Deep in-situ 3D bioprinting system and method based on spherical standing wave focused ultrasound

Abstract

This invention discloses a deep in-situ 3D bioprinting system and method based on spherical standing wave focused ultrasound. It includes a spherical focusing system for generating spherical standing wave focused ultrasound, a motion control system for controlling the focal zone position of the focused ultrasound, and a data processing system for acquiring and analyzing ultrasonic cavitation signals to obtain control signals. The spherical standing wave focused ultrasound provided by this invention has superior focusing performance compared to traveling wave focused ultrasound, enabling spherical focused ultrasound 3D printing while balancing printing depth and accuracy, and exhibiting less focal shift in complex tissues. The sphere is formed by two hemispherical transducers of the same size and opposite positions, creating a spherical standing wave focusing cavity. The focused sound field is composed of sound waves directly radiated from the spherical surface and sound waves reflected multiple times by the spherical surface. When the sound waves directly focused at the center of the sphere and the sound waves reflected in each subsequent reflection are in phase, a very high sound pressure can be generated at the center of the sphere, and the focal zone accuracy is improved.

Description

Technical Field

[0001] This invention relates to the fields of tissue engineering technology, thermosetting technology, and deep in-situ three-dimensional bioprinting technology, and in particular to a deep in-situ three-dimensional bioprinting system and method based on spherical standing wave focused ultrasound. Background Technology

[0002] Trauma, disease, and congenital malformations lead to tissue dysfunction or organ loss. With the development of tissue engineering, techniques for artificially constructing new tissues or organs in vitro or in vivo have gradually emerged, driving a new revolution in treatment. Tissue engineering involves culturing cells taken from living organisms on biocompatible, biodegradable conformal scaffolds to obtain tissues or organs with the same structure and function as the original tissue. Three-dimensional (3D) bioprinting technology is applied to the construction of conformal scaffolds with complex and intricate three-dimensional structures. Currently, 3D bioprinted tissue engineering scaffolds are still limited to in-situ printing under surgical implantation or trauma exposure. Both require exposing the tissue area to be repaired, which can cause secondary damage and increase the risk of infection and rejection. Developing non-invasive in-situ 3D bioprinting devices that can penetrate greater tissue depths and achieve high spatial accuracy is a significant clinical need.

[0003] Regarding non-invasive in-situ 3D bioprinting, previous researchers proposed 3D printing based on digital near-infrared photopolymerization. However, the penetration depth of light waves in biological tissues is limited, thus preventing non-invasive in-situ printing of deep tissues. Compared to light, ultrasound has superior tissue penetration capabilities. Mohsen Habibi et al. proposed direct acoustic printing, based on traveling wave focused ultrasound triggering acoustic-driven polymerization of polydimethylsiloxane monomers within its focal domain through energy deposition, enabling non-contact, long-distance printing. Xiao Kuang et al. achieved acoustic 3D printing of biological scaffolds by adjusting the viscosity of vinyl "bio-ink," enhancing acoustic absorption, and suppressing the acoustic microfluidic effect in traveling wave focused ultrasound, achieving a tissue penetration depth of 1.5-1.7 cm. Their research was published in Science and received widespread academic attention. In all the above studies, traveling wave focused ultrasound was used to excite cross-linking of the "ink." However, traveling wave focused ultrasound has inherent technical limitations:

[0004] (1) Its focal zone is ellipsoidal, and its axial focal zone size is generally several sound wave wavelengths (generally on the order of several millimeters to tens of millimeters), making it difficult to achieve fine printing in the axial direction. To achieve high-precision printing, it is necessary to increase the sound wave frequency and sacrifice the sound wave penetration depth;

[0005] (2) Traveling waves are prone to focal shift and focal distortion after penetrating complex biological tissues, which will further affect the accuracy of in-situ 3D bioprinting.

[0006] Therefore, achieving high spatial accuracy in printing by penetrating thick biological tissue is a major bottleneck restricting the development of focused ultrasound-based 3D bioprinting technology. Summary of the Invention

[0007] In view of this, the purpose of the present invention is to provide a deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound. This system uses spherical standing wave focused ultrasound to cause phase changes in some materials through cavitation or thermal effects, thereby penetrating deep biological tissues to achieve in-situ three-dimensional printing.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] The present invention provides a deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound, comprising a spherical focusing system, a motion control system, and a data processing system;

[0010] The spherical focusing system is used to generate spherical internal standing wave focused ultrasound, which prints the target through ultrasonic cavitation and other effects;

[0011] The motion control system is used to control the position of the focused ultrasonic focal zone of the spherical focusing system, so that the focal area can move along the designed route;

[0012] The data processing system is used to acquire ultrasonic cavitation signals and dynamic image monitoring signals of the printing process, and to analyze and process them to obtain control signals for adjusting the spherical focusing system and the motion control system.

[0013] Furthermore, the spherical focusing system includes a spherical sound collector, an ultrasonic drive power supply, a passive cavitation detector, and a B-mode ultrasound diagnostic instrument;

[0014] The spherical sound collector reflects the ultrasonic beam through the concave surface of the spherical cavity structure, forming a spherical standing wave focused ultrasound, which focuses the energy at the spherical geometric center of the spherical cavity structure.

[0015] The ultrasonic driving power supply is used to supply power to the spherical sound collector on demand to realize the excitation of different types of ultrasonic waves;

[0016] The passive cavitation detector is used to detect cavitation signals, monitor cavitation signals in real time, and feed the monitored cavitation signals back to the data processing system.

[0017] The B-mode ultrasound diagnostic instrument is used to acquire dynamic image monitoring signals during the printing process and monitor the internal image of the object in real time during printing.

[0018] Furthermore, the ultrasonic driving power supply uses adjustable frequency, duty cycle, and excitation parameters to achieve on-demand excitation of different types of ultrasonic waves.

[0019] Furthermore, the motion control system includes a high-precision robotic arm, a robotic arm controller, a sealing device, and an elastic sealing coupling medium;

[0020] The high-precision robotic arm is used to drive the spherical sound collector, so that the focal point of the spherical sound collector can move as needed;

[0021] The robotic arm controller is connected to the high-precision robotic arm and is used to send control commands to the high-precision robotic arm;

[0022] The sealing device is used to prevent the elastic coupling medium from leaking out of the cavity;

[0023] The elastic sealing coupling medium is used to transmit ultrasonic energy.

[0024] Furthermore, the data processing system includes a data acquisition unit, a control system, and a data analysis system;

[0025] The data acquisition unit is used to acquire and store cavitation signals;

[0026] The control and data analysis system is used to receive dynamic image monitoring signals and cavitation signals; and to dynamically monitor the printing process, evaluate the strength of cavitation and focus position information, so as to adjust the output power and motion speed in real time.

[0027] Furthermore, the spherical focusing system is provided with an area for placing the tissue to be printed, the area being located at the spherical geometric center of the cavity structure.

[0028] Furthermore, the tissue to be printed contains printing ink, which is a thermosetting material or a material that undergoes a phase change after a sonochemical reaction.

[0029] Furthermore, the spherical sound collector includes two hemispherical transducers forming a spherical standing wave focusing cavity. The spherical surface of the hemispherical transducer serves as the emitting surface and strong reflecting surface of the ultrasonic wave. The sound waves radiated from the spherical surface and the sound waves reflected multiple times by the spherical surface are superimposed to form a focused sound field.

[0030] Furthermore, the two hemispherical transducers are the same size and positioned opposite each other.

[0031] This invention provides a method for large-scale deep in-situ three-dimensional bioprinting using the aforementioned spherical standing wave focused ultrasound-based system, comprising the following steps:

[0032] The spherical sound collector is fixed onto a high-precision robotic arm;

[0033] The tissue to be printed, which has been injected with printing ink, is inserted into the spherical sound collector, and the remaining cavity is filled with an elastic sealing coupling medium that is sealed by a sealing device.

[0034] The motion control system sends control commands to the high-precision robotic arm, causing the high-precision robotic arm to move along a specified trajectory, thereby causing the spherical sound collector to move along a specified trajectory.

[0035] The spherical acoustic collector is driven by an ultrasonic drive power supply, and the ultrasonic drive power supply is controlled by a control and data analysis system to turn the ultrasonic output on or off and adjust the ultrasonic output power.

[0036] Control the focal zone to apply printing ink to the tissue to be printed until printing is complete.

[0037] The beneficial effects of this invention are as follows:

[0038] This invention provides a deep in-situ 3D bioprinting system based on spherical standing wave focused ultrasound. This system utilizes spherical standing wave focused ultrasound, which offers superior focusing performance compared to traveling wave focused ultrasound. It enables spherical focused ultrasound 3D printing, balancing printing depth and accuracy while minimizing focal point shift in complex tissues. It relies on a novel spherical sound collector, which forms a spherical standing wave focusing cavity using two identical, oppositely positioned hemispherical transducers (where the transducer surfaces simultaneously serve as both the emitting and strongly reflecting surfaces of the ultrasound waves). The focused sound field is composed of the superposition of sound waves directly radiated from the spherical surface and those reflected multiple times. When the sound wave directly focused at the center of the sphere and the reflected sound waves are in phase, a very high sound pressure level can be generated at the center, and the focal accuracy is improved.

[0039] The system provided by this invention balances printing depth and printing accuracy, and uses a passive cavitation detector and a B-mode ultrasound diagnostic instrument to monitor the printing process in real time. This system can penetrate complex biological tissues to precisely focus energy, forming intricate structures in the printing ink for in-situ conformal filling of defects and even for constructing tissue engineering scaffolds.

[0040] Other advantages, objectives, and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination, or may be learned from practice of the invention. The objectives and other advantages of the invention can be realized and obtained through the following description. Attached Figure Description

[0041] To make the objectives, technical solutions, and beneficial effects of this invention clearer, the following drawings are provided for illustration.

[0042] Figure 1 It is a spherical focused ultrasound 3D printing system.

[0043] Figure 2 The route for spherical standing wave focused ultrasound 3D printing.

[0044] Figure 3 This is a schematic diagram of one application scenario of this system.

[0045] Figure 4 This is an example of how this system can penetrate biological tissue for printing.

[0046] In the figure, there are: high-precision robotic arm 101, robotic arm controller 102, elastic sealing coupling medium 103, printing ink 104, passive cavitation detector 105, spherical sound collector 106, B-mode ultrasound diagnostic instrument 107, sealing device 108, tissue to be printed 109, data acquisition device 110, ultrasonic drive power supply 111, control and data analysis system 112; skin 202, fat 203, muscle 204, focused ultrasound focal zone 208, tissue area to be repaired 206, bone 205, and ultrasonic beam 207. Detailed Implementation

[0047] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0048] Example 1

[0049] like Figure 1 As shown, Figure 1 The spherical focused ultrasound 3D printing system provided in this embodiment is a deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound, including a spherical focusing system, a motion control system, and a data processing system.

[0050] The spherical focusing system is used to generate spherical standing wave focused ultrasound with adjustable frequency, power and ultrasonic mode. By concentrating energy in the focal region, it induces cavitation and thermal effects, causing the printing ink to solidify and print the target.

[0051] The motion control system is used to control the position of the focused ultrasonic focal zone of the spherical focusing system, so that the focal area can move along the designed route and accurately shape the printed part.

[0052] The data processing system is used to monitor the printed images and cavitation signals in real time, acquire B-ultrasound images and ultrasonic cavitation signals and analyze and process them to obtain control signals for adjusting the spherical focusing system and motion control system.

[0053] The spherical focusing system includes a spherical sound collector 106, an ultrasonic drive power supply 111, a passive cavitation detector 105, and a B-type ultrasonic diagnostic instrument 107.

[0054] The spherical sound collector 106 has a spherical cavity structure, which allows the ultrasonic beam to be reflected on the opposite concave surface, causing a reverse change in the beam phase, which is used to form a spherical standing wave focused ultrasound, so that the energy is precisely focused on its spherical geometric center, generating a temperature or cavitation environment that can cause ink to solidify.

[0055] The spherical sound collector in this embodiment includes two hemispherical transducers of the same size and opposite positions. The two hemispherical transducers constitute a spherical standing wave focusing cavity. The spherical surface of the hemispherical transducer is the emitting surface and strong reflecting surface of the ultrasonic wave. The sound waves radiated from the spherical surface and the sound waves reflected multiple times by the spherical surface are superimposed to form a focused sound field.

[0056] The ultrasonic drive power supply 111 is used to supply power to the spherical sound collector 106 on demand, and can adjust the frequency, duty cycle, excitation parameters, etc., to realize the on-demand excitation of different types of ultrasonic waves.

[0057] The passive cavitation detector 105 is used to detect cavitation signals, monitor the strength of cavitation signals in real time, observe the printing effect, and feed it back to the control and data analysis system 112 so as to adjust the printing parameters in real time.

[0058] The B-type ultrasound diagnostic instrument 107 is used to acquire dynamic image monitoring signals during the printing process, monitor the internal image of the object in real time during printing, observe the printing effect, and feed it back to the control and data analysis system 112 so as to adjust the printing parameters in real time.

[0059] The motion control system includes a high-precision robotic arm 101, a robotic arm controller 102, a sealing device 108, and an elastic sealing coupling medium 103.

[0060] The high-precision robotic arm 101 is used to drive the spherical sound collector 106, so that its focal point can achieve high-precision movement as needed.

[0061] The robotic arm controller 102 is connected to the high-precision robotic arm 101 and is used to send control commands to the high-precision robotic arm 101.

[0062] The sealing device 108 is used to prevent the elastic coupling medium 103 from leaking out of the cavity;

[0063] The elastic sealing coupling medium 103 is used to allow ultrasonic energy to propagate smoothly with the least possible energy attenuation.

[0064] The data processing system includes a data acquisition unit 110 and a control and data analysis system 112;

[0065] The data acquisition unit 110 is used to acquire and store cavitation signals;

[0066] The control and data analysis system 112 is used to receive dynamic image monitoring signals and cavitation signals; and to dynamically monitor the printing process, evaluate the strength of cavitation and the focus position, etc., so as to adjust the output power and the motion speed in real time to ensure printing accuracy.

[0067] The deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound provided in this embodiment works as follows:

[0068] The spherical sound collector 106 is fixed to the high-precision robotic arm 101;

[0069] The tissue 109 to be printed, which is injected with printing ink 104, is inserted into the spherical sound collector 106, and the remaining cavity is filled with the elastic sealing coupling medium 103 sealed by the sealing device 108.

[0070] The motion control system sends control commands to the high-precision robotic arm 101 to make the high-precision robotic arm 101 move along a specified trajectory, thereby causing the spherical sound collector 106 to move along a specified trajectory.

[0071] The ultrasonic drive power supply 111 drives the spherical sound collector 106, while the passive cavitation detector 105 and the B-type ultrasonic diagnostic instrument 107 observe the printing status in real time. The control and data analysis system 112 controls the ultrasonic drive power supply 111 to turn on or off the ultrasonic output and adjust the ultrasonic output parameters.

[0072] When the spherical acoustic collector 106 is driven by the ultrasonic drive power supply 111, the focal zone generated by the spherical acoustic collector 106 should be within the range of the printing ink 104 in the tissue to be printed 109. The movement of the focal zone within the printing ink 104 is controlled until printing is completed.

[0073] The motion control system in this embodiment operates as follows: a spherical acoustic collector 106 is fixed to a high-precision robotic arm 101, and the tissue 109 to be printed, injected with printing ink 104, is inserted into the spherical acoustic collector 106. The remaining cavity is filled with an elastic sealing coupling medium 103 sealed by a sealing device 108. The high-precision robotic arm 101 is jointly controlled by a robotic arm controller 102 and a control and data analysis system 112. The control and data analysis system 112 sends motion commands to the robotic arm controller 102 to make the high-precision robotic arm 101 move along a specified trajectory, thereby causing the spherical acoustic collector 106 to move along the specified trajectory. The positioning accuracy is ≤0.001 mm, and the repeatability error can reach ±0.02 mm. The tissue 109 to be printed can be a complex tissue.

[0074] Therefore, the target area printing ink 104 and the spherical sound collector 106 can achieve relative movement according to the designed path. Alternatively, in addition to using the high-precision robotic arm 101, the motion control system can use other motion mechanisms, such as a motion system composed of stepper motors or servo motors. Alternatively, the spherical sound collector 106 can also use phase control technology to adjust the focal position, ensuring that the focal area can move according to the designed path.

[0075] The spherical focusing system in this embodiment operates as follows: the spherical acoustic collector 106 is filled with an elastic sealing coupling medium 103 sealed by a sealing device 108. The spherical acoustic collector 106 is driven by an ultrasonic drive power supply 111, and the control and data analysis system 112 controls the ultrasonic drive power supply 111 to turn the ultrasonic output on or off and adjust the ultrasonic output power. The frequency of the spherical acoustic collector should be ≥20 kHz, preferably a higher frequency band; the output power is 0~1000W, selected according to actual conditions. The characteristics of spherical focused ultrasound are a small focal zone and concentrated energy, which can form a very high sound pressure at the center of the sphere and generate high temperature in the focal zone, while the temperature generated at other locations is extremely low compared to the focal zone. Therefore, the printing accuracy is extremely high. When the focal zone acts on the printing ink 104 in the tissue to be printed 109, its cured size is ≤2λ, where λ represents the wavelength of the ultrasonic wave.

[0076] For the spherical sound collector 106, a standard spherical sound collector or a phase-controlled spherical sound collector can be used. Its radius and opening size can be adjusted as needed. For the printing ink 104, thermosetting materials or materials that undergo a phase change after a sonochemical reaction can be used, including but not limited to polydimethylsiloxane (PDMS) and methacrylamide gelatin (GelMa). It can be filled into the printing position by injection or extrusion.

[0077] like Figure 2 As shown, Figure 2 For spherical focused ultrasound 3D printing technology, the control and data analysis system 112 converts the data format of the 3D model to be printed and performs slicing calculations. Path planning algorithms are used for path planning. Commonly used 3D printing path planning algorithms include Zigzag path planning algorithm, contour parallel algorithm, and global continuous filling algorithm. The planned path data is transmitted to the robotic arm controller 102 so that the high-precision robotic arm moves according to the planned path. At the same time, the ultrasonic drive power supply 111 is controlled to start printing layer by layer. Multiple layers are printed to print a complete 3D model.

[0078] In the aforementioned 3D printing system, the printing material is specifically formulated as a biocompatible thermally responsive or cavitation-responsive phase change material, and this material is placed within a complex tissue.

[0079] like Figure 3 As shown, Figure 3 In one proposed application scenario of this system in tissue printing, the printing ink 104 is delivered to the area of ​​tissue to be repaired by the injection device 201 through injection or pump extrusion. The ultrasonic beam 207 generated by the spherical sound collector 106 passes through tissues such as skin 202, fat 203, muscle 204, and bone 205 and reaches the focused ultrasound focal zone 208 for focusing. Thermal and cavitation effects occur at the focal zone, causing the material at the focal zone to permanently solidify. Combined with the motion control system, the position of the focal zone is changed to achieve in-situ 3D printing of the scaffold 209 to promote tissue repair.

[0080] like Figure 4 As shown, Figure 4 This embodiment presents an experimental sample for printing PDMS (polydimethylsiloxane) through muscle tissue. The system described herein is used for in-situ, non-invasive, precise 3D printing within complex tissues. This system utilizes a spherical standing wave focused ultrasound transducer. Regarding tissue penetration depth, ultrasound (mechanical wave) based focusing is theoretically superior to near-infrared light (electromagnetic wave) based techniques. In terms of printing accuracy, spherical standing wave focused ultrasound, by superimposing two opposite wavefronts of the standing wave, generates a smaller in-phase region than traveling waves, suppressing diffraction effects that cause focused beam distortion and creating a smaller focused area. Furthermore, the reflection of the ultrasonic beam on opposing concave surfaces causes a reverse phase change in the ultrasonic beam, which can compensate for phase and focus shifts during propagation in multi-layered media, resulting in smaller focus shifts.

[0081] In summary, this embodiment utilizes a spherical standing wave focusing ultrasonic transducer to construct non-invasive in-situ 3D printing equipment, offering advantages that existing technologies cannot replace. The 3D printing system, built upon the spherical standing wave focusing transducer, achieves a balance between printing depth and accuracy. Simultaneously, a monitoring system capable of real-time feedback monitoring is provided.

[0082] The embodiments described above are merely preferred embodiments for fully illustrating the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are all within the scope of protection of the present invention. The scope of protection of the present invention is defined by the claims.

Claims

1. A deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound, characterized in that: This includes a spherical focusing system, a motion control system, and a data processing system; The spherical focusing system is used to generate spherical internal standing wave focused ultrasound, which prints the target through ultrasonic cavitation; The motion control system is used to control the position of the focused ultrasonic focal zone of the spherical focusing system, so that the focal area can move along the designed route; The data processing system is used to acquire and analyze ultrasonic cavitation signals and dynamic image monitoring signals of the printing process. The ultrasonic cavitation signals are monitored in real time by a passive cavitation detector, and the dynamic image monitoring signals are from a B-mode ultrasound diagnostic instrument. The system is fused with the dynamic image monitoring signals to dynamically evaluate the cavitation intensity and focal position of the printing point, and to generate control signals for coordinated adjustment of the output power of the spherical focusing system and the motion speed of the motion control system.

2. The deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound as described in claim 1, characterized in that: The spherical focusing system includes a spherical sound collector, an ultrasonic drive power supply, a passive cavitation detector, and a B-mode ultrasound diagnostic instrument. The spherical sound collector reflects the ultrasonic beam through the concave surface of the spherical cavity structure, forming a spherical standing wave focused ultrasound, which focuses the energy at the spherical geometric center of the spherical cavity structure. The ultrasonic driving power supply is used to supply power to the spherical sound collector on demand to realize the excitation of different types of ultrasonic waves; The passive cavitation detector is used to detect cavitation signals, monitor cavitation signals in real time, and feed the monitored cavitation signals back to the data processing system. The B-mode ultrasound diagnostic instrument is used to acquire dynamic image monitoring signals during the printing process and monitor the internal image of the object in real time during printing.

3. The deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound as described in claim 2, characterized in that: The ultrasonic drive power supply uses adjustable frequency, duty cycle, and excitation parameters to achieve on-demand excitation of different types of ultrasonic waves.

4. The deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound as described in claim 1, characterized in that: The motion control system includes a high-precision robotic arm, a robotic arm controller, a sealing device, and an elastic sealing coupling medium; The high-precision robotic arm is used to drive the spherical sound collector, so that the focal point of the spherical sound collector can move as needed; The robotic arm controller is connected to the high-precision robotic arm and is used to send control commands to the high-precision robotic arm; The sealing device is used to prevent the elastic coupling medium from leaking out of the cavity; The elastic sealing coupling medium is used to transmit ultrasonic energy.

5. The deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound as described in claim 1, characterized in that: The spherical focusing system has an area for placing the tissue to be printed, and the area is located at the geometric center of the spherical cavity structure.

6. The deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound as described in claim 5, characterized in that: The tissue to be printed contains printing ink, which is a thermosetting material or a material that undergoes a phase change after a sonochemical reaction.

7. The deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound as described in claim 2, characterized in that: The spherical sound collector includes two hemispherical transducers forming a spherical standing wave focusing cavity. The spherical surface of the hemispherical transducer is the emitting surface and strong reflecting surface of the ultrasonic wave. The sound waves radiated from the spherical surface and the sound waves reflected multiple times by the spherical surface are superimposed to form a focused sound field.

8. The deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound as described in claim 7, characterized in that: The two hemispherical transducers are the same size and are positioned opposite each other.

9. A three-dimensional bioprinting method using the deep in-situ three-dimensional bioprinting system based on spherical standing wave focused ultrasound as described in any one of claims 1 to 8, characterized in that: Includes the following steps: The spherical sound collector is fixed onto a high-precision robotic arm; The tissue to be printed, which has been injected with printing ink, is inserted into the spherical sound collector, and the remaining cavity is filled with an elastic sealing coupling medium that is sealed by a sealing device. The motion control system sends control commands to the high-precision robotic arm, causing the high-precision robotic arm to move along a specified trajectory, thereby causing the spherical sound collector to move along a specified trajectory. The spherical sound collector is driven by an ultrasonic drive power supply. The data processing system acquires and analyzes ultrasonic cavitation signals and dynamic image monitoring signals of the printing process. The ultrasonic cavitation signals are monitored in real time by a passive cavitation detector, and the dynamic image monitoring signals are from a B-mode ultrasound diagnostic instrument. The system is fused with the dynamic image monitoring signals to dynamically evaluate the cavitation intensity and focal position of the printing point, and to generate control signals for coordinated adjustment of the output power of the spherical focusing system and the motion speed of the motion control system. The control and data analysis system controls the ultrasonic drive power supply to turn the ultrasonic output on or off and adjust the ultrasonic output power. Simultaneously, the movement speed of the robotic arm is adjusted according to the control signal; Control the focal zone to apply printing ink to the tissue to be printed until printing is complete.

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