Construction method of traumatic temporomandibular joint ankylosis animal model
An animal model of traumatic temporomandibular joint ankylosis was established by surgically removing the fibrocartilage of the mouse condyle and temporal bone fossa, removing the articular disc and carving a groove in the fossa. This solved the problem that the existing model did not fit the pathological environment of human diseases and achieved a simulation that was closer to clinical symptoms.
Patent Information
- Application Number
- CN202511125521.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-12
- Publication Date
- 2025-10-21
AI Technical Summary
Existing technologies make it difficult to provide an animal model of traumatic temporomandibular joint ankylosis that fits the complex pathological environment of human diseases for studying the pathogenesis and developing new therapies.
A surgical procedure was performed to remove part of the fibrous cartilage of the condyle and temporal fossa of mice, remove the articular disc, expose the joint space through horizontal blunt dissection, and incise the surface of the fossa to destroy the fossa cartilage. After the wound was sutured, the mice were fed soft food to establish a traumatic temporomandibular joint ankylosis model.
The established animal model is closer to the complex pathological environment of human diseases, simulates traumatic ankylosis of the joint, and is suitable for studying the pathogenesis of temporomandibular joint ankylosis and developing new therapies.
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Figure CN120814927A_ABST
Abstract
Description
Technical Field
[0001] The present disclosure relates to the technical field of methods for constructing animal models of oral diseases, and in particular to a method for constructing an animal model of traumatic temporomandibular joint ankylosis. Background Art
[0002] Temporomandibular joint ankylosis (TMJ) refers to a condition in which the temporomandibular joint (TMJ) is structurally abnormal due to intra-articular or extra-articular pathology, leading to limited or complete immobilization of the joint. This condition can cause patients to experience limited mouth opening, difficulty chewing, and speech impairment.
[0003] Animal models are important tools for studying the pathogenesis of temporomandibular joint ankylosis, developing new therapies, and optimizing clinical management. Therefore, providing an animal model that closely reflects the complex pathological environment of human disease has become a pressing technical challenge. Summary of the Invention
[0004] In order to solve the above problems, the present disclosure provides a method for constructing an animal model of traumatic temporomandibular joint ankylosis.
[0005] The present disclosure provides a method for constructing an animal model of traumatic temporomandibular joint ankylosis, the method comprising the following steps: S110: Select the preauricular area on one side of the experimental subject, avoid the superficial temporal artery and vein, and bluntly dissect along the zygomatic arch from front to back to the joint capsule, exposing the mandibular condyle neck and part of the zygomatic arch. After opening the joint capsule, separate the condyle. S120: Separate the lateral attachments of the disc through the joint space, expose the disc, cut off the peripheral attachments of the disc, and remove part or all of the disc. S130: The joint space is exposed by horizontal blunt dissection of the joint capsule, part of the fibrocartilage on the condylar head is removed, and the glenoid fossa cartilage is destroyed; S140: The wound was sutured and the animal was fed soft food after awakening to obtain an animal model of traumatic temporomandibular joint ankylosis.
[0006] In one embodiment of the present disclosure, the experimental subjects are 3-week-old mice weighing 11-13 g.
[0007] In one embodiment of the present disclosure, before step S110, step S100 is further included: performing general anesthesia and preoperative disinfection on the experimental subject.
[0008] In one embodiment of the present disclosure, the method for anesthetizing the experimental subject comprises the following steps: injecting penicillin into the experimental subject via intramuscular injection at a dose of 20,000-40,000 U / 1,000 g; The experimental subjects were injected with 1.25% tribromoethanol at a dose of 2 ml / 100 g into the abdominal cavity for general anesthesia.
[0009] In one embodiment of the present disclosure, the preoperative disinfection of the experimental subject comprises the following steps: preparing the skin of the left preauricular area of the experimental subject and then disinfecting the surgical area with povidone-iodine solution.
[0010] In one embodiment of the present disclosure, in step S120 , ophthalmic scissors are used to sever the peripheral attachments of the articular disc.
[0011] In one embodiment of the present disclosure, in step S130, fine ophthalmic forceps are used to remove part of the fibrous cartilage on the condylar head, and a minimally invasive apex elevator is used to groove the surface of the glenoid fossa to destroy the glenoid fossa cartilage.
[0012] In one embodiment of the present disclosure, in step S140, the method for suturing the wound is: tying 5-0 sutures at the mandibular ramus and the root of the zygomatic arch respectively, pulling the two sutures together, and tying a knot at the root of the zygomatic arch to close the joint space.
[0013] In one embodiment of the present disclosure, after step S140, the following step is further included: S150: performing anti-infection and analgesic treatment on the mouse.
[0014] In one embodiment of the present disclosure, the analgesic treatment method is: administering 0.01 mg / 100 g pentazocine to the experimental subject for analgesia after surgery; and / or, The anti-infection method is: giving the experimental subject the same dose of penicillin as before surgery for 2-3 days.
[0015] In one embodiment of the present disclosure, the preauricular area of the experimental subject is incised using a pretragus arc incision.
[0016] One beneficial effect of the present disclosure is that the method for constructing an animal model of traumatic temporomandibular joint ankylosis provided by the present disclosure includes selecting the preauricular area on one side of an experimental subject, avoiding the superficial temporal artery and vein, performing blunt separation from front to back along the zygomatic arch to the joint capsule and exposing the neck of the mandibular condyle and part of the zygomatic arch, separating the condyle after opening the joint capsule; separating the lateral attachments of the articular disc through the joint space, cutting off the peripheral attachments of the articular disc after exposing the articular disc, and removing part or all of the articular disc; revealing the joint space through horizontal blunt separation, removing part of the fibrocartilage on the condylar head, and destroying the articular fossa cartilage; suturing the wound, and feeding the animal with soft food after it wakes up, thereby obtaining an animal model of traumatic temporomandibular joint ankylosis.
[0017] The construction method disclosed herein uses a surgical approach to remove part of the fibrous cartilage of the condyle and the temporal bone fossa, and to remove the articular disc to create a model. The animal model established through a composite trauma approach fits the complex pathological environment of human diseases.
[0018] Other features and advantages of the present disclosure will become apparent from the following detailed description of exemplary embodiments of the present disclosure with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments of the present disclosure and, together with the description, serve to explain the principles of the present disclosure.
[0020] Figure 1 1 is a flow chart of a method for constructing an animal model of traumatic temporomandibular joint ankylosis provided by an embodiment of the present disclosure; Figure 2 This is an observation diagram of the process of constructing an animal model provided by an embodiment of the present disclosure; Figure 3 This is a measurement and observation diagram of the maximum vertical passive mouth opening of a mouse after surgery, provided by an embodiment of the present disclosure; Figure 4 This is an observation diagram of mice in the experimental group after surgery provided by an embodiment of the present disclosure; Figure 5 This is a Micro-CT image of mice in the experimental group after surgery provided by an embodiment of the present disclosure; Figure 6 This is a histological staining observation diagram of the left TMJ of the control group and the experimental group after surgery provided by an embodiment of the present disclosure. DETAILED DESCRIPTION
[0021] Various exemplary embodiments of the present disclosure will now be described in detail with reference to the accompanying drawings. It should be noted that unless otherwise specifically stated, the relative arrangement of components and steps, numerical expressions and numerical values set forth in these embodiments do not limit the scope of the present disclosure.
[0022] The following description of at least one exemplary embodiment is merely illustrative in nature and is in no way intended to limit the present disclosure, its application, or uses.
[0023] Technologies, methods, and equipment known to ordinary technicians in the relevant art may not be discussed in detail, but where appropriate, the technologies, methods, and equipment should be considered part of the specification.
[0024] It should be noted that like reference numerals and letters refer to like items in the following figures, and therefore, once an item is defined in one figure, it need not be further discussed in subsequent figures.
[0025] The specific embodiments of the present disclosure are described below with reference to the accompanying drawings.
[0026] In this document, “upper”, “lower”, “front”, “back”, “left”, “right”, etc. are only used to indicate the relative position relationship between related parts, rather than to limit the absolute positions of these related parts.
[0027] In this article, "first", "second", etc. are only used to distinguish each other, and do not indicate the importance and order, or the prerequisite for each other's existence.
[0028] In this document, “equal”, “same”, etc. are not strictly limited in a mathematical and / or geometric sense, but also include errors that can be understood by those skilled in the art and are allowed in manufacturing or use.
[0029] In order to provide an animal model that fits the complex pathological environment of human diseases, the present disclosure provides a method for constructing an animal model of traumatic temporomandibular joint ankylosis. Figure 1 、 2 As shown, the construction method includes the following steps: S110: Select the preauricular area on one side of the experimental subject, avoid the superficial temporal artery and vein, and bluntly dissect along the zygomatic arch from front to back to the joint capsule, exposing the mandibular condyle neck and part of the zygomatic arch. After opening the joint capsule, separate the condyle. S120: Separate the lateral attachments of the disc through the joint space, expose the disc, cut off the peripheral attachments of the disc, and remove part or all of the disc. S130: The joint space is exposed by horizontal blunt dissection of the joint capsule, part of the fibrocartilage on the condylar head is removed, and the glenoid fossa cartilage is destroyed; S140: The wound was sutured and the animal was fed soft food after awakening to obtain an animal model of traumatic temporomandibular joint ankylosis.
[0030] Specifically, when incising the preauricular area of the subject, the superficial temporal artery and vein were avoided to minimize bleeding. Blunt dissection along the zygomatic arch from front to back, extending to the joint capsule, exposed the mandibular condyle neck and a portion of the zygomatic arch. Blunt dissection minimizes nerve and blood vessel damage, avoiding unexpected bleeding or facial nerve branch injury that could occur with sharp dissection. Exposure of the mandibular condyle neck and a portion of the zygomatic arch exposes critical surgical areas, facilitating surgical intervention.
[0031] In step S120, the outer attachments of the disc are detached, exposing the disc so that medical personnel can perform treatment on it. Specifically, the disc is removed, or partially removed. After discectomy, hematoma and fibrous tissue growth within the joint cavity can promote adhesions, facilitating subsequent research into the pathogenesis of temporomandibular joint ankylosis.
[0032] It should be noted that the temporomandibular joint disc wraps around the condylar head and is connected to the joint capsule. The modeling surgery is performed from the outside, and the outside of the disc is seen first. After separating its lateral attachments, the instrument enters from the gap between the disc and the condyle, so that the surrounding attachments can be cut off more smoothly.
[0033] In step S130, the joint space is exposed through horizontal blunt separation, fine ophthalmic forceps are used to remove part of the fibrous cartilage on the condylar head, and a minimally invasive root apex elevator is used to groove the surface of the fossa to destroy the fossa cartilage, so as to simulate the impact trauma of the temporomandibular joint fossa.
[0034] In step S140 , the wound of the experimental subject is sutured, and the experimental subject is fed after waking up, and subsequent observation is performed.
[0035] The construction method provided in the present invention uses a surgical method to remove part of the fibrous cartilage of the condyle and the temporal bone fossa, and remove the articular disc to build a model. Specifically, by removing part of the fibrous cartilage on the condylar head and making grooves on the surface of the fossa to destroy the fossa cartilage, the joint is subjected to complex trauma. The animal model established by this method is closer to the complex pathological environment of human diseases.
[0036] Clinically, ankylosis develops when the temporomandibular joint (TMJ) disc is displaced or shattered during trauma, resulting in a loss of the barrier between the condyle and the fossa. Therefore, compared to closed-loop procedures, which only push the disc to one side, there's no way to prevent the disc from returning to its original position during subsequent movement, blocking the condyle and fossa. The surgical approach employed in this disclosure removes the disc barrier, significantly increasing the likelihood of ankylosis.
[0037] In one example provided herein, 42 3-week-old SPF-grade male C57 / BL6 mice weighing approximately 11-13 g were used. They were housed at the SPF Animal Center of the Third Affiliated Hospital of the Air Force Medical University at a room temperature of 22-24°C, a humidity of 50%-60%, with regular daily ventilation and a 12-hour light-dark cycle. Based on a preliminary study, the sample size was estimated to be 42 mice. All experimental procedures were conducted in strict accordance with animal experimental research reporting guidelines, and every effort was made to minimize animal suffering.
[0038] This disclosure uses mice as experimental subjects, as they are low-cost, easy to operate, and suitable for short-term studies. Furthermore, the 3-week-old SPF-grade male C57 / BL6 mice used in this disclosure are equivalent to 8-12-year-old children in human years, which can better simulate the temporomandibular joint trauma in childhood.
[0039] In order to facilitate the operation and reduce the pain of the experimental subject as much as possible, in one embodiment of the present disclosure, before step S110, step S100 is further included: performing general anesthesia and preoperative disinfection on the experimental subject.
[0040] The specific anesthesia method is as follows: penicillin is injected into the experimental subjects at a dose of 20,000-40,000 U / 1,000 g via intramuscular injection; 1.25% tribromoethanol is injected into the experimental subjects' abdominal cavity at a dose of 2 ml / 100 g for general anesthesia.
[0041] Before the surgery, the preauricular area of the subject was disinfected to prevent wound infection. Specifically, the left preauricular area of the subject was prepared and disinfected with povidone-iodine solution.
[0042] In one embodiment of the present disclosure, in step S120, ophthalmic scissors are used to cut off the peripheral attachments of the articular disc, and the peripheral attachments of the articular disc are precisely removed to avoid accidental damage to adjacent structures.
[0043] In one embodiment of the present disclosure, in step S130, fine ophthalmic forceps are used to remove part of the fibrous cartilage on the condylar head, and a minimally invasive root apex elevator is used to groove the surface of the fossa to destroy the fossa cartilage, so as to simulate the impact trauma of the temporomandibular joint fossa, so that the animal model established by the present disclosure is more in line with the complex pathological environment of human diseases.
[0044] In one embodiment of the present disclosure, in step S140, the method for suturing the wound is: tying 5-0 sutures at the mandibular ramus and the root of the zygomatic arch respectively, pulling the two sutures together, and tying a knot at the root of the zygomatic arch to close the joint space.
[0045] Because the distance between the condyle and the fossa is close, narrowing the joint gap is essential for the development of ankylosis. The present invention uses suture traction to shorten the distance between the condyle and the skull base without restricting the mouse's mouth movement and feeding, similar to the clinical ankylosis patients.
[0046] In addition, after step S140, the following step is also included: S150: Anti-infection and analgesic treatment is performed on the mouse. Specifically, the analgesic treatment method is: after surgery, the experimental subject is given 0.01 mg / 100 g of pentazocine for analgesia. The anti-infection method is: the experimental subject is given the same dose of penicillin as before surgery for 2-3 days, thereby reducing the pain and infection risk of the experimental subject and improving the survival rate of the experimental subject.
[0047] In one embodiment of the present disclosure, the preauricular region of the experimental subject was incised using a pretragus arcuate incision. This incision allows direct access to the joint area, exposing the joint capsule, condyle, and articular disc without excessive soft tissue retraction, thus minimizing intraoperative tissue damage.
[0048] The method for constructing the above-mentioned animal model of traumatic temporomandibular joint ankylosis is specifically described with reference to an embodiment: All mice in the experimental groups were fasted and deprived of water one day before surgery. Their body weight was measured before surgery, and they were given penicillin at a dose of 20,000-40,000 U / 1,000 g by intramuscular injection. 1.25% tribromoethanol was injected intraperitoneally at a dose of 2 ml / 100 g for general anesthesia. After anesthesia, C57 / BL6 mice were placed in the right lateral decubitus position and fixed on the animal experimental operating table. The left preauricular area was prepared and the surgical area was disinfected with povidone-iodine solution.
[0049] Using an arc-shaped incision before the tragus ( Figure 2 Avoid the superficial temporal artery and vein, and perform blunt dissection along the zygomatic arch from front to back to the joint capsule and expose the mandibular condyle neck and part of the zygomatic arch ( Figure 2 After opening the joint capsule, the condyle is separated ( Figure 2 As shown in middle C), the lateral attachment of the articular disc is separated through the joint space. After exposing the articular disc, the peripheral attachment of the articular disc is cut off with ophthalmic scissors, and part or all of the articular disc is removed ( Figure 2 D and E in the middle) to simulate the clinical injury and displacement of the articular disc.
[0050] The joint space was exposed by horizontal blunt dissection, and fine ophthalmic forceps were used to remove part of the fibrocartilage on the condylar head ( Figure 2 (as shown in F) and using a minimally invasive root apex to carve grooves on the glenoid fossa to destroy the glenoid fossa cartilage ( Figure 2 (as shown in G in the middle) to simulate the impact trauma of the temporomandibular joint. Tie 5-0 sutures at the mandibular ramus and the root of the zygomatic arch ( Figure 2 The two sutures are further pulled together and tied at the root of the zygomatic arch to close the joint space ( Figure 2 (as shown in I).
[0051] Finally, the wound was closed by sutures in layers, and the mice were fed with soft food after they woke up. Anti-infection and analgesic treatment was performed on the mice after surgery. The specific analgesic treatment method was: use analgesics (pentazocine, 0.01 mg / 100 g) for 1 day, and the anti-infection method was: give the mice the same dose of penicillin as before surgery for 2-3 days to prevent infection.
[0052] It should be noted that in Figure 2 In the figure, Za is the zygomatic arch; Co is the condyle; Gf is the glenoid fossa; Ad is the articular disc, and the position indicated by the arrow in A is the location of the surgical incision.
[0053] The construction method provided in the present invention uses a surgical method to remove part of the fibrous cartilage of the condyle and the temporal bone fossa, and remove the articular disc to build a model. Specifically, by removing part of the fibrous cartilage on the condylar head and making grooves on the surface of the fossa to destroy the fossa cartilage, the joint is subjected to complex trauma. The animal model established by this method is closer to the complex pathological environment of human diseases.
[0054] Subsequent observations such as Figures 3 to 6 As shown, Figure 3 A is the mouth opening degree of the control group mice, and B is the mouth opening degree of the experimental group mice. Figure 3 It can be seen that the mouth opening degree of the experimental group mice is smaller than that of the control group mice, that is, the mouth opening of the experimental group mice is limited compared with that of the control group mice, which is consistent with the clinical symptoms.
[0055] Figure 4 A is the left side of the experimental group mouse, B is the right side of the experimental group mouse, C is the cross section of the experimental group mouse, and the circle in C is the left joint of the mouse. Figure 4 It can be observed that compared with the right side, the left side of the mouse has irregular joint proliferation and the joint space disappears.
[0056] Depend on Figure 5 In the coronal plane of figure A, the condyle has lost its original shape and has proliferated irregularly inward and outward. The bone on the lateral side of the articular fossa has proliferated and thickened, and the original joint space has disappeared. Figure 5 The sagittal MIcro-ct image in middle B shows that the condyle and the glenoid fossa have proliferated and connected, and the joint space has disappeared.
[0057] exist Figure 6 In the control group, the joint structure, specifically the glenoid fossa, articular disc, and condyle, was clear and complete, with a distinct joint space. In the experimental group, the condyle and glenoid fossa were irregularly overgrown, with a bone bridge connecting the two on the outside of the joint, and the joint space was lost.
[0058] Depend on Figures 3 to 6 It can be concluded that in the animal model constructed by the construction method provided by the present invention, the condyle and glenoid fossa proliferate irregularly, the lateral bone bridge of the joint connects the two, and the joint space disappears, which is closer to the characteristics of human temporomandibular joint lesions.
[0059] The embodiments of the present disclosure have been described above. The above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terms used herein are selected to best explain the principles of the embodiments, their practical applications, or technical improvements in the marketplace, or to enable other persons skilled in the art to understand the embodiments disclosed herein. The scope of the present disclosure is defined by the appended claims.
Claims
1. A method for constructing an animal model of traumatic temporomandibular joint ankylosis, characterized in that: The construction method comprises the following steps: S110: Select the preauricular area on one side of the experimental subject, avoid the superficial temporal artery and vein, and bluntly dissect along the zygomatic arch from front to back to the joint capsule, exposing the mandibular condyle neck and part of the zygomatic arch. After opening the joint capsule, separate the condyle. S120: Separate the lateral attachments of the disc through the joint space, expose the disc, cut off the peripheral attachments of the disc, and remove part or all of the disc. S130: The joint space is exposed by horizontal blunt dissection of the joint capsule, part of the fibrocartilage on the condylar head is removed, and the glenoid fossa cartilage is destroyed; S140: The wound is sutured, and the experimental subject eats soft food after waking up, thereby obtaining an animal model of traumatic temporomandibular joint ankylosis.
2. The construction method according to claim 1, characterized in that The experimental subjects were 3-week-old mice weighing 11-13 g.
3. The construction method according to claim 2, characterized in that Before step S110 , the method further includes step S100 : performing general anesthesia and preoperative disinfection on the experimental subject.
4. The construction method according to claim 3, characterized in that The anesthesia method for the experimental subject comprises the following steps: injecting penicillin into the experimental subject by intramuscular injection at a dose of 20,000-40,000 U / 1,000 g; The experimental subjects were injected with 1.25% tribromoethanol at a dose of 2 ml / 100 g into the abdominal cavity for general anesthesia.
5. The construction method according to claim 3, characterized in that The preoperative disinfection of the experimental subject includes the following steps: after preparing the skin of the left preauricular area of the experimental subject, disinfecting the surgical area with povidone-iodine solution.
6. The construction method according to claim 2, characterized in that In step S120, the peripheral attachments of the articular disc are severed using ophthalmic scissors.
7. The construction method according to claim 2, characterized in that: In step S130, fine ophthalmic forceps are used to remove part of the fibrous cartilage on the condylar head, and a minimally invasive apex elevator is used to cut grooves on the surface of the glenoid fossa to destroy the glenoid fossa cartilage.
8. The construction method according to claim 2, characterized in that: In step S140 , the wound is sutured by tying 5-0 sutures at the mandibular ramus and the root of the zygomatic arch respectively, pulling the two sutures together, and tying a knot at the root of the zygomatic arch to close the joint space.
9. The construction method according to claim 4, characterized in that: After step S140, the following steps are also included: S150: performing anti-infection and analgesic treatment on the mice.
10. The construction method according to claim 9, characterized in that: The analgesic treatment method is: administering 0.01 mg / 100 g pentazocine to the experimental subject for analgesia after surgery; and / or, The anti-infection method is: giving the experimental subject the same dose of penicillin as before surgery for 2-3 days.
11. The construction method according to claim 1, characterized in that: The preauricular area of the experimental subject was incised using a pretragus arc incision.