CN111357711A - Establishment of non-compression type lumbar disc herniation nucleus pulposus animal model - Google Patents
Establishment of non-compression type lumbar disc herniation nucleus pulposus animal model Download PDFInfo
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Abstract
A method for establishing a non-compression type lumbar intervertebral disc protrusion animal model comprises the following steps: s1, cutting the rat tail at a position 1cm away from the root of the rat tail, suturing the wound and obtaining the nucleus pulposus of the intervertebral disc of the tail vertebra of the rat; s2, cutting the fibrous ring of the intervertebral disc of the caudal vertebra of the rat, wherein the nucleus pulposus is in a jelly sample, and fully mixing and diluting 5 nucleus pulposus and 50 microliters of physiological saline to form a suspension; s3, injecting 20 mul of nucleus pulposus suspension and 30 mul of 2% lidocaine injection into the lumbar epidural space to enable the suspension to be filled around nerve roots. The invention utilizes the characteristic that the tail of a rat is thick and contains enough intervertebral disc tissues, and the nucleus pulposus of the rat is in a jelly shape, and combines an epidural puncture technology to inject the autologous nucleus pulposus suspension of the rat into the epidural space of the lumbar vertebra to manufacture the nucleus pulposus tissue exposed to blood circulation and an immune system of an organism.
Description
Technical Field
The invention belongs to the technical field of animal medical animal model establishment, and particularly relates to establishment of a non-compression type lumbar disc herniation nucleus pulposus animal model.
Background
The traditional view is that the herniated nucleus pulposus of the lumbar intervertebral disc presses nerve roots to cause sciatica. In recent years, with the progress of understanding and research on lumbar intervertebral disc herniation (LDH), it has been found that sciatica occurs even if nerve roots are not pressed by the herniated nucleus pulposus, and the pathogenesis of sciatica is unclear. People hope to accurately observe the change generated in the model body by establishing a reliable and effective non-compression type lumbar disc herniation animal model and applying intervention factors, explore the disease occurrence mechanism, popularize the research result to the clinic and provide a reasonable and effective prevention and treatment means for treating the lumbar disc herniation. Therefore, establishing an ideal non-compression type animal model of the lumbar disc herniation has important significance for the mechanism research and the clinical prevention and treatment of the disease.
At present, animal models of prolapse of lumbar intervertebral disc are classified into compression type and non-compression type according to whether or not the nucleus pulposus of the lumbar intervertebral disc compresses nerve roots. The compression type lumbar disc herniation animal model is mature, but the non-compression type lumbar disc herniation animal model is rare. Scholars at home and abroad establish some animal models aiming at the mechanism characteristics of non-compression type intervertebral disc herniation, but the establishment methods of the models are complex, the animal damage is large, the success rate is low, and some models have the defects of poor repeatability and high price.
The requirements of an ideal animal model are that ① can reproduce the objective rule of intervertebral disc degenerative disease, the physiological and anatomical features of ② model animals are highly similar to those of human beings, the ③ animal model has good repeatability and high controllability, and ④ has short period, good economy, strong repeatability and high success rate.
At present, the traditional Chinese medicine is commonly used for animals of lumbar intervertebral disc protrusion models, primates such as monkeys and the like are closer to the anatomical physiological characteristics of human beings, but the animals have high price, special requirements on laboratory conditions and equipment, long experimental period and inconvenience for large-scale adoption. The large mammals of pigs and dogs have the defects of high price, long experimental period, limited experimental places and the like. In various animals, Sprague-Dawlay rats (also called SD rats) have genes close to those of human beings, and the basic spinal column constitution is similar to that of human beings, for example, the nerve roots of L4 and L5 are the main components of sciatic nerve, and the strain is complete, has strong anti-infection power and vitality, is economical and easily obtained, and can be used in large quantities.
Chinese patent application "method for establishing animal model of rupture type lumbar disc herniation", patent application No. CN201710682995.8, discloses a method for establishing animal model of rupture type lumbar disc herniation, the invention takes the nucleus pulposus of intervertebral disc fully contacting blood circulation after the rupture of the simulated posterior longitudinal ligament as an entry point, and creates the condition that the nucleus pulposus tissue is exposed in blood circulation and immune system of organism by artificially cutting the cartilage end plate to expose nucleus pulposus, and establishes an animal model capable of promoting the reabsorption of the protruded intervertebral disc. The nucleus pulposus of the intervertebral disc at the tail part of the rat is placed beside a nerve root at the waist by adopting an operation method, so that the nucleus pulposus of the intervertebral disc is pressed to the nerve root, the operation incision is long, and the lumbar vertebral plate of the rat needs to be cut to expose nerve tissues. The disadvantages of this model are: 1. the nucleus pulposus causes compression on nerve roots. 2. The operation wound is large, the death rate of rats is high, and the possibility of damaging nerves by the operation is high. 3. The influence caused by the compression of nerve roots by the traditional lumbar disc nucleus pulposus protrusion is researched.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides the establishment method of the non-compression type lumbar disc herniation animal model with simple operation, high success rate, low price, good repeatability and small interference on nerve roots.
In order to achieve the purpose, the invention provides the following technical scheme: a method for establishing an animal model of non-compression type lumbar intervertebral disc protrusion comprises the following steps:
s1, cutting the rat tail at a position 1cm away from the root of the rat tail, suturing the wound and obtaining the nucleus pulposus of the intervertebral disc of the tail vertebra of the rat;
s2, cutting the fibrous ring of the intervertebral disc of the caudal vertebra of the rat, wherein the nucleus pulposus is in a jelly sample, and fully mixing and diluting 5 nucleus pulposus and 50 microliters of physiological saline to form a suspension;
s3, injecting the 20 ul nucleus pulposus suspension and the 30 ul 2% lidocaine injection into the lumbar epidural space of the rat so that the nucleus pulposus suspension is filled around nerve roots.
Preferably, in S1, the rat is a 10-month-old SPF (Specific Pathogen Free) grade SD male rat.
Preferably, in S1, the process of cutting and obtaining the nucleus pulposus of the rat caudal vertebral disc comprises:
s11, after the rat is bred adaptively for 1 week, the rat is anesthetized;
s12, after anesthesia is successful, removing the body hair of the back and hind limbs of the rat by using a depilatory;
s13, carrying out conventional skin disinfection, cutting off rat tail at a position 1cm away from the tail root of the rat under aseptic conditions, suturing the wound, obtaining the nucleus pulposus of the intervertebral disc of the tail vertebra of the rat, mixing 5 nucleus pulposus and 50 mu l of physiological saline, and fully mixing and diluting the mixture into suspension.
Preferably, in S11, ketamine with a concentration of 1% is selected as an anesthetic, and 50 mg.Kg-1 is used for anesthetizing the rat by intraperitoneal injection.
Preferably, in S13, the 5 nuclei is mixed with 50 μ l of physiological saline, and the mixture is thoroughly mixed and diluted to form a suspension.
Preferably, the rat tail is cut off and the wound is sutured, the nucleus pulposus of the intervertebral disc of the rat caudal vertebra is obtained, 5 nucleus pulposus are taken and mixed with 50 mu l of physiological saline, and the mixture is fully mixed and diluted into a suspension.
Preferably, in the S3, the injecting the 20 μ l nucleus pulposus suspension into the lumbar epidural space by the 30 μ l2% lidocaine injection comprises: the rat is in a lateral decubitus position, epidural puncture is performed by using a No. 9 lumbar puncture needle with the interval between L4-L5 spinous processes as a puncture point, and 20 mul of autologous nucleus pulposus suspension and 30 mul of 2% lidocaine are injected respectively.
Has the advantages that:
the invention utilizes the characteristic that the tail of a rat is thick and contains enough intervertebral disc tissues, the nucleus pulposus of the rat is in a jelly shape, and combines an epidural puncture technology to inject autologous nucleus pulposus suspension of the rat into the epidural space of the lumbar vertebra to manufacture the condition that the nucleus pulposus tissues are exposed to blood circulation and an organism immune system but do not press the nerve roots, and the nerve conduction speed is measured by using an electromyography-evoked potential instrument to show that the speeds of motor nerves and sensory nerves are obviously reduced and the morphological structure of the nerve roots also obviously changes along with the time lapse, thereby proving that the function and the morphological change of the nerve roots can also occur even if the nucleus pulposus suspension positioned in the lumbar spinal canal of the rat does not press the nerve roots, and proving the effectiveness of the model.
Drawings
FIG. 1 is a table showing the change of conduction velocity of lumbar nerve roots of rats in the experimental group of the present invention.
FIG. 2 is a table showing the change of conduction velocity of lumbar nerve roots of rats in the control group according to the present invention.
FIG. 3 is a diagram showing the change of conduction velocity of lumbar nerve roots of rats in the experimental group and the control group according to the present invention.
FIG. 4 is a schematic diagram of the 10-day staining pattern of the HE of the nerve roots in the control group.
FIG. 5 is a schematic diagram of the HE staining pattern of nerve roots in the 10-day experimental group of the present invention.
FIG. 6 is a schematic flow chart of the operation of the present invention.
In fig. 3: MCV1 is the conduction velocity of lumbar motor nerves of rats in experimental groups;
SCV1 is the conduction velocity of lumbar sensory nerve of rats in experimental groups;
MCV2 is the conduction velocity of lumbar motor nerve of control rat;
SCV2 is the lumbar sensory nerve conduction velocity of control rats.
As shown in FIG. 4, in the lumbar nerve root cross section 10 days after the operation of the control group, the nerve fibers were substantially uniform in size, and the myelin sheath was circular and located in the periphery and in a red network structure, and the axon was small and located in the center (HE × 200).
As shown in FIG. 5, the lumbar nerve root cross section was observed 10 days after the operation, the nerve fibers were of different sizes, the staining was light, and the myelin sheath was mostly disintegrated, (HE × 200).
Detailed Description
The technical solution implemented by the present invention will be clearly and completely described below with reference to the accompanying drawings.
The invention discloses a method for establishing a non-compression type animal model of lumbar intervertebral disc herniation, which utilizes the characteristic that a rat tail is thick and contains sufficient amount of nucleus pulposus tissues of intervertebral disc, wherein the nucleus pulposus of the rat tail is in a jelly shape, and the epidural puncture technology is combined to inject the autologous nucleus pulposus suspension of the rat into a lumbar epidural space to manufacture the condition that the nucleus pulposus tissues are exposed in blood circulation and an organism immune system, so that the non-compression type animal model of the lumbar intervertebral disc herniation is established.
Referring to fig. 1 to 3, the change of lumbar nerve root conduction velocity of rats in experimental group and control group according to the present invention. The motor nerve conduction speed of the rats in the control group before operation is 62-74 m/s, the sensory nerve conduction speed is 60-69 m/s, and the conduction speed of the equine caudal nerve after operation has no obvious change.
The conduction speed of motor nerves and the conduction speed of sensory nerves of rats in the experimental group (nucleus pulposus transplantation group) before modeling are similar to those in the control group, the conduction speed of the motor nerves and the conduction speed of the sensory nerves after modeling are both reduced, the conduction speed of the motor nerves and the conduction speed of the sensory nerves are most obvious on the 6 th day after the operation, and the conduction speed of the sensory nerves begins to recover on the 9 th day.
Referring to fig. 6, the method for establishing the non-compression type lumbar disc herniation nucleus pulposus animal model and the operation process thereof of the present invention comprise:
s1 cutting rat tail 1cm away from the root of rat tail and suturing the wound to obtain rat tail intervertebral disc nucleus pulposus.
Specifically, 10-month-old SPF-grade SD male rats were selected as rats in the present example, and 20 rats were selected, and the body weight was 300 + -20 g. The breeding environment of the rats is that 5 rats are bred in one cage, the breeding temperature of the rats is controlled to be 20-25 ℃, and the humidity is controlled to be about 50%; the air flow is proper, the indoor air speed is controlled to be 0.1m/s, the lower limit (20 ℃) is better when cold air is opened, the upper limit (25 ℃) is better when warm air is opened, and the cage frame and the cage of the rat are required to avoid an air inlet; the illumination adopts 12-hour illumination and 12-hour darkness.
The rat is taken as an experimental animal, and has a plurality of advantages compared with large animals such as monkeys, pigs and the like: 1. the tail of the rat is thick, and the rat has more nucleus pulposus of the intervertebral disc and is easy to obtain; 2. the rat strain is mature and economical; 3. the rat has strong vitality and strong postoperative anti-infection capability.
The rats were randomly divided into two groups, one group was an experimental group, the other group was a control group, 10 rats each, and the two groups were acclimatized for 1 week.
For the experimental group, ketamine with a mass concentration of 1% was used as an anesthetic, and rats were anesthetized by intraperitoneal injection. After the anesthesia was successful, the body hair of the rat's back and hind limbs was removed with depilatory. Cutting off the rat tail at a position 1cm away from the root of the rat tail, suturing the wound, obtaining nucleus pulposus of the tail intervertebral disc of the rat, mixing 5 nucleus pulposus and 50 microliters of physiological saline, and fully stirring and diluting the mixture into suspension.
S2 method for extracting nucleus pulposus of caudal vertebra: puncturing one half of an annulus fibrosus at the side of the intervertebral disc of the caudal vertebra by using a scalpel (a sharp knife), breaking the annulus fibrosus of the intervertebral disc to expose nucleus pulposus, lightly picking out the nucleus pulposus by using a knife tip, and fully mixing and diluting 5 nucleus pulposus and 50 mu l of physiological saline into suspension. The nucleus pulposus is jelly-like and is small in quantity, so that the quality change of the nucleus pulposus is difficult to observe, and the nucleus pulposus is very suitable for being prepared into a suspension liquid to be injected around the lumbar nerve root without pressing the nerve root. The nucleus pulposus, upon contact with blood, can activate the immune response of rats, leading to the release of local inflammatory mediators, and over time, morphological and functional changes of nerve roots can be observed.
S3 formula of the depilatory comprises 3 g of ① sodium sulfide, 1 g of ② soap powder, 7g of ③ starch and a proper amount of water, wherein the mixture is mixed into paste, the rat bends at the lateral decubitus position, and epidural puncture is performed by using a No. 9 lumbar puncture needle with L4-L5 interspinous process gaps as puncture points, and 20 mul of autologous nucleus pulposus suspension and 30 mul of 2% lidocaine are injected respectively.
The judgment standard of the success of lumbar epidural puncture is as follows: the epidural space is negative pressure and can be injected with a small amount of gas; after the anesthesia recovery of the rat, the two forelimbs move freely, the two hindlimbs lose or decline in movement and sensation, and the rat recovers completely after 20-30 minutes, which indicates that the epidural puncture injection is successful.
In this embodiment, the syringe cannot be a disposable syringe made of plastic, but a syringe made of glass is required, because the resistance of the syringe made of glass after being wetted with physiological saline is very small, and the resistance of the syringe made of plastic is too large to be suitable for determining the negative pressure state of the epidural space.
In this embodiment, the purpose of injecting 30 mul 2% lidocaine at the epidural space of lumbar vertebrae is to judge the position that the pjncture needle arrived, if inject into the subarachnoid space, the limb can all be anaesthetized and can not move around the rat, even the rat appears full spinal anesthesia and dies, inject outside the lumbar vertebrae pipe, then the rat four limbs still can normally move about, only puncture the epidural space of lumbar vertebrae pipe, the rat can appear short-lived hind limb paralysis and forelimb activity is normal.
The rat can be anesthetized for 30 minutes by 50 mg.Kg-1 intraperitoneal injection of 1% ketamine, the rat spinal nerve can be blocked for 30 minutes by 30 mul 2% lidocaine by epidural injection, after the rat ketamine anesthesia is recovered, the lidocaine blocks the lumbar spinal nerve, the double forelimbs move freely, the double hind limbs move, feel is lost or subside, and the lumbar spinal nerve is completely recovered after 20-30 minutes, so that the epidural puncture and injection are proved to be successful, and therefore, the model preparation process is required to be completed within 30 minutes.
The following experimental examples prove and demonstrate that the animal model of the invention can cause the morphological and functional change of nerve roots in the non-compression type lumbar disc herniation nucleus pulposus animal model:
1. an experimental instrument: the device comprises a full-automatic paraffin embedding machine, a tissue dehydrator, a rotary slicer, a sheet spreading machine, a sheet baking machine, an electric heating blowing drying box, an optical microscope and a myoelectricity-evoked potential instrument.
2. Method for taking materials
The non-compression model group and the control group after the molding are normally raised, and rats are sacrificed on the 10 th day after the molding to cut the 4 th nerve roots and the 5 th nerve roots of the rat waists.
3. Control group: the autologous nucleus pulposus suspension was replaced with physiological saline, and the other procedures were the same as those of the experimental group.
4. HE staining
(1) Hydration of a section specimen: soaking paraffin section in xylene for 10min, replacing new xylene, and soaking for 10 min.
(2) Soaking the slices in 100% ethanol for 5min, and then replacing with new 100% ethanol and soaking for 5 min.
(3) Soaking the slices in gradient ethanol for 5min for 1 time (concentration of 90, 80, 70, and 50%), and adding pure water and PBS
Soaking and slicing for 5min respectively.
(4) Dyeing: adding hematoxylin dropwise on the slices, incubating for 15min, pouring out the hematoxylin gently, and washing with water.
(5) And dropwise adding a small amount of ammonia water on the section to turn blue, incubating for several seconds, then washing with water, observing the dyeing condition through microscopic examination, and ensuring that the dyeing depth of the hematoxylin is proper.
(6) And (3) dripping eosin on the section, incubating for 3min, washing with water, performing microscopic examination, wherein eosin staining is deep, soaking and differentiating for 5min by using 70% alcohol, washing with water, performing microscopic examination, and performing appropriate eosin staining depth.
(7) And (3) dehydrating and transparency: soaking the slices in gradient ethanol for 5min for 1 time (concentration of 70, 80, 90, and 100% respectively), soaking in xylene at room temperature for 10min, and soaking in new xylene for 10 min.
8) Microscopic examination: after the sections were taken out from xylene, the sections were mounted dropwise with a 75% glycerol solution in PBS, and observed under a microscope to take images.
5. Determination of conduction velocity of cauda equina
Carrying out intraperitoneal injection anesthesia with the dose of 30 mg.Kg-1 of 1% pentobarbital sodium, measuring the conduction velocity of cauda equina nerve roots 1 day before operation and 3, 6 and 10 days after operation by using a Dandi myoelectricity-evoked potential instrument, and repeating the measurement twice each time with the right side as a standard.
Measurement of sensory nerve conduction velocity: the stimulating electrode is arranged at the inner side of the medial malleolus of the right hind paw of the rat to stimulate the posterior tibial nerve through the skin, and the electrode is arranged at the same position every time. The recording electrode is arranged on the head side of the spinous process of L1, the reference electrode is arranged at the position 2cm on the right side of the recording electrode, the skin is cleaned, grease is removed, electrode paste is smeared, the grounding electrode is arranged at the position of gluteus maximus, the stimulation parameter is square wave pulse current, the wave width is 0.1ms, the frequency is 1.9Hz, the intensity is 3 mA-5 mA, the rear paw is preferably slightly shaken, a preamplifier of a signal recording system is provided with the sensitivity of 10 Muv/div, the filtering band is provided with the frequency of 10 Hz-5 kHz, the analysis time is 20 ms, and the average frequency is 30 times. Latency is automatically measured by the computer. The distance from the stimulation point to the recording point was input into the computer, and the conduction velocity of the right medial malleolus to the sensory nerve of the L1 spinous process was calculated.
Determination of motor nerve conduction velocity: transcutaneous electrical stimulation, the first point stimulated at the spinous process of L1 and the second point stimulated at the gluteal fold of the right gluteus maximus. The anode is arranged at the spinous process of L1, the cathode is arranged below the anode with a distance of at least 1cm, the grounding electrode is arranged at the right hind paw of a rat, the stimulation parameter is square wave pulse current, the wave width is 0.1ms, the intensity is 8 mA-10 mA, the recording electrode is a needle electrode, the Composite Muscle Action Potential (CMAP) is recorded on calf gastrocnemius muscle, the preamplifier of the recording system is recorded, the sensitivity is 10 mv/div, and the frequency is 5 Hz-10 kHz. Latency is automatically measured by the computer. The latency period after stimulation of the L1 spinous process and the gluteal fold and the distance from the L1 spinous process to the gluteal fold are measured respectively, and the motor nerve conduction velocity from the L1 spinous process to the gluteal fold is automatically calculated by a computer.
6. Statistical method
The conduction velocity difference of cauda equina before and after each group of modeling adopts paired t test, and the difference between two groups of mean values adopts grouped t test. (SPSS 11.0 statistical software). Differences are significant when p <0.05 and significant when p < 0.01. P >0.05 is indistinguishable.
7. Results
The motor nerve conduction velocity of the rat in the control group before operation is 62-74 m/s, the sensory nerve conduction velocity is 60-69 m/s, the conduction velocity of the cauda equina nerve after puncture has no obvious change, and has no difference compared with the conduction velocity before puncture (p is more than 0.05).
The motor nerve conduction velocity and the sensory nerve conduction velocity of rats in the experimental group (nucleus pulposus transplantation group) before modeling are similar to those in the control group, the motor nerve conduction velocity and the sensory nerve conduction velocity are reduced after modeling, the motor nerve conduction velocity and the sensory nerve conduction velocity are most obvious on the 6 th day after the operation, the nerve conduction velocity begins to recover on the 9 th day, the difference is very obvious compared with that before the operation (p is less than 0.01), and the difference is obvious compared with that of the control group (p is less than 0.05).
And (3) staining a nerve root HE:
general observations in the control group: the lumbar nerve root of the rat is smooth and has no adhesion with the surrounding tissues; and (3) observation by using a light mirror: the myelinated nerve fibers are uniform in size, the axons are located in the center and smaller, the nerve fibers are regularly arranged, and the myelin sheaths are located around the myelinated nerve fibers and are in a red reticular structure.
Gross observations from experimental groups: the color of the lumbar nerve root of the rat is dark, the lumbar nerve root is swollen, and the peripheral tissues are adhered; and (3) observation by using a light mirror: nerve fibers vary in size, light staining, swelling of axons, and mostly disintegration of myelin.
Through the analysis of the above experimental results, the following conclusions are drawn:
by comparing the morphology of the lumbar nerve root and the change of the nerve conduction velocity of the non-compression type lumbar disc nucleus pulposus experimental group and the control group, the model has better value in the aspect of researching the non-compression pathogenic mechanism of the lumbar disc nucleus pulposus. The interference of nerve root injury caused by nucleus pulposus pressing on nerve roots or in the process of surgical modeling is eliminated, an ideal animal model is provided for researching the non-pressing pathogenic mechanism caused by nucleus pulposus, and the method is simpler and more economic.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.
Claims (7)
1. A method for establishing a non-compression type lumbar intervertebral disc protrusion animal model is characterized by comprising the following steps:
s1, cutting a rat tail at a position 1cm away from the root of the rat tail, suturing the wound and obtaining the nucleus pulposus tissue of the intervertebral disc of the tail vertebra of the rat;
s2, cutting the fibrous ring of the intervertebral disc of the caudal vertebra of the rat, wherein the nucleus pulposus is in a jelly sample, and fully mixing and diluting 5 nucleus pulposus and 50 microliters of physiological saline to form a suspension;
s3, injecting the 20 ul nucleus pulposus suspension and the 30 ul 2% lidocaine injection into the lumbar epidural space to enable the nucleus pulposus suspension to be filled around nerve roots.
2. The method of claim 1, wherein the rat in S1 is a 10-month-old SPF SD male rat.
3. The method of claim 1, wherein the step of obtaining the rat caudal vertebral disc nucleus pulposus at S1 comprises:
s11, anesthetizing the rat;
s12, after anesthesia is successful, removing the body hair of the back and hind limbs of the rat by using a depilatory;
s13, performing conventional skin disinfection, cutting off rat tail at a position 1cm away from the tail root of the rat under aseptic conditions, suturing the wound, and obtaining the nucleus pulposus of the intervertebral disc of the tail vertebra of the rat.
4. The method for establishing an animal model of non-compression lumbar intervertebral disc protrusion according to claim 3, wherein ketamine with a concentration of 1% is selected as an anesthetic in S11, and rats are anesthetized by intraperitoneal injection at a dose of 50 mg.kg "1.
5. The method for establishing the non-compression type lumbar intervertebral disc herniation animal model according to claim 3, characterized in that 5 nuclei pulposus obtained in S13 are fully mixed with 50 μ l of physiological saline and diluted into a suspension.
6. The method for establishing the non-compression type lumbar intervertebral disc herniation animal model according to claim 1, wherein in the step S1, rat tails are cut off and wounds are sutured, 5 rat caudal intervertebral disc nuclei are obtained, and the rat caudal intervertebral disc nuclei are fully mixed with 50 μ l of physiological saline and diluted into a suspension.
7. The method for establishing the non-compression type lumbar intervertebral disc protrusion animal model according to claim 1, wherein in the step S3, the process of injecting the 20 μ l nucleus pulposus suspension and the 30 μ l2% lidocaine injection into the lumbar epidural space of the rat so that the nucleus pulposus suspension is filled around nerve roots comprises the following steps: the rat is in a lateral decubitus position, epidural puncture is performed by using a No. 9 lumbar puncture needle with the interval between L4-L5 spinous processes as a puncture point, and 20 mul of autologous nucleus pulposus suspension and 30 mul of 2% lidocaine are injected respectively.
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| PCT/CN2021/076172 WO2021185013A2 (en) | 2020-03-20 | 2021-02-09 | Establishment of non-compressive lumbar disc herniation animal model |
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| CN112825813A (en) * | 2020-12-31 | 2021-05-25 | 昆明医科大学第一附属医院 | Animal model preparation method |
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| CN112825813A (en) * | 2020-12-31 | 2021-05-25 | 昆明医科大学第一附属医院 | Animal model preparation method |
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