Device for preparing hydrogel factor concentration gradient and preparation method thereof

Abstract

The invention relates to the technical field of biological material preparation, and discloses a device for preparing hydrogel factor concentration gradient and a preparation method thereof.The device comprises a mixing container, an adapter connected with the mixing container and a guide-in device connected with the adapter; a magnetic stirring device is arranged on the adapter and comprises a magneton arranged in the adapter, a magnetic pole arranged at the top of the adapter and a power supply connected with the magnetic pole; dense holes are formed in the connecting surface of the adapter and the mixing container; the leading-in device comprises an outer pipe and at least one nesting pipe arranged in the outer pipe; one end of the nesting pipe penetrates out of the outer wall of the outer pipe; and a speed change device and a micro pump are arranged on the nesting pipe and the outer pipe. The adapter is a cylinder, at least two magnetic poles are arranged, and the magnetic poles are annularly arrayed on the top face of the adapter with the geometric center of the top face of the adapter as the circle center. A silica gel layer is arranged on the outer side of the magneton. According to the hydrogel material prepared by the device and the method, the concentration distribution of the bioactive factors in the hydrogel material shows good gradient.

Description

Technical Field

[0001] The invention relates to the technical field of biomaterial preparation, and in particular to a device for preparing a hydrogel factor concentration gradient and a preparation method thereof. Background Art

[0002] As a polymer material with a three-dimensional network structure, hydrogel has shown broad application prospects in the biomedical field due to its unique swelling, biocompatibility and controllability. Hydrogel can absorb a large amount of water in water and keep its three-dimensional structure insoluble. This property gives it unique advantages in drug delivery, tissue engineering, cell culture and regenerative medicine. As a drug carrier, hydrogel can control the release rate of drugs, achieve targeted drug delivery, improve therapeutic effects and reduce side effects. As a mimetic of the extracellular matrix, hydrogel can provide cells with a three-dimensional growth environment, promote cell proliferation, differentiation and tissue regeneration. The hydrogel matrix can simulate the microenvironment of cell growth in the body, provide an ideal platform for cell culture, and help study cell behavior, cell-cell interactions, etc. By constructing a hydrogel scaffold with a specific structure and function, tissue regeneration can be guided and damaged tissues or organs can be repaired.

[0003] At present, there are still some unresolved technical problems in the preparation technology of hydrogel factor concentration gradient: during the preparation process, the crosslinking agent has poor dispersion effect, resulting in uneven crosslinking points, which in turn makes the regularity of the polymer three-dimensional network structure poor, which will significantly affect the mechanical properties and response performance of the hydrogel; the preparation method is relatively cumbersome and requires multiple steps such as solution preparation, mixing, and centrifugation, which not only increases the difficulty of preparation, but also introduces additional errors. The existing preparation method cannot maintain the stability of the gradient for a long time, resulting in large fluctuations in the performance of the final product. Summary of the invention

[0004] In view of the above-mentioned deficiencies in the prior art, the present invention provides a device for preparing a hydrogel factor concentration gradient, which solves the problems of uneven dispersion, complicated preparation steps, poor stability and controllability of the concentration gradient in the prior art.

[0005] In order to achieve the above-mentioned invention object, the technical solution adopted by the present invention is: a device for preparing a hydrogel factor concentration gradient, comprising a mixing container, a adapter connected to the mixing container, and an introduction device connected to the adapter;

[0006] A magnetic stirring device is arranged on the adapter, and the magnetic stirring device comprises a magnet arranged inside the adapter, a magnetic pole arranged on the top of the adapter, and a power supply connected to the magnetic pole; a dense hole is arranged on the connection surface between the adapter and the mixing container;

[0007] The introduction device comprises an outer tube and at least one nested tube arranged inside the outer tube; one end of the nested tube passes through the outer wall of the outer tube; and a speed change device and a micro pump are arranged on the nested tube and the outer tube.

[0008] The invention has the following beneficial effects: the device realizes accurate introduction and flow control of multiple hydrogel factor solutions by combining the nested tubes with the speed change device and the micro pump. The speed change device can adjust the solution flow rate according to the experimental requirements, while the micro pump ensures the stable supply of the solution. This design enables the formation of a precise and controllable concentration gradient in the mixing container.

[0009] Driven by the magnetic poles, the magnets perform efficient contactless stirring inside the mixing container, avoiding impurities and contamination that may be introduced by traditional mechanical stirring. At the same time, magnetic stirring can generate uniform eddy currents, promoting the uniform distribution and full mixing of hydrogel factors in the solution, thereby further improving the accuracy and stability of the concentration gradient.

[0010] Furthermore, in the above-mentioned device for preparing a hydrogel factor concentration gradient, the adapter is a cylinder, at least two magnetic poles are provided, the magnetic poles have the geometric center of the top surface of the adapter as the center of the circle, and the annular array is on the top surface of the adapter.

[0011] The beneficial effect of adopting the above further scheme is that the magnetic pole configuration of the annular array can effectively promote the uniform mixing of the pure hydrogel precursor solution and the hydrogel precursor solution containing the bioactive factor. Under the action of the magnetic field, the magnetons will be affected by the magnetic force and move along the direction of the magnetic field. By adjusting the parameters such as the strength, spacing and rotation speed of the magnetic poles, the magnetic field gradient can be precisely controlled.

[0012] Furthermore, in the above-mentioned device for preparing a hydrogel factor concentration gradient, a silica gel layer is arranged on the outer side of the magnet.

[0013] The beneficial effects of adopting the above further solution are: the silicone layer can effectively wrap the magnet, reducing the direct contact between the magnet and the inner wall of the container during rotation or vibration, thereby avoiding debris or wear caused by friction and collision. The softness and elasticity of the silicone layer can ensure that the magnet rotates more smoothly, reducing vibration and noise.

[0014] Furthermore, in the above-mentioned device for preparing a hydrogel factor concentration gradient, a speed display screen is provided on the speed change device.

[0015] The beneficial effect of adopting the above further scheme is that a speed display screen is provided on the speed change device. The inflow speed of the pure hydrogel precursor solution and the hydrogel precursor solution containing the bioactive factor can be observed. By simultaneously monitoring the inflow speed of the pure hydrogel precursor solution and the solution containing the bioactive factor, the experimenter can more accurately control the ratio and distribution of the two solutions during the mixing process, thereby achieving more precise control of the concentration gradient formation process.

[0016] Furthermore, in the above-mentioned device for preparing a hydrogel factor concentration gradient, a scale is set on the side wall of the mixing container.

[0017] The beneficial effect of adopting the above further scheme is that the mixing container with scales on the side wall allows the experimenter to easily adjust the positions of different concentration points and flexibly set the range and spacing of the concentration gradient according to experimental requirements.

[0018] Furthermore, a method for preparing a hydrogel factor concentration gradient is prepared by the above-mentioned device for preparing a hydrogel factor concentration gradient, comprising the following steps:

[0019] S1: Connecting a container containing a pure hydrogel precursor solution to the end of the outer tube, and connecting a container containing a hydrogel precursor solution containing bioactive factors to the end of the nested tube;

[0020] S2: setting the speed change devices on the outer tube and the nested tube respectively, and then turning on the micro pump and the power supply connected to the magnetic pole;

[0021] S3: The pure hydrogel precursor solution and the hydrogel precursor solution containing bioactive factors form a gradient flow to the bottom of the adapter through the laminar flow effect to become a mixed solution;

[0022] S4: The magnetons move under the attraction or repulsion of the electromagnetic poles, stirring the mixed solution evenly. The mixed solution falls into the mixing container through the dense holes to form a hydrogel with a concentration gradient.

[0023] The beneficial effect of adopting the above further scheme is that the method mixes the pure hydrogel precursor solution and the hydrogel precursor solution containing the bioactive factor at the bottom of the adapter through the laminar flow effect to form a gradient flow. This step makes full use of the principles of fluid dynamics to ensure that the two solutions can maintain their original laminar state during the mixing process, thereby avoiding the problem of turbulent mixing unevenness that may occur in traditional methods. At the same time, by adjusting the speed change device, the flow rate ratio of the two solutions can be accurately controlled, thereby achieving precise regulation of the concentration of the bioactive factor in the final mixed solution.

[0024] The magnet moves under the attraction or repulsion of the electromagnetic poles, stirring the mixed solution to ensure that the bioactive factors in the solution are evenly distributed. This step not only speeds up the mixing process, but also improves the mixing efficiency, avoiding the unevenness and possible contamination caused by manual or mechanical stirring.

[0025] The beneficial effects of the present invention are as follows: the device realizes precise control of pure hydrogel precursor solution and hydrogel precursor solution containing bioactive factors through the outer tube and the nested tube, as well as the matching speed change device and micro pump, which not only ensures that the two solutions can flow into the adapter at the same time according to the preset ratio and speed, but also naturally forms a concentration gradient inside the adapter through the laminar flow effect, greatly improving the accuracy and repeatability of the concentration gradient formation.

[0026] The built-in magnetic stirring device, which consists of a magnet, a magnetic pole and a power supply, stirs the mixed solution inside the adapter through non-contact magnetic force. The silicone layer design on the outside of the magnet not only increases the stirring efficiency, but also reduces the bubbles and impurities that may be generated during the stirring process, ensuring the uniformity of the mixed solution. The speed change device in the device is equipped with a speed display, which is convenient for users to monitor and control the flow speed of the solution in real time to ensure the accuracy of the experimental process. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 A schematic diagram of the structure of a device for preparing a hydrogel factor concentration gradient;

[0028] Figure 2 It is a schematic diagram of the cross-sectional structure of the adapter and the introduction device;

[0029] Figure 3 Schematic diagram of cells climbing in hydrogel without factor gradient;

[0030] Figure 4 Schematic diagram of cells climbing in hydrogels with factor gradients;

[0031] Figure 5 This is the cell ingrowth diagram in the hydrogel without factor gradient;

[0032] Figure 6 This is a diagram of cell ingrowth in a hydrogel with a factor gradient;

[0033] Wherein: 1. mixing container, 2. adapter, 3. introduction device, 4. magnet, 5. magnetic pole, 6. power supply, 7. dense hole, 8. outer tube, 9. nested tube, 10. speed change device, 11. micro pump. DETAILED DESCRIPTION

[0034] The specific implementation modes of the present invention are described below so that those skilled in the art can understand the present invention. However, it should be clear that the present invention is not limited to the scope of the specific implementation modes. For those of ordinary skill in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the attached claims, these changes are obvious, and all inventions and creations utilizing the concept of the present invention are protected.

[0035] like Figures 1-2 As shown, this embodiment provides a device for preparing a hydrogel factor concentration gradient, comprising a mixing container 1, a adapter 2 connected to the mixing container 1, and an introduction device 3 connected to the adapter 2;

[0036] The mixing container 1 is a cylindrical or rectangular structure. The side wall of the container is provided with clear scale lines, so that the experimenter can easily adjust the position of different concentration points and flexibly set the range and spacing of the concentration gradient according to the experimental requirements. The top of the container is provided with an interface matching the adapter 2, and a sealing gasket is provided at the interface to ensure a tight connection without leakage.

[0037] The adapter 2 is cylindrical in shape, with a flat top for easy installation of the magnetic pole 5, and a slightly conical bottom for easy collection and discharge of the solution. The magnetic pole 5 is made of high-strength permanent magnetic material, at least two are set, and the geometric center of the top surface of the adapter 2 is used as the center of the circle, and the circular array is arranged. Each magnetic pole 5 is connected to the external power supply 6 through a wire to control the opening and closing of the magnetic field. The magnetic son 4 is located inside the adapter 2 and is made of a magnetic alloy. A soft silicone layer is wrapped on the outside to increase the friction during the stirring process and reduce the generation of bubbles. The magnetic pole 5 is installed on the top of the adapter 2. By adjusting the output voltage and current intensity of the power supply 6, the strength and direction of the magnetic field can be controlled, thereby driving the magnetic son 4 to rotate rapidly in the solution to achieve efficient stirring. The internal cavity of the adapter 2 is larger than the size of the magnetic son 4 to allow the magnetic son 4 to rotate freely without touching the wall. Densely distributed dense holes 7 are provided at the connection between the bottom of the adapter 2 and the mixing container 1, which are used to evenly drip the stirred mixed solution into the mixing container 1.

[0038] The introduction device 3 is composed of an outer tube 8 and a nested tube 9. The outer tube 8 is a straight tube structure, one end of which is connected to a container of a pure hydrogel precursor solution, and the other end is connected to the upper part of the adapter 2. The nested tube 9 is cleverly arranged inside the outer tube 8, one end of which passes through the side wall of the outer tube 8 and is connected to a container of a hydrogel precursor solution containing a bioactive factor, and the other end extends into the outer tube 8 to a certain depth. The outer tube 8 and the nested tube 9 are both equipped with a precise speed change device and a micro pump 11. By controlling the flow rate of the two, laminar mixing of the two solutions in the adapter 2 can be achieved. When multiple nested tubes 9 are provided, a variety of hydrogels containing bioactive factors can be prepared.

[0039] Preparation method:

[0040] First, ensure that all experimental equipment is clean and free of contamination. Place the pure hydrogel precursor solution and the hydrogel precursor solution containing bioactive factors into their respective containers and place them in a constant temperature environment to maintain a consistent temperature to reduce the impact of temperature differences on solution viscosity and mixing effects.

[0041] The container containing the pure hydrogel precursor solution is connected to the end of the outer tube 8 through a catheter, and the container containing the hydrogel precursor solution containing the bioactive factor is connected to the end of the nested tube 9. Ensure that all connections are tight and leak-free. Then, adjust the speed change devices on the outer tube 8 and the nested tube 9 to the predetermined flow rate, and confirm it through the speed display. At the same time, turn on the power supply 6 connected to the magnetic pole 5 and adjust it to a suitable magnetic field strength.

[0042] The micro pump 11 is started, and the pure hydrogel precursor solution and the hydrogel precursor solution containing the bioactive factors flow into the adapter 2 through the outer tube 8 and the nested tube 9 respectively under the control of their respective flow rates. Due to the difference in flow rates of the two solutions and the laminar flow effect, they gradually form a concentration gradient in the adapter 2. At the same time, the magnet 4 rotates at a high speed under the attraction or repulsion of the electromagnetic pole 5 to stir the mixed solution to ensure that the concentration gradient is evenly distributed in the solution.

[0043] The fully stirred mixed solution drips evenly into the mixing container 1 through the dense holes 7 at the bottom of the adapter 2. As the solution continues to drip and accumulate, a hydrogel precursor solution with an obvious concentration gradient is gradually formed in the mixing container 1.

[0044] The hydrogel material prepared by this method has a good gradient distribution of the concentration of the bioactive factors inside. Figures 3-4 As shown, in hydrogels without factor gradients, cells can be found to climb disorderly, while in hydrogels with factor gradients, cells tend to climb from low concentration to high concentration.

[0045] like Figures 5-6 As shown, the z-axis represents the thickness of the hydrogel, and cells grow from top to bottom. From the 3D perspective of confocal microscopy, cells have a stronger tendency to grow into the hydrogel with factor gradients, tending to grow in the direction of high concentration factors in the hydrogel.

Claims

1. A device for preparing a hydrogel factor concentration gradient, characterized in that: It comprises a mixing container (1), an adapter (2) connected to the mixing container (1), and an introduction device (3) connected to the adapter (2); The adapter (2) is provided with a magnetic stirring device, the magnetic stirring device comprising a magnet (4) arranged inside the adapter (2), a magnetic pole (5) arranged on the top of the adapter (2), and a power source (6) connected to the magnetic pole (5); a dense hole (7) is provided on the connection surface between the adapter (2) and the mixing container (1); The introduction device (3) comprises an outer tube (8) and at least one nested tube (9) arranged inside the outer tube (8); one end of the nested tube (9) passes through the outer wall of the outer tube (8); and a speed change device and a micro pump (11) are arranged on the nested tube (9) and the outer tube (8).

2. The device for preparing a hydrogel factor concentration gradient according to claim 1, characterized in that: The adapter (2) is a cylinder, and at least two magnetic poles (5) are provided. The magnetic poles (5) are arranged in an annular array on the top surface of the adapter (2) with the geometric center of the top surface of the adapter (2) as the center of the circle.

3. The device for preparing a hydrogel factor concentration gradient according to claim 1, characterized in that: A silica gel layer is arranged on the outer side of the magnet (4).

4. The device for preparing a hydrogel factor concentration gradient according to claim 1, characterized in that: A speed display screen is arranged on the speed changing device.

5. The device for preparing a hydrogel factor concentration gradient according to claim 1, characterized in that: The side wall of the mixing container (1) is provided with scales.

6. A method for preparing a hydrogel factor concentration gradient, prepared by the device for preparing a hydrogel factor concentration gradient according to any one of claims 1 to 5, characterized in that: The following steps are involved: S1: Connecting a container containing a pure hydrogel precursor solution to the end of the outer tube (8), and connecting a container containing a hydrogel precursor solution containing bioactive factors to the end of the nested tube (9); S2: setting the speed change devices on the outer tube (8) and the nested tube (9) respectively, and then turning on the micro pump (11) and the power supply (6) connected to the magnetic pole (5); S3: The pure hydrogel precursor solution and the hydrogel precursor solution containing the bioactive factors form a gradient flow to the bottom of the adapter (2) through the laminar flow effect to become a mixed solution; S4: The magnet (4) moves under the attraction or repulsion of the electromagnetic pole (5), stirring the mixed solution evenly, and the mixed solution falls into the mixing container (1) through the dense holes (7), forming a hydrogel with a concentration gradient.

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