Method for establishing rabbit knee joint cartilage injury model in plateau cold environment

Abstract

The invention provides a method for establishing a rabbit knee joint cartilage injury model in a plateau cold environment. The method comprises the following steps of: making an operative incision by taking a knee joint as a center, separating subcutaneous fascia, cutting a gap between the inner side of a patella and quadriceps femoris muscle, turning the patella outwards, and exposing a knee joint cavity and an articular surface at the far end of a femur; pore-forming is carried out on the articular surface of the distal femur until fresh blood exudation can be seen under the cartilage; and returning the patella and suturing the wound to obtain the rabbit knee joint cartilage injury model in the plateau cold environment. The model disclosed by the invention can be used for better researching a repairing mechanism of the knee joint cartilage injury in the plateau cold environment and evaluating and screening medicines and materials for treating the knee joint cartilage injury.

Description

Technical Field

[0001] This invention relates to the field of animal model technology, specifically to a method for establishing a rabbit knee joint cartilage injury model under high-altitude and cold environments. Background Technology

[0002] Knee cartilage damage is a common sports injury and degenerative disease that can lead to joint pain, limited mobility, and long-term joint dysfunction. Currently, treatment methods for knee cartilage damage mainly include medication, physical therapy, and surgery.

[0003] The cold, high-altitude environment, characterized by low oxygen and low temperature, may affect the repair process of knee cartilage damage. The efficacy of treatments for knee cartilage damage in cold, high-altitude environments cannot be assessed using pharmacological efficacy studies conducted in conventional environments. The cold, high-altitude environment may influence the repair process of knee cartilage damage. This is because the low oxygen, low pressure, and low temperature environment can promote the increased expression of IL-1, TNF-α, and IL-6 proteins, ultimately leading to the loss of articular cartilage and promoting the development of osteoarthritis.

[0004] Therefore, establishing a rabbit knee cartilage injury model under high-altitude and cold conditions is of great significance for studying the repair mechanism and treatment of knee cartilage injury under such conditions. Summary of the Invention

[0005] The purpose of this invention is to provide a method for establishing a rabbit knee cartilage injury model under high-altitude and cold conditions, thereby creating a model of knee cartilage injury in such an environment. This model aims to provide a large animal model with a high success rate and stability for research on knee cartilage injury at high altitudes.

[0006] Currently, research on rabbit knee cartilage injury models in cold, high-altitude environments is insufficient, and a reliable model is lacking for evaluating and screening drugs and materials for treating knee cartilage injury. Therefore, this invention aims to provide a method for establishing a rabbit knee cartilage injury model in cold, high-altitude environments, in order to better study the repair mechanism of knee cartilage injury in such environments and to evaluate and screen drugs and materials for treating knee cartilage injury.

[0007] The technical problem solved by this invention is achieved by the following technical solution: In a first aspect, this invention provides a method for establishing a rabbit knee joint cartilage injury model under high-altitude and cold conditions, comprising the following steps: Step 1: After raising the experimental rabbits in a simulated high-altitude environment in an artificial experimental chamber, they were anesthetized; Step 2: Make a surgical incision centered on the knee joint, separate the subcutaneous fascia, and then cut along the space between the medial side of the patella and the quadriceps femoris muscle. Lateralize the patella to expose the knee joint cavity and the articular surface of the distal femur. Step 3: A hole is made in the articular surface of the distal femur to the subchondral bone, where fresh blood can be seen seeping out. Step 4: Reposition the patella and suture the wound to obtain the rabbit knee joint cartilage injury model under the cold environment of the plateau.

[0008] In some embodiments of the present invention, in step one: The experimental rabbits were male New Zealand white rabbits weighing 2.0-4.0 kg; the artificial experimental chamber simulated a plateau environment at an altitude of 3000-7000 m and a room temperature of 0-20℃; the experimental rabbits were kept for 1-5 days.

[0009] In some embodiments of the present invention, the experimental rabbits are male New Zealand White rabbits weighing 2.5-3.0 kg; the artificial experimental chamber simulates a plateau environment at an altitude of 4500-5500 m and a room temperature of 5-15°C; the experimental rabbits are kept for 1.5-3 days. In some embodiments of the present invention, the surgical incision in step two is a longitudinal incision, and the length of the surgical incision is 2-8 cm.

[0010] In some embodiments of the present invention, the length of the surgical incision is 3-5 cm.

[0011] In some embodiments of the present invention, the diameter of the hole created in step three is 1~7mm and the depth is 1~6mm.

[0012] In some embodiments of the present invention, the diameter of the hole created in step three is 3-5 mm and the depth is 2.5-4 mm.

[0013] The present invention has the following beneficial effects: 1. For the first time, the study accurately simulated cartilage damage in rabbit knee joints under cold conditions at high altitudes, making the research results more closely reflect the actual situation of patients in high-altitude areas and improving the practicality and translational value of the research.

[0014] 2. The high-altitude environment has a significant impact on the physiological and pathological processes of organisms. This model helps to study the biological mechanisms of knee cartilage damage under this special environment, thereby improving the biological relevance of the research.

[0015] 3. This model can be used to evaluate and compare the effects of different treatment methods in the cold environment of high altitude, providing more precise and effective treatment options for patients in high-altitude areas.

[0016] 4. This model can be used to screen and evaluate the effects of novel drugs and biomaterials in the cold environment of high altitudes, thus promoting the research and development of drugs and materials suitable for high-altitude environments.

[0017] 5. The establishment of a knee cartilage injury model in cold high-altitude environments helps to gain a deeper understanding of the occurrence and development of the disease, and provides a scientific basis for the prevention and control of the disease.

[0018] 6. The establishment of this model will promote research in the field of high-altitude medicine and improve the scientific understanding and technical level of disease treatment in the special environment of high altitude.

[0019] 7. Compared with human experiments, animal models have advantages in terms of ethics and cost. The establishment of a rabbit knee cartilage injury model provides researchers with a relatively economical and easy-to-use research tool.

[0020] 8. The location of cartilage damage in this model is the femoral trochlea, which is different from the existing models that mostly use the tibial plateau and femoral groove. In addition, the existing models are all based on plains and normal temperature environments, while the construction of this model is based on high-altitude hypoxia and cold, which is more difficult to operate, but can reproduce the cartilage damage of people stationed at high altitudes. Attached Figure Description

[0021] Figure 1 This is a schematic diagram of determining the knee joint position in Example 1.

[0022] Figure 2 This is a schematic diagram of local infiltration anesthesia of the knee joint in Example 1.

[0023] Figure 3 This is a schematic diagram of the knee joint exposing the medial femoral condyle in Example 1.

[0024] Figure 4 This is a schematic diagram of the patella, an anatomical landmark of the knee joint, in Example 1.

[0025] Figure 5 This is a schematic diagram of cartilage damage caused by a manual trephine drill in Example 1.

[0026] Figure 6 This is a schematic diagram of successful cartilage damage in Example 1.

[0027] Figure 7 This is a schematic diagram showing fresh blood flowing out from the cartilage injury site in Example 1.

[0028] Figure 8 This is a schematic diagram of applying curcumin to the cartilage defect in Example 1.

[0029] Figure 9 This is a schematic diagram of suturing the knee joint cavity in Example 1.

[0030] Figure 10 This is a schematic diagram of skin suturing in Example 1.

[0031] Figure 11 This is a bar chart of IL-1 in each group in Example 2.

[0032] Figure 12 This is a bar chart of IL-6 in each group in Example 2.

[0033] Figure 13 This is a bar chart of TNF-α in each group in Example 2.

[0034] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0035] As described in this invention, the term "prevention" refers to preventing the occurrence of disease and / or preventing the recurrence of disease. Detailed Implementation

[0036] This specification provides a detailed description of specific embodiments. Those skilled in the art should recognize that the following embodiments are exemplary and should not be construed as limiting the invention. For those skilled in the art, various improvements and modifications can be made to the invention without departing from its principles; such improvements and modifications also fall within the scope of protection of the claims. The beneficial effects of the invention are illustrated below through specific examples.

[0037] Example 1 A method for establishing a rabbit knee joint cartilage injury model under high-altitude and cold environments, the steps of which are as follows: (1) Select adult male New Zealand white rabbits weighing 2.5~3.0kg and use the artificial experimental chamber in the special environment of Northwest China as the breeding ground; (2) Simulate a plateau environment with an altitude of 5000m and a room temperature of 10℃, and raise them for 2 days; (3) After entering the enclosure and maintaining the original simulated environment, the New Zealand rabbits were anesthetized and fixed in a supine position; General anesthesia: ketamine was injected intramuscularly into the buttocks (40 mg / kg) + propofol was injected into the ear vein; a micro-infusion pump was used, accurate to 1.5 mg / kg / h / min; Local infiltration: 0.5 mL of 1% lidocaine was injected around the joint capsule.

[0038] (4) Prepare the skin around the right knee joint of the New Zealand rabbit; hair removal: use an electric razor and rabbit hair removal cream to remove the hair around the knee joint. Disinfection: disinfect with povidone-iodine 3 times, and remove the povidone-iodine with 75% alcohol. Draping: cover with a sterile drape, exposing only the surgical area. Positioning: supine position, limbs fixed to the operating table, keeping the knee joint flexed at 90°.

[0039] (5) Disinfect and cover the right knee joint with a towel; (6) Make a 4cm longitudinal surgical incision with the right knee joint as the center, separate the subcutaneous fascia, and then cut along the medial side of the patella and the space between the quadriceps femoris muscle, evert the patella, and expose the knee joint cavity and the articular surface of the distal femur. (7) Use a manual trephine drill to create a hole with a diameter of 4 mm and a depth of 3 mm on the distal femoral articular surface. The standard for the depth is that fresh blood can be seen seeping out under the cartilage, so as to achieve full-thickness cartilage damage. (8) The patella was repositioned and the wound was sutured layer by layer, and finally a rabbit knee joint cartilage injury model was successfully established in the cold environment of the plateau.

[0040] Regarding the success rate, a minimally invasive incision is made at the knee joint to avoid damaging the surrounding major blood vessels and nerves. After successful modeling, the joint cavity of the New Zealand rabbit is sutured and it continues to be raised in the laboratory. The standard for successful modeling is to raise the rabbit in a high-altitude cold simulation chamber, and the modeling standard is to see fresh blood flowing out of the drill hole or to see red bone marrow.

[0041] Post-surgery, the model remains kept in a high-altitude environment (e.g., 5000 meters, 10℃) to simulate the high-altitude recovery process. Traditional models, however, are typically returned to a normal-temperature environment after surgery, failing to simulate the recovery conditions at high altitudes. This model can be used to study the effects of high-altitude hypoxia on cartilage repair and to screen for drugs suitable for high-altitude environments.

[0042] This model uses a 4cm incision in the knee joint skin (existing models use 6cm-8cm incisions, shortening this model's incision by 30-50%, which helps reduce postoperative infection rates). The inside of the knee joint is exposed, and then a 4mm diameter, 3cm deep hole is drilled into the femoral trochlear surface. This depth conforms to the International Cartilage Repair Society (ICRS) assessment of grade 4 articular cartilage damage, indicating full-thickness cartilage damage and complete cartilage body destruction. The saphenous nerve and medial inferior patellar artery are located on the tibial side; this model accesses from the medial side of the patella, effectively avoiding the neurovascular bundle. Existing models require meniscus resection and methods involving exposure of the joint cavity, making blood vessels and nerves susceptible to injury.

[0043] The construction process of the rabbit knee joint cartilage injury model in the above-mentioned high-altitude cold environment is controllable, the constructed model is stable, operable, and can well simulate clinical practice.

[0044] By using the aforementioned rabbit knee cartilage injury model, drugs or materials that promote bone growth can be effectively evaluated, thus providing reliable data support for the screening and evaluation of these drugs or materials.

[0045] Example 2 After the model was constructed, an experimental design was carried out, and the experimental groups were divided into normal cartilage group, plain normal temperature model group, high altitude cold model group, and high altitude cold model plus curcumin group. ELISA serum protein tests for IL-1, TNF-α, and IL-6 were performed. The experimental results proved that the high altitude cold environment does indeed aggravate cartilage damage and inflammation, and that curcumin has the effect of inhibiting cartilage damage and inflammation.

[0046] After successful modeling, a nano-curcumin suspension was applied to the cartilage defect. Curcumin can inhibit inflammation and promote healing. This experiment also demonstrates the model's ability to simulate the effects of various drugs on knee cartilage damage in cold, high-altitude environments.

[0047] In summary, this invention is the first to accurately simulate cartilage damage in rabbit knee joints under cold, high-altitude conditions, making the research results more closely reflect the actual situation of patients in high-altitude areas and improving the practicality and translational value of the research. Therefore, this invention effectively overcomes the various shortcomings of existing technologies and has high industrial application value.

[0048] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in the present invention should still be covered by the claims of the present invention.

Claims

1. A method for establishing a rabbit knee joint cartilage injury model under high-altitude and cold environments, characterized in that, Includes the following steps: Step 1: After raising the experimental rabbits in a simulated high-altitude environment in an artificial experimental chamber, they were anesthetized; Step 2: Make a surgical incision centered on the knee joint, separate the subcutaneous fascia, and then cut along the medial side of the patella and the space between the quadriceps femoris muscle. Turn the patella outward to expose the knee joint cavity and the articular surface of the distal femur. Step 3: A hole is made in the articular surface of the distal femur to the subchondral bone, where fresh blood can be seen seeping out. Step 4: Reposition the patella and suture the wound to obtain the rabbit knee joint cartilage injury model under the cold environment of the plateau.

2. The method for establishing a rabbit knee joint cartilage injury model under high-altitude cold environment according to claim 1, characterized in that, In step one: The experimental rabbits were male New Zealand white rabbits weighing 2.0-4.0 kg; the artificial experimental chamber simulated a plateau environment at an altitude of 3000-7000 m and a room temperature of 0-20℃; the experimental rabbits were kept for 1-5 days.

3. The method for establishing a rabbit knee joint cartilage injury model under high-altitude cold environment according to claim 2, characterized in that, The experimental rabbits were male New Zealand white rabbits weighing 2.5-3.0 kg; the artificial experimental chamber simulated a plateau environment at an altitude of 4500-5500 m and a room temperature of 5-15℃; the experimental rabbits were kept for 1.5-3 days.

4. The method for establishing a rabbit knee joint cartilage injury model under high-altitude cold environment according to claim 1, characterized in that, In step two, the surgical incision is a longitudinal incision, and the length of the surgical incision is 2-8 cm.

5. The method for establishing a rabbit knee joint cartilage injury model under high-altitude cold environment according to claim 4, characterized in that, The length of the surgical incision is 3-5 cm.

6. The method for establishing a rabbit knee joint cartilage injury model under high-altitude cold environment according to claim 1, characterized in that, In step three, the diameter of the hole is 1-7 mm and the depth is 1-6 mm.

7. The method for establishing a rabbit knee joint cartilage injury model under high-altitude cold environment according to claim 6, characterized in that, In step three, the diameter of the hole is 3-5 mm and the depth is 2.5-4 mm.

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