Oral mucosa organ chip

Abstract

The application discloses an oral mucosa organ chip, which comprises a cell culture layer, a gas driving layer and a microneedle layer. The cell culture layer is formed with a cell culture chamber; the gas driving layer is provided with a gas driving part and a gas transmission path. The gas driving part is formed with at least one expansion chamber. The expansion chamber can be injected and discharged with gas through the gas transmission path, so that the expansion chamber can be switched between expansion and contraction. The microneedle layer is provided with an elastic microneedle area corresponding to the positions of the expansion chambers. The elastic microneedle area is provided with a plurality of microneedles. The expansion chambers in the expansion movement or the contraction movement can drive the elastic microneedle area to deform, so that the microneedles contact the cells of the cell culture chamber and displace relative to the cells, so as to exert multidirectional stimulation on the cells. The chip constructs a cell culture system simulating the microenvironment of the human oral cavity. On this basis, the multidirectional and repeated physical friction stimulation that the oral cavity cells bear in the disease generation process can be accurately simulated.

Description

Technical Field

[0001] This application relates to an organ-on-a-chip, and more particularly to an oral cavity organ-on-a-chip. Specifically, it relates to an oral mucosa organ-on-a-chip. Background Technology Clinically, the occurrence of some oral diseases is closely related to long-term mechanical friction. The pathological process generally involves the oral mucosa being subjected to chronic, irregular stimulation under continuous external force, inducing abnormal repair responses and ultimately leading to pathological changes in tissue structure and function. For example, oral submucosal fibrosis is usually caused by long-term betel nut chewing; the continuous physical friction from the coarse fibers of betel nut directly causes mechanical damage to the mucosal epithelium. In addition, oral leukoplakia is also a related lesion caused by this type of friction. Therefore, in-depth investigation into the impact of irregular and repeated mechanical physical friction on the biological behavior of oral cells not only helps to reveal the pathogenesis of these diseases but also provides important theoretical basis for the screening of relevant therapeutic drugs.

[0002] Currently, research on the aforementioned oral diseases mainly relies on traditional animal models. While animal models allow for holistic in vivo studies, they suffer from limitations such as significant species differences, long modeling cycles, and high costs, making it difficult to accurately replicate the progressive pathological patterns of the oral mucosa after physical friction stimulation in humans. In recent years, organ-on-a-chip technology has provided a new approach to constructing oral disease models. This technology utilizes microfluidic systems to construct biomimetic microphysiological systems, enabling the in vitro reconstruction of the organ's microscopic tissue structure, fluid dynamics environment, and metabolic characteristics. However, existing oral microarrays still lack precise models that delve into the pathogenesis of oral mucosal cells subjected to continuous physical friction.

[0003] Therefore, there is an urgent need for an organ-on-a-chip specifically designed for studying the pathogenesis of oral mucosa-related diseases. This chip could accurately reproduce the damage to oral cells caused by continuous physical friction stimulation and simulate the dynamic inflammatory microenvironment formed during this damage process, thus comprehensively mimicking the entire process of disease progression, from initiation to carcinogenesis. This would provide an in vitro research platform for screening drugs to treat related oral diseases and for achieving early prevention and control of such diseases. Summary of the Invention

[0004] To address the aforementioned issues, this application discloses an oral mucosal organ-on-a-chip. This chip first constructs a cell culture system simulating the microenvironment of the human oral cavity. Based on this, it can accurately simulate the multidirectional, irregular, and repetitive physical friction stimulation experienced by oral cells. Under this long-term stimulation, the chip can dynamically track the gradual evolution of oral diseases caused by repeated friction. Therefore, this chip can provide a reliable in vitro experimental platform for screening drugs for treating such oral diseases and studying their pathogenesis.

[0005] To achieve the above objectives, this application adopts the following technical solution: This application provides an oral mucosal organ-on-a-chip, comprising a cell culture layer, a gas-driven layer, and a microneedle layer. The cell culture layer has cell culture chambers formed on its surface for culturing cells. The gas-driven layer has a gas driving unit and a gas transmission pathway connecting the gas driving unit. The gas driving unit has at least one expansion chamber facing the cell culture chamber. The gas transmission pathway enables gas injection and gas expulsion into the expansion chamber, allowing the expansion chamber to switch between expansion and contraction movements. The microneedle layer is disposed between the cell culture layer and the gas-driven layer. The microneedle layer has elastic microneedle regions facing the cell culture chamber at positions corresponding to the expansion chambers, and these elastic microneedles contain several microneedles. The expansion chambers, in expansion or contraction movements, can cause deformation of the elastic microneedle regions, allowing the microneedles to contact the cells in the cell culture chamber and displace relative to the cells, thereby applying stimulation to the cells in multiple directions.

[0006] This chip can visually represent the dynamic process of oral cavity cells undergoing multidirectional physical friction stimulation. Specifically, it constructs an in vivo microenvironment to support three-dimensional cell culture, thereby realistically replicating the survival state of submucosal cells in the oral cavity. The chip's advantages are twofold: First, by incorporating at least one expansion chamber with an elastic microneedle region simulating physical friction against cells, the expansion and contraction of the expansion chamber drives multiple microneedles in the elastic microneedle region to reciprocate in multiple directions to stimulate cells, thus accurately simulating the irregular and repetitive physical friction stimulation experienced by human oral cavity cells. Second, by adjusting the pressure of the injected gas, the expansion amplitude of the expansion chamber can be effectively controlled, thereby regulating the intensity of microneedle stimulation on the cells; by setting the alternating frequency of gas injection and expulsion, the periodic rhythm of the expansion and contraction of the expansion chamber can be effectively adjusted, thereby controlling the frequency of microneedle stimulation on the cells. Therefore, this chip can simulate the intensity and mode of stimulation in both force and frequency dimensions, providing a powerful tool for studying the pathogenesis of related oral diseases and a good in vitro experimental platform for screening subsequent therapeutic drugs.

[0007] In one illustrative embodiment of an oral mucosal organ-on-a-chip, the gas-driven unit further includes a base sheet with an expansion chamber formed in the center. The gas-driven layer includes a capping layer and a connecting layer. A connector is provided on the bottom surface of the capping layer, and the gas-driven unit is connected to the connector. The connecting layer is disposed on the bottom surface of the capping layer and has a channel for receiving the microneedle layer. The connector and the connecting layer can clamp and fix the base sheet of the gas-driven unit, allowing the expansion chamber to expand and contract within the receiving channel and contact the microneedle layer. This connector design ensures the installation stability of the gas-driven unit on the gas-driven layer while effectively ensuring the airtightness of the gas transmission path. Furthermore, the capping layer and the connecting layer work together to, on the one hand, clamp and fix the gas-driven unit by the base plate, ensuring the overall structure's airtightness and effectively preventing gas leakage from the expansion chamber during expansion; on the other hand, the design of the accommodating channel in the connecting layer limits the expansion degree of the expansion chamber, ensuring it expands only within a predetermined range. This avoids chamber rupture due to excessive expansion and also prevents excessive damage to cells caused by excessive expansion driving the micro-targets. Therefore, this design not only ensures the stability of the drive but also improves the reliability of long-term dynamic monitoring.

[0008] In one illustrative embodiment of the oral mucosal organ-on-a-chip, a gas transmission port is provided on the capping layer; a connecting passage is formed within the connector to communicate with the expansion chamber; and a transmission passage is formed within the connecting layer to communicate with the gas transmission port and the connecting passage. This gas transmission passage design allows the gas drive unit to inject and discharge gas while ensuring stability and sealing reliability through the connector, thereby driving the expansion chamber to perform alternating expansion and contraction movements, providing continuous driving force for the microneedle.

[0009] In one illustrative embodiment of the oral mucosa organ-on-a-chip, the gas-driven unit forms two expansion chambers, symmetrically arranged with a gap between them. This symmetrical arrangement of the two expansion chambers ensures that during gas injection, both chambers expand downwards and laterally relative to the gap, thereby simultaneously generating a downward thrust and a horizontal lateral thrust on the elastic microneedle region. Under the combined action of these two directional thrusts, the microneedle not only achieves a vertical needling effect on the cells but also simultaneously generates a horizontal relative displacement, thus more realistically simulating the physical friction stimulation experienced by cells in the oral cavity.

[0010] In one illustrative embodiment of an oral mucosa organ-on-a-chip, the gas-driven unit comprises four expansion chambers, with any two adjacent chambers symmetrically arranged and separated by a gap. This arrangement of four expansion chambers allows adjacent chambers to expand laterally relative to the gap between them, resulting in an overall expansion in two mutually perpendicular horizontal directions. This generates a downward thrust and two lateral thrusts in the elastic microneedle region, which can be understood as the X-axis and Y-axis, respectively. Under the combined action of these three main directional forces, the stimulation of cells by the microneedles more closely resembles the multidirectional physical friction stimulation experienced by cells in the oral cavity.

[0011] In one illustrative embodiment of an oral mucosal organ-on-a-chip, the gas-driven unit is provided with several expansion chambers; the gas-driven layer is provided with several gas transmission pathways corresponding to the expansion chambers, allowing independent gas injection and gas expulsion into each expansion chamber through each gas transmission pathway. This arrangement of multiple expansion chambers and gas transmission pathways enables independent control of each expansion chamber. On one hand, gases of different pressures can be injected into different expansion chambers, thereby adjusting the expansion amplitude of each chamber and achieving differentiated regulation of the stimulation received by cells in different regions of the cell culture chamber; on the other hand, the alternation frequency of gas injection and expulsion in each expansion chamber can be set independently to control the expansion rhythm of different chambers, thereby simulating the differences in physical friction stimulation received by cells in different regions of the patient's oral cavity.

[0012] In one illustrative embodiment of the oral mucosa organ-on-a-chip, the connector is provided with several barrier sections corresponding to several expansion chambers. The purpose of this barrier section design is that when the gas drive unit is connected to the connector unit, and the sealing layer and the connecting layer together clamp and fix the gas drive unit, the barrier section can effectively isolate each expansion chamber, preventing gas crosstalk during independent control, thereby ensuring that each expansion chamber maintains an independent gas path and can be controlled individually and precisely.

[0013] In one illustrative embodiment of an oral mucosal organ-on-a-chip, the cell culture layer includes a nutrient supply layer, a gel culture layer, and a porous scaffold layer. The nutrient supply layer has a reservoir and a liquid transport pathway connecting to the reservoir, through which nutrient solution can be delivered to the reservoir. The gel culture layer has a cell culture chamber and a gel injection channel connecting to the cell culture chamber, through which cell-containing gel can be delivered to the cell culture chamber. The bottom surface of the cell culture chamber has circular holes. The porous scaffold layer is disposed between the nutrient supply layer and the gel culture layer. Corresponding to the circular holes, the porous scaffold layer has several through-holes connecting to the reservoir, through which nutrient solution is supplied to the cells in the gel. This structural design of the cell culture layer enables the cell culture chamber to culture cell-containing gel, thereby achieving three-dimensional culture. This design ensures that the microneedles on the upper layer can effectively penetrate the gel and stimulate the cells inside. It also ensures that the nutrient solution below the cell culture chamber continuously delivers nutrients to the cells inside the gel, thereby enabling long-term three-dimensional cell culture. This allows the cells inside the chip to grow in a microenvironment that is closer to the human oral mucosa, providing a more realistic physiological basis for subsequent research on oral mucosa-related diseases.

[0014] In one illustrative embodiment of the oral mucosa organ-on-a-chip, the gel culture layer also forms several drug delivery pathways connecting the cell culture chambers. These drug delivery pathways can apply specific drug stimulation to the cell culture chambers, and can introduce specific chemical stimuli to study the promoting or inhibiting effects of chemical factors on disease progression; alternatively, they can directionally deliver therapeutic drugs to the three-dimensionally cultured cells to dynamically evaluate the intervention effect of the drugs on the lesions.

[0015] In one illustrative embodiment of the oral mucosal organ-on-a-chip, a gel injection port and a drug delivery port are also provided on the capping layer. The gel injection port is connected to the gel injection channel, and the drug delivery port is connected to the drug delivery pathway. A liquid transmission port is also provided on the capping layer, and the liquid transmission port is connected to the liquid transmission pathway.

[0016] The preferred embodiments will be described in a clear and easy-to-understand manner, with reference to the accompanying drawings, to further explain the above-mentioned characteristics, technical features, advantages and implementation methods of the oral mucosa organ-on-a-chip. Attached Figure Description

[0017] Various other advantages and benefits will become apparent to those skilled in the art upon reading the following detailed description of preferred embodiments. The accompanying drawings are for illustrative purposes only and are not intended to limit the scope of this application. Furthermore, the same reference numerals denote the same parts throughout the drawings. In the drawings: Figure 1An exploded view illustrating an illustrative embodiment of an oral mucosa organ-on-a-chip.

[0018] Figure 2 A schematic diagram illustrating the exploded structure of the oral mucosa organ-on-a-chip from another perspective.

[0019] Figure 3 A schematic diagram illustrating a combined structure of an illustrative embodiment of an oral mucosal organ-on-a-chip.

[0020] Figure 4 A schematic diagram illustrating a possible embodiment of a movement state of an oral mucosa organ-on-a-chip.

[0021] Figure 5 A schematic structural diagram illustrating another motion state of the oral mucosa organ-on-a-chip.

[0022] Figure 6 A schematic diagram illustrating one embodiment of the gas-driven unit.

[0023] Figure 7 A schematic diagram illustrating another illustrative embodiment of the gas-driven unit.

[0024] Figure 8 An exploded structural diagram illustrating another illustrative embodiment of the oral mucosa organ-on-a-chip.

[0025] Label Explanation 10. Cell culture layer; 11. Nutrient supply layer; 111. Liquid reservoir; 112. Liquid transport pathway; 12. Cell culture chamber; 122. Through-hole; 13. Gel culture layer; 14. Gel injection channel; 15. Porous scaffold layer; 16. Drug delivery pathway; 20. Gas-driven layer; 21. Capping layer; 211. Liquid transport port; 212. Gas transport port; 214. Gel injection port; 216. Drug delivery port; 22. Gas-driven section; 222. Expansion chamber; 224. Base sheet; 23. Connecting layer; 232. Receptacle channel; 233. Transport pathway; 25. Connector; 252. Connecting pathway; 254. Barrier section; 30. Microneedle layer; 32. Elastic microneedle region; 322. Microneedle. Detailed Implementation

[0026] To provide a clearer understanding of the technical features, objectives, and effects of this application, specific embodiments of this application are now described with reference to the accompanying drawings. In the drawings, the same reference numerals indicate components with the same or similar structures but the same function.

[0027] In this document, “illustrative” means “serving as an example, illustration or description”, and any illustration or implementation described herein as “illustrative” should not be construed as a more preferred or advantageous technical solution.

[0028] like Figure 1 , Figure 2 As shown, the oral mucosa organ-on-a-chip includes a cell culture layer 10, a gas-driven layer 20, and a microneedle layer 30. The cell culture layer 10 has a cell culture chamber 12 formed on its surface for culturing cells. In practical designs, the cell culture chamber 12 is cylindrical, with a diameter ranging from 10 mm to 12 mm. The gas-driven layer 20 is provided with a gas driving unit 22 and a gas transmission passage connected to the gas driving unit 22. The gas driving unit 22 is driven by gas delivered through the gas transmission passage. It can be located either inside or on the surface of the gas driving layer 20 to adapt to different layout requirements. The gas driving unit 22 has at least one expansion chamber 222, meaning it can have one or several expansion chambers 222, and these expansion chambers 222 are made of a flexible material. When the gas driving layer 20 is positioned above the cell culture layer 10, the expansion chambers 222 are arranged facing the direction of the cell culture chamber 12. By injecting or expelling gas into the expansion chamber 222 through the gas transmission pathway, the flexible expansion chamber 222 can expand or contract towards the cell culture chamber 12, and can switch between the two states. Figure 1 , Figure 2 As shown, a microneedle layer 30 is disposed between the cell culture layer 10 and the gas-driven layer 20. The microneedle layer 30 corresponds to the position of the expansion chamber 222 of the gas-driven layer 20, and an elastic microneedle region 32 is provided facing the cell culture chamber 12 of the cell culture layer 10. The elastic microneedle region 32 can deform under external force and can return to its initial state after the external force is removed. Several microneedles 322 are disposed on the elastic microneedle region 32. One end of the microneedle 322 is a sharp needle tip, and the other end is a needle seat fixed on the elastic microneedle region 32. In addition, the number and size of the microneedles 322 can be designed according to actual needs. In one illustrative embodiment of the elastic microneedle region 32, 61 microneedles 322 can be disposed, the length of the microneedle 322 is set to 0.8 mm, the diameter of the needle tip is set to 0.075 mm, and the diameter of the microneedle 322 needle seat is set to 0.3 mm.

[0029] like Figure 3As shown, the cell culture layer 10, microneedle layer 30, and gas-driven layer 20 are sequentially overlapped to obtain a complete chip structure. The chip has an octagonal shape, with its horizontal cross-section consisting of two parallel long sides, two parallel short sides, and a hypotenuse connecting the long and short sides. The length of the long sides ranges from 30mm to 50mm, and the length of the short sides ranges from 10mm to 25mm. Of course, those skilled in the art should understand that the overall shape and specific dimensions of the chip can be adaptively adjusted according to actual application requirements. Furthermore, the cell culture layer 10 and gas-driven layer 20 can be made of PDMS, and the microneedle layer 30 can be made of a flexible membrane material.

[0030] like Figure 4 As shown, the assembled chip structure, from top to bottom, consists of an expansion chamber 222, an elastic microneedle region 32, and a cell culture chamber 12. Taking a horizontal arrangement of two expansion chambers 222 as an example, in this case, the expansion chambers 222 are in their initial unexpanded state. Cells or cell-containing gels are cultured in the cell culture chamber 12. Several microneedles in the elastic microneedle region 32 are not fully inserted into the cell culture chamber 12, maintaining a certain distance from the cells and not making contact. Figure 5 As shown, when the operator connects a gas delivery device to the gas transmission path of the gas drive layer 20 and injects gas into the expansion chamber 222, the expansion chamber 222 continuously expands with the injection of gas. Figure 5 In the indicated state, the expansion chamber 222 expands simultaneously downwards and laterally, causing the elastic microneedle region 32 below to deform, resulting in several microneedles moving towards the cells within the cell culture chamber 12. When the expansion reaches a certain extent, the microneedles contact the cells and provide downward needle stimulation; furthermore, the lateral expansion of the expansion chamber 222 also causes the microneedles to undergo horizontal displacement relative to the cells, thus applying needle stimulation to the cells in multiple directions. When the operator controls the removal of gas from the expansion chamber 222, the expansion chamber 222 contracts back to its original position. Figure 4 The initial state is shown, and the elastic microneedle region 32 recovers synchronously. During the recovery process, the microneedles 322 displace relative to the cells in the opposite direction to the aforementioned direction, thereby applying reverse needle stimulation to the cells during the withdrawal phase as well.

[0031] In practical production design, the function of two expansion chambers 222 can also be achieved by forming one expansion chamber 222 on the gas driving part 22. For example, a long strip plate-like structure can be provided at the bottom of an expansion chamber 222. When gas is injected into the expansion chamber 222, two chambers will expand out at the bottom of the expansion chamber 222 due to the obstruction of the plate-like structure. As another example, the gas driving part 22 is a rigid structure with two openings at the bottom. An expansion chamber 222 is placed inside the gas driving part 22. As gas is injected, the expansion chamber 222 expands out two chambers from the two openings. Of course, those skilled in the art will understand that any configuration that achieves the function of multiple expansion chambers 222 through one expansion chamber 222 is within the scope of protection of this application.

[0032] Through the repeated alternation of expansion and contraction movements of the aforementioned expansion chamber 222, the microneedles can apply alternating directional complex stimulation to the cells during each alternation, thereby simulating the long-term, irregular, and multi-directional physical friction stimulation experienced by oral cells during disease progression. Furthermore, by adjusting the pressure of the injected gas, the expansion amplitude of the expansion chamber 222 can be effectively controlled, thus regulating the intensity of the microneedle stimulation to the cells. By setting the alternation frequency of gas injection and expulsion, the rhythm of expansion and contraction of the expansion chamber 222 can be effectively adjusted, thereby controlling the frequency of microneedle stimulation to the cells. Therefore, this chip can simulate stimulation intensity and stimulation mode from two dimensions: stimulation intensity and stimulation frequency. Taking oral mucosal fibrosis as an example, the stimulation of cells by several microneedles can realistically reproduce the frictional stimulation of the oral mucosa by the coarse fibers of areca nut when a patient chews areca nut. The frequency and amplitude of the expansion and contraction of the expansion chamber 222 can simulate the chewing habits of different patients, which not only provides an effective means for understanding the pathogenesis of fibrosis but also provides an experimental platform for the subsequent screening of anti-fibrotic drugs.

[0033] In one illustrative embodiment of the gas-driven layer 20, such as Figure 1 , Figure 2As shown, the gas-driven unit 22 also includes a base plate 224, with an expansion chamber 222 formed in the middle of the base plate 224. The gas-driven layer 20 can be divided into a capping layer 21 and a connecting layer 23. A connecting seat 25 is provided on the bottom surface of the capping layer 21, which is connected to the gas-driven unit 22 to ensure that the expansion chamber 222 of the gas-driven unit 22 has good airtightness. The connecting layer 23 is provided on the bottom surface of the capping layer 21 and has a receiving channel 232 for connecting the microneedle layer 30. When the capping layer 21 and the connecting layer 23 are combined, the base plate 224 of the gas-driven unit 22 is sandwiched between the two, which not only improves the stability of the gas-driven unit 22 fixed on the gas-driven layer 20, but also further ensures the airtightness of the expansion chamber 222, effectively preventing gas leakage during the expansion process. Meanwhile, the expansion chamber 222 can expand and contract within the accommodating channel 232 to limit the degree of expansion of the expansion chamber 222. This can prevent the expansion chamber 222 from rupturing due to excessive expansion, and also prevent excessive expansion from causing excessive damage to cells by the microneedles 322.

[0034] In one illustrative embodiment of the gas transport path, such as Figure 1 As shown, a gas transmission port 212 is provided on the sealing layer 21. (As indicated...) Figure 2 As shown, a connecting passage 252 communicating with the expansion chamber 222 is formed within the connecting seat 25, and combined with... Figure 1 As shown, a transmission path 233 is formed within the connecting layer 23, connecting the gas transmission port 212 and the connecting passage 252. When the capping layer 21 and the connecting layer 23 are combined, the gas transmission port 212, the transmission path 233, and the connecting passage 252 are connected to form a gas transmission path. Gas can then be injected into and discharged from the expansion chamber 222 through the gas transmission port 212, thereby driving the expansion chamber 222 to perform alternating expansion and contraction movements, providing a continuous driving force for the microneedle 322.

[0035] In one illustrative embodiment of the gas drive unit 22, such as Figure 6As shown, the gas drive unit 22 forms two expansion chambers 222, which are symmetrically arranged about the axis of symmetry L1, with a gap between them at the axis of symmetry L1. Specifically, the wall thickness of the two expansion chambers 222 is approximately 0.3 mm, and the gap distance is approximately 0.7 mm. The wall thickness and gap distance can be adjusted according to actual needs. With the two expansion chambers 222 arranged symmetrically, when gas is injected into them, both chambers exhibit a downward expansion tendency. Simultaneously, due to their symmetrical arrangement about the axis of symmetry L1 and the pre-existing gap, as gas is continuously injected, this gap is gradually filled by the expanding expansion chambers 222, causing the two expansion chambers 222 to compress each other. Under this constraint, further gas injection prevents the two expansion chambers 222 from expanding further towards the position of the axis of symmetry L1, and instead forces them to expand in the direction indicated by arrow y in the figure, i.e., along the y-axis to both sides. At this time, refer to... Figure 5 Under the combined effect of downward expansion and expansion on both sides along the y-axis, the 32 pairs of cells in the elastic microneedle region can simultaneously generate a downward needling effect and horizontal frictional stimulation on both sides along the y-axis. Of course, those skilled in the art will understand that the direction of the symmetry axis L1 is not limited to... Figure 6 As shown, it can be set to any direction according to actual needs. Accordingly, the two expansion chambers 222 will expand to both sides on a horizontal line perpendicular to the axis of symmetry L1, thereby driving the micro-needle to generate needle stimulation in the same direction to the cells.

[0036] In another illustrative embodiment of the gas drive unit 22, such as Figure 7As shown, the gas drive unit 22 has four expansion chambers 222: expansion chamber 222a, expansion chamber 222b, expansion chamber 222c, and expansion chamber 222d. Two adjacent expansion chambers 222 form a group; specifically, expansion chambers 222a and 222b can be one group, and expansion chambers 222c and 222d can be another. The expansion chambers 222 within each group are symmetrically arranged about a first axis of symmetry L2; two groups are symmetrically arranged about a second axis of symmetry L3, and the two axes of symmetry are perpendicular to each other. Gaps are formed between the four expansion chambers 222 at the first axis of symmetry L2 and the second axis of symmetry L3. With this arrangement of four expansion chambers 222, when gas is injected, all four expansion chambers 222 exhibit a downward expansion tendency, that is, they expand along the Z-axis in the figure. Simultaneously, since there are certain gaps between expansion chambers 222a and 222b, and between expansion chambers 222c and 222d, as gas is continuously injected, these gaps are gradually filled by the expanding expansion chambers 222, causing the pairs of expansion chambers 222 to compress each other. Under this constraint, further gas injection prevents the pairs of expansion chambers 222 from expanding towards the position of the first axis of symmetry L2, and instead forces them to expand in the direction indicated by arrow y in the figure, that is, along the y-axis to both sides. Similarly, since the two groups of expansion chambers are symmetrically arranged and also have certain gaps, namely between expansion chambers 222a and 222d, and between expansion chambers 222b and 222c, as gas is continuously injected, each expansion chamber 222 gradually expands, and these gaps are gradually filled, causing the pairs of expansion chambers 222 to compress each other. Further gas injection prevents the two expansion chambers 222 from expanding further towards the location of the second axis of symmetry L3. Instead, they are forced to expand in the direction indicated by arrow x in the diagram, i.e., expanding to both sides along the x-axis. At this time, under the combined action of expansion along the z-axis and expansion to both sides along the x and y axes, the elastic microneedle region 32 can simultaneously exert a downward needling effect on the cells and a horizontal frictional stimulation to both sides along the x and y axes. Under the combined action of these three main directional forces, the stimulation of the cells by the microneedles is closer to the multidirectional and irregular physical frictional stimulation experienced by cells in the oral cavity during disease development.

[0037] In another illustrative embodiment of the gas driving unit 22, the gas driving unit 22 is provided with a plurality of expansion chambers 222, and the gas driving layer 20 is provided with a plurality of gas transmission paths corresponding to the plurality of expansion chambers 222. Specifically, a plurality of gas transmission ports 212 can be provided on the capping layer 21, and a plurality of connecting paths 252 connecting each expansion chamber 222 can be provided on the connecting seat 25. Then, a plurality of transmission paths 233 are formed on the connecting layer 23, which respectively connect each gas transmission port 212 and each connecting path 252. Gas injection and gas discharge can be achieved independently through each gas transmission path. By providing a plurality of gas transmission paths corresponding to the plurality of expansion chambers 222, independent control of each expansion chamber 222 can be achieved. That is, gas at different pressures can be injected into different expansion chambers 222 to adjust the expansion amplitude of each expansion chamber 222, thereby achieving differentiated regulation of the stimulation intensity of cells in different regions of the cell culture chamber 12. Simultaneously, the alternating frequency of gas injection and discharge in each expansion chamber 222 can be set independently to control the expansion rhythm of different expansion chambers 222. Taking oral mucosal fibrosis as an example, this setting can simulate the differences in the degree of stimulation of cells in different areas of the oral cavity by areca fiber when a patient chews areca nut.

[0038] In order to effectively isolate the expansion chambers 222 and prevent gas cross-contamination during independent control, such as Figure 2 As shown, several baffles 254 are provided on the connecting seat 25 corresponding to several expansion chambers 222. The baffles 254 ensure that the air path of each expansion chamber is independent. When several baffles 254 are removed, the several expansion chambers 222 can be uniformly controlled through only one air path.

[0039] In another illustrative embodiment of the gas drive unit 22, the barrier 254 does not isolate each expansion chamber 222 from each other, but rather groups all expansion chambers 222: the barrier 254 divides them into 2, 4, or more groups of expansion chambers; the barrier 254 does not separate multiple expansion chambers 222 within the same group, and the expansion chambers 222 within the group are interconnected and allow gas exchange; the expansion chambers 222 in different groups are sealed and isolated by the barrier 254 to avoid gas crosstalk between groups. Each group of expansion chambers is configured with an independent gas transmission path, and each group can complete gas filling and discharging through its corresponding gas transmission path. In this embodiment, different ventilation pressures and filling and discharging alternation frequencies can be applied to different groups to achieve differentiated deformation control between groups, and all expansion chambers 222 within the same group maintain synchronous expansion and synchronous contraction, with the expansion amplitude and deformation rhythm being completely consistent. Taking the oral mucosal fibrosis model as an example, this setup can reproduce the stimulation differences between large areas of the oral cavity, and eliminates the need to configure a separate air path for each expansion chamber 222, thereby reducing the number of air path interfaces and lowering the complexity of the overall fluid control system.

[0040] In one illustrative embodiment of the cell culture layer 10, such as Figure 8 As shown, the cell culture layer 10 includes a nutrient supply layer 11, a gel culture layer 13, and a porous scaffold layer 15. The nutrient supply layer 11 has a thickness of 4 mm and forms a reservoir 111 and a liquid transport channel 112 connecting to the reservoir 111. The reservoir 111 is cylindrical with a diameter of 10 mm and a depth of 3 mm. Nutrient solution can be delivered to the reservoir 111 through the liquid transport channel 112. The gel culture layer 13 has a thickness of 1.6 mm and forms a cell culture chamber 12 and a gel injection channel 14 connecting to the cell culture chamber 12. The cell culture chamber 12 is cylindrical with a diameter of 10 mm to restrict the position of the gel. Cell-containing gel can be delivered to the cell culture chamber 12 through the gel injection channel 14, thereby achieving three-dimensional culture. A circular hole is also provided on the bottom surface of the cell culture chamber 12. A porous scaffold layer 15 is disposed between the nutrient supply layer 11 and the gel culture layer 13. The porous scaffold layer 15 has several through-holes 122 corresponding to the circular holes, which connect to the reservoir chambers 111. The reservoir chambers 111 supply nutrient solution to the cells in the gel through the through-holes 122. The thickness of the porous scaffold layer 15 can be 0.1 mm, and the diameter of the through-holes 122 is 0.7 mm.

[0041] In one illustrative embodiment of the gel culture layer 13, such as Figure 8As shown, the gel culture layer 13 also forms several drug delivery pathways 16 connecting the cell culture chambers 12. Specific drug stimuli can be applied to the cell culture chambers 12 through the drug delivery pathways 16. For example, specific chemical stimuli can be introduced through this pathway to study the promoting or inhibiting effects of chemical factors on disease progression; or therapeutic drugs can be directed to the three-dimensionally cultured cells to dynamically evaluate the intervention effect of the drugs on the lesions.

[0042] Furthermore, to ensure that gels, medications, and nutrient solutions can be injected into the chip from outside, such as... Figure 8 As shown, the capping layer 21 is also provided with a gel injection port 214, a drug delivery port 216, and a liquid transfer port 211. Among them, the gel injection port 214 is connected to the gel injection channel 14, the drug delivery port 216 is connected to the drug delivery passage 16, and the liquid transfer port 211 is connected to the liquid transfer passage 112.

[0043] To keep the drawings concise, only the parts relevant to this application are shown schematically in each drawing, and they do not represent the actual structure of the product. In addition, to make the drawings concise and easy to understand, in some drawings, only one of the components with the same structure or function is shown schematically, or only one of them is labeled.

[0044] It should be understood that although this specification describes various embodiments, not every embodiment contains only one independent technical solution. This way of describing the specification is only for clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

[0045] The detailed descriptions listed above are merely specific descriptions of feasible embodiments of this application and are not intended to limit the scope of protection of this application. All equivalent implementations or modifications made without departing from the spirit of the art of this application, such as combinations, divisions or repetitions of features, should be included within the scope of protection of this application.

Claims

1. An oral mucosa organ-on-a-chip, characterized in that, It includes: Cell culture layer (10), whose surface forms a cell culture chamber (12) for culturing cells; A gas-driven layer (20) is provided with a gas-driven part (22) and a gas transmission passage connecting the gas-driven part (22). The gas-driven part (22) forms at least one expansion chamber (222). The expansion chamber (222) is disposed facing the cell culture chamber (12). Gas injection and gas discharge can be realized in the expansion chamber (222) through the gas transmission passage, so that the expansion chamber (222) switches between expansion and contraction movements. A microneedle layer (30) is disposed between the cell culture layer (10) and the gas-driven layer (20). The microneedle layer (30) has an elastic microneedle region (32) facing the cell culture chamber (12) at the position corresponding to the expansion chamber (222). The elastic microneedle region (32) is provided with a number of microneedles (322). The expansion chamber (222) in the expansion or contraction motion can cause the elastic microneedle region (32) to deform, so that several microneedles (322) contact the cells in the cell culture chamber (12) and move relative to the cells to apply stimulation to the cells in multiple directions.

2. The oral mucosa organ-on-a-chip according to claim 1, characterized in that, The gas-driven unit also includes a base plate (224), and the expansion chamber (222) is formed in the middle of the base plate (224). The gas-driven layer (20) includes: The sealing layer (21) has a connecting seat (25) on its bottom surface, and the connecting seat (25) is connected to the gas driving unit (22). A connecting layer (23) is disposed on the bottom surface of the capping layer (21), and the connecting layer (23) is provided with a receiving channel (232) that connects to the microneedle layer (30). The connecting seat (25) and the communicating layer (23) can clamp and fix the base plate (224) of the gas driving part (22), so that the expansion chamber (222) can expand and contract within the accommodating channel (232) and contact the microneedle layer (30).

3. The oral mucosa organ-on-a-chip according to claim 2, characterized in that, The sealing layer (21) has a gas transmission port (212); a connecting passage (252) connecting the expansion chamber (222) is formed in the connecting seat (25); and a transmission passage (233) connecting the gas transmission port (212) and the connecting passage (252) is formed in the connecting layer (23).

4. The oral mucosa organ-on-a-chip according to claim 3, characterized in that, The gas drive unit (22) forms two expansion chambers (222), which are symmetrically arranged and have a gap between them.

5. The oral mucosa organ-on-a-chip according to claim 3, characterized in that, The gas drive unit (22) has four expansion chambers (222), and any two adjacent expansion chambers (222) are symmetrically arranged and have a gap between them.

6. The oral mucosa organ-on-a-chip according to claim 3, characterized in that, The gas drive unit (22) is provided with several expansion chambers (222); The gas-driven layer (20) is provided with several gas transmission paths corresponding to several expansion chambers (222), and gas injection and gas discharge can be achieved independently for each expansion chamber (222) through each gas transmission path.

7. The oral mucosa organ-on-a-chip according to claim 6, characterized in that, The connecting seat (25) is provided with several blocking parts (254) corresponding to several expansion chambers (222).

8. The oral mucosa organ-on-a-chip according to claim 2, characterized in that, The cell culture layer (10) includes: The nutrient supply layer (11) has a liquid storage chamber (111) and a liquid transmission passage (112) communicating with the liquid storage chamber (111), through which nutrient solution can be delivered to the liquid storage chamber (111); A gel culture layer (13) is formed therein, which has the cell culture chamber (12) and a gel injection channel (14) communicating with the cell culture chamber (12). The gel containing cells can be delivered to the cell culture chamber (12) through the gel injection channel (14). The bottom surface of the cell culture chamber (12) is provided with a round hole. A porous scaffold layer (15) is disposed between the nutrient supply layer (11) and the gel culture layer (13). The porous scaffold layer (15) has several through holes (122) corresponding to the positions of the circular holes, which connect to the liquid storage chamber (111). The liquid storage chamber (111) provides nutrient solution to the cells in the gel through the several through holes (122).

9. The oral mucosa organ-on-a-chip according to claim 8, characterized in that, The gel culture layer (13) also forms several drug delivery pathways (16) that connect the cell culture chamber (12).

10. The oral mucosa organ-on-a-chip according to claim 9, characterized in that, The capping layer (21) is also provided with a gel injection port (214) and a drug delivery port (216). The gel injection port (214) is connected to the gel injection channel (14); the drug delivery port (216) is connected to the drug delivery passage (16). The sealing layer (21) is also provided with a liquid transmission port (211), which is connected to the liquid transmission passage (112).

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