A hydrogel microstructure preparation device

Abstract

The utility model discloses a kind of hydrogel microstructure preparation devices, including liftable support assembly, motor, liquid accumulator and collection pool;The motor is installed in the lifting end of the support assembly, and the output end of the motor is connected with the liquid accumulator, and the collection pool is arranged below the liquid accumulator;The liquid accumulator includes liquid storage cylinder and one or more capillary tubes connected with the bottom of liquid storage cylinder in an inclined manner.The device provided by the utility model has simple structure, is easy to use, can prepare hydrogel microstructures of different sizes and different forms, and has advantages of low cell damage, high structural uniformity, high preparation flux and high operation stability when used for 3D cell culture and cell scaffold construction.

Description

Technical Field

[0001] This invention belongs to the field of bioengineering technology, specifically relating to a device for preparing hydrogel microstructures. Background Technology

[0002] In the field of biomedical engineering, 3D cell culture technology has become one of the core technologies in cell biology research, drug screening, and tissue engineering because it can simulate the three-dimensional microenvironment of cell growth in vivo and more realistically reflect cell physiological functions. As a key carrier for 3D cell culture, the structure and morphology of cell scaffolds (such as the uniformity of microsphere size, the diameter and pore distribution of fibers) directly affect the adhesion, proliferation and differentiation efficiency of cells. Hydrogels have become the core material for preparing microstructures (microspheres, fibers) for cell scaffolds due to their good biocompatibility, biomimetic mechanical properties and biodegradability.

[0003] Currently, various technical methods have been developed for preparing hydrogel microstructures (microspheres, fibers), but they still have significant limitations when adapted to the actual needs of 3D cell culture and cell scaffold construction, as follows: Emulsification (for hydrogel microspheres): Traditional emulsification disperses the hydrogel precursor solution in the oil phase to form droplets through mechanical stirring, which then solidify to obtain microspheres. However, this method requires the use of large amounts of organic solvents (such as paraffin and chloroform), which can easily leave residues and damage cell viability, making it difficult to meet the "non-toxic" requirement for 3D cell culture. At the same time, the uneven shear force generated by stirring leads to large differences in microsphere size (the coefficient of variation often exceeds 20%), making it impossible to form a uniform cell scaffold and affecting the consistency of cell growth.

[0004] Electrospinning (for hydrogel fibers): Electrospinning uses high voltage (usually 10-30kV) to form a jet stream from the hydrogel precursor solution, which then solidifies to obtain fibers. However, high voltage can easily cause electrical damage to sensitive cells (such as stem cells and primary cells), resulting in a cell survival rate of less than 60%. Furthermore, the diameter of fibers prepared by this method is easily affected by voltage fluctuations, with a deviation of more than 30% in the diameter of a single fiber. The unstable pore structure of the scaffold is not conducive to nutrient exchange and metabolic waste removal by cells.

[0005] Microfluidic methods (for hydrogel microspheres / fibers): Microfluidic methods precisely control the fluid interface through microchannels, enabling the fabrication of hydrogel microstructures with uniform particle size / diameter. However, this method relies on precise microchannel design, resulting in extremely low throughput per batch (typically only milligrams per hour), making it difficult to meet the needs of large-scale cell scaffold construction. Furthermore, microchannels are prone to clogging due to viscosity changes in the hydrogel precursor solution, leading to poor operational stability and increasing the reproducibility challenges of 3D cell culture experiments.

[0006] In summary, existing hydrogel microstructure preparation technologies cannot fully meet the requirements of 3D cell culture for carriers and the application scenarios of large-scale cell scaffold construction in terms of key indicators such as cell compatibility, structural uniformity, and preparation throughput. There is an urgent need for a hydrogel microstructure preparation device that can balance low cell damage, high structural uniformity, and high preparation efficiency to promote the practical application of 3D cell culture technology and tissue engineering. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of the existing technology and provide a hydrogel microstructure preparation device with simple structure and convenient use. It can prepare hydrogel microstructures of different sizes and shapes. When used for cell 3D culture and cell scaffold construction, it has the advantages of low cell damage, high structural uniformity, high preparation throughput and high operational stability.

[0008] This utility model provides the following technical solution: A hydrogel microstructure preparation device includes a liftable support assembly, a motor, a liquid reservoir, and a collection pool; The motor is installed on the lifting end of the support assembly, the output end of the motor is connected to the liquid storage tank, and the collection pool is located below the liquid storage tank; The liquid reservoir includes a liquid reservoir cylinder and one or more capillary tubes that are inclinedly connected to the bottom of the liquid reservoir cylinder.

[0009] Furthermore, the bottom of the liquid storage cylinder is provided with a built-in cone, and the connection between the cone and the liquid storage cylinder is provided with a through hole for installing a capillary tube. The inclination angle of the capillary tube is the same as the inclination angle of the cone surface of the cone.

[0010] Furthermore, the capillary includes a main body, one end of which is sealed and installed in the through hole, and the other end is connected to a tube opening with a gradually decreasing size.

[0011] Furthermore, the angle between the conical surface of the cone and the bottom surface of the liquid storage cylinder is 30°~60°.

[0012] Furthermore, the output end of the motor is connected to the liquid reservoir via a connector.

[0013] Furthermore, the support assembly includes a base, a bracket mounted on the base, and a support rod slidably sleeved on the bracket. One end of the support rod is fixed to the bracket by a knob mounted thereon, and the other end is equipped with the motor.

[0014] Furthermore, the motor is a hollow brushless motor, and the connection end of the support rod to the motor is provided with a hollow groove corresponding to the position of the hollow cavity of the motor.

[0015] Furthermore, it also includes a main control board, a motor driver, and a potentiometer. The main control board is connected to the motor driver, and the motor driver is connected to the motor and the potentiometer respectively. The potentiometer is used to adjust the motor speed.

[0016] Furthermore, it also includes a power supply, which is connected to both the main control board and the motor driver.

[0017] Compared with the prior art, the beneficial effects of this utility model are: (1) The hydrogel microstructure preparation device provided by this utility model has a simple structure. The components such as the collection pool, storage cylinder, and capillary tube can be simply connected and installed. It is easy to assemble and use, and significantly improves the efficiency of device construction. (2) In this invention, the reservoir is driven to rotate by a motor. The hydrogel precursor solution in the reservoir is subjected to the combined shearing action of centrifugal force and gravity. It flows out through the capillary and is thrown into the continuous phase solution, thereby realizing the preparation of hydrogel microstructures. Hydrogel microspheres can be prepared by controlling the capillary to be above the continuous phase solution by the support component. Fibers can be prepared by controlling the capillary opening to be in the continuous phase solution. The rotation speed of the reservoir is adjusted by the motor. Under the action of different centrifugal forces generated at different rotation speeds, the hydrogel precursor solution at the capillary opening will be subjected to different shear forces, thereby realizing the preparation of hydrogel microstructures of different sizes. (3) When the hydrogel microstructure preparation device provided by this utility model is used for 3D cell culture and cell scaffold construction, it has advantages such as low cell damage, high structural uniformity, high preparation throughput and high operational stability, as follows: Low cell damage: The formation of microstructures from hydrogel precursor solutions is driven by centrifugal force, avoiding the use of organic solvents (such as emulsification) and high voltage (such as electrospinning), eliminating residual toxicity and electrical damage to cells, providing a non-toxic and mild microstructure preparation environment for 3D cell culture, and improving cell survival rate; High structural uniformity: By utilizing the stability and controllability of centrifugal force, the particle size of hydrogel microspheres and the diameter of fibers are precisely controlled, reducing the coefficient of variation of microstructure size and ensuring that the prepared hydrogel microstructures have consistent morphology and properties, meeting the requirements of cell scaffolds for structural uniformity and ensuring the stability of the cell growth environment. High throughput and operational stability: By designing a reservoir and collection pool, the low throughput limitation of microfluidic methods is overcome, enabling efficient batch preparation of hydrogel microstructures; at the same time, the problem of microchannel blockage is avoided, enhancing the stability and repeatability of device operation and meeting the practical application needs of large-scale cell scaffold construction. Attached Figure Description

[0018] Figure 1This is a schematic diagram of the hydrogel microstructure preparation device in an embodiment of this utility model; Figure 2 This is a schematic diagram of the structure of the motor and the liquid reservoir in an embodiment of this utility model; Figure 3 This is a top view of the liquid reservoir in an embodiment of this utility model; Figure 4 yes Figure 3 Cross-sectional view along the AA direction; Figure 5 yes Figure 4 Enlarged view of the structure of section I; Figure 6 This is a schematic diagram of the hydrogel microstructure preparation process in Embodiment 2 of this utility model; Figure 7 This is a schematic diagram of the working state of the hydrogel microstructure preparation device in Embodiment 3 of this utility model; Figure 8 This is a fluorescence image of the hydrogel microspheres prepared in Example 3 of this utility model; Figure 9 This is a graph showing the change in diameter of the hydrogel microspheres prepared in Embodiment 3 of this utility model as a function of motor speed; Figure 10 This is a schematic diagram of the working state of the hydrogel microstructure preparation device in Embodiment 4 of this utility model; Figure 11 This is a fluorescence image of the hydrogel fiber prepared in Example 4 of this utility model; Figure 12 This is a graph showing the change in diameter of the hydrogel fiber prepared in Embodiment 4 of this utility model as a function of motor speed.

[0019] The following are labeled in the diagram: 1. Power supply; 2. Motor; 3. Wire; 4. Motor driver; 5. Bracket; 6. Main control board; 7. Potentiometer; 8. Collection tank; 9. Liquid reservoir; 10. Base; 11. Support rod; 12. Knob; 13. Connector; 14. Hydrogel precursor solution; 91. Liquid reservoir; 92. Capillary tube; 93. Conical body; 921. Main tube; 922. Tube opening. Detailed Implementation

[0020] The present invention will be further described below with reference to the accompanying drawings. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and should not be used to limit the scope of protection of the present invention.

[0021] It should be noted that in the description of this utility model, the terms "front," "rear," "left," "right," "upper," "lower," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. These terms are used only for the convenience of describing this utility model and do not require that this utility model be constructed and operated in a specific orientation; therefore, they should not be construed as limitations on this utility model. In the description of this utility model, unless otherwise stated, "a plurality of" means two or more.

[0022] Example 1

[0023] like Figures 1-5 As shown, this embodiment provides a hydrogel microstructure preparation device, including a liftable support assembly, a motor 2, a liquid reservoir 9, a collection pool 8, a main control board 6, a motor driver 4, a potentiometer 7, and a power supply 1.

[0024] Motor 2 is installed at the lifting end of the support assembly, and its output end is connected to the liquid reservoir 9. Specifically, the support assembly includes a base 10, a bracket 5 mounted on the base 10, and a support rod 11 slidably fitted onto the bracket 5. One end of the support rod 11 is fixed to the bracket 5 via a knob 12 mounted thereon, and the other end is fitted with motor 2. When the knob 12 is loosened, the support rod 11 can slide along the bracket 5, thereby adjusting the height of motor 2 and liquid reservoir 9. Once motor 2 and liquid reservoir 9 are adjusted to a suitable height, tightening the knob 12 can fix their positions.

[0025] In this embodiment, power is supplied to both the main control board 6 and the motor driver 4 via a power supply 1. The main control board 6 is connected to the motor driver 4, which in turn is connected to the motor 2 and the potentiometer 7. All these connections are made electrically using wires 3. The potentiometer 7 is used to adjust the speed of the motor 2, and the motor driver 4 sends the motor 2 speed signal to the main control board 6. In this embodiment, the main control board 6 is equipped with a screen that can display the motor speed in real time, facilitating the adjustment of the motor 2 speed at the potentiometer 7.

[0026] In this embodiment, motor 2 is a hollow brushless motor. The connection end of the support rod 11 to motor 2 is provided with a hollow groove corresponding to the position of the hollow cavity of motor 2, which facilitates the addition of hydrogel precursor solution through the hollow groove and the hollow cavity of motor 2. This utility model abandons the complex design of traditional multi-drive modules (such as fluid pumps and voltage regulation modules), and realizes centrifugal drive control with only a single hollow brushless DC motor. The motor can achieve precise speed adjustment through pulse width modulation (PWM) technology, and the output end of motor 2 is directly connected to the liquid reservoir 9 through connector 13. The liquid reservoir 9 is installed on connector 13 by friction, resulting in high power transmission efficiency and no energy loss or failure risk of intermediate transmission components. This single-motor drive design not only simplifies the circuit and mechanical structure of the device, but also greatly simplifies the operation process. The operator only needs to directly adjust the speed through a potentiometer 7, and motor 2 can automatically and stably output the corresponding centrifugal force without the need for additional adjustment of fluid pressure, voltage and other parameters, which significantly improves the ease of use of the equipment.

[0027] The reservoir 9 is used to store the hydrogel precursor solution, including a reservoir cylinder 91 and one or more capillary tubes 92 inclinedly connected to the bottom of the reservoir cylinder 91. Driven by the motor 2, the reservoir 9 rotates, and the hydrogel precursor solution in the reservoir 9 is subjected to the combined shearing action of centrifugal force and gravity, flowing out through the capillary tubes 92 and being thrown into the continuous phase solution, thus realizing the preparation of hydrogel microstructures. When preparing hydrogel microspheres, multiple arrayed capillary tubes 92 can be used, and the capillary tubes 92 can be positioned above the continuous phase solution by a support component, thus achieving the one-time preparation of a large number of hydrogel microspheres. When preparing hydrogel fibers, by controlling the opening of the capillary tube 92 to be in the continuous phase solution, only one capillary tube can be used, preventing the formation of multiple fibers due to excessive capillary tubes and resulting fiber entanglement.

[0028] The bottom of the liquid storage cylinder 91 is equipped with a built-in conical body 93. A through hole for installing a capillary tube 92 is provided at the connection between the conical body 93 and the liquid storage cylinder 91. The inclination angle of the capillary tube 92 is the same as the inclination angle of the conical surface of the conical body 93. The angle between the conical surface of the conical body 93 and the bottom surface of the liquid storage cylinder 91 is 30°~60°. An excessively large angle will affect the centrifugation effect; an excessively small angle will cause significant fluctuations in the surface of the continuous phase solution when preparing fibers, as the liquid storage cylinder 93 rotates.

[0029] The capillary tube 92 includes a main body 921, one end of which is sealed and installed in a through hole at the bottom of the liquid storage cylinder 91, and the other end is connected to a tube opening 922 with a gradually narrowing opening. In this embodiment, the capillary tube 92 is inserted into the through hole at the bottom of the liquid storage cylinder 91, its surface is coated with UV adhesive, and it is then irradiated with UV light to fix the capillary tube 92 and the liquid storage cylinder 91 together. In some other embodiments, an integrated liquid storage structure can also be directly fabricated.

[0030] A collection tank 8 is located below the liquid reservoir 9 and is used to store the continuous phase solution. In this embodiment, the collection tank 8 can be configured as an open container.

[0031] The hydrogel microstructure preparation device provided by this utility model has a simple structure. The components such as the collection pool 8, the liquid storage cylinder 91, and the capillary tube 92 can be simply connected and installed during use. It is easy to assemble and use, and significantly improves the efficiency of device construction.

[0032] Example 2

[0033] like Figure 6 As shown, this embodiment provides a method for preparing hydrogel microstructures, using the hydrogel microstructure preparation apparatus described in Example 1, including the following steps: S1. The collection tank, liquid storage cylinder, connector and capillary tube are made by 3D printing integrated molding process, and then assembled according to the device shown in Example 1. S2. Inject the hydrogel precursor solution into the reservoir and inject the continuous phase solution into the collection tank. S3. Adjust the height of the reservoir by using the support components to position the capillary above the continuous phase solution, and prepare microspheres of different particle sizes by adjusting the speed of the motor. Alternatively, by adjusting the height of the reservoir using the support components, the capillary orifice can be positioned in the continuous phase solution, and fibers of different diameters can be prepared by adjusting the motor speed.

[0034] Example 3

[0035] This embodiment uses the hydrogel microstructure preparation device described in Example 1 to prepare hydrogel microspheres.

[0036] (1) Design of device structure dimensions.

[0037] The reservoir has a height of 83 mm, an outer diameter of 26 mm, and an inner diameter of 24 mm. The angle α between the conical surface of the cone at the bottom of the reservoir and the bottom surface of the reservoir is 45°. There are four capillary tubes, evenly distributed at the bottom of the reservoir. The main tube has a length b of 10 mm, an outer diameter c of 1.5 mm, an inner diameter d of 1.1 mm, and an end diameter e of 0.6 mm.

[0038] (2) Sample preparation.

[0039] Preparation of hydrogel precursor solution: Sodium alginate and deionized water were mixed in a mass ratio of 0.75:99.25 in a beaker and magnetically stirred at 60°C (500 r / min) until transparent and free of bubbles to obtain an alginate solution.

[0040] Preparation of continuous phase solution: Anhydrous calcium chloride and deionized water were mixed in a beaker at a mass ratio of 3:97 and allowed to dissolve completely to obtain a calcium chloride solution.

[0041] (3) Preparation of hydrogel microspheres.

[0042] Insert the tubing through the hollow cavity of the motor into the reservoir, inject the alginate solution through the tubing, and pour the calcium chloride solution into the collection tank; Figure 7 As shown, the capillary is positioned above the calcium chloride solution by adjusting the height of the support; the motor is turned on, and the motor speed is controlled by adjusting the potentiometer. The alginate solution in the reservoir is subjected to the combined shearing action of centrifugal force and gravity, flowing out through the capillary and being thrown into the calcium chloride solution, thus achieving cross-linking to form hydrogel microspheres.

[0043] Hydrogel microspheres of different particle sizes can be prepared by adjusting a potentiometer to control the motor to drive the reservoir at different speeds. Figure 8 and Figure 9 As shown, when the rotation speed varies in the range of 0~500 r / min, the particle size of the prepared hydrogel microspheres is between 900~2700 μm.

[0044] Example 4

[0045] This embodiment uses the hydrogel microstructure preparation device described in Example 1 to prepare hydrogel fibers.

[0046] (1) Design of device structure dimensions.

[0047] The reservoir has a height of 83 mm, an outer diameter of 26 mm, and an inner diameter of 24 mm. The angle α between the conical surface of the bottom cone and the bottom surface of the reservoir is 45°. There is one capillary tube with a main body length b of 10 mm, an outer diameter c of 1.5 mm, an inner diameter d of 1.1 mm, and an end diameter e of 0.6 mm.

[0048] (2) Sample preparation.

[0049] Preparation of hydrogel precursor solution: Sodium alginate and deionized water were mixed in a mass ratio of 0.75:99.25 in a beaker and magnetically stirred at 60°C (500 r / min) until transparent and free of bubbles to obtain an alginate solution.

[0050] Preparation of continuous phase solution: Anhydrous calcium chloride and deionized water were mixed in a beaker at a mass ratio of 3:97 and allowed to dissolve completely to obtain a calcium chloride solution.

[0051] (3) Preparation of hydrogel fibers.

[0052] Insert the tubing through the hollow cavity of the motor into the reservoir, inject the alginate solution through the tubing, and pour the calcium chloride solution into the collection tank; Figure 10 As shown, the capillary is positioned below the surface of the calcium chloride solution by adjusting the height of the support; the motor is turned on, and the motor speed is controlled by adjusting the potentiometer. The alginate solution in the reservoir is subjected to the combined shearing action of centrifugal force and gravity, flowing out through the capillary and being thrown into the calcium chloride solution, thereby achieving cross-linking to form hydrogel fibers.

[0053] Hydrogel fibers of different diameters can be prepared by adjusting a potentiometer to control the motor to drive the reservoir at different speeds. For example... Figure 11 and Figure 12 As shown, when the rotation speed varies in the range of 0~500 r / min, the diameter of the prepared hydrogel fibers is between 50~150 μm.

[0054] The hydrogel microstructure preparation device provided by this invention has advantages such as low cell damage, high structural uniformity, high preparation throughput, and high operational stability when used for 3D cell culture and cell scaffold construction, as detailed below: Low cell damage: The formation of microstructures from hydrogel precursor solutions is driven by centrifugal force, avoiding the use of organic solvents (such as emulsification) and high voltage (such as electrospinning), eliminating residual toxicity and electrical damage to cells, providing a non-toxic and mild microstructure preparation environment for 3D cell culture, and improving cell survival rate; High structural uniformity: By utilizing the stability and controllability of centrifugal force, the particle size of hydrogel microspheres and the diameter of fibers are precisely controlled, reducing the coefficient of variation of microstructure size and ensuring that the prepared hydrogel microstructures have consistent morphology and properties, meeting the requirements of cell scaffolds for structural uniformity and ensuring the stability of the cell growth environment. High throughput and operational stability: By designing a reservoir and collection pool, the low throughput limitation of microfluidic methods is overcome, enabling efficient batch preparation of hydrogel microstructures; at the same time, the problem of microchannel blockage is avoided, enhancing the stability and repeatability of device operation and meeting the practical application needs of large-scale cell scaffold construction.

[0055] The above description is only a preferred embodiment of the present utility model. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present utility model, and these improvements and modifications should also be considered within the protection scope of the present utility model.

Claims

1. A device for preparing hydrogel microstructures, characterized in that, Includes a liftable support assembly, a motor, a liquid reservoir, and a collection tank; The motor is installed on the lifting end of the support assembly, the output end of the motor is connected to the liquid storage tank, and the collection pool is located below the liquid storage tank; The liquid reservoir includes a liquid reservoir cylinder and one or more capillary tubes that are inclinedly connected to the bottom of the liquid reservoir cylinder.

2. The hydrogel microstructure preparation apparatus according to claim 1, characterized in that, The bottom of the liquid storage cylinder is provided with a built-in cone, and the connection between the cone and the liquid storage cylinder is provided with a through hole for installing a capillary tube. The inclination angle of the capillary tube is the same as the inclination angle of the cone surface of the cone.

3. The hydrogel microstructure preparation apparatus according to claim 2, characterized in that, The capillary includes a main body, one end of which is sealed and installed in the through hole, and the other end is connected to a tube opening with a gradually decreasing opening. And / or, the angle between the conical surface of the cone and the bottom surface of the liquid storage cylinder is 30°~60°.

4. The hydrogel microstructure preparation apparatus according to claim 1, characterized in that, The output end of the motor is connected to the liquid reservoir via a connector.

5. The hydrogel microstructure preparation apparatus according to claim 1, characterized in that, The support assembly includes a base, a bracket on the base, and a support rod that is slidably sleeved on the bracket. One end of the support rod is fixed to the bracket by a knob installed thereon, and the other end is equipped with the motor.

6. The hydrogel microstructure preparation apparatus according to claim 5, characterized in that, The motor is a hollow brushless motor, and the support rod is provided with a hollow groove at the connection end with the motor that corresponds to the position of the hollow cavity of the motor.

7. The hydrogel microstructure preparation apparatus according to claim 1, characterized in that, It also includes a main control board, a motor driver, and a potentiometer. The main control board is connected to the motor driver, and the motor driver is connected to the motor and the potentiometer respectively. The potentiometer is used to adjust the motor speed.

8. The hydrogel microstructure preparation apparatus according to claim 7, characterized in that, It also includes a power supply, which is connected to the main control board and the motor driver respectively.

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