阅读说明：本技术 一种可调控的猕猴脊髓损伤方法 (Adjustable macaque spinal cord injury method ) 是由 邹杨鸿 周广平 张志婷 林郁 耿鑫 李经辉 徐玲 杜玲丽 余化霖 彭琴 王建红 于 2020-07-02 设计创作，主要内容包括：本发明涉及一种可调控的猕猴脊髓损伤方法,在猕猴的椎管目标脊髓节段内植入弹性球囊,球囊远端注水口埋于附近皮下；通过往球囊内注入不同体积的生理盐水,制作不同损伤程度的猕猴脊髓损伤模型。通过在猕猴椎管内使用球囊进行压迫的方法,构建可以调控损伤程度、调控损伤时程及损伤位置的猕猴脊髓损伤模型,造模效果持久,损伤渐进式,损伤程度行为可视化,对猕猴创伤小,致死率低,可用于模拟并研究脊髓外伤、脊髓内/外肿瘤、椎间盘突出、脊髓先天性畸形、脊髓感染等病变的病理损伤机制及治疗策略,对上述各类疾病的临床前试验具有重要意义。(The invention relates to an adjustable macaque spinal cord injury method, wherein an elastic balloon is implanted into a spinal cord segment of a vertebral canal target of a macaque, and a water injection port at the far end of the balloon is buried under the nearby skin; physiological saline with different volumes is injected into the saccule to manufacture the macaque spinal cord injury models with different injury degrees. The macaque spinal cord injury model which can regulate and control the injury degree, the injury time course and the injury position is constructed by using the saccule for compression in the macaque spinal canal, the modeling effect is durable, the injury is progressive, the injury degree behavior is visual, the macaque injury is small, the lethality rate is low, and the macaque spinal cord injury model can be used for simulating and researching pathological injury mechanisms and treatment strategies of pathological changes such as spinal cord trauma, spinal cord internal/external tumors, intervertebral disc herniation, spinal cord congenital malformation, spinal cord infection and the like, and has important significance for preclinical tests of various diseases.)

1. An adjustable macaque spinal cord injury method is characterized by comprising the following steps:

1) implanting an elastic balloon in a vertebral canal target spinal segment of a macaque, wherein a water injection port at the far end of the balloon is buried under the nearby skin;

2) opening a subcutaneous incision, injecting physiological saline with different volumes into the saccule to prepare macaque spinal cord injury models with different injury degrees, and evaluating the degree and development process of spinal cord injury through macaque behaviors;

3) the balloon is removed after a period of compression.

2. The method of claim 1, wherein the method comprises: the implanted saccule in the step 2) has the diameter of 1-1.5cm after being injected with the normal saline, and does not cause rejection reaction.

3. A method of regulating spinal cord injury in a macaque according to claim 1 or 2, wherein: the part implanted into the balloon in the step 1) is the 10 th thoracic vertebra.

4. The method of claim 1, wherein the method comprises: the balloon implantation part in the step 1) is cervical vertebra, thoracic vertebra or lumbar vertebra.

Technical Field

The invention belongs to the technical field of medical animal research, and particularly relates to an adjustable macaque spinal cord injury method.

Background

Spinal cord injury is a serious clinical disease, the life of a patient with spinal cord injury is less directly threatened, the life is long, but the life quality is extremely low, the life cannot be managed by oneself, and great burden is caused to individuals, families and the society. Therefore, it is one of the issues that needs to be solved urgently in biology and even medical field to prepare a spinal cord injury model by using experimental animals and study the pathological injury mechanism and treatment strategy of spinal cord injury.

Most of the existing spinal cord injury models are developed by adopting rodents and other small animals, and the problems are that:

1) rodents and other small animals have small spinal canal structures, are difficult to realize local injury, and the model is usually full spinal cord or one-side spinal cord injury, so that the injury degree cannot be finely regulated. 2) In the rehabilitation process of rodents and other small animals, symptoms such as paraplegia, bedsore, urine retention and the like after spinal cord injury are recovered quickly, and the symptoms are usually recovered slowly or difficultly after the spinal cord injury of human beings, but the symptoms are far from each other. These models are not suitable for simulating spinal cord injuries in humans, and most importantly, rodents differ greatly from humans in physiological conditions, spinal cord anatomy, and spinal cord blood supply, and therefore rodent models do not always reflect human conditions.

Rhesus macaque (rhesus monkey, hereinafter referred to as macaque) is similar to human beings in gene, anatomy, pathology, physiology, molecular regulation and behavior, the sizes of the spinal cord and the spinal canal are relatively more similar to those of the human beings, the macaque has higher cognitive level and complex behavior, and symptoms such as sensation, fine movement, limb movement and the like caused by human spinal cord injury can be expressed on the macaque, so the macaque has the irreplaceable advantages of other experimental animals on the simulation of human spinal cord diseases.

The currently known macaque spinal cord injury model still extends the modeling method of rodents: one side injury or all injuries are carried out on the spinal cord of the macaque by methods of smashing injury, transverse cutting, clamping and the like. These methods have the following problems: 1) the operation is complex, the operation wound is large, and the fatality rate is high. 2) The injury degree can not be regulated, no matter one-side injury or whole spinal cord injury, direct paraplegia of corresponding limbs can be directly caused, and symptoms such as paresis, fine movement disorder, lameness and the like of clinical mild patients can not be simulated. 3) The injury can not be reversed, but clinically, patients with spinal cord tumor and intervertebral disc herniation can obtain a certain degree of recovery after the tumor or the herniated intervertebral disc is removed by surgery, and the existing modeling method for the spinal cord injury of experimental animals cannot simulate the process.

Therefore, the existing non-human primate animal spinal cord injury models are few, the defects exist, the modeling effect is poor, the model cannot be regulated and controlled, the injury is irreversible, the fatality rate is high, and the like, so that the precious experimental animal macaque is wasted in resources.

The technical scheme of macaque spinal cord injury, which is simple and can regulate and control symptoms, is urgently needed to solve the technical problems.

Disclosure of Invention

In order to solve the problems, the invention provides a macaque spinal cord injury method which is simpler to operate and smaller in wound and can regulate and control the injury degree of the macaque spinal cord.

The specific technical scheme is as follows: a method for controlling macaque spinal cord injury comprises the following steps:

1) implanting an elastic balloon in a vertebral canal target spinal segment of a macaque, wherein a water injection port at the far end of the balloon is buried under the nearby skin;

2) opening a subcutaneous incision, injecting physiological saline with different volumes into the saccule to prepare macaque spinal cord injury models with different injury degrees, and evaluating the degree and development process of spinal cord injury through macaque behaviors;

3) the balloon is removed after a period of compression. There is some reversal of the symptoms of the macaque.

At present, various animal spinal cord injury models are almost focused on rodents or other small animals, and are rarely seen in non-human primate models. The macaque has higher cognitive level and complex behaviors, and the symptoms of the human beings in aspects of feeling, fine movement, limb movement and the like caused by spinal cord injury can be expressed on the macaque, and the spinal cord injury model manufactured by the macaque has incomparable advantages compared with other conventional experimental animals.

The spine of the macaque is divided into cervical vertebra, thoracic vertebra, lumbar vertebra, sacrum, caudal vertebra and the like, wherein 7 cervical vertebra, 12 thoracic vertebra are the same as human, the envelope of the spinal cord is yellow ligament, epidural space, dura mater, arachnoid, pia mater, spinal cord from outside to inside in sequence, the anatomical level is similar to human, and the segment of spinal innervation is also approximately the same as human. After the spinal cord of the macaque is damaged, conduction tracts of the spinal cord are affected, and the light patients are manifested as sensory disturbance, toe grasping disturbance, lower limb movement disorder and the like, and if the damage is continuously aggravated, symptoms such as unilateral paralysis, paraplegia, incontinence of urine and stool can appear, which are similar to the symptoms appearing after the spinal cord of the human being is damaged.

After the macaque is positioned before MRI operation, the position of a target cone is determined, and the diameter of the vertebral canal at the position of the target cone is measured. The method comprises the steps of performing an operation from a positioning point, sequentially cutting open skin, sequentially implanting a saccule into an intraspinal dural and extradural space of a vertebral canal of a target cone from the interspinal space, extending a water injection port at the tail end of the saccule to the subcutaneous space, determining that the saccule is arranged on the left side or the right side of a spinal cord according to which side of a required model firstly shows symptoms, and simultaneously determining the volume of physiological saline injected into the saccule according to the severity of the symptoms required by the model, wherein the control of the amount of the liquid injected into the saccule is a gradual and controllable process.

The animal model can be caused to have the following states:

1) dysesthesia of limbs below affected spinal innervating segment, dysgrasping of fingers or toes;

2) decline of limb muscle strength and movement disorder below the affected spinal cord innervation segment;

3) paralysis of the limbs below the affected spinal cord innervating segment;

4) paralysis of limbs, incontinence of urine and feces, etc. below the bilateral spinal innervation segment.

After the sacculus is implanted, the incision is sewed layer by layer, and the tail end of the sacculus is buried under the skin.

The saccule can be placed in a body for a long time to enable the symptom to be stably expressed, if the symptom needs to be adjusted from light to heavy, only a small incision needs to be opened subcutaneously, and a certain volume of normal saline is continuously injected into the saccule. If the situation of a patient with spinal cord tumor or intervertebral disc protrusion after tumor resection or protrusion intervertebral disc removal needs to be simulated clinically, only the liquid in the saccule needs to be pumped out.

Further, the implanted saccule in the step 2) has the diameter of 1-1.5cm after being injected with the normal saline, and does not cause rejection reaction.

The balloon is selected to be the type with the maximum diameter of 1-1.5cm after being filled with liquid, the value range is determined according to the diameter of the vertebral canal of the macaque, and the diameter of the vertebral canal of the macaque is found to be about 0.9-1.2cm in magnetic resonance scanning through research.

Further, the implantation balloon part in the step 1) is a 10 th thoracic vertebra.

The preferred balloon implantation position is the 10 th thoracic vertebra, and the reason is that after continuous research, the sensory and motor dysfunction of the lower limbs just below the 10 th thoracic vertebra damage section can be caused, the functions of the two upper limbs cannot be influenced, and the observation of the behavior of the lower limb dysfunction is facilitated. If the injury occurs to the segment above the 10 th thoracic vertebra, both thoracic and abdominal muscle and nerve dysfunction may occur; if the segment below the 10 th thoracic vertebra is damaged, the functional parts of the lower limbs can be remained, which is not favorable for observing typical performance. And from an anatomical perspective, the positioning and anatomical difficulty of the 10 th thoracic vertebra is easier than that of other vertebral bodies. Therefore, the 10 th thoracic vertebra is an ideal lesion site for evaluating sensory-motor dysfunction of both lower limbs.

The method for treating the spinal cord injury of the macaque is simpler to operate, has smaller wound and can regulate and control the injury degree of the spinal cord of the macaque. The degree of spinal cord injury and the corresponding symptoms can be regulated and controlled according to the requirements; can continue to adjust towards the direction of aggravating symptoms in the case of mild symptoms; can simulate the situation of human spinal cord space occupying lesion after being relieved by operation. The model is stable, and the saccule can be placed in the body for a long time to enable the symptoms to be stably expressed; the operation is simple, the injury is small, the dura mater is not damaged and seriously extruded, the influence of manual operation on the spinal cord can be hardly generated in the operation, and the animal fatality rate is low.

Further, the balloon implantation part in the step 1) is cervical vertebra, thoracic vertebra or lumbar vertebra.

The invention has the beneficial effects that: the macaque spinal cord injury model which can regulate and control the injury degree, the injury time course and the injury position is constructed by using the saccule for compression in the macaque spinal canal, the modeling effect is durable, the injury is progressive, the injury degree behavior is visual, the injury is small, the lethality rate is low, and the macaque spinal cord injury model can be used for simulating and researching pathological injury mechanisms and treatment strategies of pathological changes such as spinal cord trauma, spinal cord internal/external tumors, intervertebral disc herniation, spinal cord congenital malformation and spinal cord infection, and has important significance for preclinical tests of various diseases.

Drawings

Fig. 1 is a schematic view of the method for implanting an elastic balloon in a target spinal segment in the spinal cord injury of a macaque, which is disclosed by the invention.

Fig. 2 is a magnetic resonance image of the balloon implanted with the physiological saline solution according to the first embodiment.

Fig. 3 is a magnetic resonance image of the balloon of comparative example 1 after implantation of the balloon and injection of saline.

Fig. 4 is a magnetic resonance image of the balloon of comparative example 2 after implantation of a balloon and injection of saline.

In the upper diagram: 1-spinal canal, 2-targeted spinal segment, 3-balloon, 4-spinous process.

Detailed Description

In order to make the technical problems and technical solutions solved by the present invention more clearly apparent, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.