A bioprinting system and method for precise cell positioning

Abstract

The present application relates to a bioprinting system and method for precise cell positioning, and relates to the field of three-dimensional printing technology. The system includes: a three-dimensional motion module, which is used to achieve precise movement of the print head; a printing module, which includes a print head, which includes a printing nozzle, and the printing module is used to construct a biological scaffold and / or eject cells; an information acquisition module, which includes a camera for real-time positioning of the physical position and / or cell position of the biological scaffold; a software module, which is used to achieve motion control of the print head. The bioprinting system and method for precise cell positioning of the present application achieves precise cell positioning by precisely printing a single cell or a single row of cell sequences onto a target biological scaffold, providing the necessary foundation for the subsequent orderly growth, proliferation and signal transduction of cells.

Description

Technical Field

[0001] The present application relates to the field of three-dimensional printing technology, and in particular to a bioprinting system and method for precise cell positioning. Background Art

[0002] The bioprinting system is an upgrade based on conventional 3D printing technology. Its main function is to print and shape biomaterials or biological cells, ultimately realizing applications in human medicine or tissue engineering. Conventional bioprinting mainly consists of a 3D motion module and a multifunctional print head. The 3D motion module is mainly used to drive the high-precision 3D movement of the print head. The print head uses air pressure or a screw as a driving force to extrude the printing slurry (biomaterial) into a wire. The wire drawing path is then controlled by software to achieve two-dimensional arrangement and three-dimensional stacking of the material, ultimately achieving high-precision stacking of the biomaterial. Traditional bioprinting is only the 3D shaping of biomaterials and does not include the construction and shaping of biological cells. In clinical applications, because the bioprinted scaffold only provides mechanical support, cells can only rely on the organism's own cells to gradually erode and grow into the bioscaffold. Tissue repair is slow, and the cells are disordered and inefficient. Therefore, the application scenarios of bioprinting are very limited. With the continuous development of science and technology, the realization of cell printing will become an inevitable trend.

[0003] Since the size of general cells is around 40μm, and the accuracy of traditional bioprinting is mostly between 200-400μm, and the printing process involves mixed extrusion printing of cells and bio-hydrogels, it is impossible to achieve close arrangement of cells after actual printing. However, in the process of cell growth, proliferation and functional differentiation, signal transduction and growth connections between cells are involved, which can promote the growth and differentiation of cells into tissues or organs. The current cell printing model cannot achieve close and orderly arrangement of cells, and ultimately results in cells being able to only grow into multiple independent cell clusters nearby, and cannot be connected into tissues and functionalized.

[0004] Therefore, it is desirable to provide a bioprinting system and method for precise cell positioning, which can achieve precise cell positioning by precisely printing single cells or single rows of cell sequences onto the target biological scaffold, providing the necessary basis for the subsequent orderly growth, proliferation and signal transduction of cells, thereby better promoting cell growth and differentiation and achieving functionalization. Summary of the Invention

[0005] According to a first aspect of some embodiments of the present application, a bioprinting method for precise cell positioning is provided, the method comprising: obtaining a three-dimensional model and slicing it; generating instructions and inputting them into a three-dimensional motion module; constructing a three-dimensional bioscaffold by layer-by-layer assembly; collecting position information of the three-dimensional bioscaffold and feeding back the predetermined position; and precisely printing cells to the predetermined position of the three-dimensional bioscaffold.

[0006] In some embodiments, the three-dimensional model is imported and sliced ​​by a software module, and the generating of instructions and inputting them into the three-dimensional motion module includes: controlling the three-dimensional motion of the print head relative to the sample stage through the relative motion of the XYZ three-axis robotic arm.

[0007] In some embodiments, the extrusion print head is loaded with biomaterials, and the layer-by-layer assembly method for constructing a three-dimensional bioscaffold includes: the extrusion print head performs three-dimensional movement with the robotic arm according to the instructions, and extrudes the biomaterial by pneumatic extrusion or screw extrusion; and the three-dimensional bioscaffold is constructed by layer-by-layer assembly.

[0008] In some embodiments, the collecting of position information of a three-dimensional bioscaffold and the feedback of a predetermined position include: after each layer of the three-dimensional bioscaffold is printed, obtaining the position information of the three-dimensional bioscaffold through an upright camera, and accurately locating the physical position of each layer of the scaffold; converting the physical position into a relative position relative to the origin of the sample stage; inputting the relative position into a software module, and feeding back the predetermined position.

[0009] In some embodiments, based on the requirement for precise cell layout, constructing a three-dimensional biological scaffold includes performing a printing pause during the printing process of each layer or multiple layers.

[0010] In some embodiments, the printing ink of the thermal bubble print head includes a cell suspension, and the precise printing of cells to a predetermined position of the three-dimensional bioscaffold includes: precisely positioning the thermal bubble print head above the three-dimensional bioscaffold according to the relative position of the three-dimensional bioscaffold; and utilizing the moment of thermal bubble bursting to eject the cells and precisely print them to the predetermined position of the three-dimensional bioscaffold.

[0011] In some embodiments, the size of the print nozzle of the thermal bubble print head is adjusted so that only a single cell is present in the print nozzle, and an inverted camera is used to determine whether the cell is precisely positioned in the print nozzle. The method of ejecting the cell at the moment of thermal bubble bursting includes: ejecting the single cell quickly and orderly from the print nozzle through instantaneous heating.

[0012] In some embodiments, the precise printing to the predetermined position of the three-dimensional bioscaffold specifically includes determining the first width of the printed line by the printing nozzle size; setting the image position origin of the current printed layer of the three-dimensional bioscaffold and the second width of the printing gap; relative to the image position origin, according to the first width and the second width, calculating the theoretical coordinate value of the current printed line and the number of pixels corresponding to the horizontal axis and / or vertical axis; obtaining the real physical distance corresponding to each pixel under the current shooting conditions; determining the real physical position of the current print as the predetermined position based on the theoretical coordinate value, the number of pixels and the real physical distance corresponding to each pixel; and precisely printing the cells to the predetermined position of the three-dimensional bioscaffold.

[0013] In some embodiments, a plurality of thermal bubble print heads are arranged linearly to perform multi-cell orderly linear arrangement printing.

[0014] According to a second aspect of some embodiments of the present application, a system is provided, comprising: a memory configured to store data and instructions; a processor communicating with the memory, wherein, when executing instructions in the memory, the processor is configured to: acquire a three-dimensional model and slice it; generate instructions and input them into a three-dimensional motion module; construct a three-dimensional bioscaffold using a layer-by-layer assembly method; collect position information of the three-dimensional bioscaffold and feed back a predetermined position; and accurately print cells to a predetermined position of the three-dimensional bioscaffold.

[0015] Therefore, by accurately printing single cells or single rows of cell sequences onto the target biological scaffold, precise cell positioning can be achieved, providing the necessary basis for the subsequent orderly growth, proliferation and signal transduction of cells, thereby better promoting cell growth and differentiation and achieving functionalization. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to better understand and illustrate some embodiments of the present application, the following description of the embodiments will be made with reference to the accompanying drawings, in which like reference numerals indicate corresponding parts.

[0017] Figure 1 is an exemplary schematic diagram of a bioprinting system for precise cell positioning provided in accordance with some embodiments of the present application.

[0018] Figure 2 This is an exemplary flow chart of a bioprinting method for precise cell positioning according to some embodiments of the present application.

[0019] Figure 3 is an exemplary schematic diagram of bioprinting provided according to some embodiments of the present application.

[0020] Figure 4This is a schematic diagram of a biological scaffold captured by an upright camera according to some embodiments of the present application.

[0021] Figure 5 Schematic diagram of cell distribution after a single array ejection by a thermal bubble print head according to some embodiments of the present application.

[0022] Figure 6 This is a schematic diagram of cell distribution after multiple array injections by a thermal bubble print head according to some embodiments of the present application. DETAILED DESCRIPTION

[0023] The following description with reference to the accompanying drawings is intended to facilitate a comprehensive understanding of the various embodiments of the present application as defined by the claims and their equivalents. These embodiments include various specific details to facilitate understanding, but these are to be considered as exemplary only. Therefore, those skilled in the art will appreciate that various changes and modifications can be made to the various embodiments described herein without departing from the scope and spirit of the present application. In addition, for the sake of brevity and clarity, descriptions of well-known functions and structures will be omitted.

[0024] The terms and phrases used in the following description and claims are not limited to their literal meanings, but are intended only to enable a clear and consistent understanding of the present application. Therefore, it will be understood by those skilled in the art that the description of the various embodiments of the present application is provided for illustrative purposes only and is not intended to limit the present application as defined in the appended claims and their equivalents.

[0025] The following will be combined with the drawings in some embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the embodiments described are only some of the embodiments of the present application, not all of the embodiments. Based on the embodiments of the present application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0026] It should be noted that the terms used in the embodiments of the present application are only for the purpose of describing specific embodiments and are not intended to limit the present application. The singular forms of "one", "an", "a kind of", "said" and "the" used in the embodiments of the present application and the appended claims are also intended to include plural forms, unless the context clearly indicates otherwise. It should also be understood that the term "and / or" used in this application refers to and includes any or all possible combinations of one or more bound listed items. The expressions "first", "second", "the first" and "the second" are used to modify the corresponding elements without regard to order or importance, and are merely used to distinguish one element from another, without limiting the corresponding elements.

[0027] According to some embodiments of the present application, the terminal of the software module may be an electronic device, which may include a personal computer (PC, such as a tablet computer, desktop computer, notebook, netbook, handheld computer PDA), a user terminal device, a virtual reality device (VR), an augmented reality device (AR), a mixed reality device (MR), an XR device, a rendering machine, a smart phone, a mobile phone, an e-book reader, a portable multimedia player (PMP), an audio / video player (MP3 / MP4), a camera and a wearable device, etc. One or a combination of several. According to some embodiments of the present application, the wearable device may include an accessory type (such as a watch, a ring, a bracelet, glasses, or a head-mounted device (HMD)), an integrated type (such as electronic clothing), a decorative type (such as a skin pad, a tattoo or a built-in electronic device), etc., or a combination of several. In some embodiments of the present application, the electronic device may be flexible, not limited to the above-mentioned devices, or may be a combination of one or several of the above-mentioned various devices. In the present application, the term "user" may indicate a person using an electronic device or a device using an electronic device (such as an artificial intelligence electronic device).

[0028] The present invention provides a bioprinting system and method for precise cell positioning. To facilitate understanding of the present invention, the present invention will be described in detail below with reference to the accompanying drawings.

[0029] Figure 1 Schematic diagram of a bioprinting system for precise cell positioning according to some embodiments of the present application. Figure 1 As shown, the bioprinting system 100 for precise cell positioning may include a network 110, a control terminal 120, an apparatus terminal 130, and a server 140. Specifically, the control terminal 120 and the apparatus terminal 130 may establish communication via the network. For example, the control terminal 120 and the apparatus terminal 130 may communicate within the same local area network (e.g., a network environment with the same router). Furthermore, the control terminal 120 may be connected to the network 110 via a wired (e.g., network cable) or wireless connection, and the apparatus terminal 130 may establish a communication connection with the network 110 via a wired or wireless connection (e.g., Wi-Fi). In some embodiments, the apparatus terminal 130 may send information to the control terminal 120 and the server 140, such as image information acquired by the information acquisition module 134. Furthermore, the control terminal 120 and the server 140 may send feedback information to the apparatus terminal 130. For example, the control terminal 120 may send information such as the location of a bioscaffold to the apparatus terminal 130, and the control terminal 120 may perform feedback control based on the operation of the apparatus terminal 130. In some embodiments, the control end 120 may send a request message to the server 140 to obtain associated information of the device end 130 , for example, to obtain a three-dimensional model of biological printing, etc.

[0030] According to some embodiments of the present application, the control end 120 and the device end 130 may include various terminal devices, etc. These terminal devices may include, but are not limited to, smart terminals, mobile terminals, computers, printers, 3D printers, renderers, etc. In a 3D printing scenario, the control end 120 may include an electronic device such as a computer, which may include a software module 121, etc. The device end 130 may include a 3D motion module 132, a printing module 133, an information acquisition module 134, etc. The 3D motion module 132 may include an XYZ three-axis robotic arm, etc. The printing module 133 may include an extrusion printhead or a thermal bubble printhead, etc. The information acquisition module 134 may include an upright camera or an inverted camera, etc. As an example, the device end 130 may further include a processor 135, a display 136, a communication module 137, etc.; as another example, the various modules 132-137 of the device end 130 may be connected via a bus 131. The processor 135 may process information such as the motion trajectory sent by the control end 120 and execute the motion trajectory through the 3D motion module 132. The display 136 can display real-time images of 3D printing, etc. The communication module 137 can exchange information with the control terminal 120, server 140, etc. In some embodiments, the control terminal 120 and the device terminal 130 can be integrated into a single device, such as a 3D printer. In some embodiments, the server 140 is a type of computer that offers advantages over conventional computers, such as faster operation and higher load capacity, but correspondingly higher prices. In a network environment, the server can provide computing or application services to other clients (e.g., PCs, smartphones, ATMs, and other terminals, as well as large-scale equipment such as transportation systems). The server has high-speed CPU computing power, long-term reliable operation, strong I / O external data throughput, and improved scalability. Services that the server can provide include, but are not limited to, responding to service requests, providing services, and ensuring services. As an electronic device, the server has an extremely complex internal structure, including internal structures similar to those of conventional computers. For example, the server's internal structure may include a central processing unit (CPU), hard disk, memory, system, and system bus.

[0031] In some embodiments of the present application, the bioprinting system 100 for precise cell positioning may omit one or more elements, or may further include one or more other elements. As an example, the bioprinting system 100 for precise cell positioning may include a memory, etc., for example, for storing three-dimensional models, etc. For another example, the bioprinting system 100 may include multiple device terminals 130, which can perform three-dimensional printing separately and / or collaboratively perform three-dimensional printing. For another example, the bioprinting system 100 for precise cell positioning may include multiple control terminals 120. For another example, the bioprinting system 100 for precise cell positioning may include multiple servers 140, etc. The network 110 may be any type of communication network, which may include a computer network (e.g., a local area network (LAN) or a wide area network (WAN)), the Internet and / or a telephone network, etc., or a combination of several. In some embodiments, the network 110 may be other types of wireless communication networks. The wireless communication may include microwave communication and / or satellite communication, etc. The wireless communication may include cellular communication, for example, Global System for Mobile Communications (GSM), Code Division Multiple Access (CDMA), third generation mobile communication (3G), fourth generation mobile communication (4G), fifth generation mobile communication (5G), sixth generation mobile communication (6G), long term evolution technology (LTE), long term evolution technology upgraded version (LTE-A, LTE-Advanced), wideband code division multiple access (WCDMA), universal mobile telecommunications system (UMTS, Universal Mobile Telecommunications System), wireless broadband (WiBro, Wireless Broadband), etc., or a combination of several. In some embodiments, the device end 130 can be other electronic devices with equivalent functional modules, which may include one or a combination of three-dimensional printing equipment, virtual reality equipment (VR), rendering machine, personal computer (PC, such as tablet computer, desktop computer, notebook, netbook, handheld computer PDA), smart phone, mobile phone, e-book reader, portable multimedia player (PMP), audio / video player (MP3 / MP4), camera and wearable device.

[0032] In some embodiments, the WIFI may be other types of wireless communication technologies. According to some embodiments of the present application, the wireless communication may include wireless local area network (WiFi), Bluetooth, Bluetooth Low Energy (BLE), ZigBee, near field communication (NFC), magnetic secure transmission, radio frequency and body area network (BAN), etc., or a combination of several. According to some embodiments of the present application, the wired communication may include global navigation satellite system (Glonass / GNSS), global positioning system (GPS), Beidou navigation satellite system or Galileo (European global satellite navigation system), etc. The wired communication may include universal serial bus (USB), high-definition multimedia interface (HDMI), recommended standard 232 (RS-232), and / or plain old telephone service (POTS), etc., or a combination of several.

[0033] According to some embodiments of the present application, the control terminal 120 may be an electronic device including, for example, a software module 121. The software module 121 is configured to implement motion control of a print head. The device terminal 130 may be a bioprinting device including a bus 131, which is connected to a three-dimensional motion module 132, a printing module 133, an information acquisition module 134, and the like. In some embodiments, the three-dimensional motion module 132 is configured to achieve precise motion of the print head. After importing and slicing a three-dimensional model, the software module inputs instructions to the three-dimensional motion module, which then implements three-dimensional motion of the print head relative to the sample stage through the relative motion of the X, Y, and Z robotic arms.

[0034] In some embodiments, the printing module 133 includes a print head, each including a print nozzle, configured to construct a bioscaffold and / or eject cells. The printing module includes an extrusion print head, which is loaded with biomaterial and moves three-dimensionally with a robotic arm according to instructions, extruding the biomaterial through extrusion. The extrusion print head can extrude biomaterial using either pneumatic or screw extrusion. When extruding the biomaterial, the bioscaffold is constructed layer by layer. The printing module includes a thermal bubble print head, which uses instantaneous heating to rapidly and orderly eject printing ink from the print nozzle, achieving precise printing on the printing substrate. The thermal bubble print head is precisely positioned above the bioscaffold using a software module, ejecting cells upon the instantaneous collapse of the thermal bubble, enabling precise printing of cells to the predetermined location on the bioscaffold. The printing ink comprises a cell suspension, and the size of the print nozzle can be adjusted to ensure that only a single cell is present at the print nozzle. The printing module includes multiple thermal bubble print heads, which are arranged linearly to achieve printing of multiple cells in an orderly linear arrangement.

[0035] In some embodiments, the information acquisition module 134 includes a camera for real-time location of the bioscaffold's physical position and / or cell position. The information acquisition module includes an upright camera for precisely locating the bioscaffold's physical position. This physical position is converted into a relative position relative to the sample stage and input into the software module. The information acquisition module also includes an inverted camera for determining whether cells are precisely positioned within the printing nozzle.

[0036] It should be noted that the above description of the bioprinting system 100 for precise cell positioning is for ease of description only and does not limit the present application to the scope of the illustrated embodiments. It is understood that those skilled in the art, based on the principles of this system, may arbitrarily combine the various components, or form subsystems connected to other components, without departing from such principles, and make various modifications and changes in form and details to the application areas of the above-mentioned methods and systems. For example, the control terminal 120 and the device terminal 130 may be integrated into a single device. Such variations are within the scope of protection of this application.

[0037] Figure 2 is an exemplary flow chart of a bioprinting method for precise cell positioning according to some embodiments of the present application. Figure 2 As shown, process 200 can be implemented by a cell-precision bioprinting system 100. In some embodiments, the cell-precision bioprinting system 200 can be automatically activated or activated by an instruction. The instruction can include a system instruction, a device instruction, a user instruction, an action instruction, or a combination thereof.

[0038] At 201, a three-dimensional model is acquired and sliced. Operation 201 can be implemented by the control terminal 120 and the server 140 of the cell precise positioning bioprinting system 100. In some embodiments, the control terminal 120 and / or the server 140 can acquire the three-dimensional model and slice it.

[0039] At 202 , instructions are generated and input to a three-dimensional motion module. Operation 202 can be implemented by the control terminal 120 and the device terminal 130 of the cell precise positioning bioprinting system 100 . In some embodiments, the control terminal 120 can generate instructions and input them to the three-dimensional motion module 132 of the device terminal 130 .

[0040] According to some embodiments of the present application, the three-dimensional model is imported and sliced ​​through the software module 121, and the generation of instructions and input into the three-dimensional motion module 132 includes controlling the three-dimensional motion of the print head relative to the sample stage through the relative motion of the XYZ three-axis robotic arm.

[0041] At 203, a three-dimensional bioscaffold is constructed using a layer-by-layer assembly process. Operation 203 can be performed by the device terminal 130 of the bioprinting system 100 for precise cell positioning. In some embodiments, the device terminal 130 can construct the three-dimensional bioscaffold using a layer-by-layer assembly process based on the three-dimensional model and slice information from the control terminal 120. The device terminal 130 can transmit position information of the three-dimensional bioscaffold relative to the sample stage to the control terminal 120 via the information acquisition module 134.

[0042] According to some embodiments of the present application, the printing module 133 may include an extrusion print head and a thermal bubble print head, and the extrusion print head is loaded with biological materials. The three-dimensional biological scaffold constructed by layer-by-layer assembly can be constructed by the extrusion print head according to the instructions, performing three-dimensional movement with the robotic arm, and drawing and extrude the biological material by pneumatic extrusion or screw extrusion; the three-dimensional biological scaffold is constructed by layer-by-layer assembly.

[0043] At 204, the position information of the three-dimensional bioscaffold is collected and the predetermined position is fed back. Operation 204 can be implemented by the device terminal 130 and the control terminal 120 of the cell precision positioning bioprinting system 100. In some embodiments, the device terminal 130 can obtain the position information of the three-dimensional bioscaffold via the information collection module 134. In some embodiments, the control terminal 120 can feed back the predetermined position via the software module 121.

[0044] According to some embodiments of the present application, the information acquisition module 134 may include an upright camera and an inverted camera. The three-dimensional bioscaffold may include multiple layers, and the construction of the three-dimensional bioscaffold includes obtaining the position information of the three-dimensional bioscaffold through the upright camera after each layer of the three-dimensional bioscaffold is printed, and accurately locating the physical position of each layer of the scaffold; the device end 130 may further include a processor 135, and the processor 135 may convert the physical position into a relative position relative to the origin of the sample stage, and input the relative position to the software module 121. The software module 121 can calculate the predetermined position of the precisely printed cells based on the relative position, and feed it back to the device end 130. As an example, the upright camera may include a high-definition camera, etc., and the inverted camera may include a high-definition camera, etc.

[0045] In some embodiments, a first width of a printed line is determined by the size of a printing nozzle; an image position origin of a current printed layer of a three-dimensional bioscaffold and a second width of a printing gap are set; based on the first width and the second width, the theoretical coordinate value of the current printed line and the number of pixels corresponding to the horizontal axis and / or vertical axis are calculated; the real physical distance corresponding to each pixel under the current shooting conditions is obtained; based on the theoretical coordinate value, the number of pixels and the real physical distance corresponding to each pixel, the real physical position of the current print is determined to be a predetermined position; and the cells are precisely printed to the predetermined position of the three-dimensional bioscaffold.

[0046] Specifically, the lower left corner of the image of the current printing layer of the three-dimensional bioscaffold (at the same vertical Z-axis coordinate value) can be set as the (0,0) position origin, the first width of the extruded line (printed line) is A, and the second width of the gap (printed gap) is B. Taking the mth printed line (the current printed line) as an example (m is an integer greater than or equal to 1 and less than or equal to n). Taking the horizontal X-axis as an example (the same applies to the vertical Y-axis), the theoretical X-coordinate value of the mth printed line relative to the (0,0) position origin in the lower left corner of the image is (m-1)*(A+B)+A / 2. When executing bioprinting, the system software first determines the number of pixels corresponding to the mth printed line on the X-axis based on the theoretical X-coordinate value of the mth printed line. Furthermore, by correcting the actual physical distance corresponding to each pixel at the current shooting height, the actual X-coordinate position of the mth printed line and its corresponding actual physical position are obtained. The printing system hardware is then driven to the actual physical position (set as the predetermined position), and step 205 is then executed to accurately print the cells at the predetermined position.

[0047] At 205, cells are precisely printed at predetermined locations on the three-dimensional bioscaffold. Operation 205 can be performed by the device terminal 130 of the bioprinting system 100 for precise cell positioning. In some embodiments, the device terminal 130 can execute precise cell printing at predetermined locations on the three-dimensional bioscaffold via the printing module 133 based on the position information of the three-dimensional bioscaffold relative to the sample stage, as fed back by the control terminal 120. The printing module 133 can include a thermal bubble printhead, the printing ink of which includes a cell suspension. Precisely printing cells at predetermined locations on the three-dimensional bioscaffold can be performed by precisely positioning the thermal bubble printhead above the three-dimensional bioscaffold based on the relative position of the three-dimensional bioscaffold. The thermal bubble printhead can eject cells at the precise location of the three-dimensional bioscaffold by rupturing the thermal bubble. In some embodiments, the control terminal 120 can adjust the size of the thermal bubble printhead's print nozzle so that only a single cell is present within the print nozzle. Ejecting cells at the precise location of the three-dimensional bioscaffold by rupturing the thermal bubble includes instantaneously heating the thermal bubble printhead to rapidly and orderly eject the single cell from the print nozzle. If there is only a single cell in the printing nozzle, an inverted camera can be used to determine whether the cell is accurately positioned in the printing nozzle. In some embodiments, the device end 130 can arrange multiple thermal bubble print heads in a linear arrangement to print multiple cells in an orderly linear arrangement.

[0048] According to some embodiments of the present application, process 200 may further include executing a printing pause during each or multiple layers of printing. According to the requirement of precise cell layout, constructing a three-dimensional bioscaffold includes executing a printing pause during each or multiple layers of printing.

[0049] According to some embodiments of the present application, Figure 3 As shown, Figure 3 This is an exemplary schematic diagram of bioprinting provided according to some embodiments of the present application. Bioprinting primarily consists of a three-dimensional motion system and a multifunctional print head. The three-dimensional motion system is primarily used to drive the print head's high-precision three-dimensional motion. The print head uses a driving force such as air pressure or a screw to extrude the printing slurry (biomaterial) into a wire. Software then controls the wire drawing path to achieve two-dimensional arrangement and three-dimensional stacking of the material, ultimately achieving high-precision stacking and molding of the biomaterial.

[0050] According to some embodiments of the present application, Figure 4 As shown, Figure 4 This is a schematic diagram of a bio-scaffold captured by an upright camera according to some embodiments of the present application. The upright high-definition camera collects information about the bio-scaffold after each layer is printed, accurately locating the physical position of each scaffold within the bio-printed scaffold and converting this information into relative positions relative to the sample stage origin for input into the system. Figure 4The image of the printed stent is captured in real time by a high-definition camera. In some embodiments, the center of the image can be set as the (0,0) position origin or the lower left corner of the image can be set as the (0,0) position origin. The camera pixel is 9 μm, which can accurately measure the relative position of any point in the image of the printed stent relative to the origin and feed it back to the software module for accurate cell distribution.

[0051] As an example, Figure 4 The black center represents the biomaterial extruded by the bioprinter nozzle, and the white represents the set gap. This gap can be manually set or set by default to facilitate the flow of cell nutrients. In some embodiments, the actual positioning and recognition process of bioprinting is specifically described using the example of precise cell positioning printing in the horizontal (X-axis) direction.

[0052] The set width of the extruded line (shown in black) is A, which is determined by the material and the inner diameter of the extrusion needle (such as the print nozzle) and is accurately calibrated before printing. The set width of the gap (shown in white) is B. Parameter B can be set through software and accurately calibrated before printing. As an example, set the lower left corner of the image as the origin (0,0). When printing 1 to n lines (n is an integer greater than 1), take the mth extruded line as an example (m is an integer greater than or equal to 1 and less than or equal to n). The theoretical X coordinate value of the mth extruded line relative to the (0,0) origin in the lower left corner of the image is (m-1)*(A+B)+A / 2. At the same time, the actual X coordinate value of the mth extruded line relative to the (0,0) origin in the lower left corner of the image is calculated by the system software. For example, during calculation, the center point of the mth extruded line is taken as the reference point. The system software obtains the accurate real X-coordinate position by measuring the number of pixels from the leftmost position of the image (x=0) to the reference point (measured at the same y-axis coordinate value) and the real physical distance corresponding to each pixel point obtained by correction at the current shooting height.

[0053] When executing bioprinting, the system software first determines the corresponding mth extrusion line using the theoretical X-coordinate value of the mth extrusion line, then accurately calculates the actual X-coordinate value of the mth extrusion line and its corresponding real physical position. It then drives the printing system hardware to the actual physical position (set as the predetermined position) and executes precise cell printing, printing the cells precisely at the predetermined position. The system software searches all n extrusion lines in the X-axis direction (shown as the black lines in the figure) and accurately calculates the real physical position of each extrusion line 1 to n, then executes cell printing until the precise printing of cells on the entire bioscaffold is completed.

[0054] According to some embodiments of the present application, Figure 5 As shown, Figure 5 This is a schematic diagram of cell distribution after a single array jet from a thermal bubble printhead, according to some embodiments of the present application. A thermal bubble printhead utilizes the principle of instantaneous heating to rapidly and orderly eject printing ink from a print nozzle, accurately printing onto a printing substrate. In this application, a cell suspension is used as ink, and the nozzle size is adjusted to achieve a single cell within the nozzle before printing. During printing, the thermal bubble printhead is precisely positioned directly above the bioscaffold, and the cells are ejected instantly upon bursting the thermal bubble, allowing precise printing to the predetermined location on the bioscaffold. Figure 5 This is a schematic diagram of cell distribution after a single array injection by a thermal bubble principle print head.

[0055] According to some embodiments of the present application, Figure 6 As shown, Figure 6 Figure 1 is a schematic diagram of cell distribution after multiple array injections from a thermal bubble printhead according to some embodiments of the present application. As an example, a bioprinting system 100 for precise cell positioning can linearly arrange a series of thermal bubble printheads (e.g., 180, etc.) to achieve multi-cell printing in an orderly linear arrangement. Figure 6 This is a schematic diagram of cell distribution after multiple array jet distribution by the thermal bubble principle print head.

[0056] It should be noted that the above description of system 100 and process 200 is for convenience only and does not limit the present application to the scope of the illustrated embodiments. It is understood that those skilled in the art, based on the principles of this system, may arbitrarily combine the various operations, or construct sub-processes and combinations with other operations, without departing from such principles, and may make various modifications and changes in the form and details of the functions implementing the above-described processes and operations. For example, process 200 may further include operations such as pausing printing during the printing of each or multiple layers. Such variations are within the scope of protection of this application.

[0057] In summary, according to the bioprinting system and method for precise cell positioning of the embodiments of the present application, precise cell positioning is achieved by accurately printing single cells or single rows of cell sequences onto the target biological scaffold, providing the necessary basis for the subsequent orderly growth, proliferation and signal transduction of cells, thereby better promoting cell growth and differentiation and achieving functionalization.

[0058] It should be noted that the above embodiments are merely examples, and the present application is not limited to such examples, but may be modified in various ways.

[0059] It should be noted that, in this specification, the terms "comprises," "includes," or any other variations thereof are intended to encompass non-exclusive inclusion, such that a process, method, article, or apparatus comprising a series of elements includes not only those elements but also other elements not explicitly listed, or elements inherent to such process, method, article, or apparatus. In the absence of further limitations, an element defined by the phrase "comprising a ..." does not exclude the presence of other identical elements in the process, method, article, or apparatus comprising the element.

[0060] Finally, it should be noted that the above series of processes include not only processes executed in time series in the order described herein, but also processes executed in parallel or separately rather than in time series.

[0061] Those skilled in the art will appreciate that all or part of the processes in the above-described method embodiments can be implemented through hardware associated with computer program instructions. The program can be stored in a computer-readable storage medium, and when executed, the program can include the processes in the above-described method embodiments. The storage medium can be a magnetic disk, an optical disk, a read-only memory (ROM), or a random access memory (RAM).

[0062] The above disclosures are only some preferred embodiments of the present application and are not intended to limit the scope of the present application. A person skilled in the art will understand that all or part of the processes of the above embodiments and equivalent changes made in accordance with the claims of the present application are still within the scope of the present application.

Claims

1. A bioprinting method for precise cell positioning, characterized in that: include: Get the 3D model and slice it; Generate instructions and input them into the 3D motion module; By loading biomaterials inside the extrusion print head, a three-dimensional bioscaffold is constructed by layer-by-layer assembly. Acquiring position information of a three-dimensional biological scaffold and feeding back a predetermined position includes acquiring position information of the three-dimensional biological scaffold through an upright camera after each layer of the three-dimensional biological scaffold is printed, and accurately locating the physical position of each layer of the scaffold; converting the physical position into a relative position relative to the origin of the sample stage; inputting the relative position into a software module, and feeding back a predetermined position; The printing ink of the thermal bubble print head includes a cell suspension, and the precise printing of cells to predetermined positions of the three-dimensional biological scaffold by the thermal bubble print head includes determining a first width of the printed line by a print nozzle size; Set the second width of the printing gap; set the image position origin of the current printing layer of the three-dimensional biological scaffold; Relative to the image position origin, according to the first width and the second width, calculate the theoretical coordinate value of the current printed line and the number of pixels corresponding to the horizontal axis and / or the vertical axis; Obtaining the actual physical distance corresponding to each pixel under the current shooting conditions; determining the actual physical position of the current print as the predetermined position based on the theoretical coordinate values, the number of pixels, and the actual physical distance corresponding to each pixel; and accurately printing the cells to the predetermined position of the three-dimensional bioscaffold; Adjusting the print nozzle size of the thermal print head so that only a single cell is present in the print nozzle, and using an inverted camera to determine whether the cell is accurately positioned in the print nozzle; Using the bursting of hot bubbles to eject cells instantly includes: ejecting single cells quickly and orderly from the printing nozzle through instantaneous heating.

2. The method according to claim 1, characterized in that Importing the three-dimensional model and slicing it through the software module, generating instructions and inputting them into the three-dimensional motion module includes: The three-dimensional motion of the print head relative to the sample stage is controlled by the relative motion of the XYZ three-axis robotic arm.

3. The method according to claim 2, characterized in that The method of constructing a three-dimensional biological scaffold by layer-by-layer assembly includes: The extrusion print head performs three-dimensional movement along with the robotic arm according to the instructions, and extrudes the biomaterial by air pressure extrusion or screw extrusion; A three-dimensional biological scaffold is constructed using a layer-by-layer assembly method.

4. The method according to claim 3, characterized in that According to the requirement of precise cell layout, the construction of the three-dimensional biological scaffold includes performing a printing pause during each layer or multiple layers of printing.

5. The method according to claim 3, characterized in that The precise printing of cells to predetermined positions of the three-dimensional biological scaffold comprises: According to the relative position of the three-dimensional biological scaffold, the thermal bubble print head is accurately positioned above the three-dimensional biological scaffold; The cells are ejected at the moment of thermal bubble bursting and accurately printed to the predetermined position of the three-dimensional biological scaffold.

6. The method according to claim 5, characterized in that Arrange a plurality of the thermal bubble print heads linearly to perform multi-cell orderly linear arrangement printing.

7. A bioprinting system for precise cell positioning, characterized by: include: a memory configured to store data and instructions; A processor in communication with a memory, wherein when executing instructions in the memory, the processor is configured to: Get the 3D model and slice it; Generate instructions and input them into the 3D motion module; Constructing three-dimensional biological scaffolds by layer-by-layer assembly; Acquiring position information of a three-dimensional biological scaffold and feeding back a predetermined position includes acquiring position information of the three-dimensional biological scaffold through an upright camera after each layer of the three-dimensional biological scaffold is printed, and accurately locating the physical position of each layer of the scaffold; converting the physical position into a relative position relative to the origin of the sample stage; inputting the relative position into a software module, and feeding back a predetermined position; Precisely printing cells to a predetermined position of the three-dimensional bioscaffold includes determining a first width of a printed line by a print nozzle size; setting a second width of a print gap; setting an image position origin of a current printed layer of the three-dimensional bioscaffold; calculating, relative to the image position origin, the theoretical coordinate value of the current printed line and the number of pixels corresponding to the horizontal axis and / or vertical axis based on the first width and the second width; obtaining the real physical distance corresponding to each pixel under current shooting conditions; determining the real physical position of the current print as the predetermined position based on the theoretical coordinate value, the number of pixels and the real physical distance corresponding to each pixel; and precisely printing the cells to the predetermined position of the three-dimensional bioscaffold.

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