Preparation method and preparation device of tissue repair patch

Abstract

The invention provides a preparation method and a preparation device of a tissue repair patch, the preparation method comprises two steps of cell inoculation and culture forming, and the preparation method comprises the following specific steps: placing a culture container in a reaction tank containing a cell suspension until the cell suspension immerses the culture container; the cell suspension enters the culture container through the through holes of the culture container and immerses the culture supports, the cell suspension is stirred so that cells in the cell suspension can be attached to the culture supports, the cell suspension in the reaction tank is replaced with a cell culture medium, the culture cycle of culture forming comprises an accelerated growth period, and in the accelerated growth period, the cell culture medium is separated from the cell suspension. And moving the culture container along the first direction for at least a period of time until the extracellular matrix secreted by the cells covers each culture bracket. The preparation device comprises a reaction tank, a culture container, a plurality of culture brackets, a stirrer and a driver. According to the preparation method and the preparation device of the tissue repair patch, the preparation efficiency of the tissue repair patch can be improved.

Description

Technical Field

[0001] This application relates to the technical field of biological tissue engineering, and particularly relates to a preparation method and a preparation device for a tissue repair patch. Background Art

[0002] Currently, repairing the skin with a tissue repair patch is a commonly used clinical method. The tissue repair patch is usually composed of at least one of animal-derived collagen and polymer materials. The repair patch composed of the above two materials has poor biocompatibility and may have rejection reactions. The animal-derived collagen patch also has the risk of carrying viruses. In addition, the degradation time of the above tissue repair patch is difficult to match the repair time of human tissues, and the repair effect is poor. Human-derived collagen has excellent biocompatibility and biosafety and has a good repair effect. However, the preparation efficiency of human-derived collagen is low, and it is difficult to produce quickly in batches. Therefore, how to batch-produce human-derived collagen tissue patches with excellent biocompatibility and biosafety and improve the preparation efficiency of human-derived collagen tissue patches is an urgent problem to be solved. Summary of the Invention

[0003] This application aims to solve at least one of the technical problems existing in the prior art. For this purpose, this application proposes a preparation method for a tissue repair patch, which can improve the preparation efficiency of the tissue repair patch.

[0004] This application also proposes a preparation device for implementing the above preparation method of the tissue repair patch.

[0005] The preparation method for a tissue repair patch according to an embodiment of this application includes two steps: cell seeding and culture and shaping; Cell seeding: Place the culture container in a reaction tank containing a cell suspension until the cell suspension submerges the culture container. The cell suspension enters the culture container through the through holes of the culture container and submerges each culture support, and stir the cell suspension to make the cells in the cell suspension adhere to the culture support; Culture and shaping: Replace the cell suspension in the reaction tank with a cell culture medium. The culture period for culture and shaping includes an accelerated growth phase. During the accelerated growth phase, the culture container moves in a first direction for at least a period of time until the extracellular matrix secreted by the cells covers each culture support.

[0006] The preparation method of the tissue repair patch according to the embodiments of the present application has at least the following beneficial effects: In the cell seeding step, the cells in the cell suspension can enter the culture container through the through holes and adhere to the culture scaffold. By agitating the cell suspension, the cell suspension is caused to flow in the reaction tank, accelerating the cell seeding speed. After cell seeding, the cell suspension in the reaction tank is replaced with cell culture medium, and the cell culture medium can enter the culture container through the through holes. The cell culture medium is used to provide nutrients for the cells. The driver can drive the culture container to move in the first direction, increasing the acting force of the cell culture medium on the cells in the culture container to stimulate the cells to increase the speed of secreting extracellular matrix, thereby improving the preparation efficiency of the tissue repair patch.

[0007] According to some embodiments of the present application, the culture period of the culture and shaping step is 42 days to 56 days. The culture period also includes a static growth period before the accelerated growth period. During the static growth period, the culture container is statically placed in the reaction tank. During the accelerated growth period, the driver drives the culture container to reciprocally move in the first direction.

[0008] According to some embodiments of the present application, the culture period is 42 days. From the start of the culture and shaping step to the 14th day is the static growth period, and from the end of the static growth period to the 42nd day is the accelerated growth period.

[0009] According to some embodiments of the present application, the accelerated growth period includes multiple moving cycles. The moving cycle is 5 s to 50 s. During each moving cycle, the moving stroke of the culture container is 5 mm to 500 mm.

[0010] According to some embodiments of the present application, the accelerated growth period includes multiple sequentially performed driving cycles. Each driving cycle includes multiple sequentially performed moving cycles. Between adjacent driving cycles, the moving cycles in the latter driving cycle are less than those in the previous driving cycle, and the moving stroke in the latter driving cycle is greater than that in the previous driving cycle. The difference between the moving cycles in adjacent two driving cycles is 1 s to 20 s, and the difference between the moving strokes in adjacent two driving cycles is 5 mm to 50 mm.

[0011] According to some embodiments of the present application, the accelerated growth period includes multiple sequentially performed driving cycles. During the driving cycle, the culture container is moved in the first direction for at least a period of time. There is also a stationary cycle between adjacent two driving cycles, and the culture container is stationary relative to the reaction tank during the stationary cycle.

[0012] According to some embodiments of the present application, in the cell seeding step, multiple culture containers are placed in the reaction tank, and the cell suspension submerges each culture container. In the culture and shaping step, multiple independent drivers are connected to different culture containers, and each driver drives the corresponding culture container to reciprocally move in the first direction at different moving frequencies.

[0013] According to some embodiments of the present application, the components of the culture scaffold include at least one of polylactic acid, polydioxanone, polyglycolic acid, polycaprolactone, polycitric acid, polydopamine, sodium alginate, chitin, and chitosan.

[0014] According to some embodiments of the present application, the components of the culture container include at least one of polyvinyl alcohol, polyethylene, polyamide, polyurethane, polyethylene terephthalate, nylon, polypropylene, polyvinyl chloride, polystyrene, polylactic acid, polydioxanone, polyglycolic acid, polycaprolactone, polycitric acid, polydopamine, sodium alginate, chitin, and chitosan.

[0015] The preparation device according to an embodiment of the present application is used to prepare a tissue repair patch by the preparation method of the tissue repair patch in any of the above embodiments. The preparation device includes a reaction tank, a culture container, a plurality of culture scaffolds, a stirrer, and a driver; The reaction tank is provided with a culture chamber; The culture container is located in the culture chamber. The culture container is provided with a receiving cavity and a plurality of through holes, and each of the through holes communicates with the receiving cavity; A plurality of culture scaffolds are arranged at intervals in the receiving cavity along a first direction; The stirrer has at least a part extending into the culture chamber, and the stirrer is arranged at an interval from the culture container; The driver is connected to the culture container, and the driver is used to drive the culture container to move along the first direction.

[0016] The preparation device according to an embodiment of the present application has at least the following beneficial effects: The culture chamber is used to accommodate the culture container, and the culture container is used for the interval assembly of a plurality of culture scaffolds to ensure that cells are carried at intervals between each culture. A plurality of through holes are provided for the cell suspension or cell culture medium to enter the culture container. The stirrer is used to stir the cell suspension in the culture chamber to improve the cell seeding speed, and / or is used to stir the cell culture medium to increase the acting force of the cell culture medium on the cells. The driver is used to drive the culture container to move along the first direction, so that the cell culture medium exerts a force on the cells along the first direction to stimulate cell growth. Therefore, the preparation device in the present application can provide mechanical stimulation for cells during cell cultivation, so that the speed of cells secreting extracellular matrix is faster and the efficiency of preparing tissue repair patches is higher.

[0017] Additional aspects and advantages of the present application will be given in part in the following description, become apparent in part from the following description, or be understood through the practice of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The following further describes the present application with reference to the drawings and embodiments, where: Figure 1 Schematic structural diagram of the culture container according to an embodiment of the present application; Figure 2 Schematic assembly diagram of the culture support and the culture container according to an embodiment of the present application; Figure 3 Schematic structural diagram inside the reaction tank during the cell seeding step according to an embodiment of the present application; Figure 4 Schematic structural diagram inside the reaction tank during the culture forming step according to an embodiment of the present application; Figure 5 Process diagram of tissue patch culture according to an embodiment of the present application; Figure 6 Inclusion relationship diagram between each cycle according to an embodiment of the present application.

[0019] Reference numerals: reaction tank 100, culture chamber 110, cell suspension 120, cell culture medium 130; Culture container 200, accommodation chamber 210, through hole 220; Culture support 300; Agitator 400; Driver 500. Detailed implementation manners

[0020] The embodiments of the present application will be described in detail below. The examples of the embodiments are shown in the drawings, where the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the drawings are exemplary and are only used to explain the present application and should not be construed as a limitation to the present application.

[0021] In the description of the present application, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings. It is only for the convenience of describing the present application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as a limitation to the present application.

[0022] In the description of the present application, the meaning of several is more than one, the meaning of multiple is two or more, greater than, less than, exceeding, etc. are understood as not including the number itself, and above, below, within, etc. are understood as including the number itself. If there is a description of first and second, it is only for the purpose of distinguishing technical features and should not be understood as indicating or implying relative importance or implicitly indicating the quantity of the indicated technical features or implicitly indicating the sequence relationship of the indicated technical features.

[0023] In the description of the present application, unless otherwise clearly defined, terms such as "setting", "installation", "connection", etc. should be understood in a broad sense, and those skilled in the art can reasonably determine the specific meanings of the above terms in the present application in combination with the specific content of the technical solution.

[0024] In the description of the present application, the description with reference to terms such as "one embodiment", "some embodiments", "schematic embodiments", "examples", "specific examples", or "some examples", etc. means that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic expressions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in a suitable manner in any one or more embodiments or examples.

[0025] The embodiments of the present application are introduced below in conjunction with the accompanying drawings of the specification: Referring to Figures 1 to 5 , according to the preparation method of the tissue repair patch of the embodiment of the present application, which is used to prepare a tissue repair patch containing human-derived collagen, the preparation method of the tissue repair patch includes two sequentially performed steps of cell seeding and culturing and forming. In the cell seeding step, the culture container 200 in the seeding preparation step is placed in the reaction tank 100 containing the cell suspension until the cell suspension submerges the culture container 200. The cell suspension enters the culture container 200 through the through holes 220 and submerges each culture support 300, and the cell suspension is stirred to promote the flow of the cell suspension in the culture container 200. Then, the cells in the cell suspension adhere to the culture support 300 to achieve cell seeding. Stirring the cell suspension can promote the movement of the cells in the cell suspension, making it easier for the cells in the cell suspension to come into contact with the culture support 300, avoiding local deposition of cells in the reaction tank 100, and being beneficial to improving the efficiency of cell seeding.

[0026] Among them, a plurality of culture supports 300 are assembled at intervals along the first direction in the culture container 200, and the culture container 200 has a plurality of through holes 220. The culture support 300 has pores. As the cell suspension is stirred, the cells in the cell suspension can enter the pores of the culture support 300 and finally adhere to the pores to complete cell seeding.

[0027] In the culturing and shaping step, the cell suspension in the reaction tank 100 is replaced with a cell culture medium, which may include components such as glucose, amino acids, inorganic salts, and cytokines. The cell culture medium is used to provide essential nutrients for the cultured cells after inoculation. The culturing cycle for shaping includes an accelerated growth phase. During the accelerated growth phase, the culture container 200 is connected to the driver 500, and the driver 500 drives the culture container 200 to move in a first direction for at least a period of time, so as to increase the interaction force between the cells attached to each culture scaffold 300 and the cell culture medium, stimulate the cells, promote cell growth, which is beneficial to shortening the culturing cycle for cell shaping and improving the speed of cell culturing and shaping. In the culturing and shaping step, until the extracellular matrix secreted by the cells covers each culture scaffold 300, a shaped tissue repair patch is prepared.

[0028] Reference Figure 5 , A is a microstructural diagram of the culture scaffold 300, B is a microscopic schematic diagram of the culturing process of cells on the culture scaffold 300, and C is a microstructural diagram of the culture scaffold 300 completely covered by cells. Therefore, the preparation process of the tissue repair patch can be judged by an optical device such as a microscope that can observe the microstructure.

[0029] Specifically, the first direction can be a vertical direction, a horizontal direction, a circumferential direction, or other directions. Taking the first direction as the vertical direction as an example, the culture scaffolds 300 are arranged at intervals in the first direction to ensure that the cell suspension is in full contact with each culture scaffold 300, enabling faster cell inoculation. Also, it can prevent adhesion between cells cultured on different culture scaffolds 300, facilitating the separation of each tissue repair patch after cell culturing and shaping. In the culturing and shaping step, the driver 500 drives the culture container 200 to move in the first direction. The culture scaffold 300 includes an upper surface and a lower surface perpendicular to the first direction. The culture scaffold 300 moves with the culture container 200, and the extracellular matrix can uniformly stimulate the upper surface and the lower surface of the culture scaffold 300, making the stimulation received by different cells closer, so that the tissue repair patch is more uniform and dense.

[0030] Also, compared with stimulating cells through other rigid solid structures, it can avoid cell damage and ensure the integrity of cell shaping.

[0031] The cell suspension contains cells for preparing tissue repair patches. The cells in the cell suspension include, but are not limited to, at least one of induced pluripotent stem cells, mesenchymal stem cells, epidermal stem cells, fibroblasts, smooth muscle cells, endothelial cells, and human cells enhanced by gene editing technology. Compared with tissue repair patches prepared based on animal-derived collagen and polymer materials, the above cells have excellent biocompatibility and biosafety, avoid rejection reactions with the human body, reduce the risk of virus infection, and can also match the repair time of human tissues, with good repair effects. The number of cells in the cell suspension can be 5×10 5 cells / ml to 5×10 7 cells / ml.

[0032] It should be noted that when preparing the repair patch, to ensure the normal growth of cells, the inside of the reaction tank 100 is a sterile environment. And the temperature inside the reaction tank 100 can be 37°C, the pH value can be 7, and the oxygen content ratio can be 90% to 95%. The control of the above parameters can be monitored through a temperature sensor, a pH sensor, and a dissolved oxygen sensor.

[0033] Reference Figures 1 to 4 In some other embodiments, referring to

[0034] Reference Figures 1 to 4 In some other embodiments, in the inoculation preparation step, multiple culture containers 200 can be assembled. For example, four culture containers 200 are assembled in the inoculation preparation step. Six culture scaffolds 300 are assembled at intervals along the first direction in each culture container 200. Furthermore, in the cell inoculation step, four culture containers 200 are placed in the reaction tank 100 at the same time. The increase in the number of culture containers 200 and culture scaffolds 300 is beneficial to further improving the preparation speed of tissue repair.

[0035] It should be noted that the number of culture containers 200 and the number of culture scaffolds 300 in each culture container 200 can be adjusted according to requirements.

[0036] Reference Figures 1 to 4, in some embodiments, the culture period of the culturing and forming step is 42 days to 56 days (including 42 days and 56 days). The culture period includes a static growth period before the accelerated growth period, that is, the static growth period and the accelerated growth period are carried out in sequence. During the static growth period, the culture container 200 is statically placed in the reaction tank 100, that is, the driver 500 does not drive the culture container 200 to move during the static growth period. During the accelerated growth period, the driver 500 drives the culture container 200 to reciprocate in the first direction, so that an interaction is generated between each culture support 300 in the culture container 200 and the cell culture medium to stimulate the cells to secrete extracellular matrix. Among them, the setting of the static growth period is used for the cells to adapt to the growth environment. The cells adhere to the culture support 300 during the static growth period, avoiding generating an interaction force with the extracellular matrix and detaching from the culture support 300, which is convenient for more stably culturing and forming the cells.

[0037] It should be noted that the time lengths of the above-mentioned culture period, static growth period and accelerated growth period can be adjusted according to needs. For example, the culture period can be 42 days, 49 days or 56 days, etc. The time length of the static growth period can be any length from 1 day to 14 days (including 1 day and 14 days), and the time length of the accelerated growth period can be any length from 28 days to 42 days (including 28 days here, not including 42 days), that is, the static growth period and the accelerated growth period together constitute the culture period of the culturing and forming step.

[0038] Reference Figures 1 to 4 , in some embodiments, in the culturing and forming step, the culture period of the cells is 42 days. From the start of the culturing and forming step to the 14th day (including the 14th day) is the static growth period, and from the end of the static growth period to the 42nd day (including the 42nd day) is the accelerated growth period. The static growth period is used to provide an adaptation time for the growth of the cells and ensure the stable adhesion between the cells and the culture support 300. The accelerated growth period is after the static growth period. During the accelerated growth period, the driver 500 drives the culture container 200 to reciprocate in the first direction, so that a certain force is generated between the cells adhered to each culture support 300 and the cell culture medium, thereby realizing the stimulation of the cells, which is beneficial to promoting the secretion of extracellular matrix and improving the preparation efficiency of the tissue repair patch.

[0039] It should be noted that the main component of the extracellular matrix is collagen. Collagen is the main structural protein of human tissues, has excellent biocompatibility and biosafety for patients, and the degradation time matches the repair time of human tissues, and excellent repair effects can be achieved.

[0040] Specifically, in the culturing and shaping step, there is a culturing cycle of 42 days. From the start of the culturing and shaping step to the 14th day (including the 14th day) is the static growth period. During the static growth period, the culture container 200 is immersed in the cell culture medium, so that each culture support 300 is static relative to the cell culture medium to allow the cells to grow adaptively, which is beneficial to ensuring the normal cell morphology and reducing cell damage or apoptosis during the growth and proliferation stage. From the end of the static growth period to the 42nd day is the accelerated growth period. The driver 500 drives the culture container 200 to reciprocate along the first direction, and each culture support 300 moves relative to the cell culture medium. Due to the relative movement between the cells and the cell culture medium, a force different from the static state is generated to stimulate the cells to accelerate the secretion rate of the extracellular matrix.

[0041] Reference Figures 1 to 5 , in some embodiments, the accelerated growth period includes a plurality of movement cycles. The movement cycle is 5 s to 50 s. Within each movement cycle, the driver 500 drives the culture container 200 to reciprocate along the first direction, and the movement stroke of the culture container 200 is 5 mm to 500 mm. By limiting the movement stroke of the culture container 200, it can not only avoid insufficient stimulation of the cells due to too small a movement stroke, but also avoid too large a movement stroke resulting in too large a volume of the reaction tank 100, so that the reaction tank 100 can accommodate more culture containers 200 within a limited volume and ensure the mechanical stimulation force on the cells, which is convenient for the batch production of tissue repair patches.

[0042] For example, the movement stroke of the culture container 200 can be any length among 5 mm, 50 mm, 100 mm, 150 mm, 200 mm, 250 mm, 300 mm, 350 mm, 400 mm, 450 mm or 500 mm, and the movement cycle can be any duration among 5 s, 10 s, 15 s, 20 s, 25 s, 30 s, 35 s, 40 s, 45 s or 50 s.

[0043] It should be noted that the above movement stroke is the total length of the movement of the culture container 200 within a single movement cycle. The time required for a single movement stroke, that is, the movement speed range of the culture container 200 is 0.1 mm / s to 100 mm / s. Limiting the upper limit of the movement speed of the culture container 200 is used to avoid too high a degradation speed due to too high a movement speed of the culture support 300 relative to the cell culture medium, which affects the subsequent cell culture process, and is beneficial to ensuring the stability of the cell morphology. Also, limiting the lower limit of the movement speed of the culture container 200 is used to avoid insufficient mechanical stimulation and ensure that the cells secrete a sufficient amount of extracellular matrix during the culture cycle.

[0044] Reference Figures 1 to 5, in some embodiments, the accelerated growth period includes a plurality of sequentially performed driving cycles, each driving cycle includes a plurality of sequentially performed moving cycles. Between adjacent driving cycles, the moving cycles in the subsequent driving cycle are less than those in the previous driving cycle, and the moving stroke in the subsequent driving cycle is greater than that in the previous driving cycle. The difference in the moving cycles between two adjacent driving cycles is 1 s to 20 s, and the difference in the moving strokes between two adjacent driving cycles is 5 mm to 50 mm. Thus, in different driving cycles, the moving speed of the culture vessel 200 is different. Furthermore, it avoids the decrease in the speed of secreting extracellular matrix caused by the cells adapting to external force stimulation, enables the cells to maintain an excellent speed of secreting extracellular matrix in each driving cycle, and makes the performance of the tissue repair patch better.

[0045] It should be understood that in each driving cycle of the present application, the moving speed of the culture vessel 200 gradually increases, that is, the degree of stimulation to the cells increases sequentially. Selecting a smaller stimulation intensity at the initial stage of cultivation and applying it to the cells can ensure the stability of the cells in the culture scaffold 300. As the extracellular matrix is secreted, the stability of the cells on the culture scaffold 300 gradually increases, and they can withstand a greater intensity of mechanical stimulation. Then, by applying a larger stimulation intensity to the cells, it can not only ensure that the cells maintain the speed of secreting extracellular matrix, but also avoid cell detachment.

[0046] Specifically, the driving cycle can be adjusted according to requirements. In one driving cycle, the driver 500 drives the culture vessel 200 to move along the first direction with a fixed moving stroke, and each moving cycle is the same. When entering the next driving cycle, the driver 500 drives the culture vessel 200 to increase the moving stroke along the first direction by 5 mm to 50 mm, and shorten the moving cycle by 1 s to 20 s, so as to gradually increase the stimulation intensity to the cells, ensure the ability of the cells to secrete extracellular matrix, be beneficial to shortening the preparation cycle of the tissue repair patch, improving the preparation efficiency of the tissue repair patch, and facilitating the batch production of the tissue repair patch.

[0047] Reference Figures 1 to 5, in some embodiments, the accelerated growth phase includes a plurality of driving cycles that are carried out sequentially. Within a single driving cycle, the culture vessel is moved in a first direction for at least a period of time. There is also a stationary period between adjacent driving cycles. During the stationary period, the culture vessel is stationary relative to the reaction tank. It should be understood that the stationary period is part of the accelerated growth phase. Thus, within a certain period of time, the culture vessel 200 is driven to move in the first direction. After moving for a certain period of time, the culture vessel 200 stops moving to provide a buffer for the cells to secrete the extracellular matrix and to prevent the cells from adapting to the stimulation brought about by the movement of the culture vessel 200. After being stationary for a certain period of time, the culture vessel 200 is driven to continue moving in the first direction, and this cycle repeats. That is, during the driving growth phase, the culture vessel 200 reciprocates between moving in the first direction and being stationary, which is beneficial to maintaining the rate of extracellular matrix secretion by the cells.

[0048] Specifically, the driving cycle can be 1 day. Within 1 day, the culture vessel 200 moves in the first direction for a duration of 5 h to 18 h and is stationary for a duration of 6 h to 19 h. For example, the culture vessel 200 moves in the first direction for 5 h and is stationary for 19 h, or moves for 12 h and is stationary for 12 h, or moves for 18 h and is stationary for 6 h. Among them, the first direction can be a two-way or circumferential direction to ensure the reciprocating movement of the culture vessel 200. The moving stimulation duration and the stationary duration per day can be adaptively adjusted according to the actual situation.

[0049] Reference Figure 6 , in any embodiment of the present application, the culture cycle includes a stationary growth phase and an accelerated growth phase that are carried out sequentially. The accelerated growth phase includes a plurality of driving cycles and a plurality of stationary periods. The stationary periods can be located between adjacent driving cycles, and can also be located at the end of the accelerated growth phase. Each driving cycle includes a plurality of moving cycles.

[0050] It should be noted that Figure 6 is only used to illustrate the relationship between the various cycles in the present application. The number of each cycle is not limited to the number in Figure 6 the figure.

[0051] Reference Figures 1 to 5, in some embodiments, in the cell seeding step, a plurality of culture containers 200 are placed in the reaction tank 100, and the cell suspension submerges each culture container 200. And in the culture and shaping step, a plurality of independent drivers 500 are connected to different culture containers 200, and each driver 500 drives each culture container 200 to reciprocate along the first direction at different moving frequencies. Since the moving frequencies of the culture containers 200 are different, the cells attached to the culture scaffold 300 in the culture container 200 are stimulated differently. Furthermore, the amounts of extracellular matrix secreted by the cells in different culture containers 200 are different. During the same cultivation period, tissue repair patches with different thicknesses can be prepared simultaneously, which is beneficial to meeting different requirements for the thickness of tissue repair patches.

[0052] For example, for parts with frequent activities and requirements for wear resistance, such as joints, hands and feet, etc., thicker tissue repair patches can be used for repair. For parts with higher appearance requirements such as the face and neck, the thickness of the patch needs to be controlled more precisely, and thinner tissue repair patches can be used for repair.

[0053] Reference Figures 1 to 5 , in some embodiments, the components of the culture scaffold 300 include at least one of polylactic acid, polydioxanone, polyglycolic acid, polycaprolactone, polycitric acid, polydopamine, sodium alginate, chitin, and chitosan, and have good biocompatibility and degradability. Thus, as the culture carrier of the tissue repair patch, the culture scaffold 300 is used to provide support for the culture and shaping of the tissue repair patch on the one hand. On the other hand, the interaction between the culture scaffold 300 and human tissues is more gentle and harmless, which helps to reduce the rejection reaction that may be caused after the patch is implanted, and can also be gradually absorbed and metabolized by the human body, reducing the complications caused by residues.

[0054] Reference Figures 1 to 5 , in some embodiments, the components of the culture container 200 include at least one of polyvinyl alcohol, polyethylene, polyamide, polyurethane, polyethylene terephthalate, nylon, polypropylene, polyvinyl chloride, polystyrene, polylactic acid, polydioxanone, polyglycolic acid, polycaprolactone, polycitric acid, polydopamine, sodium alginate, chitin, and chitosan. The above components all have good biocompatibility to avoid the porous container from affecting the normal physiological functions of cells.

[0055] Reference Figures 1 to 4, The preparation device according to the embodiments of the present application is used to prepare tissue repair patches by the preparation method of tissue repair patches in any of the above embodiments. The preparation device includes a reaction tank 100, a culture container 200, a plurality of culture scaffolds 300, a stirrer 400, and a driver 500. The reaction tank 100 is provided with a culture chamber 110, and the inside of the culture chamber 110 is a sterile environment to avoid cell contamination. In the two steps of cell seeding and culture molding, the culture container 200 is located in the culture chamber 110. The culture container 200 is provided with a receiving cavity 210 and a plurality of through holes 220, and each through hole 220 communicates with the receiving cavity 210. Along the first direction, each culture scaffold 300 is arranged at intervals in each receiving cavity 210. In the cell seeding step, a cell suspension is contained in the culture chamber 110, and the cell suspension submerges the culture container 200. Through the through holes 220 of the culture container 200, the cell suspension can enter the receiving cavity 210. Moreover, due to the spaced arrangement of each culture scaffold 300, both opposite sides of the culture scaffold 300 can be in contact with the cell suspension, and the cell suspension deposits and adheres in the pores of the culture scaffold 300 to achieve cell seeding.

[0056] In addition, at least a part of the stirrer 400 extends into the culture chamber 110, and the stirrer 400 is arranged at intervals from the culture container 200 to avoid collision between the stirrer 400 and the culture container 200 during stirring. The stirrer 400 is used to stir the cell suspension in the cell seeding step to promote the flow of the cell suspension, make the cells in the cell suspension more evenly distributed, and is beneficial to improving the cell seeding speed. In addition, the stirrer 400 is also used to continue stirring the cell culture medium in the culture molding step to promote the flow of the cell culture medium, increase the interaction force between the cells and the cell culture medium, stimulate the cells to secrete extracellular matrix, and is beneficial to improving the preparation efficiency of tissue repair patches. The driver 500 is connected to the culture container 200 and is used to drive the culture container 200 to move along the first direction. As the culture container 200 moves, the interaction force between the cells on each culture scaffold 300 and the cell culture medium increases, realizing mechanical stimulation of the cells, which is beneficial to promoting cell growth.

[0057] Specifically, the first direction can be the vertical direction. The stirrer 400 rotates with the first direction as the rotation axis to stir the cell suspension and extracellular matrix, making the flow direction of the cell suspension and extracellular matrix tend to be the horizontal direction. The driver 500 drives the culture container 200 to move along the first direction to generate a vertical force between the cells and the cell culture medium. The two cooperate with each other to be able to apply impact stimulation to the cells in multiple directions, which is beneficial to improving the cell growth efficiency.

[0058] Reference Figures 1 to 5, in some other embodiments, the preparation device may include a plurality of culture containers 200. Along the first direction, a plurality of culture scaffolds 300 are assembled at intervals in each culture container 200. According to requirements, a plurality of culture containers 200 can be placed in the culture cavity 110 at the same time, which is convenient for batch production of tissue repair patches.

[0059] The embodiments of the present application have been described in detail above in conjunction with the accompanying drawings. However, the present application is not limited to the above embodiments. Various changes can be made without departing from the spirit of the present application within the scope of knowledge possessed by those of ordinary skill in the art. In addition, the embodiments of the present application and the features in the embodiments can be combined with each other without conflict.

Claims

1. A preparation method of a tissue repair patch, characterized in that, Including: Cell seeding: Place the culture container in a reaction tank containing a cell suspension until the cell suspension submerges the culture container. The cell suspension enters the culture container through the through-holes of the culture container and submerges each culture support. Stir the cell suspension to enable the cells in the cell suspension to adhere to the culture support. Culturing and forming: Replace the cell suspension in the reaction tank with a cell culture medium. The culturing period for forming includes an accelerated growth phase. During the accelerated growth phase, the culture container moves in the first direction for at least a certain period of time until the extracellular matrix secreted by the cells covers each culture support.

2. The preparation method of the tissue repair patch according to claim 1, characterized in that, The culturing period of the culturing and forming step is 42 to 56 days. The culturing period also includes a static growth phase before the accelerated growth phase. During the static growth phase, the culture container is static in the reaction tank. During the accelerated growth phase, a driver drives the culture container to reciprocate in the first direction.

3. The preparation method of the tissue repair patch according to claim 2, characterized in that, The culturing period is 42 days. From the start of the culturing and forming step to the 14th day is the static growth phase, and from the end of the static growth phase to the 42nd day is the accelerated growth phase.

4. The preparation method of the tissue repair patch according to any one of claims 2 to 4, characterized in that, The accelerated growth phase includes multiple moving cycles. The moving cycle is 5 s to 50 s. During each moving cycle, the moving stroke of the culture container is 5 mm to 500 mm.

5. The preparation method of the tissue repair patch according to claim 4, wherein The accelerated growth phase includes multiple sequentially performed driving cycles. Each driving cycle includes multiple sequentially performed moving cycles. Between adjacent driving cycles, the moving cycles in the latter driving cycle are shorter than those in the former driving cycle, and the moving stroke in the latter driving cycle is greater than that in the former driving cycle. The difference between the moving cycles in adjacent driving cycles is 1 s to 20 s, and the difference between the moving strokes in adjacent driving cycles is 5 mm to 50 mm.

6. The preparation method of the tissue repair patch according to claim 1, characterized in that, The accelerated growth phase includes multiple sequentially performed driving cycles. During the driving cycle, the culture container is moved in the first direction for at least a certain period of time. There is also a stationary cycle between adjacent driving cycles, during which the culture container is stationary relative to the reaction tank.

7. The preparation method of the tissue repair patch according to claim 1, characterized in that, In the cell seeding step, multiple culture containers are placed in the reaction tank, and the cell suspension submerges each culture container. In the culturing and forming step, multiple independent drivers are connected to different culture containers, and each driver drives the corresponding culture container to reciprocate in the first direction at different moving frequencies.

8. The preparation method of the tissue repair patch according to claim 1, wherein, The components of the culture support include at least one of polylactic acid, polydioxanone, polyglycolic acid, polycaprolactone, polycitric acid, polydopamine, sodium alginate, chitin, and chitosan.

9. The preparation method of the tissue repair patch according to claim 1, wherein, The components of the culture container include at least one of polyvinyl alcohol, polyethylene, polyamide, polyurethane, polyethylene terephthalate, nylon, polypropylene, polyvinyl chloride, polystyrene, polylactic acid, polydioxanone, polyglycolic acid, polycaprolactone, polycitric acid, polydopamine, sodium alginate, chitin, and chitosan.

10. A preparation device for preparing a tissue repair patch by the method for preparing a tissue repair patch according to any one of claims 1 to 9, characterized in that, Comprising: A reaction tank provided with a culture chamber; A culture container located in the culture chamber, the culture container being provided with a receiving cavity and a plurality of through holes, each of the through holes communicating with the receiving cavity; A plurality of culture supports, arranged at intervals in the receiving cavity along a first direction; A stirrer, at least a part of which extends into the culture chamber, the stirrer being arranged at an interval from the culture container; A driver connected to the culture container, the driver being used to drive the culture container to move along the first direction.

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