Tissue engineering ligament in-vitro culture device for simulating abdominal pressure biomechanical environment
By designing a culture device including a weighing device and a motor-driven transmission system, the problem of insufficient pelvic floor ligament load simulation in the prior art is solved, and efficient cell culture is achieved that simulates the biomechanical environment of abdominal pressure under sterile conditions, and the effect of tissue performance is improved close to the real human environment.
Patent Information
- Application Number
- CN202510757581.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-09
- Publication Date
- 2025-08-19
AI Technical Summary
The prior art lacks fine estimates of pelvic floor ligament load, fails to simulate the time-degeneration and versatility of real abdominal pressure, and the existing devices have shortcomings in sterile environment and biocompatibility, resulting in a large gap between the performance of cultured tissue and the original tissue.
A device including an outer box and a culture mechanism is designed. The outer box top plate is equipped with a hydrophobic PTEE filter membrane and a gas filter. The culture mechanism includes a weighing device, a motor-driven transmission system and an elastic cushion, which can simulate the biomechanical environment of abdominal pressure, provide static or dynamic pressure stimulation, and perform cell culture under sterile conditions.
It realizes precise simulation of abdominal pressure conditions in a sterile environment, provides a variety of stress loading modes, improves the effect of cultured tissues close to the real human environment, reduces manufacturing costs, and is suitable for long-term cell culture.
Smart Images
Figure CN120505202A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the field of medicine, in particular to an in vitro culture device for tissue engineering ligaments that simulates an abdominal pressure biomechanical environment. Background Art
[0002] In vitro culture of pelvic floor ligaments holds promise as the most effective sling for 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 cycle. However, current studies generally lack precise estimates of the load on the pelvic floor ligaments. These studies fail to consider the time-varying nature (e.g., abdominal pressure) and multidirectional nature (uniaxial and biaxial stress) of the actual stress environment. Studies have focused solely on observing changes in cell proliferation and tissue stiffness under a single stress stimulus. Few culture methods or devices can directly apply stress to the cell-scaffold complex, resulting in a significant gap between the serviceability of the cultured tissue and that of the original tissue.
[0003] Furthermore, the few devices currently available on the market that can stress cell-scaffold complexes often suffer from design flaws, such as insufficient control over stress amplitude, frequency, and duration, an inability to maintain a sterile environment for long-term culture, and insufficient biocompatibility of the contact surfaces. Therefore, in the study of 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] The present invention aims to address the above-mentioned deficiencies in the prior art and proposes an in vitro tissue engineering ligament culture device with a simple structure, ingenious design, and reasonable layout, which can strictly ensure that the culture process is carried out in a sterile environment and truly simulate the biomechanical environment under abdominal pressure conditions.
[0005] The technical solution of the present invention is: an in vitro culture device for tissue engineering ligaments that simulates the biomechanical environment of abdominal pressure, characterized in that: the culture device comprises an outer box 1 and a culture mechanism 2 located inside the outer box 1, wherein the outer box 1 is made of a transparent plate. The top plate of the outer box 1 is provided with a plurality of double-sided hydrophobic PTFE filter membranes 3 distributed in an array, and a box cover 5 is also provided at the opening of the outer box 1. The culture 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 the turntable 10. A connecting shaft 11 is provided at an eccentric position of the turntable 10. The connecting shaft 11 is rotatably connected to the top of a linkage member 12, and the bottom end of the linkage member 12 is hinged to the top of a driving rod 13. The driving rod 13 is movably connected in a guide sleeve 14 fixed on the support plate 8. At the same time, the bottom end of the driving rod 13 is fixedly connected to the driving shaft 15. The bottom end of the driving shaft 15 is connected to an elastic pressure block 16, which is located in the inner cavity of a culture vessel 17. The culture vessel 17 is located on the weighing device 7. At the same time, a ring member 18 located below the elastic pressure block 16 is also provided in the culture vessel 17. The outer wall of the annular member 18 is provided with an annular groove 19 , and the cell-scaffold complex 21 can be fixed on the surface of the annular member 18 by an elastic sleeve 20 connected to the annular groove 19 . A control box 22 and a display screen 23 are also provided on the base 6. A control module and a battery are provided in the control box 22. The battery provides power to the weighing device 7, the motor 9, the display screen 23 and the control module, while 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.
[0006] The elastic pressing block 16 is made of elastic silicone and has a spherical bottom surface.
[0007] The elastic pressing block 16 is made of elastic silicone and has a flat bottom surface.
[0008] Needle-type gas filters 4 are also provided at the four corners of the top plate of the outer box 1 .
[0009] Compared with the prior art, the present invention has the following advantages: This type of tissue engineering ligament in vitro culture device, which simulates the abdominal pressure biomechanical environment, has a simple structure, ingenious design, and reasonable layout. It uses a motor to drive the transmission mechanism to drive the elastic pressure block to move back and forth, applying static pressure or dynamic cyclic pressure to the cell scaffold complex stretched on the surface of the ring. 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 the pelvic floor ligaments and tissues caused by abdominal pressure in different physiological states in the body (such as standing, coughing, defecation, jumping, etc.).
[0010] This culture device features a semi-enclosed box housing the ligament culture mechanism. This box incorporates a gas filtration system, ensuring sterile cell culture at a specific CO2 concentration, ensuring a suitable environment for long-term cell culture. The culture mechanism can provide stress loading modes of varying magnitude and frequency, surpassing the traditional single-step method of simulating abdominal pressure, allowing the pressure stimulation to more closely resemble the real human environment. Furthermore, the elastic pressure blocks are made of silicone, which is non-toxic and harmless, preventing contamination of ligaments and cell tissue. Furthermore, they effectively simulate the elastic contact state of engineered ligaments with pelvic floor tissue in vivo. This culture device also boasts a simple manufacturing process and low manufacturing cost, offering numerous advantages, making it particularly suitable for widespread application in this field and promising a promising market. BRIEF DESCRIPTION OF THE DRAWINGS
[0011] Figure 1 It is a schematic diagram of the overall structure of an embodiment of the present invention.
[0012] Figure 2 It is a schematic diagram of the three-dimensional structure of the culture mechanism part in an embodiment of the present invention.
[0013] Figure 3 It is a cross-sectional view of the culture mechanism portion in an embodiment of the present invention.
[0014] Figure 4 yes Figure 3 Enlarged view of part A in . DETAILED DESCRIPTION
[0015] The specific embodiments of the present invention will be described below with reference to the accompanying drawings. Figures 1 to 4 As shown: A tissue engineering ligament in vitro culture device that simulates the biomechanical environment of abdominal pressure, which includes an outer box 1 and a culture mechanism 2 located inside the outer box 1. The outer box 1 is made of a transparent plate. The top plate of the outer box 1 is provided with a plurality of double-sided hydrophobic PTFE filter membranes 3 distributed in an array, and a box cover 5 is also provided at the opening of the outer box 1. The culture 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 the turntable 10. A connecting shaft 11 is provided at an eccentric position of the turntable 10. The connecting shaft 11 is rotatably connected to the top of a linkage member 12, and the bottom end of the linkage member 12 is hinged to the top of a driving rod 13. The driving rod 13 is movably connected in a guide sleeve 14 fixed on the support plate 8. At the same time, the bottom end of the driving rod 13 is fixedly connected to the driving shaft 15. The bottom end of the driving shaft 15 is connected to an elastic pressure block 16, which is located in the inner cavity of a culture vessel 17. The culture vessel 17 is located on the weighing device 7. At the same time, a ring member 18 located below the elastic pressure block 16 is also provided in the culture vessel 17. The outer wall of the annular member 18 is provided with an annular groove 19 , and the cell-scaffold complex 21 can be fixed on the surface of the annular member 18 by an elastic sleeve 20 connected to the annular groove 19 . A control box 22 and a display screen 23 are also provided on the base 6. A control module and a battery are provided in the control box 22. The battery provides power to the weighing device 7, the motor 9, the display screen 23 and the control module, while 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.
[0016] The elastic pressing block 16 is made of elastic silicone and has a spherical bottom surface.
[0017] Needle-type gas filters 4 are also provided at the four corners of the top plate of the outer box 1 .
[0018] The working process of the in vitro tissue-engineered ligament culture device for simulating an abdominal pressure biomechanical environment according to an embodiment of the present invention is as follows: first, the cell-scaffold complex 21 is placed on the annular member 18, and then the elastic sleeve 20 is inserted into the annular groove 19. That is, the elastic sleeve 20 is used to fix the edge of the cell-scaffold complex 21 in the annular groove 19, thereby fixing the cell-scaffold complex 21 on the annular member 18. At this time, the cell-scaffold complex 21 and the annular member 18 form a drum-like structure, with the cell-scaffold complex 21 acting as the drum skin and the annular member 18 acting as the drum body. The middle portion of the cell-scaffold complex 21 is suspended in the air and stretched. Place the ring 18 in the culture vessel 17, close the upper cover of the culture vessel 17, and ensure that the elastic pressing block 16 is also located in the culture vessel 17 and above the cell-scaffold complex 21. Then, the culture mechanism 2 is placed as a whole into the outer box 1 (the operation process needs to be carried out in a sterile environment), the box lid 5 is closed, and the outer box 1 is placed as a whole into the cell culture incubator. The high-concentration CO2 gas in the cell culture incubator can enter the outer box 1 through the double-sided hydrophobic PTFE filter membrane 3 and the needle gas filter 4 to ensure that the CO2 gas content during the culture process meets the requirements; and 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 taken out of the cell culture incubator for observation and 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 is contaminated with the cells planted on the cell scaffold complex 21, its growth may be affected. However, if the culture mechanism 2 is placed as a whole 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. When the culture mechanism 2 is working, the control module will control the motor 9 to drive the turntable 10 to rotate according to a pre-set program. When the turntable 10 rotates, it will drive the linkage member 12 to swing in space through the connecting shaft 11 on it, thereby driving the driving rod 13 connected to the bottom of the linkage member 12 to move. Since the driving rod 13 is movably connected in the guide sleeve 14, under the guidance of the guide sleeve 14, the driving rod 13 will drive the driving shaft 15 at its bottom end to make reciprocating linear motion, thereby realizing the reciprocating downward pressing action of the elastic pressure block 16; each time it presses 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; By controlling the output speed of the motor 9, the speed of the downward pressing action of the elastic pressing block 16 can be adjusted; by controlling the working off time of the motor 9, the duration of the downward pressing action of the elastic pressing block 16 and the duration of the state in which the elastic pressing block 16 does not press down on the cell-scaffold complex 21 (i.e., the two are not in contact) can be adjusted; at the same time, by cooperating with the downward pressing action and the weighing device 7, the pressure applied by the elastic pressing block 16 can be accurately controlled. For example, during the downward pressing process, the pressure gradually increases. When the weighing device 7 detects that the current pressure reaches a preset trigger value, it sends a signal to the control system, and the control system controls the motor 9 to stop working and maintain the current pressure, or controls the motor 9 to rotate in the opposite direction, and the elastic pressing block 16 moves upward to wait for the next pressure operation; During the culture process, the display screen 23 will display the current culture conditions and progress, such as the culture duration, the number of times the elastic pressing 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 transparent side wall of the box body 1; During the downward pressure of the elastic pressing block 16, since there is no support underneath the cell-scaffold complex 21, this structure simulates the real environment of tissue ligaments in the human body, thereby better culturing the cells planted on the cell-scaffold complex 21; After the culture is completed, the outer box 1 is taken out of the cell culture incubator as a whole, and the outer box 1 is opened. The annular member 18 is taken out of the culture vessel 17 , the elastic sleeve 20 is removed, and the cell-scaffold complex 21 is removed from the annular member 18 .
Claims
1. An in vitro culture device for tissue-engineered ligaments that simulates an abdominal pressure biomechanical environment, characterized by: The culture device comprises an outer box (1) and a culture mechanism (2) located inside the outer box (1); the outer box (1) is made of a transparent plate. A plurality of double-sided hydrophobic PTFE filter membranes (3) distributed in an array are provided on the top plate of the outer box (1), and a box cover (5) is also provided at the opening of the outer box (1). The culture mechanism (2) includes a base (6), a weighing device (7) and a support plate (8) are provided on the base (6), a motor (9) is provided on the top of the support plate (8), a working end of the motor (9) is connected to a turntable (10), a connecting shaft (11) is provided at an eccentric position of the turntable (10), the connecting shaft (11) is rotatably connected to the top end of a linkage member (12), and the bottom end of the linkage member (12) is hinged to the top end of a driving rod (13), and the driving rod (13) is movably connected in a guide sleeve (14) fixed on the support plate (8), and the bottom end of the driving rod (13) is fixedly connected to the driving shaft (15), and the bottom end of the driving shaft (15) is connected to an elastic pressure block (16), and the elastic pressure block (16) is located in the inner cavity of the culture vessel (17), and the culture vessel (17) is located on the weighing device (7). At the same time, the culture vessel (17) is also provided with a ring member (18) located below the elastic pressure block (16). The outer wall of the annular member (18) is provided with an annular groove (19), and the cell scaffold complex (21) can be fixed on the surface of the annular member (18) by an elastic sleeve (20) connected in the annular groove (19). A control box (22) and a display screen (23) are also provided on the base (6). A control module and a battery are provided in the control box (22). 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).
2. The in vitro tissue engineering ligament culture device for simulating an abdominal pressure biomechanical environment according to claim 1, characterized in that: The elastic pressing block (16) is made of elastic silicone and has a spherical bottom surface.
3. The in vitro culture device for tissue engineering ligaments simulating an abdominal pressure biomechanical environment according to claim 1, characterized in that: The elastic pressing block (16) is made of elastic silicone and has a flat bottom surface.
4. The in vitro culture device for tissue engineering ligaments simulating an abdominal pressure biomechanical environment according to claim 1, characterized in that: Needle-type gas filters (4) are also provided at the four corners of the top plate of the outer box (1).