the first fluorescent agent may be fluorochrome DiOC6, fluorochrome FITC, or a mixture of fluorochrome DiOC6 and fluorochrome FITC, wherein the concentration of fluorochrome DiOC6 may be 10 μ g/ml, the concentration of fluorochrome FITC may be 10 μ g/ml, fluorochrome DiOC6 and blood may be mixed according to a volume ratio of 1:100, fluorochrome FITC and blood may be mixed according to a volume ratio of 1:100, and when the fluorochrome DiOC6 and the fluorochrome FITC are used simultaneously, the ratio of fluorochrome DiOC6 to blood and the ratio of fluorochrome FITC to blood are both 1:100, so that the labeling effect of the first fluorescent agent is good.
The first fluorescent agent and the blood are fully and uniformly mixed in a water bath at 37 ℃, so that the blood recovers physiological temperature and is favorable for quickly recovering activity of the blood, and insoluble fibrin or platelets in the blood are fully fluorescently labeled by the first fluorescent agent. In order to mix the first fluorescent agent and the blood sufficiently in the water bath at 37 ℃, the mixing in the water bath needs to last for a period of time, which may be 10min, for example.
The first fluorescent agent and the blood are uniformly mixed and then injected into the circulation loop, after the medical implant intervention body 103 works for a period of time, the blood containing the first fluorescent agent can form a fluorescence-dyed thrombus on the surface of the medical implant intervention body 103, so that the observation module 4 can capture a fluorescence image of the thrombus and further can quantitatively observe the thrombus area of the medical implant intervention body 103.
The second fluorescent agent is directly coated on the thrombus so that the observation module 4 can capture a fluorescence image on the thrombus and can quantitatively observe the thrombus area of the medical implant intervention body 103. In a specific experiment, the second fluorescent agent may be DiOC6, FITC, or a mixture of DiOC6 and FITC, wherein the concentration of DiOC6 may be 10 μ g/ml, and the concentration of FITC may be 10 μ g/ml, so that the labeling effect of the second fluorescent agent is good.
It should be noted that the blood can be collected as follows: selecting animals with normal body temperature and no disease characteristics, and fasting for 12 h;
collecting animal blood into blood bag containing anticoagulant by venipuncture with large-caliber needle (14G or larger), wherein the blood can naturally flow out by gravity or be collected by pressure difference principle with negative pressure not exceeding 100 mmHg;
the blood is used within 24h after fresh human blood is collected, and is used within 48h after fresh blood of other animals except human is collected, preferably within one hour after blood is collected, and the collected blood is transported and stored at the temperature of 2-8 ℃.
The blood collection step can be used to obtain blood required by experimental tests, and the state of the blood is close to the state of the blood in the animal body.
In addition, blood can be replaced by plasma according to different experimental requirements so as to obtain a clearer observation field.
Step S2: the observation module 4 is used for quantitatively observing the area of thrombus on the surface of the medical implant intervention body 103. That is to say, the observation module 4 can capture the fluorescence image of the thrombus on the surface of the medical implant intervention body 103, so as to realize quantitative observation of the thrombus growth condition on the medical implant intervention body 103 and the area of the thrombus on the surface of the medical implant intervention body 103, detect whether the thrombolysis performance of the medical implant intervention body 103 meets the clinical use requirement, and provide more observation means for the design optimization of the medical implant intervention body 103.
According to the experimental method of the extracorporeal circulation experiment table for medical implant intervention body observation in the embodiment of the second aspect of the invention, the extracorporeal circulation experiment table 1000 for medical implant intervention body observation in the embodiment of the first aspect of the invention can truly and better simulate the human body internal circulation environment in the aspects of pressure, flow, temperature and the like, and provides a good experimental basis for a fluorescence observation experimental method. According to the fluorescence observation experiment method provided by the embodiment of the second aspect of the invention, firstly, the first fluorescent dye is mixed with blood to form a fluorescence-dyed thrombus on the surface of the medical implant intervention body 103, or the second fluorescent agent is directly coated on the surface of the medical implant intervention body 103 with the thrombus formed, so that the fluorescence dyeing of the thrombus is realized, the observation module 4 can capture the fluorescence image of the thrombus on the surface of the medical implant intervention body 103, the growth condition of the thrombus on the medical implant intervention body 103 and the area of the thrombus on the surface of the medical implant intervention body 103 are quantitatively observed, whether the blood-dissolving coagulation performance of the medical implant intervention body 103 meets the clinical use requirement is detected, more observation means are provided for the design optimization of the medical implant intervention body 103, and the design optimization of the medical implant intervention body 103 can be effectively guided.
According to an embodiment of the second aspect of the invention, the first fluorescent agent is a fluorescent agent comprising DiOC6 or/and FITC, the marking effect is good; the second fluorescent agent is DiOC6 or/and FITC, and the marking effect is good.
According to a further embodiment of the second aspect of the present invention, in step S2, the observation module 4 includes the fluorescence observation system 401 for implementing fluorescence observation function in an embodiment of the first aspect of the present invention, and the fluorescence observation system 401 includes the laser 403, the first optical filter 4011, and the single lens reflex camera 4012; the laser 403 is used for emitting continuous laser to irradiate on the experimental section module 1, so that the fluorescent reagent on the thrombus on the surface of the medical implant intervention body 103 excites fluorescence; the first optical filter 4011 is arranged between the experimental segment module 1 and the single lens reflex 4012; the single lens reflex 4012 is used for shooting a first fluorescence image obtained by filtering fluorescence from a fluorescence reagent on thrombus on the surface of the medical implant intervention body 103 through the first optical filter 4011.
Step S2 specifically includes the following substeps:
step S201: the blue laser with the wavelength of 490-515nm provided by the laser 403 is used to irradiate the experimental segment module 1, so that the fluorescent reagent on the thrombus on the surface of the medical implant 103 excites green fluorescence.
The blue excitation light with the wavelength of 490-515nm provided by the laser 403 is used for exciting the first fluorescent agent or the second fluorescent agent to generate fluorescence, and the blue excitation light with the wavelength of 490-515nm is adopted because the excitation wavelengths of the first fluorescent agent and the second fluorescent agent are located in the wavelength range of 490-515nm, and is irradiated on the experimental section module 1, that is, the surface of the medical implant 103 in the experimental section module 1 is irradiated through the transparent tube 104, so that the fluorescent reagent on the thrombus on the surface of the medical implant 103 is excited to emit green fluorescence, and thus the single lens reflex 4012 can capture the fluorescence image of the thrombus;
step S202: a green band-pass filter with the wavelength of 550nm-570nm is selected as a first filter 4011 and is additionally arranged in front of a single lens reflex 4012.
It should be noted that, a green band-pass filter with a wavelength of 550nm to 570nm is used to filter the laser light emitted by the laser 403, because the wavelength of the laser light emitted by the laser 403 is closer to the wavelength of the fluorescence generated by the first fluorescent agent and the second fluorescent agent, the presence of the excitation light interferes with the observation of the thrombus, the first filter 4011 is arranged to filter out the light rays in other bands, and only the light with a wavelength of 550nm to 570nm is left, so that the picture shot by the single lens reflex 4012 is clearer and easier to distinguish, and the information such as the area of the thrombus is recorded more accurately and clearly.
Step S203: a first fluorescence image of the thrombus on the surface of the medical implant intervention body 103 is shot by using the single lens reflex 4012 under the condition of a total dark field, and the thrombus area is observed quantitatively according to the first fluorescence image.
That is to say, the single lens reflex 4012 is under the condition of the whole dark field when taking the thrombus on the surface of the medical implant intervention body 103, and under the condition of the whole dark field, the light intensity contrast is increased, so that the single lens reflex 4012 is favorable for obtaining clearer and easily distinguished thrombus fluorescence images when taking the thrombus, thereby being favorable for observing information such as fluorescence area of the thrombus, and taking the thrombus for many times according to a certain time interval, and being capable of observing the generation process of the thrombus.
The third aspect of the invention also provides an experimental method of the extracorporeal circulation experiment table 1000 for medical implant interventional body observation.
As shown in fig. 3, the experimental method of the extracorporeal circulation laboratory table for medical implant interventional body observation according to the third aspect of the invention is the extracorporeal circulation laboratory table 1000 for medical implant interventional body observation according to any one of the embodiments of the first aspect of the invention. The experimental method is a PIV flow field experimental method and comprises the following steps:
step S6: and adding the third fluorescent particles into the transparent liquid to obtain a fluorescent particle solution. Here, the third fluorescent particle is suitable for flow field observation.
It should be noted that the third fluorescent particle may be a fluorescent particle with a diameter of about 10-20 microns, the fluorescent particle with this size range is suitable for performing flow field observation, the flow field observation method is to first shoot position pictures of 2 fluorescent particles in a flow field at a time interval (about 100 microseconds), then automatically analyze the position change of the same fluorescent particle through software, and finally obtain the velocity distribution in the flow field by dividing the position change value by the time interval to obtain the flow field condition, the fluorescent particle with this size can better track the flow condition of the fluid when flowing along with the liquid, and is also easier to observe, therefore, the fluorescent particle with a diameter of about 10-20 microns is adopted. Preferably, the third fluorescent particle is a Fluo-610 fluorescent particle, the diameter of the fluorescent particle is in the range of 10-20 microns, and the excitation wavelength is in the range of 490-515nm wavelength provided by the laser 403, so that when fluid observation is performed by using the Fluo-610 fluorescent particle, the lens of the laser 403 does not need to be replaced, and the observation is convenient to perform. Preferably, the concentration of the third fluorescent particle in the transparent liquid is 0.1g/L, wherein the transparent solution may be normal saline, tap water or transparent plasma, depending on experimental scenarios and requirements.
Step S7: the fluorescent particle solution is injected into the circulation loop and mixed thoroughly.
Specifically, the injection volume of the fluorescent particle solution is 200 ± 20mL, because the inner diameter of the hose 21 in the circulation loop module 2 is 3/8 feet, and the total length of the circulation loop is 200 ± 20cm, the injection of 200mL of the fluorescent particle solution in the circulation loop can better simulate the circulation in the human body, and the circulation loop module 2 is started to fully mix the fluorescent particle solution, so that the fluorescent particles are uniformly dispersed in the circulation loop module 2, which is beneficial to more accurately observing the condition of the flow field.
Step S8: the observation module 4 is used to observe the flow field in the experimental section module 1. According to the result of the flow field observation, the flowing condition of the peripheral blood when the medical implant intervention body 103 is in the human body can be judged, so that the design optimization of the medical implant intervention body 103 can be effectively guided, and a new observation means is provided for the research of the medical implant intervention body 103.
According to the experimental method of the extracorporeal circulation experiment table 1000 for medical implant intervention body observation in the embodiment of the third aspect of the present invention, the third fluorescent particles are added into the transparent liquid to obtain the fluorescent particle solution, the fluorescent particle solution is injected into the circulation loop, the flow field in the experiment section module 1 is observed by using the flow observation function of the observation module 4, and the flow condition of the peripheral blood when the medical implant intervention body 103 is in the human body can be judged according to the result of the flow field observation, so that the design optimization of the medical implant intervention body 103 can be effectively guided, and a new observation means is provided for the research of the medical implant intervention body 103.
According to an embodiment of the third aspect of the invention, the third fluorescent particle is a Fluo-610 fluorescent particle.
It can be understood that the diameter of the Fluo-610 fluorescent particle is in the range of 10-20 microns, which is beneficial for smooth observation, and the excitation wavelength is in the range of 490-515nm wavelength provided by the laser 403, so that the lens of the laser 403 does not need to be replaced during observation, and the observation is more convenient.
According to a further embodiment of the third aspect of the present invention, the observation module 4 includes a PIV flow field observation system 402 in some embodiments according to the first aspect of the present invention, the PIV flow field observation system 402 includes a laser 403, a CCD camera 4021 and a second filter 4022, the laser 403 is used for exciting pulsed laser light to irradiate on the experiment section module 1 so as to excite fluorescent particles in the experiment liquid in the experiment section module 1 to emit fluorescent light; the second optical filter 4022 is arranged between the experimental segment module 1 and the CCD camera 4021; the CCD camera 4021 is configured to capture a second fluorescence image obtained by filtering fluorescence from fluorescent particles in the test liquid in the test section module 1 through the second optical filter 4022, and the capturing frequency of the CCD camera 4021 is synchronized with the pulse frequency of the pulse laser. The step 8 specifically comprises the following substeps:
step S801: the experimental section module 1 is irradiated with the pulsed laser light provided by the laser 403, so that the fluorescent particles in the fluorescent particle solution in the experimental section module 1 are excited to fluoresce.
The laser 403 is configured to provide pulsed laser to excite the third fluorescent particles in the experiment segment module 1 to generate fluorescence, so as to observe the flow field in the experiment segment module 1.
Step S802: a second fluorescence image of fluorescence excited by the third fluorescent particle in the experimental section module 1 after passing through the second optical filter 4022 is photographed by the CCD camera 4021 under a full dark field condition, and the photographing frequency of the CCD camera 4021 is synchronized with the pulse frequency of the pulse laser.
As shown in fig. 2, the CCD camera 4021 may implement high-frequency shooting, may record the position of the third fluorescent particle for multiple times in a short time, and is favorable for observing the flow field, the second optical filter 4022 is used to filter the laser emitted by the laser 403, so that the image shot by the CCD camera 4021 is clearer, easy to distinguish, and more accurately and clearly obtains the flow field condition, the intensity of the pulse laser emitted by the laser 403 is high, and the instantaneous pulse energy is high, so the laser 403 cannot be in a working state for a long time, and after the emitted instantaneous laser excites the third fluorescent particle in the experimental liquid to excite fluorescence, instantaneous shooting needs to be performed at the same time, so the shooting frequency of the CCD camera 4021 needs to be synchronized with the pulse frequency of the pulse laser.
Step S803: the obtained second fluorescence image is processed by using special processing software, and the flow field condition of the drainage basin near the medical implant intervention body 103 is obtained.
Specifically, the special processing software is dynamic studio v3.41, the special processing software can automatically analyze the position change of the same fluorescent particle, and finally, the position change value is divided by the time interval to obtain the velocity distribution in the flow field, so that the flow field condition is obtained.
The following describes in detail the extracorporeal circulation test table and test method for medical implant interventional body observation according to the present invention with specific examples.
Example 1: extracorporeal circulation experiment table for medical implant intervention body observation
In example 1, the extracorporeal circulation experiment table 1000 for medical implant interventional body observation comprises an experiment section module 1, a circulation loop module 2, a temperature control module 3 and an observation module 4,
the experimental segment module 1 comprises a medical implant body 103 with a power pump and a transparent tube 104 with biocompatibility, wherein the medical implant body 103 is arranged in the transparent tube 104, and the transparent tube 104 is provided with an inlet end 101 and an outlet end 102.
One end of the circulation loop module 2 is connected with the inlet end 101 of the transparent pipe 104, and the other end is connected with the outlet end 102 of the transparent pipe 104, so that the circulation loop module 2 and the experiment section module 1 together form a circulation loop for the circulation flow of the experiment liquid; the circulation loop module 2 is used to monitor and control the pressure and flow of the test liquid circulating in the circulation loop.
The temperature control module 3 is used to control the temperature of the test liquid in the circulation loop.
The observation module 4 has a fluorescence observation function and realizes quantitative observation of the area of thrombus on the surface of the medical implant; the observation module 4 also has a flow field observation function, and is used for observing the flow field in the experiment section module 1.
Specifically, the circulation loop module 2 includes a hose 21 having biocompatibility, a first pressure sensor 22, a first throttle 23, an ultrasonic flow sensor 24, a reservoir 25, a first injection port, a second injection port, and a second pressure sensor 26; one end of the hose 21 is connected to the inlet end 101 of the transparent tube 104 and the other end is connected to the other end of the transparent tube 104; a first pressure sensor 22, a first throttle 23, an ultrasonic flow sensor 24, a liquid storage container 25, a first injection port, a second injection port, and a second pressure sensor 26 are sequentially disposed on the hose 21 in a direction from the other end of the hose 21 to one end of the hose 21;
wherein the first pressure sensor 22 is close to the outlet end 102, and is used for monitoring the outlet pressure of the experimental liquid which circularly flows in the circulating loop after flowing out of the experimental section module 1; a second pressure sensor 26 is close to the inlet end 101 for monitoring the inlet pressure of the test liquid circulating in the circulation loop before entering the test section module 1; the first throttle valve 23 and the reservoir 25 are used for regulating the flow rate and pressure of the test liquid circulating in the circulation circuit, and the reservoir 25 is also used for discharging gas in the test liquid circulating in the circulation circuit; the ultrasonic flow sensor 24 is used for monitoring the flow rate of the test liquid circulating in the circulation loop; the first injection port and the second injection port are used for respectively injecting corresponding experimental liquid according to experimental requirements, and the hose 21 is a medical PVC hose.
The circulation loop module 2 further comprises an injection pump 27 and a second throttle valve 28, the injection pump 27 is connected with the second injection port through a connecting pipe, and the second throttle valve 28 is arranged on the connecting pipe; the syringe pump 27 is used to inject the pharmaceutical agent.
The temperature control module 3 comprises a heating resistor 31, a temperature controller 33 and an infrared temperature detector 32; the heating resistor 31 is wrapped on the liquid storage container 25 and used for heating the liquid storage container 25; the temperature controller 33 is used for controlling the temperature of the heating resistor 31 so as to maintain the experiment liquid circulating in the circulating loop at a constant physiological temperature; the infrared temperature detector 32 is used for measuring the temperature of the experimental section module 1.
The observation module 4 comprises a fluorescence observation system 401 for realizing a fluorescence observation function, wherein the fluorescence observation system 401 comprises a laser 403, a first optical filter 4011 and a single lens reflex 4012; the laser 403 is used for emitting continuous laser to irradiate on the experimental section module 1, so that the fluorescent reagent on the thrombus on the surface of the medical implant intervention body 103 excites fluorescence; the first optical filter 4011 is arranged between the experimental segment module 1 and the single lens reflex 4012; the single lens reflex 4012 is configured to capture a first fluorescence image obtained by filtering fluorescence from the fluorescent reagent on the surface thrombus of the medical implant intervention body 103 through the first optical filter 4011.
The observation module 4 further comprises a PIV flow field observation system 402 for realizing a flow field observation function, the PIV flow field observation system 402 comprises a laser 403, a CCD camera 4021 and a second optical filter 4022, and the laser 403 is used for exciting pulsed laser to irradiate the experimental section module 1 so as to excite fluorescent particles in the experimental liquid in the experimental section module 1 to emit fluorescence; the second optical filter 4022 is arranged between the experimental segment module 1 and the CCD camera 4021; the CCD camera 4021 is configured to capture a second fluorescence image obtained by filtering fluorescence from fluorescent particles in the test liquid in the test section module 1 through the second optical filter 4022, and the capturing frequency of the CCD camera 4021 is synchronized with the pulse frequency of the pulse laser.
The PIV flow field observation system 402 further includes a signal synchronizer 4023, and the signal synchronizer 4023 is electrically connected to the laser 403 and the CCD camera 4021, respectively, to synchronize the pulse frequency of the pulsed laser and the shooting frequency of the CCD camera 4021.
The functions and effects of the functional modules in this example are the same as those of the corresponding functional modules in the foregoing, and are not described again here.
Example 2: a fluorescence observation experiment of the artificial heart pump thrombosis process was performed using the extracorporeal circulation test table 1000 for medical implant interventional body observation of example 1.
In example 2, the medical implant access body 103 in the experimental segment module 1 is an artificial heart pump, and the experimental fluid in the circulation loop is a fluorescent-stained transparent plasma.
The experiment included the following steps:
fully and uniformly mixing a fluorescent dye DiOC6 with the concentration of 10 mu g/ml and a fluorescent dye FITC with the concentration of 10 mu g/ml with freshly collected blood in a water bath at 37 ℃ for 10min, wherein the volume ratio of the fluorescent dye DiOC6 to the freshly collected blood is 1:100, and the volume ratio of the fluorescent dye FITC to the freshly collected blood is 1: 100;
when thrombus exists on the surface of the medical implant intervention body, the surface of the thrombus on the medical implant intervention body 103 is directly coated with a fluorescent dye DiOC6 with the concentration of 10 mu g/ml and a fluorescent dye FITC with the concentration of 10 mu g/ml, and the fluorescent dye DiOC6 and the fluorescent dye FITC can be coated respectively or coated after being mixed;
a green band-pass filter with the wavelength of 550nm-570nm is additionally arranged in front of the single lens reflex 4012;
the laser 403 emits blue excitation light with the wavelength of 490-515nm to irradiate the surface of the artificial heart pump, and the thrombus on the surface of the artificial heart pump is dyed by a fluorescent dye DiOC6 and a fluorescent dye FITC and then excited by the blue excitation light to emit green fluorescence;
under the condition of a whole dark field, a fluorescence image of the thrombus is shot by using the single-lens reflex 4012, and the process of thrombus formation can be obtained by shooting for a plurality of times at certain time intervals.
Example 3: an experiment of PIV flow field observation at the anterior guide vane of an artificial heart pump was performed using the extracorporeal circulation test rig 1000 for medical implant body observation of example 1.
In example 3, the medical implant access body 103 in the experimental segment module 1 is an artificial heart pump, and the transparent liquid in the circulation loop is physiological saline. The experiment comprises the following steps:
adding Fluo-610 fluorescent particles into normal saline, and fully mixing to obtain a transparent solution containing 0.1g/L Fluo-610 fluorescent particles;
injecting 200mL of transparent solution containing Fluo-610 fluorescent particles into the circulation loop, and starting the circulation loop module 2 to fully mix the transparent solution containing Fluo-610 fluorescent particles;
under the condition of a full dark field, the PIV flow field observation system 402 is opened, the laser 403 emits pulse laser to excite Fluo-610 fluorescent particles to generate fluorescence, the fluorescence is filtered by the second optical filter 4022 and then captured by the CCD camera 4021, and the signal synchronizer 4023 synchronizes the pulse laser frequency of the laser 403 and the shooting frequency of the CCD camera 4021;
the obtained image is processed by using special processing software to obtain the flow field condition of the drainage basin near the medical implant intervention body 103.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an illustrative embodiment," "an example," "a specific example," or "some examples" or the like mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
While embodiments of the invention have been shown and described, it will be understood by those of ordinary skill in the art that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.

Claims (12)

1. An extracorporeal circulation laboratory table for medical implant body observation, comprising:
the experimental section module comprises a medical implant intervention body with a power pump and a transparent tube with biocompatibility, wherein the medical implant intervention body is arranged in the transparent tube, and the transparent tube is provided with an inlet end and an outlet end;
a circulation loop module, wherein one end of the circulation loop module is connected with the inlet end of the transparent pipe, and the other end of the circulation loop module is connected with the outlet end of the transparent pipe, so that the circulation loop module and the experiment section module jointly form a circulation loop for the experimental liquid to circularly flow; the circulation loop module is used for monitoring and controlling the pressure and the flow of the experimental liquid circulating in the circulation loop; the circulation loop module comprises a hose with biocompatibility, a first pressure sensor, a first throttling valve, an ultrasonic flow sensor, a liquid storage container, a first injection port, a second injection port and a second pressure sensor; one end of the hose is connected with the inlet end of the transparent pipe, and the other end of the hose is connected with the other end of the transparent pipe; the first pressure sensor, the first throttle valve, the ultrasonic flow sensor, the liquid storage container, the first injection port, the second injection port and the second pressure sensor are sequentially arranged on the hose in the direction from the other end of the hose to one end of the hose;
a temperature control module for controlling the temperature of the test liquid in the circulation loop; the temperature control module comprises a heating resistor, a temperature controller and an infrared temperature detector; the heating resistor wraps the liquid storage container and is used for heating the liquid storage container; the temperature controller is used for controlling the temperature of the heating resistor so as to maintain the experiment liquid circulating in the circulating loop at a constant physiological temperature; the infrared temperature detector is used for measuring the temperature of the experimental section module;
the observation module has a fluorescence observation function, and realizes real-time observation of thrombus growth on the medical implant intervention body and quantitative observation of the area of thrombus on the surface of the medical implant intervention body; the observation module also has a flow field observation function and is used for observing the flow field in the experiment section module;
the first injection port and the second injection port are used for respectively injecting corresponding experimental liquid according to experimental requirements, the circulation loop module further comprises an injection pump and a second throttle valve, the injection pump is connected with the second injection port through a connecting pipe, and the second throttle valve is arranged on the connecting pipe; the syringe pump is for injecting a pharmaceutical agent.
2. The extracorporeal circulation laboratory table for medical implant interventional body observation according to claim 1,
the first pressure sensor is close to the outlet end and used for monitoring the outlet pressure of the experiment liquid which circulates in the circulation loop and flows out of the experiment section module; the second pressure sensor is close to the inlet end and used for monitoring the inlet pressure of the experimental liquid which circulates in the circulation loop before entering the experimental section module; the first throttle valve and the liquid storage container are used for regulating the flow rate and the pressure of the experimental liquid circulating in the circulation loop, and the liquid storage container is also used for discharging gas in the experimental liquid circulating in the circulation loop; the ultrasonic flow sensor is used for monitoring the flow of the test liquid circulating in the circulation loop.
3. The extracorporeal circulation laboratory table for medical implant intervention body observation as recited in claim 2, wherein the hose is a medical PVC hose.
4. The extracorporeal circulation laboratory table for medical implant intervention body observation according to any one of claims 1-3, wherein the observation module comprises a fluorescence observation system for realizing the fluorescence observation function, the fluorescence observation system comprises a laser, a first optical filter and a single lens reflex; the laser is used for emitting continuous laser to irradiate the experimental section module so as to enable the fluorescent reagent on the thrombus on the surface of the medical implant body to excite fluorescence; the first optical filter is arranged between the experimental section module and the single lens reflex; the single lens reflex is used for shooting a first fluorescence image obtained by filtering fluorescence of a fluorescence reagent on thrombus on the surface of the medical implant body through the first optical filter.
5. The extracorporeal circulation test table for medical implant intervention body observation according to claim 4, wherein the observation module further comprises a PIV flow field observation system for realizing a flow field observation function, the PIV flow field observation system comprises the laser, a CCD camera and a second filter, the laser is used for exciting pulsed laser to irradiate on the experiment section module so as to excite fluorescence from fluorescent particles in an experiment liquid in the experiment section module; the second optical filter is arranged between the experimental section module and the CCD camera; the CCD camera is used for shooting a second fluorescence image obtained by filtering fluorescence of fluorescent particles in the experimental liquid in the experimental section module through the second optical filter, and the shooting frequency of the CCD camera is synchronous with the pulse frequency of the pulse laser.
6. The extracorporeal circulation laboratory table for medical implant intervention body observation according to claim 5, wherein the PIV flow field observation system further comprises a signal synchronizer electrically connected with the laser and the CCD camera, respectively, to synchronize the pulse frequency of the pulsed laser and the photographing frequency of the CCD camera.
7. An experimental method using an extracorporeal circulation laboratory table for medical implant interventional body observation according to any one of claims 1-6, wherein the experimental method is a fluorescence observation experimental method, comprising the steps of:
step S1: mixing the first fluorescent agent and the collected blood in proportion, fully and uniformly mixing the mixture in a water bath at 37 ℃, injecting the mixture into the circulation loop, and discharging the blood after the medical implant intervention body works for a period of time to form thrombus; or a second fluorescent agent is directly coated on the thrombus on the surface of the medical implant intervention body as an observation object and is placed in the transparent tube;
step S2: and quantitatively observing the area of the thrombus on the surface of the medical implant body by utilizing the observation module.
8. The experimental procedure for an extracorporeal circulation laboratory table for medical implant interventional observation according to claim 7, characterised in that the first fluorescent agent is a fluorescent agent comprising DiOC6 or/and FITC; the second fluorescent agent is DiOC6 or/and FITC.
9. The experimental method of an in vitro cycling laboratory bench for medical implant interventional body observation according to the claim 8, characterized in that in the step S2, the observation module comprises the fluorescence observation system according to the claim 4, the step S2 comprises the following sub-steps:
step S201: irradiating the experimental section module by using blue laser with the wavelength of 490-515nm provided by the laser to enable the fluorescent reagent on the thrombus on the surface of the medical implant body to excite green fluorescence;
step S202: a green band-pass filter with the wavelength of 550nm-570nm is selected as the first filter and is additionally arranged in front of the single lens reflex;
step S203: and shooting a first fluorescence image of the thrombus on the surface of the medical implant interventional body by using the single-lens reflex camera under the condition of a full dark field, and quantitatively observing the area of the thrombus according to the first fluorescence image.
10. An experimental method using an extracorporeal circulation laboratory table for medical implant interventional body observation according to any one of claims 7-9, wherein the experimental method is a PIV flow field experimental method, comprising the steps of:
step S6: adding the third fluorescent particles into the transparent liquid to obtain a fluorescent particle solution;
step S7: injecting the fluorescent particle solution into the circulation loop, and fully mixing;
step S8: and the observation module is used for observing the flow field in the experiment section module.
11. The experimental procedure for an extracorporeal circulation laboratory table for medical implant intervention body visualization as recited in claim 10, wherein the third fluorescent particle is a Fluo-610 fluorescent particle.
12. The experimental method of an extracorporeal circulation laboratory table for medical implant interventional observation as set forth in claim 11, wherein the observation module comprises the PIV flow field observation system according to claim 5 or 6, and the step S8 specifically comprises the following sub-steps:
step S801: irradiating the experiment section module with pulsed laser provided by the laser to enable third fluorescent particles in the fluorescent particle solution in the experiment section module to excite fluorescence;
step S802: shooting a second fluorescence image of fluorescence excited by a third fluorescence particle in the experimental section module through the second optical filter under a full dark field condition by using the CCD camera, wherein the shooting frequency of the CCD camera is synchronous with the pulse frequency of the pulse laser:
step S803: and processing the obtained second fluorescence image by using special processing software to obtain the flow field condition of the watershed near the medical implant.
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