An in-vitro culture device for tissue engineered ligament in a simulated abdominal pressure biomechanical environment

Abstract

This utility model discloses an in vitro culture device for tissue-engineered ligaments simulating an abdominal pressure biomechanical environment. The device comprises an outer box and a culture mechanism located within the outer box. The top plate of the outer box is provided with multiple arrayed double-sided hydrophobic PTEE filter membranes, and needle-type gas filters are also provided at the four corners of the top plate. A lid is provided at the opening of the outer box. The culture mechanism includes a base, on which a weighing device and a support plate are mounted. A motor is mounted on the top of the support plate, and the working end of the motor is connected to a turntable. A connecting shaft is located at the eccentric position of the turntable, and the connecting shaft is rotatably connected to the top end of a linkage component. The bottom end of the linkage component is hinged to the top end of a drive rod. The drive rod is movably connected within a guide sleeve fixed to the support plate, and the bottom end of the drive rod is fixedly connected to a drive shaft. An elastic pressure block is connected to the bottom end of the drive shaft.

Description

Technical Field

[0001] This invention relates to the medical field, and in particular to an in vitro culture device for tissue-engineered ligaments that simulates the biomechanical environment of abdominal pressure. Background Technology

[0002] In vitro culture of pelvic floor ligaments holds promise as the most effective means of providing suspensions for the surgical treatment of stress urinary incontinence. During in vitro culture, the stress environment plays a crucial role in improving the mechanical properties of ligament tissue and shortening the culture period. However, current research generally lacks precise estimation of pelvic floor ligament loads, failing to consider the time-varying nature (e.g., abdominal pressure) and multidirectional nature (uniaxial, biaxial stress) of the actual stress environment. Studies have only observed changes in cell proliferation and tissue stiffness under single stress stimuli, and few culture methods or devices can directly apply stress loads to the cell-scaffold complex. This results in a significant difference in the usability of the cultured tissue compared to the original tissue.

[0003] Furthermore, many of the few devices currently available on the market that can apply stress loads to cell-scaffold complexes suffer from design flaws, such as insufficient control over parameters like stress amplitude, frequency, and time, inability to maintain a sterile environment for long-term culture, and insufficient biocompatibility of the contact surfaces. Therefore, in research on pelvic floor dysfunction disorders (such as stress urinary incontinence), there is an urgent need for an in vitro culture device that can simulate the biomechanical environment of real abdominal pressure under strictly sterile conditions. Summary of the Invention

[0004] This invention addresses the aforementioned shortcomings of existing technologies by proposing an in vitro culture device for tissue engineering ligaments that is simple in structure, ingeniously designed, rationally laid out, and capable of strictly ensuring that the culture process is carried out in a sterile environment and realistically simulating the biomechanical environment under abdominal pressure conditions.

[0005] The technical solution of this utility model is: an in vitro culture device for tissue-engineered ligaments simulating the biomechanical environment of abdominal pressure, characterized in that: the culture device includes an outer box 1 and a culture mechanism 2 located inside the outer box 1, the outer box 1 being made of transparent sheet material.

[0006] The top plate of the outer box 1 is provided with multiple double-sided hydrophobic PTEE filter membranes 3 arranged in an array, and the opening of the outer box 1 is also provided with a box cover 5.

[0007] The culture mechanism 2 includes a base 6, on which a weighing device 7 and a support plate 8 are mounted. A motor 9 is mounted on the top of the support plate 8, and the working end of the motor 9 is connected to a turntable 10. A connecting shaft 11 is located at an eccentric position on the turntable 10. The connecting shaft 11 is rotatably connected to the top end of a linkage 12, while the bottom end of the linkage 12 is hinged to the top end of a drive rod 13. The drive rod 13 is movably connected within a guide sleeve 14 fixed to the support plate 8, and its bottom end is fixedly connected to a drive shaft 15. An elastic pressure block 16 is connected to the bottom end of the drive shaft 15. The elastic pressure block 16 is located inside the cavity of a culture vessel 17, which is situated on the weighing device 7. An annular component 18 is also located inside the culture vessel 17 below the elastic pressure block 16.

[0008] The outer wall of the annular component 18 is provided with an annular groove 19. The cell scaffold composite 21 can be fixed to the surface of the annular component 18 by the elastic sleeve 20 connected in the annular groove 19.

[0009] The base 6 is also equipped with a control box 22 and a display screen 23. The control box 22 contains a control module and a battery. The battery provides power to the weighing device 7, the motor 9, the display screen 23 and the control module. The control module controls the weighing device 7 and the motor 9 in a unified manner. The control module is also electrically connected to the display screen 23.

[0010] The elastic pressure block 16 is made of elastic silicone, and its bottom surface is spherical.

[0011] The elastic pressure block 16 is made of elastic silicone, and its bottom surface is flat.

[0012] Needle-type gas filters 4 are also installed at the four corners of the top plate of the outer box 1.

[0013] Compared with the prior art, this utility model has the following advantages:

[0014] This tissue-engineered ligament in vitro culture device, which simulates the biomechanical environment of abdominal pressure, has a simple structure, ingenious design, and reasonable layout. It uses a motor to drive a transmission mechanism to drive the elastic pressure block to reciprocate, applying static pressure or dynamic cyclic pressure to the cell scaffold complex taut on the surface of the annular component. At the same time, it can also adjust parameters such as pressure application time and frequency during dynamic pressure operation to simulate the mechanical stimulation of abdominal pressure on the pelvic floor ligaments and tissues under different physiological states in the body (such as standing, coughing, defecation, jumping, etc.).

[0015] This culture device features a semi-enclosed box containing the ligament culture mechanism. This box has a gas filtration function, ensuring sterile cell culture at a specific CO2 concentration, guaranteeing a suitable environment for long-term cell culture. The culture mechanism offers stress loading modes of varying sizes and frequencies, breaking through the traditional method of simply simulating abdominal pressure and making the simulated abdominal pressure stimulation more closely resemble the real human body environment. Furthermore, the elastic pressure blocks are made of silicone, which is non-toxic, harmless, and will not contaminate ligaments or cell tissues, while also effectively simulating the elastic contact between engineered ligaments and pelvic floor tissues in vivo. Moreover, this culture device is simple to manufacture and inexpensive, possessing numerous advantages and making it particularly suitable for widespread application in this field, with a very broad market prospect. Attached Figure Description

[0016] Figure 1 This is a schematic diagram of the overall structure of an embodiment of the present utility model.

[0017] Figure 2 This is a three-dimensional structural diagram of the culture mechanism in an embodiment of this utility model.

[0018] Figure 3 This is a cross-sectional view of the culture mechanism portion in an embodiment of this utility model.

[0019] Figure 4 yes Figure 3 Enlarged view of part A in the image. Detailed Implementation

[0020] The specific embodiments of this utility model will be described below with reference to the accompanying drawings. Figures 1 to 4 As shown: An in vitro culture device for tissue-engineered ligaments simulating the biomechanical environment of abdominal pressure, comprising an outer box 1 and a culture mechanism 2 located within the outer box 1, wherein the outer box 1 is made of transparent sheet material.

[0021] The top plate of the outer box 1 is provided with multiple double-sided hydrophobic PTEE filter membranes 3 arranged in an array, and the opening of the outer box 1 is also provided with a box cover 5.

[0022] The culture mechanism 2 includes a base 6, on which a weighing device 7 and a support plate 8 are mounted. A motor 9 is mounted on the top of the support plate 8, and the working end of the motor 9 is connected to a turntable 10. A connecting shaft 11 is located at an eccentric position on the turntable 10. The connecting shaft 11 is rotatably connected to the top end of a linkage 12, while the bottom end of the linkage 12 is hinged to the top end of a drive rod 13. The drive rod 13 is movably connected within a guide sleeve 14 fixed to the support plate 8, and its bottom end is fixedly connected to a drive shaft 15. An elastic pressure block 16 is connected to the bottom end of the drive shaft 15. The elastic pressure block 16 is located inside the cavity of a culture vessel 17, which is situated on the weighing device 7. An annular component 18 is also located inside the culture vessel 17 below the elastic pressure block 16.

[0023] The outer wall of the annular component 18 is provided with an annular groove 19. The cell scaffold composite 21 can be fixed to the surface of the annular component 18 by the elastic sleeve 20 connected in the annular groove 19.

[0024] The base 6 is also equipped with a control box 22 and a display screen 23. The control box 22 contains a control module and a battery. The battery provides power to the weighing device 7, the motor 9, the display screen 23 and the control module. The control module controls the weighing device 7 and the motor 9 in a unified manner. The control module is also electrically connected to the display screen 23.

[0025] The elastic pressure block 16 is made of elastic silicone, and its bottom surface is spherical.

[0026] Needle-type gas filters 4 are also installed at the four corners of the top plate of the outer box 1.

[0027] The working process of the tissue-engineered ligament in vitro culture device simulating the biomechanical environment of abdominal pressure in this embodiment of the present invention is as follows: First, the cell scaffold composite 21 is placed on the annular part 18, and then the elastic sleeve 20 is fitted into the annular groove 19. That is, the edge part of the cell scaffold composite 21 is fixed in the annular groove 19 by the elastic sleeve 20, thereby realizing the fixation of the cell scaffold composite 21 on the annular part 18. At this time, the cell scaffold composite 21 and the annular part 18 form a drum-like structure. The cell scaffold composite 21 is equivalent to the drum skin, and the annular part 18 is equivalent to the drum body. The middle part of the cell scaffold composite 21 is suspended and taut.

[0028] Place the ring-shaped piece 18 inside the culture dish 17, close the top cover of the culture dish 17, and ensure that the elastic pressure block 16 is also inside the culture dish 17 and above the cell scaffold complex 21.

[0029] Then, the entire culture unit 2 is placed into the outer box 1 (the operation must be performed in a sterile environment), the lid 5 is closed, and the entire outer box 1 is placed into the cell culture incubator. The high concentration of CO2 gas in the cell culture incubator can enter the interior of the outer box 1 through the double-sided hydrophobic PTEE filter membrane 3 and the needle gas filter 4 to ensure that the CO2 gas content during the culture process meets the requirements. This design with the outer box 1 can ensure that the cultured cell scaffold complex 21 is completely isolated from other samples in the cell culture incubator, and there will be no cross-contamination. At the same time, when the current cell scaffold complex 21 needs to be removed from the cell culture incubator for observation or other operations, it needs to be disinfected again before being put back into the cell culture incubator. However, during the conventional alcohol disinfection process, if the alcohol comes into contact with the cells planted on the cell scaffold complex 21, it may affect their growth. However, by placing the entire culture unit 2 into the outer box 1, only the outer surface of the outer box 1 needs to be disinfected, which will not affect the in vitro culture process.

[0030] When the culture device 2 is working, the control module will control the motor 9 to drive the turntable 10 to rotate according to the preset program. When the turntable 10 rotates, it will drive the linkage 12 to swing in space through the connecting shaft 11, thereby driving the drive rod 13 connected below the linkage 12 to move. Since the drive rod 13 is movably connected in the guide sleeve 14, under the guidance of the guide sleeve 14, the drive rod 13 will drive the drive shaft 15 at its bottom to make reciprocating linear motion, thereby realizing the reciprocating downward action of the elastic pressure block 16. Each time it is pressed down, the weighing device 7 located below the culture vessel 17 will detect the current pressure value and feed it back to the control system, which will record it.

[0031] By controlling the output speed of motor 9, the speed of the downward pressing action of elastic block 16 can be adjusted; by controlling the working and resting time of motor 9, the duration of the downward pressing action of elastic block 16 can be adjusted, as well as the duration of the state in which elastic block 16 does not press down on cell scaffold composite 21 (i.e., the two are not in contact); at the same time, by cooperating with the pressing action and weighing device 7, the pressure applied by elastic block 16 can be precisely controlled. For example, during the pressing process, the pressure gradually increases. When the weighing device 7 detects that the current pressure has reached the preset trigger value, it sends a signal to the control system. The control system controls motor 9 to stop working and maintain the current pressure, or controls motor 9 to rotate in the opposite direction, and elastic block 16 moves upward, waiting for the next pressing operation.

[0032] During the cultivation process, the display screen 23 will show the current cultivation conditions and progress, such as the cultivation duration, the number of times the elastic pressure block 16 is pressed down, the pressure range, and other parameters. The staff can directly observe the content displayed on the display screen 23 through the side wall of the transparent box 1.

[0033] During the pressing of the elastic block 16, since there is no support below the cell scaffold composite 21, this structure simulates the real environment of tissue ligaments in the human body, thus allowing for better culture of cells planted on the cell scaffold composite 21.

[0034] After the culture is complete, remove the outer box 1 from the cell culture incubator as a whole, open the outer box 1, remove the ring part 18 from the culture dish 17, remove the elastic sleeve 20, and remove the cell scaffold complex 21 from the ring part 18.

Claims

1. A device for in vitro culture of tissue engineered ligament simulating the biomechanical environment of abdominal pressure, characterized in that: The culture device includes an outer box (1) and a culture mechanism (2) located inside the outer box (1). The outer box (1) is made of transparent sheet material. The top plate of the outer box (1) is provided with multiple double-sided hydrophobic PTEE filter membranes (3) arranged in an array, and the opening of the outer box (1) is also provided with a box cover (5). The cultivation mechanism (2) includes a base (6), on which a weighing device (7) and a support plate (8) are provided. A motor (9) is provided on the top of the support plate (8). The working end of the motor (9) is connected to a turntable (10). A connecting shaft (11) is provided at the eccentric position of the turntable (10). The connecting shaft (11) is rotatably connected to the top end of a linkage (12), while the bottom end of the linkage (12) is hinged to the top end of a drive rod (13). (13) is movably connected to the guide sleeve (14) fixed on the support plate (8), and the bottom end of the drive rod (13) is fixedly connected to the drive shaft (15). The bottom end of the drive shaft (15) is connected to an elastic pressure block (16). The elastic pressure block (16) is located in the inner cavity of the culture vessel (17). The culture vessel (17) is located on the weighing device (7). At the same time, an annular part (18) located below the elastic pressure block (16) is also provided in the culture vessel (17). The outer wall of the annular component (18) is provided with an annular groove (19). The cell scaffold composite (21) can be fixed to the surface of the annular component (18) by the elastic sleeve (20) connected in the annular groove (19). The base (6) is also provided with a control box (22) and a display screen (23). The control box (22) contains a control module and a battery. The battery provides power to the weighing device (7), motor (9), display screen (23) and control module. The control module controls the weighing device (7) and motor (9) in a unified manner. The control module is also electrically connected to the display screen (23).

2. The tissue engineered ligament in-vitro culturing device that simulates abdominal pressure biomechanical environment according to claim 1, characterized in that: The elastic block (16) is made of elastic silicone and its bottom surface is spherical.

3. The in vitro culture device for tissue-engineered ligaments simulating abdominal pressure biomechanical environment according to claim 1, characterized in that: The elastic block (16) is made of elastic silicone and its bottom surface is flat.

4. The tissue engineered ligament in-vitro culturing device that simulates abdominal pressure biomechanical environment according to claim 1, characterized in that: Needle gas filters (4) are also provided at the four corners of the top plate of the outer box (1).

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