A sphincter in-vitro culture environment construction device and application thereof

Abstract

The application relates to the technical field of cell in-vitro culture, and discloses a sphincter in-vitro culture environment construction device which comprises a culture device and a control host computer, the culture device is internally provided with a contraction member, the control host computer is connected with the culture device in a matched mode, and is used for regulating and controlling the action of the contraction member, so as to construct a physiological contraction environment. The application can simulate the annular contraction and the physiological contraction characteristics of the high and low pressure of the human sphincter, restore the dynamic contraction process, adopt a detachable structure, adapt to different culture cavities, has strong experimental flexibility, combines a perfusion design with a three-dimensional culture layer, simulates in-vivo nutrition transportation and metabolism discharge, provides a bionic growth environment for cells, integrates strain force detection and temperature control closed-loop regulation, is accurate in parameters and reliable in data, makes up for the short board of traditional static culture and animal experiments, provides a stable and efficient experimental platform for sphincter disease mechanism, mechanical response research and drug screening.

Description

Technical Field

[0001] This invention relates to the field of cell culture technology, and in particular to a device for constructing an in vitro culture environment for sphincter muscles and its application. Background Technology

[0002] The lower esophageal sphincter (LES) is a crucial physiological barrier in the digestive tract. Relying on sustained tension and rhythmic circular contractions, it maintains pressure balance between the esophagus and stomach, preventing the reflux of gastric contents and ensuring the normal function of the digestive system. LES dysfunction is a core contributing factor to digestive diseases such as gastroesophageal reflux disease (GERD). In-depth research into the biological behavior of sphincter cells under physiological and mechanical stress is essential for understanding disease mechanisms, drug development, and regenerative medicine repair.

[0003] Currently, research on sphincter function heavily relies on animal experiments and clinical observations. Animal models suffer from problems such as large species differences, long experimental cycles, strict ethical constraints, and high costs, making it difficult to accurately replicate the true physiological response of the human sphincter. Clinical research is limited by individual differences, invasive risks, and difficulties in obtaining samples, making it impossible to achieve controllable adjustment and long-term dynamic monitoring of mechanical stimulation parameters. Therefore, developing in vitro culture devices that can simulate the physiological contraction characteristics of the sphincter is a key path to overcome the limitations of in vivo research and to carry out basic research and functional verification. Existing in vitro cell cultures are mostly static, only providing a basic nutritional environment, and cannot reproduce the unique contraction and relaxation dynamic process of the sphincter, nor can they provide cells with dynamic mechanical stimulation close to that in vivo. The few devices that can apply mechanical stimulation can only provide simple non-specific stimulation, which does not match the physiological contraction characteristics of the sphincter. They are unable to truly reflect the physiological state of cells, nor can they induce cells to produce physiological responses that conform to the in vivo environment, thus limiting the reliability and reference value of experiments. Summary of the Invention

[0004] This invention addresses the shortcomings of existing technologies by providing a device for constructing an in vitro culture environment for sphincter muscles and its application, thus solving the problems of existing technologies being unable to reproduce simulated physiological contractions of sphincter muscles, lacking zonal stimulation, and having an unstable three-dimensional culture environment.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: A device for constructing an in vitro culture environment for sphincter muscles includes a culture device and a control host. The culture device is equipped with a contractile element, and the control host is connected to the culture device to regulate the movement of the contractile element and construct a physiological contraction environment.

[0006] Preferably, the culture device includes a support cylinder, an outer covering layer, an annular cavity, and a placement chamber. The outer covering layer covers the outside of the support cylinder, and a connecting end is provided on the inside of the support cylinder. The annular cavity is detachably installed inside the support cylinder through the connecting end, forming the placement chamber between the two. The shrink-fit component is located in the placement chamber. The detachable design facilitates the replacement of the annular cavity and equipment maintenance. The support cylinder and the outer covering layer ensure the overall structural stability, and the placement chamber provides a regular installation space for the shrink-fit component, adapting to different culture scenarios.

[0007] Preferably, the placement chamber is further provided with a limiting cover, and the contractile component is an airbag ring. The limiting cover covers the contractile component so that the contractile component can only be squeezed in the direction of the annular cavity. The annular cavity is provided with a detection module for detecting strain force at the position of the contractile component. The limiting cover restricts the directional deformation of the airbag and accurately replicates the annular contraction action of the sphincter muscle. The detection module collects strain force data in real time, providing a basis for closed-loop control of contraction force.

[0008] Preferably, the shrinkable part is provided with an air inlet and an air outlet. The air inlet is connected to an air guide pipe, and the air outlet is connected to an air outlet pipe. The end of the air guide pipe away from the shrinkable part is provided with a connector, and the end of the air outlet pipe away from the shrinkable part is provided with an air outlet valve. The air path interfaces are clearly divided, the flexible air guide pipe can be better adapted for use, and the electrically controlled air outlet valve can accurately control the exhaust, ensuring smooth and controllable inflation and deflation.

[0009] Preferably, the control host is equipped with an interactive module, and the control host is equipped with an air pump. The air pump is equipped with a connecting pipe, which is connected to a connector to communicate with the shrink fitting. The interactive module facilitates parameter setting and data viewing, the micro air pump provides a stable air source, and the quick connector is convenient for docking, ensuring reliable air circuit sealing and stable air supply.

[0010] Preferably, the control host is equipped with a control module, which is connected to the interaction module, the detection module and the vent valve signal to realize closed-loop control of parameter reception, strain feedback and valve regulation, so as to ensure accurate and stable contraction frequency and force and improve simulation accuracy.

[0011] Preferably, at least two sets of contractile elements are arranged axially in the placement chamber, namely the contractile elements in the upper region and the contractile elements in the lower region, forming a zoned contraction structure to distinguish between high and low pressure contraction areas, simulating the physiological differences between low pressure in the esophageal body and high pressure in the sphincter, and meeting the needs of differentiated contraction simulation.

[0012] Preferably, the connecting pipe connected to the output end of the air pump is equipped with a three-way connector, which branches into two independent air paths. Each air path is equipped with an air inlet valve. The two air inlet valves are then connected to the connectors on the two sets of contraction components through pipes. The air inlet valves are connected to the control module for signal transmission. The independent air paths control the two sets of contraction components respectively, and the inflation pressure of each zone can be adjusted independently to accurately match the zone contraction control requirements.

[0013] Preferably, the inner wall of the annular cavity is provided with a support structure, and a culture layer is formed on the inner side. The culture layer is provided with a culture channel, and the two ends of the culture channel are provided with pipe joints. The support structure fixes the three-dimensional culture layer, the gel scaffold is adapted to cell growth, and the channel can be connected to an external perfusion system to simulate the in vivo nutrient delivery and metabolic waste excretion.

[0014] The application of the aforementioned sphincter in vitro culture environment construction device is used for three-dimensional in vitro culture of sphincter cells, physiological contraction simulation, mechanical stimulation response research, and drug screening. This application meets the research needs related to sphincter cells, makes up for the shortcomings of traditional culture and in vivo experiments, and provides stable and reliable experimental support for disease mechanism, functional verification, and drug screening.

[0015] Compared with the prior art, the present invention has the following beneficial effects: This invention can simulate the circular contraction and regional high and low pressure physiological contraction characteristics of human sphincter muscles, reproducing the dynamic contraction process. It adopts a detachable structure to adapt to different culture chambers, providing high experimental flexibility. The three-dimensional culture layer combined with perfusion design simulates the in vivo nutrient transport and metabolic excretion, providing a biomimetic growth environment for cells. It integrates strain detection and closed-loop temperature control, ensuring precise parameters and reliable data. It overcomes the shortcomings of traditional static culture and animal experiments, providing a stable and efficient experimental platform for sphincter disease mechanism, mechanical response research, and drug screening. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1 This is a schematic diagram of the overall structure of the present invention; Figure 2 This is a cross-sectional view of the culture apparatus of the present invention; Figure 3 This is a view of the annular cavity structure of the present invention; Figure 4 This is a schematic diagram of the control principle of the single-zone control host of the present invention; Figure 5This is a schematic diagram of the control principle of the partition control host of the present invention; Figure 6 This is a schematic diagram of the air passage connection of the partitioned shrinkage component of the present invention.

[0018] Figure number explanation: 1. Culture device; 11. Outer covering layer; 12. Support cylinder; 212. Connecting end; 13. Shrinkage component; 131. Air inlet; 132. Air guide pipe; 133. Connector; 14. Limiting cover; 15. Annular cavity; 151. Support structure; 16. Culture layer; 17. Culture channel; 18. Placement chamber; 19. Pipe connector; 2. Control host; 21. Interaction module; 3. Connecting pipe; 4. Control module; 5. Air pump; 6. Vent valve; 7. Heating component; 8. Temperature sensor; 9. Detection module; 10. Air inlet valve; 101. T-connector. Detailed Implementation

[0019] The present invention will now be described in further detail with reference to the accompanying drawings.

[0020] The following description is intended to disclose the invention so that those skilled in the art can implement it. The preferred embodiments described below are merely examples, and other obvious modifications will be apparent to those skilled in the art. The basic principles of the invention defined in the following description can be used in other embodiments, modifications, improvements, equivalents, and other technical solutions that do not depart from the spirit and scope of the invention.

[0021] Those skilled in the art should understand that, in the disclosure of this invention, the terms "longitudinal," "lateral," "upper," "lower," "left," "right," "front," "rear," "vertical," "horizontal," "top," "bottom," "inner," and "outer," etc., indicate the orientation or position based on the orientation or positional relationship shown in the accompanying drawings. They are merely simplified descriptions for the convenience of describing this invention and do not indicate or imply that the device or component referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, the above terms should not be construed as limitations on this invention.

[0022] It is understood that the term "a" should be understood as "at least one" or "one or more", that is, in one embodiment, the number of an element can be one, while in another embodiment, the number of the element can be multiple, and the term "a" should not be understood as a limitation on the number.

[0023] Example: Please see Figure 1-6A device for constructing an in vitro culture environment for sphincter muscles is provided, which consists of two parts: a culture device 1 and a control host 2. The culture device 1 is a culture and contraction execution component, and is equipped with a contraction component 13 inside. The control host 2 is a control and power output component, which is connected to the culture device 1 through air pipes and signal lines. It is used to regulate the contraction component 13 to complete the inflation and deflation contraction actions, and can construct a physiological contraction environment that simulates the human sphincter muscle, thus meeting the needs of in vitro culture and contraction simulation.

[0024] The basic structure of the culture device 1 includes a support cylinder 12, an outer covering layer 11, an annular cavity 15, and a placement chamber 18. The outer covering layer 11 is made of highly elastic, biocompatible medical-grade silicone rubber and is entirely wrapped around the outside of the support cylinder 12 for protection. The support cylinder 12 is an annular rigid cylinder with an integrally formed or fixedly installed connecting end 212 on the inner side. The connecting end 212 may be equipped with a magnetic attraction structure. The annular cavity 15 is a cylindrical flexible cavity that can be detachably installed inside the support cylinder 12 through the connecting end 212 with a magnetic attraction structure. The structure can consist of two magnets, one located at the connecting end 212 and the other at the corresponding position on the annular cavity 15. The detachable nature allows the inner cavity to be replaced with different shaped culture cavities according to experimental needs, such as a straight tube to simulate the peristalsis of the esophageal body, a U-shaped or curved shape to simulate the peristalsis or specific anatomical structures required in the digestive tract, and an annular concave standard cavity specifically for the annular contraction stimulation of the sphincter. After installation, the inner wall of the support cylinder 12 and the outer wall of the annular cavity 15 enclose each other to form an annular placement chamber 18, and the contractile element 13 is installed in the placement chamber 18.

[0025] A limiting cover 14 is installed inside the placement chamber 18. The contraction component 13 adopts an annular airbag ring. The two work together to achieve directional contraction. The limiting cover 14 is an annular rigid cover with an opening on the inner side. When it cooperates with the support cylinder 12, it forms a closure on the outer side and the upper and lower sides of the contraction component 13. The limiting cover 14 completely covers the airbag ring, leaving only the inner opening facing the annular cavity 15. When the airbag ring inflates, it is constrained by the limiting cover 14 and can only be directionally squeezed from the inner opening towards the annular cavity 15, avoiding deformation in other radial directions. This accurately simulates the annular contraction of the sphincter muscle. The outer wall of the annular cavity 15, corresponding to the airbag ring, is the compression area of ​​the contraction component 13. A detection module 9 can be fixedly installed thereon. Pressure sensors or strain gauges can be used to fit against the outer wall of the annular cavity 15 to detect the strain force generated by the compression of the airbag ring in real time. The force signal is converted into an electrical signal for output, which is used to control the contraction force. The detection module 9 is detachably installed as an integral part of the annular cavity 15. Its signal output end is equipped with a signal male connector via a signal line. The corresponding position of the control host 2 is equipped with a matching signal female connector. The signal female connector is connected to the detection module 9. After the annular cavity 15 is installed, the signal male connector is connected to the signal female connector, and the strain force data of the detection module 9 can be directly transmitted to the control module 4. When disassembling the annular cavity 15, the signal connector can be separated, which is convenient for disassembly and assembly and avoids damage to the signal line by pulling.

[0026] The shrinking component 13 is provided with an air passage interface including an air inlet 131 and an air vent. The air inlet 131 is used for inflation, and the air vent is used for deflation. The air inlet 131 is sealed and connected to a flexible air guide tube 132. The air guide tube 132 extends out of the culture device 1, and a connector 133 is fixedly installed at the end away from the shrinking component 13. The connector 133 can be a quick connector to facilitate connection with the air passage on the control host 2. The air vent is sealed and connected to a flexible air outlet tube. The air outlet tube also extends out of the culture device 1, and an air vent valve 6 is installed at the end away from the shrinking component 13. The air vent valve 6 is an electrically controlled solenoid valve, which is controlled by the control host to achieve automatic air venting.

[0027] The basic structure of the control host 2 includes an interactive module 21, which is installed on the control host 2 panel. It is a touch interactive panel that integrates parameter setting buttons and a status display screen. It can set parameters such as contraction frequency, contraction force, and temperature, and displays strain, temperature, and working status in real time. The air pump 5 is built into the housing of the control host 2. It can be a miniature silent air pump to provide a stable air source for the contraction component 13. The output end of the air pump 5 is sealed to the connecting pipe 3. The end of the connecting pipe 3 can be a quick-connect fitting, which can be quickly and sealed to the connector 133 of the culture device 1 to realize the air path connection between the air pump 5 and the contraction component 13, and provide power for the inflation of the airbag ring.

[0028] The control host 2 is equipped with a control module 4, which is the core control unit and can be a microcontroller. The control module 4 is bidirectionally connected to the interaction module 21 via a signal line, receives the control parameters input by the interaction module 21, and feeds back the working status and detection data to the interaction module 21 for display. The control module 4 is connected to the detection module 9 via a signal line, and receives the strain force data fed back by the detection module 9 in real time. The control module 4 is connected to the vent valve 6 via a signal line, and controls the vent valve 6 to open or close according to the strain force data and preset parameters, thereby regulating the inflation pressure of the airbag ring.

[0029] At least two sets of contraction elements 13 are arranged along the axial direction in the placement chamber 18 to form a zoned contraction structure. The two sets of contraction elements 13 are the upper zone contraction element 13 and the lower zone contraction element 13, respectively, and are fitted with independent limiting covers 14 to prevent interference between them. The lower zone contraction element 13 is used to simulate the high pressure contraction characteristics of the lower esophageal sphincter, and the upper zone contraction element 13 is used to simulate the low pressure contraction characteristics of the esophageal body. Both sets of contraction elements 13 are equipped with independent air inlet ports 131, air outlet ports, air guide tubes 132 and connectors 133, which can realize independent operation of a single zone or synchronous operation of two zones to meet the requirements of zoned differentiated contraction simulation.

[0030] To achieve independent inflation control of the two sets of shrinkable parts 13, a three-way connector 101 and an air inlet valve 10 are added to the air circuit. The three-way connector 101 is installed in the middle of the connecting pipe 3 at the output end of the air pump 5. The three-way connector 101 divides the main air circuit into two independent branch air circuits. An air inlet valve 10 is installed in series on each branch air circuit. The air inlet valve 10 can be an electrically controlled proportional flow solenoid valve. The output ends of the two air inlet valves 10 are respectively connected to the connectors 133 of the two sets of shrinkable parts 13 through independent pipes. Both air inlet valves 10 are connected to the control module 4 through signal lines. The control module 4 can independently control the opening, closing and opening degree of each air inlet valve 10, so as to realize independent regulation of the inflation volume and inflation pressure of the two sets of shrinkable parts 13, and adapt to the differentiated shrinkage requirements of different zones.

[0031] The culture-related structures of the annular cavity 15 include a support structure 151 and a culture layer 16. The support structure 151 is integrally formed on the inner wall of the annular cavity 15. The support structure 151 is annular depressions, grooves, or micropillar arrays, used to help fix the culture layer 16 and prevent it from falling off or deforming. The culture layer 16 is made of biocompatible gelling materials, such as gelatin, gelMA, sodium alginate, PEGDA, etc. When in use, the liquid gelling solution is injected into the interior of the annular cavity 15 and cured by thermal cross-linking or photocross-linking to form an annular three-dimensional gel scaffold that fits the inner wall, providing a three-dimensional growth environment for sphincter cells. After curing, a through culture channel 17 is formed inside the culture layer 16, which runs through the entire culture layer 16 axially. The two ends of the culture channel 17 are sealed with pipe connectors 19, one end is a liquid inlet connector and the other end is a liquid outlet connector, which can be connected to a peristaltic pump or a culture medium circulation pump to realize the circulation and perfusion of culture medium in the culture channel 17, simulating the in vivo nutrient delivery, metabolic waste removal, and mucus dynamic environment.

[0032] A heating element 7 and a temperature sensor 8 are fixedly installed inside the placement chamber 18. The heating element 7 can be a miniature flexible heating plate, which is placed on the inner wall of the placement chamber 18 and can be attached to the annular cavity 15 for uniform heat dissipation. The temperature sensor 8 is a high-precision thermistor that detects the culture environment temperature in real time. The temperature sensor 8 is connected to the control module 4 through a signal line. The control module 4 adjusts the start and stop of the heating element 7 according to the preset temperature and real-time temperature data to maintain a stable culture environment temperature. All components that come into contact with cells and culture medium are made of medical-grade biocompatible materials, which are free from deformation, degradation and the release of harmful substances. The annular cavity 15 has a thin-walled structure with good thermal conductivity, which can evenly transfer heat to the culture layer 16 to ensure normal cell growth.

[0033] Heating component 7 and temperature sensor 8 are fixedly installed in placement chamber 18. Heating component 7 can be a miniature flexible heating element, which is placed on the inner wall of placement chamber 18 and can be attached to the annular cavity for uniform heat dissipation. Temperature sensor 8 is a high-precision thermistor, which detects the temperature of the culture environment in real time. Temperature sensor 8 is connected to control module 4 through signal line. Control module 4 adjusts the start and stop of heating component 7 according to preset temperature and real-time temperature data to maintain the stability of culture environment temperature. All parts that come into contact with cells and culture medium are made of medical-grade biocompatible materials, without deformation, degradation and release of harmful substances. The annular cavity is a thin-walled structure with good thermal conductivity, which can evenly transfer heat to the culture layer to ensure normal cell growth. At the same time, culture channel (17) can be connected to a conventional culture medium perfusion system. Carbon dioxide is introduced through the storage bottle of the perfusion system to pre-balance the pH of the culture medium. With a conventional carbon dioxide concentration detection device, the gas supply concentration can be monitored in real time to ensure the stability of the gas in the culture environment. Conventional cell culture equipment for carbon dioxide supply can be directly configured for use.

[0034] During the preparation phase of using this device, prepare the biocompatible gel solution, sphincter cell suspension, cell culture medium, and PBS buffer. Prepare the peristaltic pump and silicone tubing for perfusion. Install the sterilized annular cavity 15 into the support cylinder 12 via the magnetic connector 212. Fix the limiting cover 14 and the contraction element 13, ensuring that the contraction element 13 faces the annular cavity 15. Pass the gas inlet tube 132 and the outlet tube out of the support cylinder 12, with the connector 133 exposed. The vent valve 6 is located on the outside of the support cylinder 12. Connect the connecting tube 3 of the control host 2 to the connecting tube 1 of the culture device 1. 33. Quickly seal and connect to ensure no gas leakage in the gas path. Insert the signal male connector of the detection module 9 into the signal female connector of the control host 2 to complete the signal connection. Connect the pipe connectors 19 at both ends of the culture channel 17 to the peristaltic pump and culture medium storage bottle through silicone tubing to form a circulating perfusion loop. During the cell culture stage, slowly inject the liquid gel solution containing sphincter cells into the annular cavity 15, avoiding the generation of air bubbles. Depending on the type of gel material, perform thermal crosslinking or photocrosslinking curing to form a three-dimensional culture layer 16 containing cells. At the same time, introduce carbon dioxide to fully balance the pH. A conventional carbon dioxide concentration detection device monitors the concentration in real time. After maintaining a stable gas environment, a peristaltic pump is started to inject culture medium preheated to 37°C into culture channel 17. After removing air bubbles, the medium is circulated and perfused at a flow rate of 0.5-1 mL / min to maintain cell nutrient supply. The temperature is set via interactive module 21, and the heating component 7 is adjusted by control module 4. After the temperature stabilizes, the cells are incubated statically for 24 hours to allow them to adhere to the culture layer 16. During the contraction simulation phase, contraction parameters are set via interactive module 21, such as a contraction frequency of 6-12 times per minute and upper zone contraction. The force is 5-15 mmHg, the lower zone is 20-45 mmHg, the contraction time is 1-3 seconds, the relaxation time is 2-5 seconds, and the temperature is 37℃. Then, the contraction simulation is started. The control module 4 controls the air pump 5 to work. The air inlet valve 10 independently controls the inflation of the two sets of contraction components 13 according to the parameters. The air bladder ring directionally squeezes the annular cavity 15 to achieve annular contraction. The detection module 9 provides real-time feedback of strain data. The control module 4 controls the vent valve 6 in a closed loop to maintain the preset contraction force. The culture is continuously perfused for 24-72 hours under the contraction simulation state. During this period, the contraction parameters can be adjusted according to the experimental requirements. The data such as strain, temperature, contraction frequency, and inflation pressure can be viewed in real time through the interactive module 21. After the culture is completed, the air pump 5 and peristaltic pump are turned off, the power is disconnected, the annular cavity 15 is removed, the culture layer 16 is peeled off, and experiments such as cell viability detection, morphological observation, gene expression analysis, and protein detection are performed. The experimental data stored in the interactive module 21 is exported for subsequent statistical analysis. During the disassembly and maintenance stage, the gas path and perfusion line are disconnected, the power is turned off, the signal connector of the detection module 9 is unplugged, the annular cavity 15 is removed from the support cylinder 12, and detachable parts such as the contraction part 13, the limiting cover 14, and the air duct 132 are disassembled. Residual gel, cells, and culture medium can be removed by rinsing with PBS buffer. After disinfection again, the components are stored aseptically. After the control host 2 is powered off, it is stored in a dry, ventilated, and light-proof environment.

[0035] Those skilled in the art should understand that the embodiments of the present invention described above and shown in the accompanying drawings are merely examples and do not limit the present invention. The objectives of the present invention have been fully and effectively achieved. The functions and structural principles of the present invention have been shown and explained in the embodiments, and any modifications or variations of the embodiments of the present invention may be made without departing from the stated principles.

Claims

1. A device for constructing an in vitro culture environment for sphincter muscles, characterized in that, It includes a culture device (1) and a control host (2). The culture device (1) is equipped with a contraction component (13). The control host (2) is connected to the culture device (1) to regulate the movement of the contraction component (13) and construct a physiological contraction environment.

2. The sphincter in vitro culture environment construction device according to claim 1, characterized in that, The culture device (1) includes a support cylinder (12), an outer covering layer (11), an annular cavity (15), and a placement chamber (18). The outer covering layer (11) covers the outside of the support cylinder (12). A connecting end (212) is provided on the inside of the support cylinder (12). The annular cavity (15) is detachably disposed inside the support cylinder (12) through the connecting end (212), and the two form the placement chamber (18). The shrinking member (13) is disposed in the placement chamber (18).

3. The sphincter in vitro culture environment construction device according to claim 2, characterized in that, The placement chamber (18) is also provided with a limiting cover (14). The shrinking member (13) is an airbag ring. The limiting cover (14) covers the shrinking member (13) so that the shrinking member (13) can only be squeezed towards the annular cavity (15). The annular cavity (15) is provided with a detection module (9) for detecting strain force at the position corresponding to the shrinking member (13).

4. The sphincter in vitro culture environment construction device according to claim 3, characterized in that, The shrinkable part (13) is provided with an air inlet (131) and an air vent. An air guide pipe (132) is connected to the air inlet (131), and an air outlet pipe is connected to the air vent. A connector (133) is provided at the end of the air guide pipe (132) away from the shrinkable part (13), and an air vent valve (6) is provided at the end of the air outlet pipe away from the shrinkable part (13).

5. The sphincter in vitro culture environment construction device according to claim 4, characterized in that, The control host (2) is provided with an interactive module (21), and the control host (2) is provided with an air pump (5). The air pump (5) is provided with a connecting pipe (3). The connecting pipe (3) is connected to the connector (133) so that it communicates with the shrink member (13).

6. The sphincter in vitro culture environment construction device according to claim 5, characterized in that, The control host (2) is equipped with a control module (4), which is connected to the interaction module (21), the detection module (9) and the vent valve (6).

7. The sphincter in vitro culture environment construction device according to claim 5, characterized in that, At least two sets of contraction elements (13) are arranged along the axial direction in the placement chamber (18), namely the contraction element (13) of the upper region and the contraction element (13) of the lower region, forming a partitioned contraction structure.

8. The apparatus for constructing an in vitro culture environment for sphincter muscle according to claim 7, characterized in that, The air pump (5) output end is connected to the connecting pipe (3) with a three-way connector (101). The three-way connector (101) splits into two independent air paths. Each air path is equipped with an air inlet valve (10). The two air inlet valves (10) are then connected to the connectors (133) connected to the two sets of shrink fittings (13) through pipes. The air inlet valves (10) are connected to the control module (4) via signal.

9. The apparatus for constructing an in vitro culture environment for sphincter muscle according to claim 2, characterized in that, The inner wall of the annular cavity (15) is provided with a support structure (151), and a culture layer (16) is formed on the inner side. The culture layer (16) is provided with a culture channel (17), and the two ends of the culture channel (17) are provided with pipe joints (19).

10. The application of a sphincter in vitro culture environment construction device as described in any one of claims 1-9, characterized in that, Used for in vitro three-dimensional culture of sphincter cells, simulation of physiological contraction, study of mechanical stimulation response, and drug screening.

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