CN110760441B - Method and device for generating ultrasonic radiation force field for in vitro culture of engineered cartilage - Google Patents

Method and device for generating ultrasonic radiation force field for in vitro culture of engineered cartilage Download PDF

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CN110760441B
CN110760441B CN201910911890.4A CN201910911890A CN110760441B CN 110760441 B CN110760441 B CN 110760441B CN 201910911890 A CN201910911890 A CN 201910911890A CN 110760441 B CN110760441 B CN 110760441B
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ultrasonic transducer
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cartilage
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孙安玉
赖蛟娇
居冰峰
戴霖
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Zhejiang University ZJU
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Abstract

本发明提供一种工程软骨体外培养超声辐射力场产生方法及装置。本发明装置包括超声换能器、连接架、升降台、固定底座、导向环和工程软骨体外培养装置、超声发射接收装置、信号采集处理模块和示波器。本发明方法首先调节升降台的内置齿轮控制使超声换能器的发射头浸入工程软骨体外培养装置的培养基中。通过信号采集处理模块和示波器确定培养基中超声换能器的发射头的焦距，使工程软骨处于超声换能器的发射头的焦点处；通过旋转超声换能器，确定超声辐射力场方向；最后启动超声发射接收装置，激发超声换能器。本发明方法与装置产生的超声辐射力场可对体外培养工程软骨进行力学调控，获得正常的、生物性好的工程软骨。

Figure 201910911890

The invention provides a method and a device for generating an ultrasonic radiation force field for in vitro culture of engineered cartilage. The device of the invention includes an ultrasonic transducer, a connecting frame, a lifting platform, a fixed base, a guide ring and an in vitro culturing device for engineering cartilage, an ultrasonic transmitting and receiving device, a signal acquisition and processing module and an oscilloscope. The method of the invention firstly adjusts the built-in gear control of the lifting platform so that the transmitting head of the ultrasonic transducer is immersed in the culture medium of the in vitro culturing device of the engineered cartilage. The focal length of the transmitting head of the ultrasonic transducer in the culture medium is determined by the signal acquisition and processing module and the oscilloscope, so that the engineered cartilage is at the focal point of the transmitting head of the ultrasonic transducer; by rotating the ultrasonic transducer, the direction of the ultrasonic radiation force field is determined; Finally, start the ultrasonic transmitting and receiving device to excite the ultrasonic transducer. The ultrasonic radiation force field generated by the method and the device of the invention can perform mechanical regulation on the in vitro cultured engineered cartilage to obtain normal and biologically good engineered cartilage.

Figure 201910911890

Description

Method and device for generating ultrasonic radiation force field for in-vitro culture of engineering cartilage
Technical Field
The invention belongs to the field of ultrasonic regulation, articular cartilage repair and biomedical tissue engineering, and particularly relates to a method and a device for generating an ultrasonic radiation force field for in-vitro culture of an engineered cartilage.
Background
In joint motion, normal cartilage is an indispensable tissue. However, with the development of modern technology and the change of working and living habits, more and more people tend to maintain an action posture for a long time, and the problem of articular cartilage damage is easy to occur. Meanwhile, the articular cartilage has poor self-healing capability and can not be regenerated, so that the problems of joint stiffness, pain aggravation and the like are easily caused after the articular cartilage is damaged, and the life quality of a patient is seriously influenced. At present, the treatment schemes of articular cartilage damage comprise joint lavage, microfracture operation, periosteum transplantation, cartilage transplantation and the like, but all have defects. Clinical research shows that the engineering cartilage transplantation is a limited way for repairing articular cartilage damage and realizing functional reconstruction at present, and is expected to become a brand new treatment mode for repairing bone defects.
The engineering cartilage transplantation technology is a technology for culturing cartilage cells in vitro by a method of planting the cartilage cells on a degradable biological material bracket, constructing cartilage tissues and then implanting the bracket of composite cells into a defect to form bone tissues with physiological functions. The engineered cartilage transplantation combines the operability and the stability of tissue engineered cartilage suture, and the interface healing capacity is improved by using biological materials as cell carriers. However, the mechanical properties of the engineered cartilage produced by the prior art are far different from those of normal cartilage, and cannot meet the requirements of clinical transplantation. At present, the engineering cartilage tissue with good biocompatibility and mechanical property is difficult to obtain by an in vitro culture technology. How to construct normal tissue engineering cartilage under the in vitro condition is used for repairing cartilage damage, provides a better choice for the treatment of cartilage defect, and has good clinical significance.
Williamson et al have shown that the micro-environment of cartilage growth influences its mechanical properties. The cartilage is always in the mechanical environment of static and dynamic pressure alternate activity in vivo, so that the metabolic activity of extracellular matrix is continuously adjusted, collagen and pyridine crosslinking is promoted, and the mechanical property and shape structure of the cartilage are changed. When the engineering cartilage is statically cultured in vitro, the hollow phenomenon of the engineering cartilage is easy to form, and the mechanical property of the finally generated engineering cartilage is insufficient. Most of the existing bioreactors can only realize constant shearing force and dynamic loading pressure. The combination of the two acting forces is single, which is not favorable for culturing the articular cartilage with good structure and function. Therefore, when the engineered cartilage is cultured in vitro, in order to obtain the structural shape and the functional tissue similar to those of the natural cartilage, it is necessary to design a dynamic microenvironment suitable for the growth of the engineered cartilage tissue.
Research in recent years on dynamic culture of engineered cartilage shows that low-intensity ultrasonic waves can regulate and control the mechanical properties of the engineered cartilage by accelerating cartilage formation. The ultrasound waves, as an alternating pressure wave, can create a stable cavity and transmit beamlets within the tissue. Therapeutic intensity ultrasound can produce shear forces on cell membranes, and this effect may directly alter gene expression by affecting the cytoskeleton. Ultrasound also allows for the transport of transmembrane ions and subsequent cellular responses to be altered through transmembrane channels in the cell membrane. The ultrasonic radiation force changes the collagen correlation condition in the cells, so that the ultrasonic regulation and control of the mechanical property of the constructed engineering cartilage become possible. Therefore, in the process of culturing the engineering cartilage in vitro, the structural shape and the mechanical property of the engineering cartilage can be regulated and controlled through the ultrasonic radiation force, and the normal engineering cartilage is obtained.
The invention aims at the requirements, and designs a method and a device for generating an ultrasonic radiation force field for in vitro culture of engineering cartilage through innovative designs such as an ultrasonic transducer working mode, an up-down displacement module and the like. The ultrasonic radiation force field generated by the method and the device can perform mechanical regulation and control on the in vitro culture engineering cartilage to obtain the normal engineering cartilage with good biological property. Realizing the anisotropy of the engineering cartilage, carrying out the chondrocyte transplantation and finally realizing the articular cartilage repair.
Disclosure of Invention
The invention provides a method and a device for generating an ultrasonic radiation force field for in vitro culture of an engineering cartilage, aiming at the application requirements of the occasions such as anisotropy of tissue engineering, mechanical property regulation and control in the process of in vitro culture of the engineering cartilage and the like on specific ultrasonic radiation force. The method can also improve the mechanical property of the engineering cartilage and realize the rapid repair of the cartilage and the comprehensive healing of the bone.
An ultrasonic radiation force field generating device for in-vitro culture of engineering cartilage comprises an ultrasonic transducer (1), a connecting frame (2), a lifting table (3), a fixing base (4), a guide ring (5), an in-vitro culture device (6) of the engineering cartilage, an ultrasonic transmitting and receiving device (7), a signal acquisition and processing module (8) and an oscilloscope (9).
Elevating platform (3) include stiff end and motion end, elevating platform (3) are fixed on unable adjustment base (4) through the stiff end, unable adjustment base (4) are for the square platform that contains cylindrical indent, link (2) are fixed on the motion end of elevating platform (3), ultrasonic transducer (1) are fixed on link (2), guide ring (5) are fixed in the cylindrical indent of unable adjustment base (4), engineering cartilage in vitro culture device (6) are fixed in through the restriction location of guide ring (5) and unable adjustment base (4) fixing device in the cylindrical indent, ultrasonic transducer (1) is connected in ultrasonic emission receiving arrangement (7), signal acquisition processing module (8) are connected ultrasonic transducer (1), oscilloscope (9) are connected signal acquisition processing module (8).
The lifting platform (3) is a gear and rack meshing lifting platform, the fixed end is a T-shaped fixed plate, two symmetrical U-shaped grooves are formed in the length direction of a transverse plate of the T-shaped fixed plate, the U-shaped grooves are located in the middle of the width direction of the transverse plate and can be used for adjusting the position of the lifting platform (3) in a left-right mode within a certain range, the U-shaped grooves are matched with threaded holes in the fixed base plate (4) and fixedly lock the lifting platform (3) through threaded connection, a rack is installed at the vertical end of the T-shaped fixed plate, and the rack is installed at the central position of the vertical end of the T-shaped fixed plate; the movable end is a square block containing a built-in gear, the built-in gear is meshed with the rack at the fixed end, and the built-in gear is manually adjusted to realize the up-and-down movement of the square block.
The connecting frame (2) is a T-shaped connecting piece, the transverse end of the T-shaped connecting piece is installed on the moving end of the lifting platform (3) through a threaded structure, a cylindrical hole used for fixing the ultrasonic transducer (1) is formed in the vertical direction of the long end of the T-shaped connecting piece, a through groove is formed in the bottom surface of the cylindrical hole and the bottom surface of the long end of the T-shaped connecting piece, a threaded through hole is transversely formed in one side of the through groove, a matched threaded hole is formed in the other side of the through groove, the size of the cylindrical hole is slightly changed through threaded connection, and the ultrasonic transducer (1) is locked.
The ultrasonic transducer (1) is a line focus ultrasonic transducer.
The guide ring (5) is a hollow ring with a micro gap, guide teeth are arranged on an inner ring of the micro gap which is symmetrical about the circle center of the guide ring (5), an outer ring of the guide ring (5) is matched with the cylindrical inner recess on the fixed base (4), the inner ring of the guide ring (5) is matched with a culture dish of an engineering cartilage in-vitro culture device (6), and the guide teeth of the guide ring (5) are matched with a base structure of the culture dish to limit the rotation of the culture dish; the center of the micro gap of the guide ring (5) is in a straight line with the circle center of the guide ring (5) and the center of the rack of the lifting platform (3), and the guide ring (5) is fixed in the cylindrical inner recess of the fixed base (4) through fixing glue.
There is the fixed block at the both ends of unable adjustment base (4), and the symmetric center of fixed block is on same straight line with the axial center of cylindrical indent, the fixed block transversely has the relative screw hole that sets up, fixes through helicitic texture engineering cartilage in vitro culture apparatus (6), the axial center of the cylindrical indent of unable adjustment base (4) is on same straight line with the axial center in link cylinder hole.
The engineering cartilage in-vitro culture device (6) comprises a culture dish, a culture medium, engineering cartilage cells and a culture box, and is used for in-vitro culture of the engineering cartilage cells; the height of the culture solution in the engineering cartilage in-vitro culture device (6) is larger than the focal length of the ultrasonic transducer (1).
The ultrasonic transmitting and receiving device (7) is connected with the ultrasonic transducer (1) through a cable and is used for exciting the ultrasonic transducer (1).
The signal acquisition and processing module (8) is based on a high-speed operational amplifier, the specific model is LMH6629, and the signal acquisition and processing module (8) is matched with an oscilloscope (9) to determine the position of the transmitting head of the ultrasonic transducer (1).
A method for generating an ultrasonic radiation force field for in vitro culture of engineering cartilage comprises the following steps:
step (1), connecting an ultrasonic transducer (1) with an ultrasonic transmitting and receiving device (7) through a cable;
and (2) controlling the vertical distance between the emitting head of the ultrasonic transducer (1) and the culture device by manually adjusting the built-in gear of the lifting table (3) so that the emitting head of the ultrasonic transducer (1) is immersed in the culture medium of the engineered cartilage in-vitro culture device (6).
Determining the focal distance of the emitting head of the ultrasonic transducer (1) in the culture medium through a signal acquisition processing module (8), so that the engineering cartilage is positioned at the focal point of the emitting head of the ultrasonic transducer (1);
obtaining the distance L between the transmitting head of the ultrasonic transducer (1) and the engineering cartilage through an oscilloscope (9) according to the product of the distance equal to the period T of the waveform and the sound velocity c in the culture medium; the focus position at the peak maximum value is determined according to the peak amplitude by adjusting the displacement of the emitting head of the ultrasonic transducer (1) up and down;
step (4), the direction of an ultrasonic radiation force field is determined by rotating the ultrasonic transducer (1) to be matched with the guide ring (5) and the engineering cartilage culture device (6);
uploading the waveform of the oscilloscope (9) to a computer upper computer, calculating by the computer upper computer to obtain the sound intensity, and regulating and controlling the position of the transmitting head of the ultrasonic transducer (1) in real time by the obtained sound intensity;
and (6) setting corresponding frequency, emission energy and repetition frequency, starting an ultrasonic emission and receiving device (7), and exciting an ultrasonic transducer (1) to generate an ultrasonic radiation force field.
The invention has the following beneficial effects:
the invention can generate an ultrasonic radiation force field suitable for the in vitro culture of the engineering cartilage, and regulate and control the chondrocyte cultured in vitro to construct a normal engineering cartilage; the ultrasonic radiation force field generated by the invention can change the size of the ultrasonic radiation force by adjusting the distance of the focusing probe up and down. The method can be used for adjusting and controlling the mechanical property of the engineering cartilage, can also be used for realizing the anisotropy of the engineering cartilage tissue, greatly improves the mechanical property of the constructed engineering cartilage, and is used for repairing the articular cartilage.
Drawings
FIG. 1 is a schematic diagram of the structure of the apparatus of the present invention;
FIG. 2 is a flow chart of the method of the present invention;
figure 3 is a top view of the apparatus of the present invention.
In the figure: 1. ultrasonic transducer 2, link 3, elevating platform 4, unable adjustment base 5, guide ring 6, engineering cartilage in vitro culture device 7, ultrasonic emission receiving arrangement 8, signal acquisition processing module 9, oscilloscope.
Detailed Description
The invention is further illustrated with reference to the following figures and examples, without however being limited to the scope of the invention as described below.
As shown in fig. 1, an ultrasonic radiation force field generating device for in vitro culture of engineering cartilage comprises an ultrasonic transducer 1, a connecting frame 2, a lifting table 3, a fixing base 4, a guide ring 5, an engineering cartilage in vitro culture device 6, an ultrasonic transmitting and receiving device 7, a signal acquisition and processing module 8 and an oscilloscope 9.
Elevating platform 3 includes stiff end and motion end, elevating platform 3 passes through the stiff end to be fixed on unable adjustment base 4, unable adjustment base 4 is for the square platform who contains cylindrical indent, link 2 is fixed on elevating platform 3's motion end, ultrasonic transducer 1 is fixed on link 2, guide ring 5 is fixed in unable adjustment base 4's cylindrical indent, engineering cartilage in vitro culture apparatus 6 passes through the restriction location of guide ring 5 and unable adjustment base 4 fixing device fixes in cylindrical indent, ultrasonic transmitting and receiving device 7 connects ultrasonic transducer 1, ultrasonic transducer 1 is connected to signal acquisition processing module 8, oscilloscope 9 connects signal acquisition processing module 8.
Figure 3 is a top view of the apparatus of the present invention.
The lifting platform 3 is a gear rack meshing lifting platform, the fixed end is a T-shaped fixed plate, two symmetrical U-shaped grooves are formed in the transverse end of the T-shaped fixed plate in the length direction of the transverse plate, the U-shaped grooves are located in the middle of the transverse plate in the width direction, and the position of the lifting platform 3 can be adjusted left and right within a certain range. The diameter of the U-shaped groove is 3mm, the length of the U-shaped groove is 8mm, the U-shaped groove is matched with a threaded hole in the fixed bottom plate 4, the lifting platform 3 is fixedly locked through threaded connection, a rack is installed at the vertical end of the T-shaped fixed plate, and the rack is installed at the central position of the vertical end of the T-shaped fixed plate; the moving end is a square block containing a built-in gear, the built-in gear is meshed with the rack at the fixed end, and the built-in gear is manually adjusted to realize the up-and-down movement of the square block. The range of the lifting platform 3 is +/-40 mm, and the precision is 0.1 mm.
The link 2 is T shape connecting piece, the horizontal end of T shape connecting piece is installed on elevating platform 3's motion end through helicitic texture, the cylinder hole that is used for fixed ultrasonic transducer 1 is offered to the vertical direction of the long end of T shape connecting piece, the size of cylinder hole is 18mm the same with ultrasonic transducer 1 external diameter, logical groove has been offered with the bottom surface of the long end of T shape connecting piece to the cylinder hole, the size that leads to the groove is 2 x 5 x 3mm, logical groove one side transversely is provided with the screw through-hole, the screw through-hole diameter is 2mm, the opposite side is provided with supporting screw hole, the screw hole specification is M1.5, through the small size that changes the cylinder hole of threaded connection, locking ultrasonic transducer 1.
The ultrasonic transducer 1 is a line focus ultrasonic transducer, the line focus ultrasonic transducer is cylindrical, and the emitting head of the line focus ultrasonic transducer is a concave hemisphere. The outer diameter of the line focus ultrasonic transducer is 18mm, and the length of the line focus ultrasonic transducer is 78 mm. The piezoelectric material of the line focus ultrasonic transducer is PET crystal, the self frequency is 2.5MHz, the focal length is 18mm, and the size of the crystal is 14 x 14 mm.
The guide ring 5 is a hollow ring with a micro gap, guide teeth are arranged on an inner ring of the micro gap which is symmetrical about the circle center of the guide ring 5, an outer ring of the guide ring 5 is matched with a cylindrical inner recess on the fixed base 4, the inner ring of the guide ring 5 is matched with a culture dish of the engineering cartilage in-vitro culture device 6, and the guide teeth of the guide ring 5 are matched with a base structure of the culture dish to limit the rotation of the culture dish; the center of the micro gap of the guide ring 5, the center of the circle of the guide ring 5 and the center of the rack of the lifting platform 3 form a straight line, and the guide ring 5 is fixed in the cylindrical inner recess of the fixed base 4 through fixing glue.
There is the fixed block at unable adjustment base 4's both ends, and the symmetric center of fixed block and the axial center of cylindrical indent are on the same straight line, and the fixed block transversely has the relative screw hole that sets up, through the fixed engineering cartilage in vitro culture apparatus 6 of helicitic texture, and the axial center of the cylindrical indent of unable adjustment base 4 and the axial center of link cylinder hole are on the same straight line.
The engineering cartilage in-vitro culture device 6 comprises a culture dish, a culture medium, engineering cartilage cells and a culture box and is used for in-vitro culture of the engineering cartilage cells; the height of the culture solution in the engineered cartilage in-vitro culture device 6 is larger than the focal length of the ultrasonic transducer 1.
The ultrasonic transmitting and receiving device 7 is connected with the ultrasonic transducer 1 through a cable and is used for exciting the ultrasonic transducer 1.
The signal acquisition and processing module 8 is based on a high-speed operational amplifier, the specific model is LMH6629, and the signal acquisition and processing module 8 is matched with the oscilloscope 9 to determine the position of the transmitting head of the ultrasonic transducer 1.
As shown in fig. 2, a method for generating an ultrasonic radiation force field for in vitro culture of engineered cartilage:
step (1), connecting an ultrasonic transducer 1 with an ultrasonic transmitting and receiving device 7 through a cable;
and (2) controlling the vertical distance between the emitting head of the ultrasonic transducer 1 and the culture device by manually adjusting the built-in gear of the lifting platform 3, so that the emitting head of the ultrasonic transducer 1 is immersed in the culture medium of the engineered cartilage in-vitro culture device 6.
Step (3), determining the focal distance of the emitting head of the ultrasonic transducer 1 in the culture medium through the signal acquisition processing module 8, so that the engineering cartilage is positioned at the focal point of the emitting head of the ultrasonic transducer 1;
obtaining the distance L between the emitting head of the ultrasonic transducer 1 and the engineering cartilage through an oscilloscope 9 according to the product of the distance equal to the period T of the waveform and the sound velocity c in the culture medium; determining the focal position at the maximum value of the wave crest according to the amplitude of the wave crest by adjusting the displacement of the emitting head of the ultrasonic transducer 1 up and down;
step (4), the direction of an ultrasonic radiation force field is determined by rotating the ultrasonic transducer 1 to be matched with the guide ring 5 and the engineering cartilage culture device 6;
uploading the waveform of the oscilloscope 9 to a computer upper computer, calculating by the computer upper computer to obtain the sound intensity, and regulating and controlling the position of the transmitting head of the ultrasonic transducer 1 in real time through the obtained sound intensity;
and (6) setting corresponding frequency, emission energy and repetition frequency, starting the ultrasonic emission and receiving device 7, and exciting the ultrasonic transducer 1 to generate an ultrasonic radiation force field.
Example 1:
firstly, the lifting platform 3 is arranged at a designated position of the fixed base 1, the moving end of the lifting platform 3 is adjusted to a zero position, and the position of the lifting platform is manually locked. The connecting frame 2 is then mounted to the moving end of the lifting table 3 and secured using a threaded structure. And then the line focus ultrasonic transducer 1 is arranged on the connecting frame, and the line focus ultrasonic transducer 1 is locked on the connecting frame 2 by matching the fixing hole on the connecting frame 2 with the thread. The line focus ultrasonic transducer 1 is connected to the ultrasonic transmitting and receiving device 7 through a cable. The prepared engineering cartilage in-vitro culture device 6 is placed on the fixed base 4, the engineering cartilage in-vitro culture device 6 is rotated to determine the direction of the applied ultrasonic radiation force, and the engineering cartilage in-vitro culture device 6 is fixedly clamped through the fixed block of the fixed base 1. The up-down distance between the emitting head of the ultrasonic transducer 1 and the culture device is manually adjusted by using the lifting platform 3, so that the emitting head of the ultrasonic transducer 1 is immersed in the culture medium of the culture device. The focal distance of the emitting head of the ultrasonic transducer 1 in the culture medium is determined through the signal acquisition processing module 8, so that the engineering cartilage is positioned at the focus of the emitting head of the ultrasonic transducer 1. And displaying the distance L between the emitting head of the ultrasonic transducer 1 and the engineering cartilage through an oscilloscope 9 according to the product of the distance equal to the period T of the waveform and the sound velocity c in the culture medium. And determining the focal position at the maximum value of the wave crest according to the amplitude of the wave crest by adjusting the displacement of the emitting head of the ultrasonic transducer 1 up and down. And the final ultrasonic radiation force field direction is determined by the cooperation of the rotary ultrasonic transducer 1, the guide ring 5 and the engineering cartilage culture device 6. Uploading the waveform of the oscilloscope 9 to a computer upper computer, calculating by the computer upper computer to obtain the sound intensity, and regulating and controlling the position of the transmitting head of the ultrasonic transducer 1 in real time through the obtained sound intensity; the frequency of ultrasonic emission is set to be 2.5MHz, the energy emitted by a probe is 32uJ, the repetition frequency is 1KHz, and a pulse type ultrasonic emission receiver is adopted to generate an alternate ultrasonic sound field. The sound waves propagate in the culture medium, generating a certain pressure p. When encountering the engineering cartilage tissue, the engineering cartilage tissue collides with the engineering cartilage tissue to form ultrasonic radiation force F. The ultrasonic radiation force up-and-down displacement module and the engineering cartilage in-vitro culture device are placed in an incubator and used for in-vitro culture of the engineering cartilage. The ultrasonic transmitting and receiving device is of a pulse negative pressure type, can generate dynamic alternating ultrasonic radiation force F and simulates a tissue culture microenvironment in a human body. The entire device was placed in an incubator and stimulated continuously for 3 days. And taking out the engineering cartilage tissue, and testing the mechanical property of the engineering cartilage tissue. Observing the regulation and control of the mechanical property of the engineering cartilage by the ultrasonic radiation force field.

Claims (6)

1.一种工程软骨体外培养超声辐射力场产生装置，其特征在于，该装置包括超声换能器(1)、连接架(2)、升降台(3)、固定底座(4)、导向环(5)和工程软骨体外培养装置(6)、超声发射接收装置(7)、信号采集处理模块(8)和示波器(9)；1. a device for producing an ultrasonic radiation force field for in vitro culture of engineered cartilage, is characterized in that, the device comprises an ultrasonic transducer (1), a connecting frame (2), a lifting platform (3), a fixed base (4), a guide ring (5) and an engineering cartilage in vitro culture device (6), an ultrasonic transmitting and receiving device (7), a signal acquisition and processing module (8) and an oscilloscope (9); 升降台(3)包括固定端与运动端，升降台(3)通过固定端固定在固定底座(4)上，固定底座(4)为包含圆柱形内凹的方形平台，连接架(2)固定在升降台(3)的运动端上，超声换能器(1)固定在连接架(2)上，导向环(5)固定在固定底座(4)的圆柱形内凹内，工程软骨体外培养装置(6)通过导向环(5)的限制定位和固定底座(4)固定装置固定在所述圆柱形内凹内，超声发射接收装置(7)连接超声换能器(1)，信号采集处理模块(8)连接所述超声换能器(1)，所述示波器(9)连接信号采集处理模块(8)；The lifting platform (3) includes a fixed end and a moving end, the lifting platform (3) is fixed on a fixed base (4) through the fixed end, the fixed base (4) is a square platform with a cylindrical concave, and the connecting frame (2) is fixed On the moving end of the lifting platform (3), the ultrasonic transducer (1) is fixed on the connecting frame (2), the guide ring (5) is fixed in the cylindrical concave of the fixing base (4), and the engineered cartilage is cultured in vitro. The device (6) is fixed in the cylindrical concave by the limiting positioning of the guide ring (5) and the fixing device of the fixing base (4), the ultrasonic transmitting and receiving device (7) is connected to the ultrasonic transducer (1), and the signal is collected and processed. The module (8) is connected to the ultrasonic transducer (1), and the oscilloscope (9) is connected to the signal acquisition and processing module (8); 所述的升降台(3)为齿轮齿条啮合升降台，所述的固定端为T形固定板，所述T形固定板横板的长度方向有对称的两个U形槽，U形槽位于横板宽度方向的中间位置，可在一定范围内左右调节升降台(3)的位置，U形槽与固定底板4上的螺纹孔配合，通过螺纹连接固定锁紧升降台(3)，所述T形固定板的竖直端安装有齿条，齿条安装于T形固定板竖直端的中心位置；所述的运动端为包含内置齿轮的方形块，所述的内置齿轮与固定端的齿条啮合，通过手动调节内置齿轮，实现方形块的上下运动；The lifting platform (3) is a gear rack meshing lifting platform, the fixed end is a T-shaped fixing plate, and the length direction of the horizontal plate of the T-shaped fixing plate has two symmetrical U-shaped grooves, the U-shaped groove Located in the middle of the width direction of the horizontal plate, the position of the lifting platform (3) can be adjusted left and right within a certain range. The U-shaped groove cooperates with the threaded holes on the fixed base plate 4, and the lifting platform (3) is fixed and locked by threaded connection. The vertical end of the T-shaped fixed plate is installed with a rack, and the rack is installed at the center of the vertical end of the T-shaped fixed plate; the moving end is a square block containing a built-in gear, the built-in gear and the teeth of the fixed end The bars are meshed, and the up and down movement of the square block is realized by manually adjusting the built-in gear; 所述的连接架(2)为T形连接件，所述T形连接件的横端通过螺纹结构安装在所述升降台(3)的运动端上，所述T形连接件的长端的竖直方向开设有用于固定超声换能器(1)的圆柱孔，所述圆柱孔与所述T形连接件的长端的底面开设有通槽，所述通槽一侧横向设置有螺纹通孔，另一侧设置有配套的螺纹孔，通过螺纹连接微小改变圆柱孔的大小，锁紧超声换能器(1)；The connecting frame (2) is a T-shaped connecting piece, the horizontal end of the T-shaped connecting piece is mounted on the moving end of the lifting platform (3) through a threaded structure, and the vertical end of the long end of the T-shaped connecting piece is mounted on the moving end of the lifting platform (3). A cylindrical hole for fixing the ultrasonic transducer (1) is provided in the straight direction, a through groove is formed between the cylindrical hole and the bottom surface of the long end of the T-shaped connecting piece, and a threaded through hole is laterally provided on one side of the through groove, The other side is provided with a matching threaded hole, and the size of the cylindrical hole is slightly changed through the threaded connection to lock the ultrasonic transducer (1); 所述超声换能器(1)为线聚焦超声换能器；The ultrasonic transducer (1) is a line focused ultrasonic transducer; 所述导向环(5)为含微小缺口的空心环，所述微小缺口关于导向环(5)圆心对称的内环处有导向齿，所述导向环(5)的外环与所述固定底座(4)上的圆柱形内凹内相配合，所述导向环(5)的内环与工程软骨体外培养装置(6)的培养皿配合，所述导向环(5)的导向齿与培养皿的基底结构配合，限制培养皿的旋转；所述导向环(5)的微小缺口中心与导向环(5)圆心以及升降台(3)的齿条中心成一条直线，所述导向环(5)通过固定胶固定在固定底座(4)的圆柱形内凹内。The guide ring (5) is a hollow ring with a small gap, and the small gap is provided with guide teeth at the inner ring symmetrical about the center of the guide ring (5), and the outer ring of the guide ring (5) is connected to the fixed base. The cylindrical concave on (4) is matched with the inner ring of the guide ring (5), the inner ring of the guide ring (5) is matched with the culture dish of the engineered cartilage in vitro culture device (6), and the guide teeth of the guide ring (5) are matched with the culture dish. The base structure of the guide ring (5) is matched with the base structure to limit the rotation of the culture dish; the center of the small gap of the guide ring (5) is in a straight line with the center of the guide ring (5) and the center of the rack of the lifting platform (3). It is fixed in the cylindrical concave of the fixing base (4) by fixing glue. 2.根据权利要求1所述的一种工程软骨体外培养超声辐射力场产生装置，其特征在于，所述固定底座(4)的两端有固定块，固定块的对称中心与圆柱形内凹的轴向中心在同一直线上，所述固定块横向有相对设置的螺纹孔，通过螺纹结构固定所述工程软骨体外培养装置(6)，所述固定底座(4)的圆柱形内凹的轴向中心与连接架圆柱孔的轴向中心在同一直线上。2. a kind of engineering cartilage in vitro culture ultrasonic radiation force field generating device according to claim 1, is characterized in that, the two ends of described fixing base (4) have fixing blocks, and the symmetrical center of the fixing block and the cylindrical concave The axial center of the fixed block is on the same straight line, and the fixing block has oppositely arranged threaded holes in the transverse direction, and the engineered cartilage in vitro culture device (6) is fixed through the threaded structure. The cylindrical concave shaft of the fixing base (4) The center is on the same line as the axial center of the cylindrical hole of the connecting frame. 3.根据权利要求2所述的一种工程软骨体外培养超声辐射力场产生装置，其特征在于，所述工程软骨体外培养装置(6)包括培养皿、培养基、工程软骨细胞、培养箱，用于体外培养工程软骨细胞；所述工程软骨体外培养装置(6)中的培养液的高度大于所述超声换能器(1)的焦距。3. The device for generating an ultrasonic radiation force field for in vitro culture of engineered cartilage according to claim 2, wherein the device for in vitro culture of engineered cartilage (6) comprises a petri dish, a culture medium, an engineered chondrocyte, and an incubator, It is used for culturing engineered chondrocytes in vitro; the height of the culture solution in the in vitro culturing device for engineered cartilage (6) is greater than the focal length of the ultrasonic transducer (1). 4.根据权利要求3所述的一种工程软骨体外培养超声辐射力场产生装置，其特征在于，所述的超声发射接收装置(7)通过电缆连接所述超声换能器(1)，用于激发超声换能器(1)。4. a kind of engineering cartilage in vitro culture ultrasonic radiation force field generating device according to claim 3, is characterized in that, described ultrasonic transmitting and receiving device (7) connects described ultrasonic transducer (1) by cable, uses for exciting the ultrasonic transducer (1). 5.根据权利要求4所述的一种工程软骨体外培养超声辐射力场产生装置，其特征在于，所述的信号采集处理模块(8)基于高速运算放大器，具体型号为LMH6629，所述的信号采集处理模块(8)与示波器(9)进行配合，确定超声换能器(1)发射头的位置。5. a kind of engineering cartilage in vitro culture ultrasonic radiation force field generating device according to claim 4, is characterized in that, described signal acquisition and processing module (8) is based on high-speed operational amplifier, and concrete model is LMH6629, and described signal The acquisition and processing module (8) cooperates with the oscilloscope (9) to determine the position of the transmitting head of the ultrasonic transducer (1). 6.根据权利要求1-5任意所述的一种工程软骨体外培养超声辐射力场产生方法，其特征在于，具体步骤如下：6. according to any described a kind of engineering cartilage in vitro culture ultrasonic radiation force field generation method described arbitrarily, it is characterized in that, concrete steps are as follows: 步骤(1).通过电缆连接超声换能器(1)与超声发射接收装置(7)；Step (1). Connect the ultrasonic transducer (1) and the ultrasonic transmitting and receiving device (7) through a cable; 步骤(2).通过手动调节升降台(3)的内置齿轮控制超声换能器(1)的发射头与培养装置的上下距离，使超声换能器(1)的发射头浸入工程软骨体外培养装置(6)的培养基中；Step (2). The distance between the transmitter of the ultrasonic transducer (1) and the culture device is controlled by manually adjusting the built-in gear of the lift table (3), so that the transmitter of the ultrasonic transducer (1) is immersed in the engineered cartilage for in vitro culture in the culture medium of the device (6); 步骤(3).通过信号采集处理模块(8)确定培养基中超声换能器(1)的发射头的焦距，使工程软骨处于超声换能器(1)的发射头的焦点处；Step (3). Determine the focal length of the transmitting head of the ultrasonic transducer (1) in the culture medium through the signal acquisition and processing module (8), so that the engineered cartilage is at the focal point of the transmitting head of the ultrasonic transducer (1); 根据距离等于波形的周期T与培养基中声速c的乘积，通过示波器(9)获得超声换能器(1)的发射头与工程软骨的距离L；通过上下调节超声换能器(1)的发射头的位移，根据波峰幅值的大小，确定处于波峰最大值处的焦点位置；According to the product of the distance equal to the period T of the waveform and the speed of sound c in the medium, the distance L between the transmitter head of the ultrasonic transducer (1) and the engineered cartilage is obtained through the oscilloscope (9); The displacement of the transmitting head determines the focus position at the maximum value of the wave peak according to the amplitude of the wave peak; 步骤(4).通过旋转超声换能器(1)，与导向环(5)及工程软骨培养装置(6)相配合，确定超声辐射力场方向；Step (4). By rotating the ultrasonic transducer (1), in cooperation with the guide ring (5) and the engineering cartilage culture device (6), the direction of the ultrasonic radiation force field is determined; 步骤(5).将示波器(9)的波形上传至电脑上位机，通过电脑上位机计算获得声强，通过获得的声强对超声换能器(1)的发射头位置进行实时调控；Step (5). The waveform of the oscilloscope (9) is uploaded to the computer host computer, the sound intensity is obtained through the computer host computer calculation, and the transmitter position of the ultrasonic transducer (1) is regulated in real time by the obtained sound intensity; 步骤(6).设置相应的频率、发射能量大小及重复频率大小，启动超声发射接收装置(7)，激发超声换能器(1)产生超声辐射力场。Step (6). Set the corresponding frequency, transmitting energy and repetition frequency, start the ultrasonic transmitting and receiving device (7), and excite the ultrasonic transducer (1) to generate the ultrasonic radiation force field.
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