Rat caudal vertebra intervertebral disc degeneration modeling device and method

Abstract

The invention discloses a rat caudal vertebra intervertebral disc degeneration molding device which comprises an anesthesia box, a tail outlet hole is formed in the anesthesia box, a bearing table is arranged on the outer side of the anesthesia box, an arc-shaped locking plate is slidably installed on the bearing table, a containing groove is formed between the arc-shaped locking plate and the bearing table, and a sleeve is slidably installed on the arc-shaped locking plate in the circumferential direction; a puncture cylinder is in threaded connection with the interior of the sleeve, a graduated scale is axially arranged on the outer side wall of the puncture cylinder, a puncture needle is installed in the puncture cylinder and comprises a base and a needle head connected to the base, the base is installed in the puncture cylinder, the needle head sequentially penetrates through the puncture cylinder, the sleeve and the arc-shaped locking plate and then extends into the containing groove, and a bandage is arranged above the bearing table. An X-ray machine is arranged on the periphery of the bearing plate. The invention further discloses a rat caudal vertebra intervertebral disc degeneration modeling method using the rat caudal vertebra intervertebral disc degeneration modeling device. According to the rat caudal vertebra intervertebral disc degeneration modeling device and method, few tools are used, the process is simple, and application and popularization are convenient.

Description

Technical Field

[0001] The present invention relates to the technical field of medical experimental techniques, and particularly to a rat caudal vertebra intervertebral disc degeneration modeling device and method. Background Art

[0002] The intervertebral disc is composed of structures such as the nucleus pulposus tissue, annulus fibrosus, and cartilage endplate. There is a high positive correlation between intervertebral disc degeneration and low back pain. Intervertebral disc degeneration refers to the process of gradual deterioration and damage of the structure and function of the intervertebral disc. In the early stage of intervertebral disc degeneration, the extracellular matrix homeostasis in the nucleus pulposus is damaged due to a series of reasons, the hydration decreases, the compressive capacity weakens, the annulus fibrosus bears too much load and generates cracks. With the progress of degeneration, the cracks increase, and some nerves and blood vessels grow in from the damaged cracks of the annulus fibrosus, and the nucleus pulposus tissue protrudes from the damaged annulus fibrosus. The abnormal local microenvironment in the degenerated intervertebral disc will stimulate the pain-sensing nerves to generate discogenic pain, and the sciatic nerve will also be compressed by the protruding nucleus pulposus tissue, causing the patient to have radiating pain and numbness in the lower body. At present, for the clinical treatment of intervertebral disc degeneration, such as minimally invasive nucleus pulposus resection, etc., it mainly relieves pain and cannot cure the disease.

[0003] With the development of tissue engineering, it provides new possibilities for the treatment and regeneration of intervertebral disc degeneration. Selecting a suitable animal intervertebral disc degeneration model can better simulate human degeneration and explore its treatment methods. The degeneration model induced by annulus fibrosus puncture can largely simulate human intervertebral disc herniation, and rats have been widely used in intervertebral disc degeneration research due to their low price, fast growth cycle, easy positioning of caudal intervertebral discs, and simple degeneration induction. However, the tools used in the existing rat caudal vertebra intervertebral disc modeling surgery are relatively complex, and the process is also more cumbersome, which is not conducive to popularization and application. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a rat caudal vertebra intervertebral disc degeneration modeling device and method that use fewer tools, have a relatively simple process, and are convenient for popularization and application.

[0005] The rat caudal vertebra intervertebral disc degeneration modeling device in the present invention includes an anesthesia box. An exit tail hole is provided on the anesthesia box. A horizontally arranged bearing plate is provided outside the anesthesia box. A bearing platform opposite to the exit tail hole is provided on the upper side surface of the bearing plate. An arc-shaped locking plate is slidably installed on the bearing platform. Two ends of the arc-shaped locking plate are respectively slidably connected to opposite sides of the bearing platform. An accommodation groove is formed between the arc-shaped locking plate and the bearing platform. A sleeve is slidably installed on the arc-shaped locking plate along the circumferential direction. The axis of the sleeve is arranged along the radial direction of the arc-shaped locking plate. A puncture cylinder is threadedly connected inside the sleeve. A scale arranged axially is provided on the outer side wall of the puncture cylinder. A puncture needle is installed inside the puncture cylinder. The puncture needle includes a base and a needle head connected to the base. The base is installed inside the puncture cylinder. The needle head extends into the accommodation groove after passing through the puncture cylinder, the sleeve and the arc-shaped locking plate in sequence. Above the bearing platform, a first strap and a second strap are provided. The first strap and the second strap are respectively located on opposite sides of the arc-shaped locking plate. Two ends of the first strap and the second strap are respectively connected to the upper side surface of the bearing plate on opposite sides of the bearing platform. An X-ray machine for determining the puncture position of the needle head by irradiation imaging is provided on the periphery of the bearing plate.

[0006] The rat caudal vertebra intervertebral disc degeneration modeling device in the present invention, wherein the anesthesia box includes a box body and a box cover. The box body includes a box side wall arranged vertically in a rectangular cylindrical structure. A box bottom wall is fixedly provided at the lower barrel opening of the box side wall. The box cover covers the upper barrel opening of the box side wall. The exit tail hole is provided on the box side wall. An air injection hole arranged opposite to the exit tail hole is provided on the box side wall. A ventilation hole is also provided on the box side wall.

[0007] The rat caudal vertebra intervertebral disc degeneration modeling device in the present invention, wherein the bearing platform is strip-shaped. One end of the bearing platform is arranged close to the anesthesia box, and the other end of the bearing platform is arranged far from the anesthesia box. The lower side surface of the bearing platform is fixedly provided on the bearing plate. The upper side surface of the bearing platform is a bearing surface. The bearing surface is an arc-shaped surface sunken downward. Two ends of the arc-shaped locking plate are respectively slidably connected to opposite sides of the bearing platform along the length direction of the bearing platform.

[0008] The rat caudal vertebra intervertebral disc degeneration modeling device in the present invention, wherein the arc-shaped locking plate is an arc-shaped plate bulging upward. A circumferentially arranged first sliding groove is provided on the outer peripheral surface of the arc-shaped locking plate. The first sliding groove communicates with the accommodation groove. A slider is fixedly provided at one end of the sleeve close to the arc-shaped locking plate. The slider is slidably installed in the first sliding groove. The needle head extends into the accommodation groove after passing through the puncture cylinder, the sleeve, the slider and the first sliding groove on the arc-shaped locking plate in sequence.

[0009] The rat caudal vertebra intervertebral disc degeneration modeling device in the present invention, wherein a locking member is inserted through the puncture cylinder along the radial direction. The locking member abuts against the end of the base away from the needle head.

[0010] The rat caudal vertebra intervertebral disc degeneration modeling device in the present invention, wherein an axially arranged second sliding groove is provided on the arc-shaped locking plate, one end of the second sliding groove communicates with the first sliding groove, and the other end of the second sliding groove penetrates through the arc-shaped locking plate. Both ends of the first strap and the second strap are respectively connected to the upper sides of the bearing plates on opposite sides of the bearing table through adhesives or bolts.

[0011] The rat caudal vertebra intervertebral disc degeneration modeling method using the above device includes the following steps:

[0012] Step 1: Open the lid of the anesthesia box, place the rat in the box, and then cover the lid again.

[0013] Step 2: Pass anesthesia gas into the box through the air injection hole, observe the condition of the rat through the air permeable holes on the box body. After the rat is completely anesthetized, open the lid of the box, and pass the rat's tail through the tail hole and extend it from inside the box to outside the box.

[0014] Step 3: Move the first strap and the second strap away from above the bearing table, and slide the arc-shaped locking plate along the bearing table to the end of the bearing table far from the anesthesia box. Lay the rat's tail along the length direction of the bearing table on the bearing surface. Then, start palpating from the root of the rat's tail towards the tail end to determine the positions of the third vertebra to the seventh vertebra counted from the root of the rat's tail towards the tail end. The gap between any two adjacent vertebrae among the third vertebra to the seventh vertebra is the intervertebral disc. Take at least one intervertebral disc between the third vertebra and the seventh vertebra as the intervertebral disc to be punctured. Then, slide the arc-shaped locking plate towards the direction close to the anesthesia box along the bearing table and at the same time place the rat's tail in the receiving groove. Slide the arc-shaped locking plate to the position of the intervertebral disc to be punctured on the rat's tail and make the needle tip of the puncture needle face the intervertebral disc to be punctured. While the arc-shaped locking plate presses the rat's tail, press the first strap and the second strap on the rat's tail, and connect both ends of the first strap and the second strap to the upper sides of the bearing plates on opposite sides of the bearing table respectively to fix the rat's tail to give a certain tension to the intervertebral disc to be punctured.

[0015] Step 4: Move the sleeve circumferentially along the arc-shaped locking plate. When the sleeve moves to the puncture position of the intervertebral disc to be punctured on the rat's tail, rotate the puncture cylinder relative to the sleeve to make the puncture needle move closer to the rat's tail. When the needle tip of the puncture needle touches the rat's tail, stop rotating the puncture cylinder. Image the rat's tail and the needle tip through an X-ray machine to determine the puncture position of the needle. If the puncture position of the needle does not meet the requirements, correct the puncture position of the needle until the needle tip is aligned with the puncture position of the intervertebral disc to be punctured on the rat's tail.

[0016] Step 5: Continue to rotate the puncture cylinder relative to the sleeve, and insert the needle tip of the puncture needle into the rat tail. After the needle tip penetrates 5 mm deep into the rat tail, stop the penetration. At this time, rotate the needle tip 360 degrees for 5 times and stay for 30 seconds. Then, rotate the puncture cylinder in the reverse direction relative to the sleeve until the needle tip of the puncture needle is withdrawn from the rat tail.

[0017] Step 6: Remove the first strap and the second strap, and slide the arc-shaped lock plate along the carrier platform away from the anesthesia box to separate the arc-shaped lock plate from the rat tail. Then, disinfect the puncture needle opening on the rat tail, and then transfer the rat from the anesthesia box to the rat cage for continued feeding.

[0018] Step 7: Verify the intervertebral disc degeneration condition of the rat within the preset time. When obvious changes occur in the observation indexes of the intervertebral disc of the rat, the puncture causes intervertebral disc degeneration and the model establishment is successful.

[0019] In the method for establishing a rat caudal vertebra intervertebral disc degeneration model in the present invention, in the seventh step, after the preset time, the tail of the rat is imaged by an X-ray machine, and the observation index is the gap between two adjacent vertebral bodies of the punctured intervertebral disc. When the gap becomes narrower relative to the initial state, the puncture causes intervertebral disc degeneration and the model establishment is successful.

[0020] In the method for establishing a rat caudal vertebra intervertebral disc degeneration model in the present invention, in the seventh step, after the preset time, the tail of the rat is imaged by a nuclear magnetic resonance device, and the observation index is the T2-weighted image of the punctured intervertebral disc. The T2-weighted image of the intervertebral disc of a normal rat is a high signal. If the T2-weighted image of the punctured intervertebral disc is a low signal, the puncture causes intervertebral disc degeneration and the model establishment is successful.

[0021] In the method for establishing a rat caudal vertebra intervertebral disc degeneration model in the present invention, in the seventh step, after the preset time, the rat is sacrificed, the punctured intervertebral disc is dissected, placed in 4% paraformaldehyde for fixation for 24 hours, and then decalcified in 15% ethylenediaminetetraacetic acid (EDTA) for 30 days. After the decalcified intervertebral disc is dehydrated and cleared, it is immersed in melted paraffin for embedding. Sections are made of the embedded intervertebral disc, and the thickness of the sections does not exceed 5 μm.

[0022] The sections of the intervertebral disc are stained with hematoxylin and eosin (HE), and the observation index is the histological structure of the stained intervertebral disc. If the histological structure is damaged, the puncture causes intervertebral disc degeneration and the model establishment is successful, and / or

[0023] The sections of the intervertebral disc are stained with safranin-fast green, and the observation indexes are the histological structure, proteoglycan content, and type II collagen content of the stained intervertebral disc. If the histological structure is damaged and the proteoglycan content and type II collagen content decrease, the puncture causes intervertebral disc degeneration and the model establishment is successful.

[0024] The difference between the rat caudal vertebra intervertebral disc degeneration modeling device and method of the present invention and the prior art lies in that the present invention anesthetizes the rat through an anesthesia box, and then fixes the rat's tail through a carrier table, a binding strap and an arc-shaped locking plate. Then, a puncture operation is performed on the intervertebral disc of the rat's tail through a sleeve, a puncture cylinder and a puncture needle. And during the puncture process, the position of the needle tip of the puncture needle can be corrected by an X-ray machine to facilitate accurate puncture. Thus, it can be seen that the present invention integrates an anesthesia box, an X-ray machine, a carrier table, a binding strap, an arc-shaped locking plate, a sleeve and a puncture cylinder. Only by using the above-mentioned few tools can a modeling operation be performed on the rat caudal vertebra intervertebral disc, and the process is relatively simple and convenient for popularization and application.

[0025] The present invention will be further described below with reference to the accompanying drawings. Description of the Drawings

[0026] Figure 1 is a schematic structural diagram of the rat caudal vertebra intervertebral disc degeneration modeling device of the present invention (the X-ray machine is not shown in the figure);

[0027] Figure 2 is the A-direction view of the anesthesia box in the present invention along Figure 1 in;

[0028] Figure 3 is a schematic structural diagram of the carrier plate, carrier table, binding strap, arc-shaped locking plate, sleeve, puncture cylinder, puncture needle and locking member of the present invention;

[0029] Figure 4 is a schematic structural diagram of the puncture cylinder of the present invention;

[0030] Figure 5 is a schematic structural diagram of the locking member of the present invention;

[0031] Figure 6 is a schematic structural diagram of the sleeve of the present invention;

[0032] Figure 7 is a schematic structural diagram of the arc-shaped locking plate of the present invention;

[0033] Figure 8 is an external view after puncturing the rat caudal vertebra intervertebral disc by using the modeling device of the present invention;

[0034] Figure 9 is an X-ray diagram of the rat's tail and the needle tip in the modeling method of the present invention;

[0035] Figure 10 is an X-ray diagram of the rat caudal vertebra intervertebral disc 8 weeks after puncture in the modeling method of the present invention;

[0036] Figure 11MRI (Magnetic Resonance Imaging) image of the rat caudal vertebral intervertebral disc 8 weeks after puncture in the modeling method of the present invention;

[0037] Figure 12 HE staining image of the normal rat caudal vertebral intervertebral disc;

[0038] Figure 13 HE staining image of the rat caudal vertebral intervertebral disc 8 weeks after puncture in the modeling method of the present invention;

[0039] Figure 14 Safranin-fast green staining image of the normal rat caudal vertebral intervertebral disc;

[0040] Figure 15 Safranin-fast green staining image of the rat caudal vertebral intervertebral disc 8 weeks after puncture in the modeling method of the present invention. Detailed implementation manners

[0041] As Figure 1 shown, and in combination with Figures 2-7 shown, the rat caudal vertebral intervertebral disc degeneration modeling device in the present invention includes an anesthesia box 1. An out-tail hole 6 is provided on the anesthesia box 1. A horizontally arranged bearing plate 7 is provided outside the anesthesia box 1. A bearing platform 8 opposite to the out-tail hole 6 is provided on the upper side of the bearing plate 7. An arc-shaped locking plate 11 is slidably installed on the bearing platform 8. The two ends of the arc-shaped locking plate 11 are respectively slidably connected to the opposite sides of the bearing platform 8. An accommodation groove is formed between the arc-shaped locking plate 11 and the bearing platform 8. A sleeve 14 is slidably installed on the arc-shaped locking plate 11 along the circumferential direction. The axis of the sleeve 14 is arranged along the radial direction of the arc-shaped locking plate 11. A puncture cylinder 15 is threadedly connected inside the sleeve 14. A scale arranged axially is provided on the outer side wall of the puncture cylinder 15. A puncture needle 19 is installed inside the puncture cylinder 15. The puncture needle 19 includes a base 17 and a needle head 18 connected to the base 17. The base 17 is installed inside the puncture cylinder 15. The needle head 18 extends into the accommodation groove after passing through the puncture cylinder 15, the sleeve 14 and the arc-shaped locking plate 11 in sequence. A first strap 9 and a second strap 10 are provided above the bearing platform 8. The first strap 9 and the second strap 10 are respectively located on the opposite sides of the arc-shaped locking plate 11. The two ends of the first strap 9 and the second strap 10 are respectively connected to the upper side of the bearing plate 7 on the opposite sides of the bearing platform 8. An X-ray machine (not shown in the figure) for determining the puncture position of the needle head 18 by irradiation imaging is provided outside the bearing plate 7.

[0042] As Figure 1 、 2As shown, the rat caudal vertebra intervertebral disc degeneration modeling device in the present invention, wherein the anesthesia box 1 includes a box body 3 and a box cover 2. The box body 3 includes a box side wall arranged vertically in a rectangular cylindrical structure. The lower barrel opening of the box side wall is fixedly provided with a box bottom wall. The upper barrel opening of the box side wall is covered with the box cover 2. The tail outlet hole 6 is arranged on the box side wall. The box side wall is provided with an air injection hole 4 arranged opposite to the tail outlet hole 6. The box side wall is also provided with a ventilation hole 5.

[0043] The anesthesia box 1 is used to place rats and anesthetize them. When anesthetizing a rat, open the box cover 2, put the rat into the anesthesia box 1, cover the box cover 2, and then inject anesthetic gas (such as isoflurane) into the box body 3 through the air injection hole 4. After a certain period of time, the rat can be anesthetized. During the anesthesia process, external air can enter the anesthesia box 1 through the ventilation hole 5 for the rat to breathe normally. When specifically setting, the ventilation hole 5 is set to an appropriate size so that it can not only meet the normal breathing of the rat but also not leak a large amount of anesthetic gas to affect the anesthesia of the rat.

[0044] As Figure 1 、 3 As shown, the loading platform 8 in the rat caudal vertebra intervertebral disc degeneration modeling device of the present invention is in a long strip shape. One end of the loading platform 8 is arranged close to the anesthesia box 1, and the other end of the loading platform 8 is arranged far from the anesthesia box 1. The lower side surface of the loading platform 8 is fixedly arranged on the loading plate 7. The loading platform 8 and the loading plate 7 are integrally formed structures. The upper side surface of the loading platform 8 is a loading surface, and the loading surface is a downward concave arc surface, which is convenient for placing the rat's tail. The two ends of the arc-shaped locking plate 11 are respectively slidably connected to the opposite sides of the loading platform 8 along the length direction of the loading platform 8.

[0045] The specific way that the arc-shaped locking plate 11 is slidably connected to the loading platform 8 is as follows: The opposite sides of the loading platform 8 for slidably connecting the arc-shaped locking plate 11 are respectively provided with chutes arranged along the length direction. The two ends of the arc-shaped locking plate 11 are respectively provided with sliders. The arc-shaped locking plate 11 is slidably connected to the loading platform 8 through the sliders and the chutes. Of course, the positions of the chutes and the sliders can also be swapped, that is, the opposite sides of the loading platform 8 for slidably connecting the arc-shaped locking plate 11 are respectively provided with sliders arranged along the length direction, and the two ends of the arc-shaped locking plate 11 are respectively provided with chutes. In this way, the arc-shaped locking plate 11 can also be slidably connected to the loading platform 8 through the sliders and the chutes.

[0046] The air injection hole 4 and the tail outlet hole 6 are respectively arranged on two opposite side walls of the box body 3 of the anesthesia box 1. The loading platform 8 is arranged opposite to the tail outlet hole 6, and the length direction of the loading platform 8 is the same as the direction from the air injection hole 4 to the tail outlet hole 6. In this way, when the arc-shaped locking plate 11 slides along the loading platform 8, the arc-shaped locking plate 11 slides closer to or farther away from the anesthesia box 1.

[0047] AsFigure 1 , 3 As shown in FIGS. 6 and 7, in the rat caudal vertebra intervertebral disc degeneration modeling device of the present invention, the arc-shaped locking plate 11 is an arc-shaped plate protruding upward. A first chute 12 arranged circumferentially is provided on the outer peripheral surface of the arc-shaped locking plate 11. The first chute 12 communicates with the receiving groove. A slider 26 is fixedly provided at one end of the sleeve 14 close to the arc-shaped locking plate 11. The slider 26 is slidably installed in the first chute 12. The needle 18 passes through the puncture cylinder 15, the sleeve 14, the slider 26 and the first chute 12 on the arc-shaped locking plate 11 in sequence and then extends into the receiving groove.

[0048] As Figure 4 shown, a first baffle 20 is fixedly provided at one end of the puncture cylinder 15 connected to the sleeve 14. A first needle hole 21 for the needle 18 to pass through is provided on the first baffle 20. The base 17 of the puncture needle 19 is installed in the barrel cavity of the puncture cylinder 15, and the base 17 abuts against the first baffle 20. The needle 18 of the puncture needle 19 extends out of the puncture cylinder 15 through the first needle hole 21. As Figure 6 shown, when the slider 26 is provided on the sleeve 14, a second baffle 24 is fixedly provided at the barrel opening at one end of the sleeve 14 close to the arc-shaped locking plate 11. A sleeve 25 communicating with the barrel cavity of the sleeve 14 is fixedly provided on the second baffle 24. The slider 26 is fixedly provided on the sleeve 25. A second needle hole 27 communicating with the sleeve 25 is provided on the slider 26. That is to say, the barrel cavity of the sleeve 14, the lumen of the sleeve 25 and the second needle hole 27 of the slider 26 communicate in sequence. In this way, the needle 18 extending out of the puncture cylinder 15 can pass through the barrel cavity of the sleeve 14, the lumen of the sleeve 25 and the second needle hole 27 in sequence, and then penetrate into the receiving groove through the connection between the first chute 12 and the receiving groove, thus realizing that the needle 18 passes through the puncture cylinder 15, the sleeve 14, the slider 26 and the first chute 12 on the arc-shaped locking plate 11 in sequence and then extends into the receiving groove.

[0049] In order to realize the communication between the first chute 12 and the receiving groove, a strip-shaped hole is opened on the bottom wall of the first chute 12 along the length direction of the first chute 12. The needle 18 penetrates into the receiving groove from the first chute 12 through this strip-shaped hole.

[0050] As Figure 1 , 3As shown in FIGS. 4 and 5, in the rat caudal vertebral intervertebral disc degeneration modeling device of the present invention, a locking member 16 is inserted through the piercing cylinder 15 in the radial direction, and the locking member 16 abuts against the end of the base 17 away from the needle 18. A lock hole 22 for accommodating the locking member 16 is provided on the wall of the piercing cylinder 15. The locking member 16 is a rectangular rod-shaped structure. When the locking rod is inserted into the lock hole 22, the locking member 16 abuts against the end of the base 17 away from the needle 18. In this way, the piercing needle 19 cannot rotate relative to the piercing cylinder 15. When the piercing cylinder 15 is rotated relative to the sleeve 14, the piercing cylinder 15 drives the piercing needle 19 to rotate together. Since the piercing cylinder 15 and the sleeve 14 are threadedly connected, the piercing cylinder 15 drives the piercing needle 19 to move inwardly into the sleeve 14 until the piercing needle 19 passes through the cavity of the sleeve 14, the lumen of the sleeve 25, the second needle hole 27 of the slider 26, and the communication portion between the first chute 12 and the receiving groove in sequence and extends into the receiving groove. When the piercing cylinder 15 is rotated in the reverse direction relative to the sleeve 14, the piercing cylinder 15 can drive the piercing needle 19 to move outwardly from the sleeve 14, that is, the piercing needle 19 can be withdrawn from the receiving groove, the communication portion between the first chute 12 and the receiving groove, the second needle hole 27 of the slider 26, and the lumen of the sleeve 25 in sequence and enter the cavity of the sleeve 14. Of course, when the locking member 16 is removed from the lock hole 22 of the piercing cylinder 15, the piercing needle 19 can rotate relative to the piercing cylinder 15.

[0051] To facilitate observing the state of the piercing needle 19 in the piercing cylinder 15, an observation port 23 is opened on the wall of the piercing cylinder 15, and the observation port 23 communicates with the lock hole 22.

[0052] As Figure 1 、 3 、7, in the rat caudal vertebral intervertebral disc degeneration modeling device of the present invention, an axially arranged second chute 13 is provided on the arc-shaped lock plate 11. One end of the second chute 13 communicates with the first chute 12, and the other end of the second chute 13 penetrates through the arc-shaped lock plate 11. The slider 26 on the sleeve 14 first slides into the second chute 13 and then slides into the first chute 12 from the second chute 13. The shape of the slider 26 is adapted to the shapes of both the first chute 12 and the second chute 13, that is, both the first chute 12 and the second chute 13 are arc-shaped chutes, and the slider 26 is an arc-shaped slider.

[0053] As Figure 1 、 3 shown, both ends of the first strap 9 and the second strap 10 are respectively connected to the upper sides of the bearing plates 7 on the opposite sides of the carrier 8 by Velcro or bolts. Connecting the straps 9 and 10 to the bearing plate 7 by Velcro or bolts facilitates the disassembly and assembly of the straps 9 and 10.

[0054] As Figures 1-7 shown and in combination with Figures 8-15As shown, the method for modeling rat caudal vertebral disc degeneration using the above device in the present invention includes the following steps:

[0055] Step 1: Open the lid 2 of the anesthesia box 1, place the rat in the box body 3, and then cover the lid 2 again.

[0056] Step 2: Inject anesthetic gas (such as isoflurane) into the box body 3 through the air injection hole 4, observe the condition of the rat through the air permeable hole 5 on the box body 3. After the rat is completely anesthetized, open the lid 2, pass the rat's tail through the tail outlet hole 6 and extend it from the box body 3 to the outside of the box body 3.

[0057] Step 3: Move the first strap 9 and the second strap 10 away from above the carrier 8, and slide the arc-shaped locking plate 11 along the carrier 8 to the end of the carrier 8 away from the anesthesia box 1. Lay the rat's tail along the length direction of the carrier 8 on the bearing surface. Then, start palpating from the root of the rat's tail towards the tail end to determine the positions of the third vertebra to the seventh vertebra counted from the root of the rat's tail towards the tail end. The gap between any two adjacent vertebrae among the third vertebra to the seventh vertebra is the intervertebral disc. Take at least one intervertebral disc between the third vertebra and the seventh vertebra as the intervertebral disc to be punctured. Then, slide the arc-shaped locking plate 11 towards the direction close to the anesthesia box 1 along the carrier 8 and at the same time place the rat's tail in the receiving groove. Slide the arc-shaped locking plate 11 to the position of the intervertebral disc to be punctured on the rat's tail and make the needle tip 18 of the puncture needle 19 face the intervertebral disc to be punctured. While the arc-shaped locking plate 11 presses the rat's tail, press the first strap 9 and the second strap 10 on the rat's tail, and connect both ends of the first strap 9 and the second strap 10 to the upper sides of the bearing plates 7 on the opposite sides of the carrier 8 respectively to fix the rat's tail to give a certain tension to the intervertebral disc to be punctured, that is, the first strap 9 and the second strap 10 can fix the rat's tail and give a certain tension to the intervertebral disc to be punctured, facilitating the puncture operation.

[0058] Step 4: Move the sleeve 14 along the circumferential direction of the arc-shaped locking plate 11. When the sleeve 14 moves to the puncture position of the intervertebral disc to be punctured on the rat's tail, rotate the puncture cylinder 15 relative to the sleeve 14 to make the puncture needle 19 move closer to the rat's tail. When the needle tip 18 of the puncture needle 19 contacts the rat's tail, stop rotating the puncture cylinder 15. Image the rat's tail and the needle tip 18 through the X-ray machine to determine the puncture position of the needle tip 18. If the puncture position of the needle tip 18 does not meet the requirements, correct the puncture position of the needle tip 18 until the needle tip 18 is aligned with the puncture position of the intervertebral disc to be punctured on the rat's tail.

[0059] Step 5: Continue to rotate the puncture cylinder 15 relative to the sleeve 14. The needle tip 18 of the puncture needle 19 pierces into the rat tail. After piercing to a depth of 5 mm into the rat tail, stop piercing. At this time, rotate the needle tip 18 360 degrees for 5 times and stay for 30 seconds. Then rotate the puncture cylinder 15 in the reverse direction relative to the sleeve 14 until the needle tip 18 of the puncture needle 19 is withdrawn from the rat tail.

[0060] Step 6: Remove the first strap 9 and the second strap 10, and slide the arc-shaped lock plate 11 along the carrier 8 in the direction away from the anesthesia box 1 to separate the arc-shaped lock plate 11 from the rat tail. Then disinfect the puncture needle 19 opening on the rat tail. After that, transfer the rat from the anesthesia box 1 to the rat cage for continued feeding.

[0061] Step 7: Verify the intervertebral disc degeneration condition of the rat within the preset time. When obvious changes occur in the observation indexes of the intervertebral disc of the rat, the puncture causes intervertebral disc degeneration and the model establishment is successful.

[0062] In the above Step 5, when rotating the puncture cylinder 15 relative to the sleeve 14, the puncture cylinder 15 moves into the interior of the sleeve 14. The depth of the needle tip 18 piercing into the rat tail can be determined by the scale on the outer wall of the puncture cylinder 15. When rotating the needle tip 18, it is necessary to first remove the locking member 16 from the puncture cylinder 15. At this time, the puncture needle 19 can rotate relative to the puncture cylinder 15, that is, the puncture needle 19 can be rotated relative to the puncture cylinder 15, so that the needle tip 18 can make a rotational movement. After the rotation of the needle tip 18 is completed, insert the locking member 16 into the lock hole 22 of the puncture cylinder 15 again. At this time, the puncture needle 19 can follow the puncture cylinder 15 to rotate relative to the sleeve 14 again. Thus, when rotating the puncture cylinder 15 in the reverse direction relative to the sleeve 14, the puncture needle 19 follows the puncture cylinder 15 to move out of the sleeve 14 together, so that the needle tip 18 of the puncture needle 19 can be withdrawn from the rat tail.

[0063] In the method for establishing a rat caudal intervertebral disc degeneration model of the present invention, in the above Step 7, after the preset time, image the rat tail through an X-ray machine. The observation index is the gap between two adjacent vertebral bodies of the punctured intervertebral disc. When the gap becomes narrower relative to the initial state, the puncture causes intervertebral disc degeneration and the model establishment is successful. Set the preset time to 8 weeks. After this 8-week preset time, image the rat tail through an X-ray machine. The X-ray image formed is as Figure 10 shown. The position B in the figure is the gap between two adjacent vertebral bodies of a normal intervertebral disc, and the position C is the gap between two adjacent vertebral bodies of the punctured intervertebral disc. Relative to the gap at position B, the gap at position C is significantly narrowed.

[0064] The method for modeling rat caudal intervertebral disc degeneration in the present invention. In step seven, after a preset time, the tail of the rat is imaged by a nuclear magnetic resonance device. The observation index is the T2-weighted image of the punctured intervertebral disc. The T2-weighted image of the intervertebral disc of a normal rat is a high signal. If the T2-weighted image of the punctured intervertebral disc is a low signal, then the puncture causes intervertebral disc degeneration and the modeling is successful. The preset time is set to 8 weeks. After this 8-week preset time, magnetic resonance imaging (MRI) is performed on the rat's tail using a nuclear magnetic resonance device, and an MRI image as shown in Figure 11 is obtained. In the figure, the area D is a normal intervertebral disc, showing a high signal, which means that the gap between the adjacent two vertebral bodies of the normal intervertebral disc is larger, while the area E is the punctured intervertebral disc, showing a low signal, which means that the gap between the adjacent two vertebral bodies of the punctured intervertebral disc is smaller.

[0065] The method for modeling rat caudal intervertebral disc degeneration in the present invention. In step seven, after a preset time, the rat is sacrificed, the punctured intervertebral disc is dissected, placed in 4% paraformaldehyde for fixation for 24 hours, and then decalcified in 15% ethylenediaminetetraacetic acid (EDTA) for 30 days. After dehydration and clearing of the decalcified intervertebral disc, it is immersed in melted paraffin for embedding. Sections are made of the embedded intervertebral disc, and the thickness of the sections does not exceed 5 μm.

[0066] Hematoxylin and eosin (HE) staining is performed on the sectioned intervertebral disc. The observation index is the histological structure of the stained intervertebral disc. If the histological structure is damaged, then the puncture causes intervertebral disc degeneration and the modeling is successful, and / or

[0067] Safranin-fast green staining is performed on the sectioned intervertebral disc. The observation indexes are the histological structure, proteoglycan content, and type II collagen content of the stained intervertebral disc. If the histological structure is damaged and the proteoglycan content and type II collagen content decrease, then the puncture causes intervertebral disc degeneration and the modeling is successful.

[0068] In step seven above, the preset time is set to 8 weeks. After this 8-week preset time, HE staining is performed on the sectioned intervertebral disc, and a stained image as shown in Figure 13 is obtained. It can be concluded from the figure that the histological structure of the punctured intervertebral disc has been damaged. Of course, for comparison, HE staining can also be performed on a normal intervertebral disc (the staining method is the same as that of the punctured intervertebral disc and will not be elaborated), and a stained image as shown in Figure 12 is obtained. It can be concluded from the figure that the histological structure of the normal intervertebral disc has not been damaged.

[0069] In step seven above, the preset time is set to 8 weeks. After this 8-week preset time, safranin-fast green staining is performed on the sectioned intervertebral disc, and a stained image as shown in Figure 15The stained image shows that the histological structure of the punctured intervertebral disc has been damaged, and the contents of proteoglycan and type II collagen have decreased. Of course, for comparison, a normal intervertebral disc can also be stained with safranin-fast green (the staining method is the same as that of the punctured intervertebral disc and will not be elaborated here), and the stained image as shown in Figure 14 is obtained. It can be seen from the figure that the histological structure of the normal intervertebral disc has not been damaged, and the contents of proteoglycan and type II collagen have not decreased.

[0070] The preset time in the above step seven can also be set to 4 weeks. In addition, in the above step four, an X-ray machine can be used to image the rat tail and the needle 18 to determine the puncture position of the needle 18 and correct it. In fact, in the above step five, after the needle 18 is inserted into the rat tail, the X-ray machine can also be used to image the rat tail and the needle 18 to check whether the puncture position of the needle 18 is accurate. As shown in Figure 9 is the X-ray image of the needle 18 inserted into the rat tail. Figure 8 is the appearance image of the rat caudal intervertebral disc obtained after the puncture surgery.

[0071] In the present invention, the rat is anesthetized by the anesthesia box 1, and then the rat tail is fixed by the carrier table 8, the binding band and the arc-shaped locking plate 11. Then, the intervertebral disc on the rat tail is punctured by the sleeve 14, the puncture cylinder 15 and the puncture needle 19. During the puncture process, the position of the needle 18 of the puncture needle 19 can be corrected by the X-ray machine for accurate puncture. It can be seen that the present invention integrates the anesthesia box 1, the X-ray machine, the carrier table 8, the binding band, the arc-shaped locking plate 11, the sleeve 14 and the puncture cylinder 15. Only a few of the above tools are needed to perform the modeling surgery on the rat caudal intervertebral disc, and the process is relatively simple, which is convenient for popularization and application.

[0072] It should be noted that the orientation or positional relationship indicated by the terms "center", "upper", "lower", "front", "rear", "left", "right", "middle", etc. is based on the orientation or positional relationship shown in the drawings. It is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, so it should not be construed as a limitation of the present invention.

[0073] In the description of the present invention, it should be noted that unless otherwise clearly specified and limited, the terms "connected" and "connected" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific situations.

[0074] The embodiments described above are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the spirit of the present invention's design, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A rat coccygeal disc degeneration modeling device, characterized in that: The invention comprises an anesthesia box, wherein the anesthesia box is provided with a tail hole, a horizontally arranged bearing plate is provided on the outer side of the anesthesia box, a bearing platform opposite to the tail hole is provided on the upper side of the bearing plate, an arc-shaped locking plate is slidably mounted on the bearing platform, two ends of the arc-shaped locking plate are respectively slidably connected to the opposite sides of the bearing platform, a receiving groove is formed between the arc-shaped locking plate and the bearing platform, a sleeve is slidably mounted on the arc-shaped locking plate along the circumferential direction, the axis of the sleeve is arranged along the radial direction of the arc-shaped locking plate, a puncture tube is connected to the inner thread of the sleeve, and an axially arranged engraved groove is provided on the outer side wall of the puncture tube. A ruler, a puncture needle is installed in the puncture tube, the puncture needle includes a base and a needle connected to the base, the base is installed in the puncture tube, the needle passes through the puncture tube, the sleeve and the arc-shaped locking plate in sequence and then extends into the accommodating groove, a first strap and a second strap are provided above the supporting platform, the first strap and the second strap are respectively located on the opposite sides of the arc-shaped locking plate, both ends of the first strap and the second strap are respectively connected to the side surfaces of the supporting plate on the opposite sides of the supporting platform, and an X-ray machine is provided on the periphery of the supporting plate for determining the puncture position of the needle by irradiation imaging.

2. The rat coccygeal disc degeneration modeling device according to claim 1, characterized in that: The anesthesia box includes a box body and a box cover, the box body includes a box side wall arranged vertically and in a rectangular cylindrical structure, the lower cylinder opening of the box side wall is fixedly provided with a box bottom wall, the upper cylinder opening of the box side wall is covered with a box cover, the tail hole is arranged on the box side wall, the box side wall is provided with an air injection hole arranged opposite to the tail hole, and the box side wall is also provided with an air vent.

3. The rat coccygeal disc degeneration modeling device according to claim 2, characterized in that: The supporting platform is in the shape of an elongated strip, with one end of the supporting platform being arranged close to the anesthesia box, and the other end of the supporting platform being arranged away from the anesthesia box. The lower side surface of the supporting platform is fixedly arranged on the supporting plate, and the upper side surface of the supporting platform is the supporting surface, which is a downwardly concave arc surface, and the two ends of the arc-shaped locking plate are respectively slidably connected to the opposite sides of the supporting platform along the length direction of the supporting platform.

4. The rat coccygeal disc degeneration modeling device according to claim 3, characterized in that: The arc-shaped locking plate is an upwardly protruding arc-shaped plate, and a circumferentially arranged first slide groove is provided on the outer peripheral surface of the arc-shaped locking plate, and the first slide groove is communicated with the accommodating groove. A slider is fixedly provided at one end of the sleeve close to the arc-shaped locking plate, and the slider is slidably installed in the first slide groove. The needle passes through the puncture tube, the sleeve, the slider and the first slide groove on the arc-shaped locking plate in sequence and then extends into the accommodating groove.

5. The rat coccygeal disc degeneration modeling device according to claim 4, characterized in that: A locking piece is inserted radially on the puncture tube, and the locking piece abuts against an end of the base away from the needle.

6. The rat coccygeal disc degeneration modeling device according to claim 5, characterized in that: The arc-shaped locking plate is provided with an axially arranged second slide groove, one end of the second slide groove is connected to the first slide groove, and the other end of the second slide groove passes through the arc-shaped locking plate, and both ends of the first strap and the second strap are respectively connected to the side surfaces of the supporting plate on opposite sides of the supporting platform by Velcro or bolts.

7. A method for modeling rat coccygeal intervertebral disc degeneration using the device of claim 6, characterized in that: The following steps are involved: Step 1: Open the lid of the anesthesia box, place the rat in the box, and then close the lid. Step 2: Inject anesthetic gas into the box through the air injection hole, observe the rat's condition through the air vents on the box, and when the rat is completely anesthetized, open the box cover and extend the rat's tail through the tail outlet hole from inside the box to outside the box. Step 3: Remove the first strap and the second strap from above the support platform, and slide the arc-shaped locking plate along the support platform to the end of the support platform away from the anesthesia box, lay the rat's tail on the support surface along the length direction of the support platform, and then palpate from the root of the rat's tail to the tail end to determine the position of the third vertebra to the seventh vertebra counting from the root of the rat's tail to the tail end. The gap between any two adjacent vertebrae from the third vertebra to the seventh vertebra is the intervertebral disc, and at least one intervertebral disc between the third vertebra to the seventh vertebra is the intervertebral disc to be penetrated. The intervertebral disc is punctured, and then the arc-shaped locking plate is slid along the bearing platform toward the direction close to the anesthesia box and the rat tail is placed in the receiving groove at the same time. The arc-shaped locking plate is slid to the intervertebral disc position to be punctured of the rat tail and the needle head of the puncture needle is opposite to the intervertebral disc to be punctured. While the arc-shaped locking plate presses the rat tail, the first strap and the second strap are pressed on the rat tail, and both ends of the first strap and the second strap are respectively connected to the side surfaces of the bearing plate on the opposite sides of the bearing platform, and the rat tail is fixed to give a certain tension to the intervertebral disc to be punctured. Step 4: Move the sleeve along the circumference of the arc-shaped locking plate. When the sleeve moves to the puncture position of the intervertebral disc to be punctured in the rat's tail, rotate the puncture tube relative to the sleeve to move the puncture needle close to the rat's tail. When the needle tip of the puncture needle contacts the rat's tail, stop rotating the puncture tube, image the rat's tail and the needle tip through an X-ray machine, and determine the puncture position of the needle tip. If the puncture position of the needle tip does not meet the requirements, correct the puncture position of the needle tip until the needle tip is aligned with the puncture position of the intervertebral disc to be punctured in the rat's tail. Step 5: Continue to rotate the puncture tube relative to the sleeve, and the needle tip of the puncture needle will penetrate the rat's tail. After the needle tip penetrates 5 mm into the rat's tail, stop puncturing. At this time, rotate the needle tip 360 degrees 5 times, stay for 30 seconds, and then rotate the puncture tube in the opposite direction relative to the sleeve until the needle tip of the puncture needle is pulled out of the rat's tail. Step 6: Remove the first and second straps, and slide the arc-shaped locking plate along the support platform away from the anesthesia box to separate the arc-shaped locking plate from the rat's tail. Then disinfect the puncture needle hole on the rat's tail, and then transfer the rat from the anesthesia box to the mouse cage for continued feeding. Step 7: Verify the intervertebral disc degeneration of the rats within the preset time. When the observed indicators of the intervertebral disc of the rats show obvious changes, the puncture causes intervertebral disc degeneration and the model is successful.

8. The rat coccygeal disc degeneration modeling method according to claim 7, characterized in that: In step seven, after a preset time, the rat's tail is imaged by an X-ray machine, and the observation index is the gap between two adjacent vertebrae of the punctured intervertebral disc. When the gap becomes narrower than the initial state, the puncture causes intervertebral disc degeneration and the model is successful.

9. The rat coccygeal disc degeneration modeling method according to claim 7, characterized in that: In the step seven, after a preset time, the rat's tail is imaged by a nuclear magnetic resonance device, and the observation index is the T2-weighted image of the punctured intervertebral disc. The T2-weighted image of the intervertebral disc of a normal rat is a high signal. If the T2-weighted image of the punctured intervertebral disc is a low signal, the puncture causes intervertebral disc degeneration and the model is successful.

10. The rat coccygeal disc degeneration modeling method according to claim 7, characterized in that: In step 7, after a preset time, the rats are killed, the punctured intervertebral discs are dissected, fixed in 4% paraformaldehyde for 24 hours, and then decalcified in 15% ethylenediaminetetraacetic acid (EDTA) for 30 days. The decalcified intervertebral discs are dehydrated and transparent, and then immersed in melted paraffin for embedding. The embedded intervertebral discs are sliced, and the thickness of the slices does not exceed 5 μm. The sliced ​​intervertebral disc is stained with hematoxylin and eosin (HE). The observation index is the histological structure of the stained intervertebral disc. If the histological structure is destroyed, the puncture causes intervertebral disc degeneration and the model is successful, and / or The sliced ​​intervertebral disc was stained with safranin-fast green, and the observation indicators were the histological structure, proteoglycan content and type II collagen content of the stained intervertebral disc. If the histological structure was destroyed and the proteoglycan content and type II collagen content decreased, the puncture caused intervertebral disc degeneration and the model was successful.

Images