A mobile injection apparatus for non-human primate brain region AAV injection

Abstract

The application relates to the fields of gene therapy and neuroscientific experimental technology, and particularly relates to a mobile injection device for non-human primate brain area AAV injection, which comprises a ring, a mounting frame rotatably connected to the top of the ring, a first driving structure installed between the ring and the mounting frame and used for driving the mounting frame to rotate on the ring, a sliding plate slidably connected to one side of the mounting frame and located in the interior of the ring, a second driving structure installed on the mounting frame and used for driving the sliding plate to move up and down, a swing plate assembled at the end of the sliding plate away from the mounting frame, an injection needle installed at the top of the swing plate, a pushing structure installed at the top of the swing plate, and an assembling structure also installed at the top of the swing plate. The injection device is provided with a structure for synchronous injection during the needle withdrawal process, can complete virus delivery of the target brain area through a single needle channel, reduces the number of puncture needle channels, and reduces mechanical damage to the brain tissue.

Description

Technical Field

[0001] This invention belongs to the field of gene therapy and neuroscience experimental technology, specifically a mobile injection device for AAV injection in the brain regions of non-human primates. Background Technology

[0002] In preclinical studies of gene therapy for central nervous system diseases, adeno-associated virus (AAV) is a widely used gene delivery vector. Non-human primates, due to their high similarity to human brain structures, are important experimental animal models in this field. Currently, AAV injection into the brain regions of non-human primates is mostly performed using a stereotactic approach with multiple injection points. Because the target brain regions in non-human primates are relatively large, researchers need to pre-plan multiple injection coordinate points, sequentially performing needle insertion, quantitative injection, and needle withdrawal at each point. Coverage of the target brain region is achieved through the superposition of viral diffusion at multiple points. This injection method has several shortcomings. The calculation and positioning of multiple injection points is cumbersome, requiring high skill levels from operators and resulting in a long overall experimental cycle. Multiple punctures can create multiple needle tracks in the brain tissue, causing multiple mechanical injuries and potentially affecting the normal function of the target brain region, which is detrimental to animal welfare and the reliability of experimental data. The repeated disassembly and reassembly of syringes or replacement of needles increases the probability of virus sample contamination. The natural spread of the virus under the point injection mode is limited, and to cover a large brain region, it is often necessary to increase the number of injection points or the volume of a single injection, which can easily lead to problems such as increased local brain tissue pressure. Summary of the Invention

[0003] The purpose of this invention is to provide a mobile injection device for AAV injection in the brain region of non-human primates. Through the structure of simultaneous injection during needle withdrawal, the virus can be delivered to the target brain region through a single needle channel, reducing the number of puncture needle channels and minimizing mechanical damage to brain tissue. By continuously outputting viral fluid throughout the needle withdrawal process, the virus can form a continuous distribution along the needle channel, improving the uniformity of virus distribution in the target brain region, increasing the surface area for virus diffusion, and facilitating viral transduction in the brain parenchyma.

[0004] The technical solution adopted in this invention is as follows: A mobile injection device for AAV injection in the brain region of non-human primates includes a ring, a mounting frame rotatably connected to the top of the ring, a first drive structure between the ring and the mounting frame for driving the mounting frame to rotate on the ring, a sliding plate slidably connected to one side of the mounting frame and inside the ring, a second drive structure mounted on the mounting frame for driving the sliding plate to move up and down, a swing plate mounted at the end of the sliding plate away from the mounting frame, an injection needle mounted on the top of the swing plate, a pushing structure mounted on the top of the swing plate for pushing the injection needle to inject, and an assembly structure mounted on the top of the swing plate for assembling the injection needle and changing the position of the injection needle.

[0005] The first drive structure includes a first motor, which is installed inside the mounting bracket on the side away from the annular ring. A gear is fixed to the output end of the first motor, and a rack is installed on the outer side of the annular ring, with the gear meshing with the rack.

[0006] The second drive structure includes a mounting plate, which is fixed to the side of the mounting bracket near the sliding plate and is disposed on the top of the sliding plate. A first threaded rod is rotatably connected to the bottom of the mounting plate and is threadedly connected to the sliding plate. A second motor is mounted on the top of the mounting plate, and the output end of the second motor is vertically downward connected to the first threaded rod.

[0007] A third motor is installed on one side of the sliding plate. The output end of the third motor passes through the sliding plate and is connected to the swing plate. The swing plate is rotatably connected to the sliding plate.

[0008] The pushing structure includes two assembly pieces, which are fixed to the top of the swing plate and disposed on one side of the injection needle. A second threaded rod is rotatably connected between the two assembly pieces, and a pushing plate is threadedly connected to the outer side of the second threaded rod. The pushing plate is slidably connected to the swing plate, and one end of the pushing plate abuts against the piston of the injection needle.

[0009] The assembly structure includes a fixing block, which is fixed to the top of the swing plate. An electric push rod is installed inside the fixing block. An elastic retaining frame is fixed on the stroke rod of the electric push rod, and the elastic retaining frame is sleeved on the outside of the injection needle.

[0010] This invention, through a structure that synchronizes injection during needle withdrawal, enables virus delivery to the target brain region via a single needle path, reducing the number of puncture needle paths and minimizing mechanical damage to brain tissue. Continuous viral output throughout the needle withdrawal process ensures a continuous viral distribution along the needle path, improving the uniformity of viral distribution within the target brain region, increasing the surface area for viral diffusion, and facilitating viral transduction within the brain parenchyma. It eliminates the need for planning multiple injection sites and repeated positioning and clamping, simplifying the experimental procedure and improving its repeatability. Furthermore, by controlling the needle withdrawal speed and injection flow rate in tandem, the amount of virus deposited per unit length of needle path remains stable, enhancing the precision and consistency of the injection operation. Attached Figure Description

[0011] Figure 1 This is a schematic diagram of the structure of the present invention; Figure 2 This is a schematic diagram of the structure between the first threaded rod, the injection needle, and the rack in this invention; Figure 3 In this invention Figure 2 Enlarged view of point A in the middle; Figure 4 This is a schematic diagram of the structure between the second threaded rod, the electric push rod, and the injection needle in this invention; In the attached diagram, the components represented by each number are as follows: 1. Ring; 2. Mounting bracket; 3. Swing plate; 4. Injection needle; 5. First motor; 6. Gear; 7. Rack; 8. Second motor; 9. Sliding plate; 10. First threaded rod; 11. Third motor; 12. Assembly piece; 13. Fourth motor; 14. Second threaded rod; 15. Push plate; 16. Fixing block; 17. Electric push rod; 18. Elastic clip frame; 19. Mounting piece. Detailed Implementation

[0012] To make the objectives and advantages of this invention clearer, the invention will be specifically described below with reference to embodiments. It should be understood that the following text is merely used to describe one or more specific embodiments of the invention and does not strictly limit the scope of protection specifically claimed by the invention.

[0013] like Figures 1-4As shown, a mobile injection device for AAV injection in the brain region of non-human primates includes a ring 1. A mounting frame 2 is rotatably connected to the top of the ring 1. A first drive structure is installed between the ring 1 and the mounting frame 2 to drive the mounting frame 2 to rotate on the ring 1. A sliding plate 9 is slidably connected to one side of the mounting frame 2 and inside the ring 1. A second drive structure is installed on the mounting frame 2 to drive the sliding plate 9 to move up and down. A swing plate 3 is assembled at the end of the sliding plate 9 away from the mounting frame 2. An injection needle 4 is installed on the top of the swing plate 3. A push structure is installed on the top of the swing plate 3. An assembly structure is also installed on the top of the swing plate 3. The push structure is used to drive the piston movement of the injection needle 4 to complete the injection of the drug solution. The assembly structure is used to fix the injection needle 4 and adjust the radial position of the injection needle 4.

[0014] See attached document Figures 1-3 The first drive structure includes a first motor 5, which is installed inside the mounting frame 2 on the side away from the annular ring 1. A gear 6 is fixed to the output end of the first motor 5, and a rack 7 is installed on the outer side of the annular ring 1. The gear 6 and the rack 7 are meshed and connected. When the first motor 5 is running, its output end drives the gear 6 to rotate. Through the meshing transmission of the gear 6 and the rack 7, the mounting frame 2 slides along the circumference of the annular ring 1, thereby adjusting the circumferential angle of the injection needle 4 to adapt to puncture paths in different directions.

[0015] See attached document Figure 2 The second drive structure includes a mounting plate 19, which is fixed to the side of the mounting frame 2 near the sliding plate 9 and is located on the top of the sliding plate 9. The bottom of the mounting plate 19 is rotatably connected to a first threaded rod 10, which is threadedly connected to the sliding plate 9. A second motor 8 is mounted on the top of the mounting plate 19. The output end of the second motor 8 is vertically downward and connected to the first threaded rod 10. When the second motor 8 is running, the first threaded rod 10 rotates synchronously, driving the sliding plate 9 to slide vertically along the side wall of the mounting frame 2 through threaded transmission, thereby realizing the up and down movement of the injection needle 4.

[0016] A third motor 11 is installed on one side of the sliding plate 9. The output end of the third motor 11 passes through the sliding plate 9 and is connected to the swing plate 3. The swing plate 3 is rotatably connected to the sliding plate 9. The swing plate 3 is driven to deflect by the third motor 11, which can adjust the puncture angle of the injection needle 4 to adapt to different needle insertion angles.

[0017] See attached document Figure 4The pushing structure includes two assembly pieces 12, which are fixed to the top of the swing plate 3. The two assembly pieces 12 are located on one side of the injection needle 4. A second threaded rod 14 is rotatably connected between the two assembly pieces 12. A push plate 15 is threadedly connected to the outer side of the second threaded rod 14. The push plate 15 is slidably connected to the swing plate 3. One end of the push plate 15 abuts against the piston of the injection needle 4. When the fourth motor 13 is running, the second threaded rod 14 rotates synchronously. Through the threaded transmission, the push plate 15 slides along the surface of the swing plate 3, thereby pushing the piston of the injection needle 4 forward to achieve uniform injection of the drug liquid.

[0018] See attached document Figure 4 The assembly structure includes a fixing block 16, which is fixed to the top of the swing plate 3. An electric push rod 17 is installed inside the fixing block 16. An elastic retaining frame 18 is fixed on the stroke rod of the electric push rod 17, and the elastic retaining frame 18 is sleeved on the outside of the injection needle 4. When the electric push rod 17 is running, it can drive the elastic retaining frame 18 and the injection needle 4 to move radially together, finely adjust the needle tip position of the injection needle 4, and improve the positioning accuracy.

[0019] The working principle of this invention is as follows: Before the injection, the ring 1 is fixed and engaged with the external stereoscopic positioning device to complete the overall positioning of the instrument. The injection needle 4, which is loaded with AAV virus solution, is clamped into the elastic frame 18. According to the puncture path of the target brain region, the first motor 5, the third motor 11 and the electric push rod 17 are driven to adjust the circumferential position, deflection angle and radial position of the injection needle 4 so that the needle tip is aligned with the preset puncture point. Then the second motor 8 is driven to move the sliding plate 9 downward, so that the injection needle 4 moves up and down. During injection, the second motor 8 and the fourth motor 13 are started simultaneously. The second motor 8 drives the sliding plate 9 to rise at a set speed to withdraw the needle. At the same time, the fourth motor 13 drives the push plate 15 to push the piston of the injection needle 4 forward at a constant speed, piercing the brain tissue until the needle tip reaches the deepest part of the target brain region, and the virus solution is continuously output along the needle withdrawal path to form a continuous drug solution distribution along the needle channel. The steps are as follows: Anesthetize and fix the head of non-human primates such as rhesus monkeys, determine the stereotactic coordinates of the target brain region, such as the subthalamic nucleus, through MRI or CT scans, and plan a single puncture needle path that runs through the anterior-posterior or dorsoventral range of the entire target brain region. Connect the injection needle 4, which contains AAV virus at a titer of 1×10^12 vg / mL, to a 33G needle.

[0020] The needle withdrawal speed is set to 0.3 mm / s in the control unit, corresponding to an injection flow rate of 1.2 μL / min, so that the injection volume in each millimeter needle channel is 0.066 μL.

[0021] The stereotactic instrument slowly inserts the needle along the preset needle path until the needle tip reaches the deepest point of the target brain region, at the coordinates furthest from the starting injection point.

[0022] Initiate the injection procedure: The infusion pump starts injecting at a constant rate of 1.2 μL / min, while the needle withdrawal mechanism withdraws the needle at a uniform speed of 0.3 mm / s, continuously moving the needle from the deepest point to the superficial layer until the needle tip leaves the upper boundary of the target brain region.

[0023] Stop the injection, completely withdraw the needle, and complete the single-needle linear injection.

[0024] This application requires only one needle tract to achieve viral coverage of a large brain region, reducing the number of puncture needle tracts from multiple to one, greatly reducing the risk of mechanical damage to non-human primate brain functional areas. Furthermore, by injecting while withdrawing the needle, the AAV virus is continuously and linearly distributed throughout the needle tract, avoiding the problems of local virus aggregation or uneven diffusion caused by traditional point injection. This facilitates the widespread and uniform transduction of AAV in the target brain region, and eliminates the need to calculate and switch multiple injection points, greatly simplifying the operation steps, improving the success rate and reproducibility of the experiment. Moreover, the viral source line formed by linear injection has a larger diffusion surface area than a single point source. Under the same injection volume, the effective diffusion range of the virus in the brain parenchyma is significantly increased, and the transduction efficiency is higher.

[0025] The above description is merely a preferred embodiment of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention. Structures, devices, and operating methods not specifically described or explained in this invention are implemented according to conventional methods in the art unless otherwise specified or limited.

Claims

1. A mobile injection device for AAV injection in the brain region of non-human primates, comprising a ring coil (1), characterized in that: A mounting bracket (2) is rotatably connected to the top of the annular ring (1) around the annular ring (1). A first driving structure is installed between the annular ring (1) and the mounting bracket (2) to drive the mounting bracket (2) to rotate on the annular ring (1). A sliding plate (9) is slidably connected to one side of the mounting bracket (2) and inside the annular ring (1). A second driving structure is installed on the mounting bracket (2) to drive the sliding plate (9) to move up and down. A swing plate (3) is mounted on the end of the sliding plate (9) away from the mounting bracket (2). An injection needle (4) is mounted on the top of the swing plate (3). A pushing structure is installed on the top of the swing plate (3) to push the injection needle (4) to inject. An assembly structure is also installed on the top of the swing plate (3) to assemble the injection needle (4) and change the position of the injection needle (4).

2. The mobile injection device for AAV injection in the brain region of non-human primates according to claim 1, characterized in that: The first drive structure includes a first motor (5), which is installed inside the mounting bracket (2) on the side away from the annular ring (1). A gear (6) is fixed at the output end of the first motor (5), and a rack (7) is installed on the outer side of the annular ring (1), and the gear (6) meshes with the rack (7).

3. A mobile injection device for AAV injection in the brain region of non-human primates according to claim 1, characterized in that: The second drive structure includes a mounting plate (19), which is fixed to the side of the mounting bracket (2) near the sliding plate (9) and is located on the top of the sliding plate (9). A first threaded rod (10) is rotatably connected to the bottom of the mounting plate (19) and is threadedly connected to the sliding plate (9). A second motor (8) is mounted on the top of the mounting plate (19), and the output end of the second motor (8) is vertically downward connected to the first threaded rod (10).

4. A mobile injection device for AAV injection in the brain region of non-human primates according to claim 1, characterized in that: A third motor (11) is installed on one side of the sliding plate (9). The output end of the third motor (11) passes through the sliding plate (9) and is connected to the swing plate (3). The swing plate (3) is rotatably connected to the sliding plate (9).

5. A mobile injection device for AAV injection in the brain region of non-human primates according to claim 1, characterized in that: The pushing structure includes two assembly pieces (12), and the two assembly pieces (12) are fixed on the top of the swing plate (3). The two assembly pieces (12) are disposed on one side of the injection needle (4). A second threaded rod (14) is rotatably connected between the two assembly pieces (12), and a push plate (15) is threadedly connected to the outer side of the second threaded rod (14). The push plate (15) is slidably connected to the swing plate (3), and one end of the push plate (15) abuts against the piston of the injection needle (4).

6. A mobile injection device for AAV injection in the brain region of non-human primates according to claim 1, characterized in that: The assembly structure includes a fixing block (16), which is fixed to the top of the swing plate (3). An electric push rod (17) is installed inside the fixing block (16). An elastic clip (18) is fixed on the stroke rod of the electric push rod (17), and the elastic clip (18) is sleeved on the outside of the injection needle (4).

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