Construction method of lumbar spinal stenosis animal model

Abstract

The invention discloses a method for constructing a lumbar spinal stenosis animal model, which comprises the following steps of: fixing an experimental rabbit at a prone position, and then forming a 2-3cm incision in the back side of the waist along the longitudinal direction so as to completely expose an intervertebral plate window between a first lumbar vertebra and a second lumbar vertebra; a disposable venous indwelling needle is used for puncturing the ligamentum flavum at the interlaminal window and then puncturing the disposable venous indwelling needle into the spinal canal, and then a metal needle core of the venous indwelling needle is pulled out, so that the soft catheter part of the venous indwelling needle is indwelled in the spinal canal; uniformly mixing PMMA bone cement powder and liquid to prepare fluid bone cement, extracting the bone cement by using an injector, slowly injecting the bone cement into the spinal canal, and removing the soft catheter after injection is completed; the injected bone cement is adhered to the inner wall of the spinal canal after being solidified, so that the inner diameter and the volume of the spinal canal are reduced in physical space, compression on spinal nerves is formed, and the animal model with lumbar spinal stenosis is obtained. The problem that in the preparation process of an animal model, nerves are prone to being damaged, and research test requirements cannot be met is solved.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of constructing animal models, and particularly relates to a construction method of a lumbar spinal stenosis animal model. BACKGROUND

[0002] Lumbar spinal stenosis is a common disease in orthopedics, and specifically refers to a kind of disease that various causes cause the shortening of each diameter line of the spinal canal and the decrease of the effective space of the canal lumen, causes the compression of the dural sac, the spinal cord or the nerve root, and thus causes the corresponding nerve dysfunction. The causes of lumbar spinal stenosis are very complex, and are caused by congenital and degenerative diseases of the spine. The research on lumbar spinal stenosis is increasing, and researchers simulate and observe the process of lumbar spinal stenosis in the living body of experimental animals (mice, rabbits and the like) by making a lumbar spinal stenosis animal model, analyze various accompanying symptoms, provide a reference for the research of etiology and pathology, and can also evaluate the expected effect of the treatment method and the drug for lumbar spinal stenosis.

[0003] At present, the construction method of the lumbar spinal stenosis animal model mostly adopts the balloon catheter compression method and the cerclage method, and the lumbar spinal stenosis is formed by compression through a balloon or the like. However, such compression by external force through a balloon or the like needs to be accurately controlled, and nerve damage is easily caused in the operation process due to the external force. In the later observation and research, it is unable to distinguish whether the various symptoms generated are caused by nerve damage or the lumbar spinal stenosis, and the requirement of accurately simulating lumbar spinal stenosis cannot be met, which affects the accuracy of the corresponding research. SUMMARY

[0004] In view of the above problems, the application discloses a construction method of a lumbar spinal stenosis animal model, which solves the problem that the lumbar spinal stenosis animal model is easily damaged in the preparation process and cannot meet the research requirements.

[0005] The application is implemented by adopting the following technical scheme: A construction method of a lumbar spinal stenosis animal model, comprising the following steps: (1) Take an experimental rabbit in a prone position, then make a 2-3 cm incision on the dorsal side of the waist of the experimental rabbit in a longitudinal direction, and completely expose the interlaminar window between the first lumbar vertebra and the second lumbar vertebra of the experimental rabbit; (2) Take a disposable intravenous indwelling needle, puncture the yellow ligament at the interlaminar window of the first lumbar vertebra and the second lumbar vertebra of the experimental rabbit, then puncture into the spinal canal, and then remove the metal needle core of the intravenous indwelling needle, and retain the soft catheter part of the intravenous indwelling needle in the spinal canal; (3) the PMMA bone cement powder and liquid are mixed evenly to form a fluid bone cement, then 0.5-0.8 mL of the bone cement is extracted by a syringe and slowly injected into the spinal canal through a soft catheter, the soft catheter is removed after 10-12 minutes of waiting after the injection is completed; (4) the bone cement injected in step (3) is solidified and adheres to the inner wall of the spinal canal, which reduces the diameter and volume of the spinal canal in physical space and forms compression on the spinal cord nerves, thus obtaining an animal model of lumbar spinal stenosis.

[0006] Further, in step (1), the experimental rabbit is an adult New Zealand white rabbit.

[0007] Further, in step (2), the first lumbar spine and the second lumbar spine are longitudinally cut 2-3 cm and then separated layer by layer to expose the interlaminar window of the first lumbar spine and the second lumbar spine, respectively, with the first lumbar spine and the second lumbar spine as reference mark points.

[0008] Further, in step (2), after the intravenous indwelling needle is inserted into the spinal canal, the metal needle core of the intravenous indwelling needle is removed after no cerebrospinal fluid flows out after back suction, and the soft catheter part of the intravenous indwelling needle is retained in the spinal canal.

[0009] Further, in step (3), the PMMA bone cement powder and liquid are mixed evenly to form a fluid bone cement, then 0.5-0.8 mL of the bone cement is extracted by a syringe and slowly injected into the spinal canal through a soft catheter, the soft catheter is removed after 10-12 minutes of waiting after the injection is completed;

[0010] Further, in step (3), 0.5-0.8 mL of the bone cement is extracted by a syringe and slowly injected into the spinal canal through a soft catheter at a uniform speed, and the injection time is not less than 1 min.

[0011] The technical solution has the following beneficial effects compared with the prior art: The method is simple, efficient, high success rate and good expansibility, and fills the blank of the construction of the lumbar spinal stenosis animal model. The New Zealand white rabbits are selected as the experimental rabbits, a 2-3 cm incision is made in the waist of the experimental rabbit in the longitudinal direction, and the interlaminar window between the first and second lumbar vertebrae of the experimental rabbit is completely exposed, then a disposable intravenous puncture needle is used to pierce the yellow ligament in the interlaminar window and reach the intraspinal canal, then the PMMA is prepared to obtain a fluid-like bone cement and immediately injected into the lumbar spinal canal, and after the solidification forms an intraspinal canal occupation, the soft catheter is removed, the solidified PMMA adheres to the intraspinal canal wall, reduces the intraspinal canal diameter and the intraspinal canal volume, forms compression to the spinal cord nerve, forms a lumbar spinal stenosis animal model (LSS animal model), and the process of solidification of the solidified bone cement from liquid to solid is slow, which will not cause external force damage to the nerve, and solves the problem that the lumbar spinal stenosis animal model is easy to damage the nerve during preparation and cannot meet the research test requirements. BRIEF DESCRIPTION OF DRAWINGS

[0012] Figure 1 is the picture of the operation described in step (1) in Example 1.

[0013] Figure 2 is the picture of the interlaminar window after the operation described in step (1) in Example 1.

[0014] Figure 3 is the picture of the operation described in step (2) in Example 1.

[0015] Figure 4 is the picture of the operation described in step (4) in Example 1.

[0016] Figure 5 is the picture of the operation described in step (5) in Example 1.

[0017] Figure 6 is the vertebral body cross-sectional view of the lumbar spinal stenosis animal model obtained by the method described in Examples 1-4.

[0018] Figure 7 is the X-ray perspective image of the vertebral body of the lumbar spinal stenosis animal model obtained by the method described in Examples 1-4. DETAILED DESCRIPTION

[0019] The present application is further illustrated by the following examples, but not as a limitation to the present application. The specific experimental conditions and methods not specified in the following examples are generally conventional means known to those skilled in the art.

[0020] Example 1: A construction method of a lumbar spinal stenosis animal model, comprising the following steps: (1) Take adult New Zealand white rabbits as experimental rabbits and fix them in a prone position. Then, make a 3 cm incision on the dorsal side of the experimental rabbit's waist in the longitudinal direction and completely expose the interlaminar window between the first and second lumbar vertebrae of the experimental rabbit. Specifically, as shown in Figure 1 , take the first and second lumbar spinous processes of the experimental rabbit as reference marker points, make a 3 cm longitudinal incision, and then, as shown in Figure 2 , separate and expose the interlaminar window of the first and second lumbar vertebrae layer by layer; (2) As shown in Figure 3 , take a disposable intravenous indwelling needle to puncture the yellow ligament at the interlaminar window of the first and second lumbar vertebrae of the experimental rabbit, and then insert it into the spinal canal. After inserting the intravenous indwelling needle into the spinal canal, back suction so that no cerebrospinal fluid flows out, remove the metal needle core of the intravenous indwelling needle, and leave the soft catheter part of the intravenous indwelling needle in the spinal canal; (3) Take the powder and liquid of PMMA bone cement and mix them evenly to make a fluid-like bone cement. Then, as shown in Figure 4 , use a syringe to draw 0.5 mL of bone cement and inject it into the spinal canal through the soft catheter at a constant speed, with an injection time of 1 min. After 10 minutes of waiting after injection is completed, remove the soft catheter; (4) As shown in Figure 5 , after the bone cement injected in step (3) is solidified, it adheres to the inner wall of the spinal canal, reducing the diameter and volume of the spinal canal in physical space and forming compression on the spinal cord nerves, obtaining a lumbar spinal stenosis animal model (see Figure 6 and Figure 7 A).

[0021] Example 2: A method for constructing a lumbar spinal stenosis animal model, comprising the following steps: (1) Take adult New Zealand white rabbits as experimental rabbits and fix them in a prone position. Then, make a 2 cm incision on the dorsal side of the experimental rabbit's waist in the longitudinal direction and completely expose the first and second lumbar vertebrae of the experimental rabbit. Specifically, take the first and second lumbar spinous processes of the experimental rabbit as reference marker points, make a 2 cm longitudinal incision, and then separate and expose the interlaminar window of the first and second lumbar vertebrae layer by layer; (2) Take a disposable intravenous indwelling needle to puncture the yellow ligament at the interlaminar window of the first and second lumbar vertebrae of the experimental rabbit, and then insert it into the spinal canal. After inserting the intravenous indwelling needle into the spinal canal, back suction so that no cerebrospinal fluid flows out, remove the metal needle core of the intravenous indwelling needle, and leave the soft catheter part of the intravenous indwelling needle in the spinal canal; (3) Take the powder and liquid of PMMA bone cement and mix them evenly to make a fluid-like bone cement, then add physiological saline to the bone cement and mix, draw 0.6 mL of the bone cement with a syringe and slowly inject it into the spinal canal through the soft catheter, the injection time is 2 min, the volume ratio of physiological saline to bone cement is 1:10, after the injection is completed and waiting for 12 min, remove the soft catheter; (4) The injected bone cement in step (3) is solidified and adheres to the inner wall of the spinal canal, which reduces the inner diameter and volume of the spinal canal in physical space, forming compression on the spinal cord nerve, obtaining a lumbar spinal stenosis animal model (see Figure 6 and Figure 7 B).

[0022] Example 3: A method for constructing a lumbar spinal stenosis animal model, comprising the following steps: (1) Take adult New Zealand white rabbits as experimental rabbits and fix them in a prone position, then make a 2 cm longitudinal incision on the dorsal side of the experimental rabbit's waist and completely expose the interlaminar window between the first and second lumbar vertebrae of the experimental rabbit, specifically, take the first and second lumbar spinous processes of the experimental rabbit as reference marker points, make a 2 cm longitudinal incision, then separate and expose the interlaminar window of the first and second lumbar vertebrae layer by layer; (2) Take a disposable intravenous indwelling needle, pierce the yellow ligament at the interlaminar window of the first and second lumbar vertebrae of the experimental rabbit, then pierce into the spinal canal, after the needle is pierced into the spinal canal, back suction is performed so that no cerebrospinal fluid flows out, then the metal needle core of the intravenous indwelling needle is removed, and the soft catheter part of the intravenous indwelling needle is retained in the spinal canal; (3) Take the powder and liquid of PMMA bone cement and mix them evenly to make a fluid-like bone cement, then add physiological saline to the bone cement and mix, draw 0.7 mL of the bone cement with a syringe and slowly inject it into the spinal canal through the soft catheter, the injection time is 1.5 min, the volume ratio of physiological saline to bone cement is 1:8, after the injection is completed and waiting for 11 min, remove the soft catheter; (4) The injected bone cement in step (3) is solidified and adheres to the inner wall of the spinal canal, which reduces the inner diameter and volume of the spinal canal in physical space, forming compression on the spinal cord nerve, obtaining a lumbar spinal stenosis animal model (see Figure 6 and Figure 7 C).

[0023] Example 4: A method for constructing a lumbar spinal stenosis animal model, comprising the following steps: (1) Take adult New Zealand white rabbits as experimental rabbits and take prone position fixation, then make a 3cm incision on the dorsal side of the experimental rabbit's waist in the longitudinal direction and completely expose the interlaminar window between the first and second lumbar vertebrae of the experimental rabbit, that is, take the first lumbar vertebra and the second lumbar vertebra of the experimental rabbit as reference marker points, cut 3cm longitudinally, and then separate and expose the interlaminar window of the first and second lumbar vertebrae layer by layer; (2) Take a disposable intravenous catheter to puncture the yellow ligament at the interlaminar window of the first and second lumbar vertebrae of the experimental rabbit, then puncture into the spinal canal, and after the intravenous catheter is inserted into the spinal canal, back suction is performed so that no cerebrospinal fluid flows out, then the metal needle core of the intravenous catheter is removed, and the soft catheter part of the intravenous catheter is retained in the spinal canal; (3) Take the powder and liquid of PMMA bone cement and mix them evenly to make the bone cement into a fluid state, then add physiological saline to the bone cement, draw 0.8mL of bone cement with a syringe, and slowly inject it into the spinal canal through the soft catheter, and the injection time is 2min, the volume ratio of physiological saline and bone cement is 1:9, and the soft catheter is removed after 12min of injection; (4) The bone cement injected in step (3) is solidified and adheres to the inner wall of the spinal canal, reducing the diameter and volume of the spinal canal in physical space, forming compression on the spinal cord nerves, and obtaining a lumbar spinal stenosis animal model (see Figure 6 and Figure 7 D).

[0024] In addition, it should be understood that although the present specification is described in terms of embodiments, not every embodiment contains only one independent technical solution, and the description of the specification is only for the sake of clarity, and those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can be combined appropriately to form other embodiments that those skilled in the art can understand.

Claims

1. A method for constructing an animal model of lumbar spinal stenosis, characterized by: The method comprises the following steps: (1) Take the experimental rabbit to adopt prone position fixation, then open 2-3 cm along the longitudinal direction on the dorsal side of the experimental rabbit's waist, completely expose the interlaminar window between the first lumbar vertebra and the second lumbar vertebra of the experimental rabbit; (2) Take a disposable intravenous catheter to puncture the yellow ligament at the interlaminar window of the first lumbar vertebra and the second lumbar vertebra of the experimental rabbit, then puncture into the spinal canal, then remove the metal needle core of the intravenous catheter, and leave the soft catheter part of the intravenous catheter in the spinal canal; (3) Take the powder and liquid of PMMA bone cement and mix them evenly to make the bone cement into a fluid, then use a syringe to extract 0.5-0.8 mL of bone cement and slowly inject it into the spinal canal through the soft catheter, remove the soft catheter after waiting for 10-12 minutes after injection is completed; (4) After the bone cement injected in step (3) is solidified, it adheres to the inner wall of the spinal canal, which reduces the diameter and volume of the spinal canal in physical space, forms compression on the spinal cord nerve, and finally obtains the animal model of lumbar spinal stenosis.

2. The method for constructing an animal model of lumbar spinal stenosis according to claim 1, characterized in that: In step (1), the experimental rabbit is an adult New Zealand white rabbit.

3. The method for constructing an animal model of lumbar spinal stenosis according to claim 1, characterized in that: In step (2), the first lumbar vertebra and the second lumbar vertebra of the experimental rabbit are taken as reference mark points, and the interlaminar window of the first lumbar vertebra and the second lumbar vertebra is exposed layer by layer after being cut open longitudinally for 2-3 cm.

4. The method for constructing an animal model of lumbar spinal stenosis according to claim 1, characterized in that: In step (2), after the intravenous catheter is punctured into the spinal canal, the metal needle core of the intravenous catheter is removed after back suction to ensure that no cerebrospinal fluid flows out, so that the soft catheter part of the intravenous catheter remains in the spinal canal.

5. The method for constructing an animal model of lumbar spinal stenosis according to claim 1, characterized in that: In step (3), the powder and liquid of PMMA bone cement are mixed evenly to make the bone cement into a fluid, then physiological saline is added to the bone cement, and 0.5-0.8 mL of bone cement is extracted by a syringe and slowly injected into the spinal canal through the soft catheter, the volume ratio of physiological saline to bone cement is 1: (8-10).

6. The method for constructing an animal model of lumbar spinal stenosis according to claim 1, characterized in that: In step (3), 0.5-0.8 mL of bone cement is extracted by a syringe and slowly injected into the spinal canal through the soft catheter at a uniform speed, and the injection time is not less than 1 min.

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