Biomimetic motor organ chip system and method of use
By constructing a biomimetic motor organ-on-a-chip system, a highly biomimetic simulation of the movement process was achieved, which solved the shortcomings of existing models in simulating the function of the overall motor system, improved research efficiency and human-derived models, revealed new mechanisms of movement, and provided new research methods for personalized exercise prescriptions.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- ZHUZHOU CENT HOSPITAL
- Filing Date
- 2025-08-21
- Publication Date
- 2026-06-23
Smart Images
Figure CN121022586B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of organ-on-a-chip and biomimetic biological experimental model construction technology, and in particular to a biomimetic movement organ-on-a-chip system and its usage method. Background Technology
[0002] Exercise, as a core non-pharmacological intervention, has been widely proven to have multiple pathophysiological benefits. Different types of exercise offer varying benefits to different organ systems, demonstrating high intervention specificity. Exercise intervention involves the synergistic regulation of multiple organs and systems, and its molecular mechanisms are complex and difficult to elucidate systematically. Therefore, in-depth analysis of the molecular mechanisms underlying exercise benefits not only helps optimize the clinical efficacy of personalized exercise prescriptions but also provides a theoretical basis for the development of novel therapeutic targets. With the rapid development of systems biology and multi-omics technologies, the biological effects induced by exercise have gradually transitioned from phenomenological observation to mechanistic analysis. However, unlike other molecular mechanism studies, existing research models of exercise-related mechanisms still have significant limitations. While in vivo animal models can better simulate the real exercise environment and reveal its regulatory effects on the whole body, they are limited by stress response interference and the complexity of macroscopic regulation, making it difficult to accurately verify the causal relationships between specific molecular pathways. In vitro models can achieve local observation at the cellular and molecular levels, but they cannot simulate the dynamic interactions of multiple systems induced by exercise and the systemic adaptive regulatory mechanisms under pathological conditions. Therefore, constructing composite research models with higher physiological relevance and cross-system integration capabilities is key to promoting in-depth research on movement mechanisms.
[0003] Organ-on-a-chip (OAS) is a biomimetic in vitro model based on microfluidic technology. It utilizes physiologically functional human cells to construct a three-dimensional microenvironment, simulating organ structure, function, and dynamic physiological processes within microscale channels. Compared to traditional two-dimensional cell culture and animal models, OAS can more realistically reproduce the in vivo microenvironment, such as shear force, mechanical stress, and gas-liquid interfaces. It offers high controllability and reproducibility, accurately simulating the molecular responses and signal transduction processes of specific tissues under disease states, and is widely used in drug screening, toxicology evaluation, and disease mechanism research. Its greatest advantage lies in its ability to achieve multi-organ cascade and multi-factor regulation, providing an experimental platform for studying complex cross-organ and cross-system signaling networks, particularly suitable for elucidating systemic pathological mechanisms. Furthermore, OAS combines the advantages of both in vivo and in vitro models, achieving higher humanization and reproducibility, which helps improve experimental interpretability and clinical translation efficiency, making it a crucial support platform for precision medicine research.
[0004] Significant progress has been made in experimental research using various organ-on-a-chip models, including heart-on-a-chip, lung-on-a-chip, brain-on-a-chip, and skeletal muscle-on-a-chip. For example, lung-on-a-chips achieve alveolar gas exchange (such as the exchange of oxygen and carbon dioxide) by embedding biological membranes; heart-on-a-chips simulate heart tissue and its functions by constructing three-dimensional hydrogel tissue and embedding cardiomyocytes and fibroblasts; skeletal muscle-on-a-chips construct mature skeletal muscle bundles through anchoring structures; and multi-organ-on-a-chips connect different organs through vascular channels to simulate the exchange of substances between organs in the body. Therefore, organ-on-a-chip technology has shown significant advantages in overcoming the current challenges in studying molecular mechanisms of movement. However, current research on biomimetic movement organ-on-a-chips is still in its early exploratory stages, and a truly meaningful chip platform capable of simulating the function of the entire musculoskeletal system is still lacking. Summary of the Invention
[0005] The purpose of this invention is to provide a biomimetic motor organ-on-a-chip system and its usage method, which constructs in vivo and in vitro models that can mimic the movement process, improving research efficiency, making the models more human-like, shortening the process from basic research to clinical application, and increasing the possibility of translational applications; constructing highly biomimetic in vivo and in vitro research models, effectively realizing crosstalk regulation between different cells / organs during movement, which helps to reveal new mechanisms of movement; realizing biomimetic processes of different exercise intensities, providing new research methods for personalized application of exercise prescriptions; applicable to movement process research in different scenarios and dimensions, with strong scalability.
[0006] This invention provides a biomimetic motor organ-on-a-chip system and its usage method, comprising a biomimetic motor organ-on-a-chip, a dynamic culture medium supply system, a closed-loop motion control system, and a data display terminal. The wiring terminals of the strain sensor on the top cover of the biomimetic motor organ-on-a-chip are connected to the sensor signal acquisition and analysis module in the closed-loop motion control system. The wiring terminals of the electrical stimulation electrodes on the bottom electrical stimulation layer of the biomimetic motor organ-on-a-chip are connected to the electrical stimulation generation module in the closed-loop motion control system. The inlet, outlet, skeletal muscle bundle culture chamber, and outlet of the co-culture cell chamber on the top cover of the biomimetic motor organ-on-a-chip are connected to the cell culture medium and recovery storage tank in the dynamic culture medium supply system. Data interaction occurs between the closed-loop motion control system, the dynamic culture medium supply system, and the data display terminal. The sensor signal acquisition and analysis module monitors the skeletal muscle bundles and feeds back the data to the electrical stimulation generation module.
[0007] Preferably, the biomimetic motor organ chip includes a top cover, an upper cell culture chamber, a cell chamber partition layer, a lower cell culture chamber, and a bottom electrical stimulation layer; the top cover, upper cell culture chamber, cell chamber partition layer, lower cell culture chamber, and bottom electrical stimulation layer are arranged sequentially from top to bottom, and the cell chamber partition layer and the upper cell culture chamber are stacked in multiple layers.
[0008] Preferably, the dynamic culture medium supply system includes a pressure source, a microfluidic pressure pump, and a recovery reservoir. The pressure source is connected to the microfluidic pressure pump, and the microfluidic pressure pump is connected to the recovery reservoir and the cell culture medium.
[0009] Preferably, the closed-loop motion control system includes a sensor signal acquisition and analysis module and an electrical stimulation generation module. The sensor signal acquisition module is connected to the wiring terminals of the strain sensor, and the electrical stimulation generation module is connected to the wiring terminals of the electrical stimulation electrode.
[0010] Preferably, the top cover includes an anchor point, a strain sensor, a terminal block of the strain sensor, a co-culture cell chamber inlet, a co-culture cell chamber outlet, a skeletal muscle bundle culture chamber inlet, and a skeletal muscle bundle culture chamber outlet. The top cover is provided with an anchor point, and the strain sensor is built into the top cover. The anchor point is made of flexible material and is mechanically coupled to the strain sensor. The co-culture cell chamber inlet, co-culture cell chamber outlet, skeletal muscle bundle culture chamber inlet, and skeletal muscle bundle culture chamber outlet are located around the surface of the top cover.
[0011] Preferably, the upper cell culture chamber includes a first co-culture cell chamber, a second co-culture cell chamber, a skeletal muscle bundle culture chamber, a porous membrane, microchannels for the first and second co-culture cell chambers, a liquid inlet for the co-culture cell chamber, a liquid outlet for the co-culture cell chamber, a liquid inlet for the skeletal muscle bundle culture chamber, and a liquid outlet for the skeletal muscle bundle culture chamber. The skeletal muscle bundle culture chamber is located at the center of the upper cell culture chamber. The skeletal muscle bundle culture chamber is separated from the first and second co-culture cell chambers on both sides by a porous membrane. The upper cell culture chamber is provided with microchannels for the first and second co-culture cell chambers. The porous membrane contains a silver nanowire mesh structure.
[0012] The cell chamber partition layer includes a connecting hole for the skeletal muscle bundle culture chamber, an outlet for the co-cultured cell chamber, an inlet for the co-cultured cell chamber, an outlet for the skeletal muscle bundle culture chamber, and an inlet for the skeletal muscle bundle culture chamber. The connecting hole for the skeletal muscle bundle culture chamber is located at the center of the cell chamber partition layer.
[0013] Preferably, the lower cell culture chamber includes a third co-culture cell chamber, a skeletal muscle bundle culture chamber, a fourth co-culture cell chamber, and a skeletal muscle bundle culture chamber microchannel. The third co-culture cell chamber microchannel, the fourth co-culture cell chamber microchannel, the electrode terminal groove, the co-culture cell chamber outlet, and the co-culture cell chamber inlet are located in the center of the lower cell culture chamber. The third and fourth co-culture cell chambers are located on both sides of the skeletal muscle bundle culture chamber. The lower cell culture chamber contains the skeletal muscle bundle culture chamber microchannel, the third co-culture cell chamber microchannel, and the fourth co-culture chamber microchannel.
[0014] The bottom electrical stimulation layer includes the terminals of the electrical stimulation electrodes and the electrode ends of the electrical stimulation electrodes. The electrode ends of the electrical stimulation electrodes directly contact the skeletal muscle bundles. The terminals of the electrical stimulation electrodes are connected to the electrical stimulation generation module.
[0015] Preferably, the co-culture cell chamber inlets on the top cover include a first co-culture cell chamber inlet, a second co-culture cell chamber inlet, a third co-culture cell chamber inlet, and a fourth co-culture cell chamber inlet; the co-culture cell chamber outlets on the top cover include a first co-culture cell chamber outlet, a second co-culture cell chamber outlet, a third co-culture cell chamber outlet, and a fourth co-culture cell chamber outlet; the structures of the co-culture cell chamber inlets and outlets of the upper cell culture chamber are the same as those on the top cover; the co-culture cell chamber inlets on the cell chamber partition layer include... The third and fourth co-culture cell chambers have liquid inlets; the co-culture cell chamber outlets on the cell chamber partition layer include the third and fourth co-culture cell chamber outlets; the lower cell culture chambers have co-culture cell chamber inlets including the third and fourth co-culture cell chamber inlets, and co-culture cell chamber outlets including the third and fourth co-culture cell chamber outlets; the skeletal muscle bundle culture chamber is equipped with a skeletal muscle bundle culture chamber inlet and a skeletal muscle bundle culture chamber outlet.
[0016] Preferably, a method of using a biomimetic musculoskeletal chip system includes the following steps: Step S1, constructing skeletal muscle bundles;
[0017] A mixture of skeletal muscle cells and hydrogel is injected into the chamber of a conventional skeletal muscle chip. Anchors on the top cover are inserted into the matching chamber of the conventional skeletal muscle chip. The top cover with integrated anchors is then closed, and skeletal muscle bundles are cultured.
[0018] The special structure of the anchor point is used to guide the skeletal muscle cells to grow in a bundle-like structure.
[0019] Step S2: Asynchronous co-culture of multiple cells;
[0020] During skeletal muscle bundle culture, other cells are seeded in the biomimetic musculoskeletal chip; different cell types are cultured independently through cell compartment septa.
[0021] Other cells include endothelial cells, neurons, or immune cells; the porous membrane's silver nanowires shield against electrical stimulation interference;
[0022] Step S3: Integration of bionic musculoskeletal chip;
[0023] The top cover containing skeletal muscle bundles was moved into the skeletal muscle bundle culture chamber of the bionic musculoskeletal organ chip and connected to the closed-loop motion control system and the dynamic culture medium supply system.
[0024] Step S4: Closed-loop motion intervention;
[0025] In the closed-loop motion control system, motion parameters are set, electrical stimulation and culture medium circulation are initiated, and the intensity of skeletal muscle bundle contraction is monitored in real time and the intensity of electrical stimulation is dynamically adjusted.
[0026] Exercise parameter settings include setting the exercise mode and exercise intervention time; the exercise mode includes low-intensity endurance, high-intensity explosive, or scalable mode; when the skeletal muscle fasciculation intensity is between 3% and 10%, it is low-intensity endurance mode, and when the skeletal muscle fasciculation intensity is not less than 18%, it is high-intensity explosive mode; the scalable mode generates new exercise type decision rules by inputting a custom contraction intensity threshold through the user interface; the custom contraction intensity threshold includes the skeletal muscle contraction intensity range and the upper limit of the electrical stimulation frequency;
[0027] The strain sensor collects skeletal muscle bundle contraction data in real time and analyzes the current electrical stimulation contraction intensity and frequency. If the current state does not belong to the set exercise mode, the electrical stimulation parameters are dynamically adjusted. If the current state belongs to the set exercise mode, the current stimulation intensity is maintained and the exercise intervention continues.
[0028] When a low-intensity endurance mode is set but a high-intensity burst mode is detected, reduce the electrical stimulation amplitude until the contraction intensity of the skeletal muscle bundles drops back to 3%–10%; when a high-intensity burst mode is set but the contraction intensity of the skeletal muscle bundles does not reach 18%, increase the electrical stimulation frequency until the target intensity is reached.
[0029] If the exercise intervention time has not been reached, the intervention will continue with skeletal muscle status monitoring. If the predetermined time has been reached, the experiment will be terminated. The intervention process triggers a dual decision-making mechanism, prioritizing the matching of exercise patterns and monitoring time thresholds.
[0030] Step S5: Multimodal analysis;
[0031] Cell samples can be obtained by disassembling the bionic movement organ chip layer for immunofluorescence staining, Western blotting, or other related studies.
[0032] Therefore, this invention employs the aforementioned biomimetic motor organ-on-a-chip system and its usage method to construct in vivo and in vitro models that mimic the movement process, improving research efficiency. The models are more human-like, shortening the process from basic research to clinical application and increasing the possibility of translational applications. The highly biomimetic in vivo and in vitro research models effectively achieve crosstalk regulation between different cells / organs during movement, helping to reveal new mechanisms of movement. It realizes biomimetic processes of different exercise intensities, providing new research methods for personalized exercise prescriptions. It is applicable to movement process research in different scenarios and dimensions, exhibiting strong scalability.
[0033] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description
[0034] Figure 1 This is an overall system block diagram of a biomimetic musculoskeletal chip system of the present invention;
[0035] Figure 2 This is a schematic diagram showing the exploded layers of a biomimetic motor organ chip system according to the present invention;
[0036] Figure 3 This is a schematic diagram of the top cover structure in a biomimetic musculoskeletal chip system of the present invention;
[0037] Figure 4 This is a schematic diagram of the wiring terminal of the strain sensor in the top cover of a biomimetic kinetic organ chip system of the present invention;
[0038] Figure 5 This is a schematic diagram of the strain sensor in a biomimetic kinetic organ-on-a-chip system of the present invention;
[0039] Figure 6 This is a schematic diagram of the upper cell culture chamber layer in a biomimetic musculoskeletal chip system of the present invention;
[0040] Figure 7 This is a schematic diagram of the structure of the lower surface of the upper cell culture chamber in a biomimetic musculoskeletal chip system of the present invention;
[0041] Figure 8 This is a schematic diagram of the structure of the cell cavity layer separator in a biomimetic musculoskeletal chip system of the present invention;
[0042] Figure 9 This is a schematic diagram of the lower cell culture chamber layer in a biomimetic musculoskeletal chip system of the present invention;
[0043] Figure 10 This is a schematic diagram of the structure of the lower surface of the cell culture chamber layer in a biomimetic musculoskeletal chip system of the present invention;
[0044] Figure 11 This is a schematic diagram of the bottom electrical stimulation layer of a biomimetic motor organ chip system of the present invention;
[0045] Figure 12 This is a flowchart illustrating the usage method of the biomimetic motor organ chip system of the present invention;
[0046] Figure 13 This is a schematic diagram of the motion control process in the usage method of a biomimetic motor organ chip system of the present invention.
[0047] Figure Labels
[0048] 1. Bionic motion organ-on-a-chip; 2. Dynamic culture medium supply system; 3. Closed-loop motion control system; 4. Data display terminal; 11. Top cover; 12. Upper cell culture chamber; 13. Cell chamber partition layer; 14. Lower cell culture chamber; 15. Bottom electrical stimulation layer; 21. Air pressure source; 22. Microfluidic pressure pump; 23. Cell culture medium and recovery reservoir; 31. Sensor signal acquisition and analysis module; 32. Electrical stimulation generation module; 111. Anchor point; 112. Strain sensor; 113. Strain sensor wiring terminal; 114. Skeletal muscle bundle culture chamber inlet; 115. Skeletal muscle bundle culture chamber outlet; 121. First co-culture cell chamber; 122. Second co-culture cell chamber; 123. Skeletal muscle bundle culture chamber; 124. Porous membrane; 125. Microchannel of the first co-culture cell chamber; 12 6. Microchannel of the second co-culture cell chamber; 131. Connecting hole of the skeletal muscle bundle culture chamber; 141. Third co-culture cell chamber; 142. Fourth co-culture cell chamber; 143. Microchannel of the skeletal muscle bundle culture chamber; 144. Microchannel of the third co-culture cell chamber; 145. Microchannel of the fourth co-culture cell chamber; 146. Groove of electrode terminal; 151. Terminal of the electrical stimulation electrode; 152. Electrode end of the electrical stimulation electrode; 161. Inlet of the first co-culture cell chamber; 162. Inlet of the second co-culture cell chamber; 163. Inlet of the third co-culture cell chamber; 164. Inlet of the fourth co-culture cell chamber; 165. Outlet of the first co-culture cell chamber; 166. Outlet of the second co-culture cell chamber; 167. Outlet of the third co-culture cell chamber; 168. Outlet of the fourth co-culture cell chamber. Detailed Implementation
[0049] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.
[0050] Unless otherwise defined, the technical or scientific terms used in this invention shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.
[0051] The terms "first," "second," and similar words used in this invention do not indicate any order, quantity, or importance, but are merely used to distinguish different components. Terms such as "comprising" or "including" mean that the element or object preceding the word encompasses the elements or objects listed after the word and their equivalents, without excluding other elements or objects. Terms such as "connected" or "linked" are not limited to physical or mechanical connections, but can include electrical connections, whether direct or indirect. Terms such as "upper," "lower," "left," and "right" are used only to indicate relative positional relationships; when the absolute position of the described object changes, the relative positional relationship may also change accordingly.
[0052] Example 1
[0053] like Figures 1-13 As shown, the present invention discloses a bionic motor organ-on-a-chip system and its usage method, comprising a bionic motor organ-on-a-chip 1, a dynamic culture medium supply system 2, a closed-loop motion control system 3, and a data display terminal 4. The wiring terminal 113 of the strain sensor on the top cover 11 of the bionic motor organ-on-a-chip 1 is connected to the sensor signal acquisition and analysis module 31 in the closed-loop motion control system 3, and the wiring terminal 151 of the electrical stimulation electrode on the bottom electrical stimulation layer 15 of the bionic motor organ-on-a-chip 1 is connected to the electrical stimulation generation module 32 in the closed-loop motion control system 3.
[0054] The inlet and outlet of the co-culture cell chamber, the inlet 114 of the skeletal muscle bundle culture chamber, and the outlet 115 of the skeletal muscle bundle culture chamber on the top cover 11 of the biomimetic kinetic organ chip 1 are connected to the cell culture medium and the recovery storage tank 23 in the dynamic culture medium supply system 2. Data interaction occurs between the closed-loop motion control system 3, the dynamic culture medium supply system 2, and the data display terminal 4. The sensor signal acquisition and analysis module 31 monitors the contraction intensity of the skeletal muscle bundles in real time and provides feedback to adjust the output intensity of the electrical stimulation generation module 32.
[0055] The biomimetic motor organ chip 1 includes a top cover 11, an upper cell culture chamber 12, a cell chamber partition layer 13, a lower cell culture chamber 14, and a bottom electrical stimulation layer 15. These components are arranged sequentially from top to bottom. The top cover 11 seals the chip. The cell chamber partition layer 13 and the upper cell culture chamber 12 are stacked in multiple layers, allowing for an expansion of the number of co-culture chambers. The cell chamber partition layer 13 is used to divide the upper cell culture chamber 12 and the lower cell culture chamber 14.
[0056] The dynamic culture medium supply system 2 includes a pressure source 21, a microfluidic pressure pump 22, a cell culture medium, and a recovery reservoir 23. The pressure source 21 is connected to the microfluidic pressure pump 22, and the microfluidic pressure pump 22 is connected to the cell culture medium and the recovery reservoir 23.
[0057] The closed-loop motion control system 3 includes a sensor signal acquisition and analysis module 31 and an electrical stimulation generation module 32. The sensor signal acquisition and analysis module 31 is connected to the terminal 113 of the strain sensor, and the electrical stimulation generation module 32 is connected to the terminal 151 of the electrical stimulation electrode.
[0058] The top cover includes an anchor point 111, a strain sensor 112, a terminal block 113 for the strain sensor, a co-culture cell chamber inlet, a co-culture cell chamber outlet, a skeletal muscle bundle culture chamber inlet 114, and a skeletal muscle bundle culture chamber outlet 115. The top cover 11 has an anchor point 111 and a built-in strain sensor 112. The strain sensor 112 is used to monitor the amplitude of muscle bundle contraction in real time and provide feedback to adjust the electrical stimulation level. The terminal block 113 of the strain sensor is connected to the strain sensor 112 to collect the electrical signal from the strain sensor 112. The anchor point 111 is mechanically coupled to the strain sensor 112, transmitting the muscle bundle contraction force to the strain sensor 112. The co-culture cell chamber inlet, co-culture cell chamber outlet, skeletal muscle bundle culture chamber inlet 114, and skeletal muscle bundle culture chamber outlet 115 are located around the surface of the top cover 11.
[0059] The upper cell culture chamber 12 includes a first co-culture cell chamber 121, a second co-culture cell chamber 122, a skeletal muscle bundle culture chamber 123, a porous membrane 124, a microchannel 125 for the first co-culture cell chamber, a microchannel 126 for the second co-culture cell chamber, a liquid inlet for the co-culture cell chamber, a liquid outlet for the co-culture cell chamber, a liquid inlet 114 for the skeletal muscle bundle culture chamber, and a liquid outlet 115 for the skeletal muscle bundle culture chamber. The skeletal muscle bundle culture chamber 123 is located at the center of the upper cell culture chamber 12. The first co-culture cell chamber 121 provides a relatively independent cell culture environment for the co-cultured cells. The second co-culture cell chamber 122 is another relatively complete chamber, with a functionalized porous membrane 124 on one side, providing a relatively independent cell culture environment for the co-cultured cells. The skeletal muscle bundle culture chamber 123 is separated from the first co-culture cell chamber 121 and the second co-culture cell chamber 122 on both sides by the porous membrane 124. The porous membrane 124 has through-holes, enabling the co-culture of skeletal muscle bundles and other cells. Its internal structure contains a network of silver nanowires, which prevents interference with the electric field of other co-cultured cells during electrical stimulation of the skeletal muscle bundles. A porous membrane 124 is located on one side of the second co-culture cell chamber 122. The upper cell culture chamber 12 contains a first co-culture cell chamber microchannel 125 and a second co-culture cell chamber microchannel 126. The porous membrane 124 contains a silver nanowire network. The first co-culture cell chamber microchannel 125 provides a microchannel for the entry of fresh culture medium and the exit of waste liquid in the first co-culture cell chamber 121, facilitating the entry of fresh culture medium and the exit of waste liquid. The second co-culture cell chamber microchannel 126 provides a microchannel for the entry of fresh culture medium and the exit of waste liquid in the second co-culture cell chamber 122, facilitating the entry of fresh culture medium and the exit of waste liquid.
[0060] The cell chamber partition layer 13 includes a connecting hole 131 for the skeletal muscle bundle culture chamber, a liquid outlet for the co-cultured cell chamber, a liquid inlet for the co-cultured cell chamber, a liquid outlet 114 for the skeletal muscle bundle culture chamber, and a liquid inlet 115 for the skeletal muscle bundle culture chamber. The connecting hole 131 for the skeletal muscle bundle culture chamber is located at the center of the cell chamber partition layer 13. The connecting hole 131 for the skeletal muscle bundle culture chamber allows the skeletal muscle bundle culture chambers in the cell culture chamber extension layer and the cell culture chamber layer to be merged into a single skeletal muscle bundle culture chamber 123.
[0061] The lower cell culture chamber 14 includes a third co-culture cell chamber 141, a skeletal muscle bundle culture chamber 123, a fourth co-culture cell chamber 142, a skeletal muscle bundle culture chamber microchannel 143, a third co-culture cell chamber microchannel 144, a fourth co-culture cell chamber microchannel 145, an electrode terminal groove 146, a co-culture cell chamber outlet, and a co-culture cell chamber inlet. The third and fourth co-culture cell chambers 141 and 142 provide relatively independent cell culture environments for the co-cultured cells. The skeletal muscle bundle culture chamber 123 is located at the center of the lower cell culture chamber 14, and the third and fourth co-culture cell chambers 141 and 142 are located on either side of the skeletal muscle bundle culture chamber 123. The lower cell culture chamber 14 contains the skeletal muscle bundle culture chamber microchannel 143, the third co-culture cell chamber microchannel 144, and the fourth co-culture chamber microchannel 145. The electrode terminal groove 146 is used to achieve complete assembly of skeletal muscle bundles, and the bottom is equipped with platinum electrodes to achieve electrical stimulation of the muscle bundles, so that it can simulate movement.
[0062] The bottom electrical stimulation layer 15 includes a terminal 151 and an electrode tip 152 of an electrical stimulation electrode. The electrode tip 152 directly contacts the skeletal muscle bundles. The terminal 151 of the electrical stimulation electrode is connected to the electrical stimulation generation module 32. The electrode tip 152 is in direct contact with the culture medium of the skeletal muscle bundles, providing stable electrical stimulation to simulate movement. The terminal 151 of the electrical stimulation electrode is connected to the electrical stimulation generation module 32 at the control end, guiding the electrical stimulation current generated by the electrical stimulation generation module 32 into the skeletal muscle bundles, allowing them to be electrically stimulated to simulate movement.
[0063] The co-culture cell chamber inlets on the top cover 11 include a first co-culture cell chamber inlet 161, a second co-culture cell chamber inlet 162, a third co-culture cell chamber inlet 163, and a fourth co-culture cell chamber inlet 164. The co-culture cell chamber outlets on the top cover 11 include a first co-culture cell chamber outlet 165, a second co-culture cell chamber outlet 166, a third co-culture cell chamber outlet 167, and a fourth co-culture cell chamber outlet 168. The co-culture cell chamber inlets and outlets of the upper cell culture chamber 12 have the same structure as those on the top cover 11; the co-culture cell chamber inlets on the cell chamber separator layer 13 include a third co-culture cell chamber inlet 163 and a fourth co-culture cell chamber inlet 164.
[0064] The co-culture cell chamber outlets on the cell chamber partition layer 13 include a third co-culture cell chamber outlet 167 and a fourth co-culture cell chamber outlet 168. The co-culture cell chamber inlets of the lower cell culture chamber 14 include a third co-culture cell chamber inlet 163 and a fourth co-culture cell chamber inlet 164, and the co-culture cell chamber outlets of the lower cell culture chamber 14 include a third co-culture cell chamber outlet 167 and a fourth co-culture cell chamber outlet 168. The skeletal muscle bundle culture chamber is equipped with a skeletal muscle bundle culture chamber inlet 114 and a skeletal muscle bundle culture chamber outlet 115. The co-culture cell chamber inlets and the first co-culture cell chamber outlet facilitate connection to external culture medium supply and waste liquid removal devices, providing fresh culture medium to the co-culture cell chambers.
[0065] A method for using a biomimetic motor organ-on-a-chip system includes the following steps:
[0066] Step S1: System construction and skeletal muscle bundle guidance;
[0067] A mixture of skeletal muscle cells and hydrogel is injected into the chamber of a conventional skeletal muscle chip. Anchor points 111 on the top cover 11 are inserted into the chamber of the mixed conventional skeletal muscle chip. The top cover 111 with integrated anchor points 111 is then closed, and skeletal muscle bundles are cultured. The special structure of the anchor points 111 is used to guide the skeletal muscle cells to grow in a bundle-like structure.
[0068] The skeletal muscle bundle culture chamber inlet 114, the skeletal muscle bundle culture chamber outlet 115, the co-culture cell chamber inlet, and the co-culture cell chamber outlet are connected to the dynamic culture medium supply system 2. The strain sensor terminal 113 is connected to the sensor signal acquisition and analysis module 31, and the electrical stimulation electrode terminal 151 is connected to the electrical stimulation generation module 32; the data display terminal 4 is connected to the dynamic culture medium control system 2.
[0069] Step S2: Asynchronous co-culture of multiple cells;
[0070] During skeletal muscle bundle culture, other cells are seeded in the biomimetic musculoskeletal chip 1; different cell types are independently cultured through cell chamber separators 13, and other cells are seeded in the first co-culture cell chamber 121, the second co-culture cell chamber 122, the third co-culture cell chamber 141, and the fourth co-culture cell chamber 142, respectively. These other cells include endothelial cells, neurons, or immune cells; silver nanowires in the porous membrane 124 shield against electrical stimulation interference; and a microfluidic pressure pump 22 controls the flow rate.
[0071] Step S3: Integration of the bionic kinetic organ chip 1;
[0072] The top cover 11 containing skeletal muscle bundles is moved into the skeletal muscle bundle culture chamber of the bionic kinetic organ chip 1, and connected to the closed-loop motion control system 3 and the dynamic culture medium supply system 2.
[0073] Step S4: Closed-loop motion intervention;
[0074] In the closed-loop motion control system 3, motion parameters are set, electrical stimulation and culture medium circulation are initiated, and the intensity of skeletal muscle bundle contraction is monitored in real time and the intensity of electrical stimulation is dynamically adjusted.
[0075] Exercise parameter settings include setting the exercise mode and exercise intervention time; exercise modes include low-intensity endurance, high-intensity explosive, or scalable modes; when the skeletal muscle fasciculation intensity is between 3% and 10%, it indicates that the muscle is in a suitable exercise state, which is low-intensity endurance mode; when the skeletal muscle fasciculation intensity is not less than 18%, it indicates that the muscle may be in an over-exercise state, which is high-intensity explosive mode; scalable mode generates new exercise type decision rules by inputting a custom contraction intensity threshold through the user interface; the custom contraction intensity threshold includes the skeletal muscle contraction intensity range and the upper limit of the electrical stimulation frequency;
[0076] The strain sensor 112 collects skeletal muscle bundle contraction data in real time, analyzing the current electrical stimulation contraction intensity and frequency. If the current state does not belong to the set exercise mode, the electrical stimulation parameters are dynamically adjusted; if the current state belongs to the set exercise mode, the current stimulation intensity is maintained and exercise intervention continues. When a low-intensity endurance mode is set but a high-intensity explosive mode is detected, the electrical stimulation amplitude is reduced until the skeletal muscle bundle contraction intensity drops back to 3%–10%. When a high-intensity explosive mode is set but the skeletal muscle bundle contraction intensity does not reach 18%, the electrical stimulation frequency is increased until the target intensity is reached.
[0077] If the exercise intervention time is not reached, the experiment returns to skeletal muscle status monitoring. If the intervention continues, the experiment terminates when the predetermined time is reached. The intervention process triggers a dual decision-making mechanism, prioritizing ensuring the matching of exercise patterns and secondarily monitoring time thresholds.
[0078] Step S5: Multimodal analysis;
[0079] Cell samples were obtained by disassembling the first layer of the bionic movement organ chip for immunofluorescence staining, Western blotting, or other related studies.
[0080] Therefore, this invention employs the aforementioned biomimetic motor organ-on-a-chip system and its usage method to construct in vivo and in vitro models that mimic the movement process, improving research efficiency. The models are more human-like, shortening the process from basic research to clinical application and increasing the possibility of translational applications. The highly biomimetic in vivo and in vitro research models effectively achieve crosstalk regulation between different cells / organs during movement, helping to reveal new mechanisms of movement. It realizes biomimetic processes of different exercise intensities, providing new research methods for personalized exercise prescriptions. It is applicable to movement process research in different scenarios and dimensions, exhibiting strong scalability.
[0081] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.
Claims
1. A biomimetic motor organ-on-a-chip system, characterized in that, The device includes a bionic motion organ-on-a-chip, a dynamic culture medium supply system, a closed-loop motion control system, and a data display terminal. The strain sensor on the top cover of the bionic motion organ-on-a-chip is connected to the sensor signal acquisition and analysis module in the closed-loop motion control system. The electrical stimulation electrodes on the bottom electrical stimulation layer of the bionic motion organ-on-a-chip are connected to the electrical stimulation generation module in the closed-loop motion control system. The inlet, outlet, skeletal muscle bundle culture chamber, and outlet of the co-culture cell chamber on the top cover of the bionic motion organ-on-a-chip are connected to the cell culture medium and recovery reservoir in the dynamic culture medium supply system. Data exchange occurs between the closed-loop motion control system, the dynamic culture medium supply system, and the data display terminal. The sensor signal acquisition and analysis module monitors the skeletal muscle bundles and feeds back the data to the electrical stimulation generation module. The bionic motor organ chip includes a top cover, an upper cell culture chamber, a cell chamber partition layer, a lower cell culture chamber, and a bottom electrical stimulation layer; the top cover, upper cell culture chamber, cell chamber partition layer, lower cell culture chamber, and bottom electrical stimulation layer are arranged sequentially from top to bottom, and the cell chamber partition layer and upper cell culture chamber are superimposed in multiple layers; The closed-loop motion control system includes a sensor signal acquisition and analysis module and an electrical stimulation generation module. The sensor signal acquisition module is connected to the terminals of the strain sensor, and the electrical stimulation generation module is connected to the terminals of the electrical stimulation electrodes. The top cover includes anchor points, strain sensors, terminals of the strain sensors, inlet and outlet of the co-culture cell chamber, inlet and outlet of the skeletal muscle bundle culture chamber, and outlet of the skeletal muscle bundle culture chamber. The top cover is equipped with anchor points and has built-in strain sensors. The anchor points are made of flexible material and are mechanically coupled to the strain sensors. The inlet and outlet of the co-culture cell chamber, the inlet and outlet of the skeletal muscle bundle culture chamber are located around the surface of the top cover. The upper cell culture chamber includes a first co-culture cell chamber, a second co-culture cell chamber, a skeletal muscle bundle culture chamber, a porous membrane, microchannels for the first and second co-culture cell chambers, a liquid inlet for the co-culture cell chamber, a liquid outlet for the co-culture cell chamber, a liquid inlet for the skeletal muscle bundle culture chamber, and a liquid outlet for the skeletal muscle bundle culture chamber. The skeletal muscle bundle culture chamber is located at the center of the upper cell culture chamber. The skeletal muscle bundle culture chamber is separated from the first and second co-culture cell chambers on both sides by a porous membrane. The upper cell culture chamber contains microchannels for the first and second co-culture cell chambers. The porous membrane contains a silver nanowire mesh structure. The cell compartment partition layer includes a connecting hole for the skeletal muscle bundle culture chamber, a liquid outlet for the co-cultured cell chamber, a liquid inlet for the co-cultured cell chamber, a liquid outlet for the skeletal muscle bundle culture chamber, and a liquid inlet for the skeletal muscle bundle culture chamber. The connecting hole for the skeletal muscle bundle culture chamber is located at the center of the cell compartment partition layer. The lower cell culture chamber includes a third co-culture cell chamber, a skeletal muscle bundle culture chamber, a fourth co-culture cell chamber, and a skeletal muscle bundle culture chamber microchannel. The lower cell culture chamber also includes an electrode terminal groove, a co-culture cell chamber outlet, and a co-culture cell chamber inlet. The skeletal muscle bundle culture chamber is located in the center of the lower cell culture chamber. The third and fourth co-culture cell chambers are located on either side of the skeletal muscle bundle culture chamber. The lower cell culture chamber contains the skeletal muscle bundle culture chamber microchannel, the third co-culture cell chamber microchannel, and the fourth co-culture chamber microchannel. The bottom electrical stimulation layer includes the terminals of the electrical stimulation electrodes and the electrode ends of the electrical stimulation electrodes. The electrode ends of the electrical stimulation electrodes directly contact the skeletal muscle bundles. The terminals of the electrical stimulation electrodes are connected to the electrical stimulation generation module.
2. The bionic musculoskeletal chip system according to claim 1, characterized in that, The dynamic culture medium supply system includes a gas pressure source, a microfluidic pressure pump, and a recovery storage tank. The gas pressure source is connected to the microfluidic pressure pump, and the microfluidic pressure pump is connected to the recovery storage tank and the cell culture medium.
3. The bionic musculoskeletal chip system according to claim 1, characterized in that, The co-culture cell chamber inlets on the top cover include a first co-culture cell chamber inlet, a second co-culture cell chamber inlet, a third co-culture cell chamber inlet, and a fourth co-culture cell chamber inlet; the co-culture cell chamber outlets on the top cover also include a first co-culture cell chamber outlet, a second co-culture cell chamber outlet, a third co-culture cell chamber outlet, and a fourth co-culture cell chamber outlet; the structures of the co-culture cell chamber inlets and outlets in the upper cell culture chamber are the same as those on the top cover; the co-culture cell chamber inlets on the cell chamber partition layer include a third... The co-culture cell chamber has a liquid inlet and a fourth co-culture cell chamber inlet; the co-culture cell chamber outlets on the cell chamber partition layer include the third co-culture cell chamber outlet and the fourth co-culture cell chamber outlet; the lower cell culture chamber has co-culture cell chamber inlets including the third co-culture cell chamber inlet and the fourth co-culture cell chamber outlet, and the lower cell culture chamber has co-culture cell chamber outlets including the third co-culture cell chamber outlet and the fourth co-culture cell chamber outlet; the skeletal muscle bundle culture chamber is equipped with a skeletal muscle bundle culture chamber inlet and a skeletal muscle bundle culture chamber outlet.
4. A method of using a biomimetic musculoskeletal-on-a-chip system as described in any one of claims 1-3, the method being used for non-therapeutic and diagnostic purposes, characterized in that... Includes the following steps: Step S1: Construct skeletal muscle bundles; A mixture of skeletal muscle cells and hydrogel is injected into the chamber of a conventional skeletal muscle chip. Anchors on the top cover are inserted into the matching chamber of the conventional skeletal muscle chip. The top cover with integrated anchors is then closed, and skeletal muscle bundles are cultured. The structure of the anchor point is used to guide the skeletal muscle cells to grow in a bundle-like structure. Step S2: Asynchronous co-culture of multiple cells; During skeletal muscle bundle culture, other cells are seeded in the biomimetic musculoskeletal chip; different cell types are cultured independently through cell compartment septa. Other cells include endothelial cells, neurons, or immune cells; the porous membrane's silver nanowires shield against electrical stimulation interference; Step S3: Integration of bionic musculoskeletal chip; The top cover containing skeletal muscle bundles was moved into the skeletal muscle bundle culture chamber of the bionic musculoskeletal organ chip and connected to the closed-loop motion control system and the dynamic culture medium supply system. Step S4: Closed-loop motion intervention; In the closed-loop motion control system, motion parameters are set, electrical stimulation and culture medium circulation are initiated, and the intensity of skeletal muscle bundle contraction is monitored in real time and the intensity of electrical stimulation is dynamically adjusted. Exercise parameter settings include setting the exercise mode and exercise intervention time; the exercise mode includes low-intensity endurance, high-intensity explosive, or scalable mode; when the skeletal muscle fasciculation intensity is between 3% and 10%, it is low-intensity endurance mode, and when the skeletal muscle fasciculation intensity is not less than 18%, it is high-intensity explosive mode; the scalable mode generates new exercise type decision rules by inputting a custom contraction intensity threshold through the user interface; the custom contraction intensity threshold includes the skeletal muscle contraction intensity range and the upper limit of the electrical stimulation frequency; Real-time data on skeletal muscle bundle contraction is collected using strain sensors to analyze the current electrical stimulation contraction intensity and frequency. If the current state does not belong to the set exercise mode, the electrical stimulation parameters are dynamically adjusted; If the current state belongs to the set exercise mode, then maintain the current stimulus intensity and continue the exercise intervention; When a low-intensity endurance mode is set but a high-intensity burst mode is detected, the electrical stimulation amplitude is reduced until the contraction intensity of the skeletal muscle bundles drops back to 3%~10%; when a high-intensity burst mode is set but the contraction intensity of the skeletal muscle bundles does not reach 18%, the electrical stimulation frequency is increased until the target intensity is reached. If the exercise intervention time has not been reached, the intervention will continue with skeletal muscle status monitoring. If the predetermined time has been reached, the experiment will be terminated. The intervention process triggers a dual decision-making mechanism, prioritizing the matching of exercise patterns and monitoring time thresholds. Step S5: Multimodal analysis; Cell samples were obtained by disassembling the bionic movement organ chip layer for immunofluorescence staining, Western blotting, or other related research analyses.