Rat molar transplantation experimental model for autologous tooth transplantation research, construction method and application

Abstract

The invention discloses a rat molar transplantation experimental model for autologous tooth transplantation research, a construction method and application, and belongs to the technical field of oral basic medical experiments. The method comprises the following specific steps that firstly, a to-be-processed transplanted rat model in which a third molar is inserted into a second molar alveolar socket in advance is obtained, and the distal root, the mesial cheek root and the palate root of the third molar in the model are correspondingly inserted into the mesial palate root, the distal cheek and the mesial cheek root alveolar socket of the second molar; then carrying out acid etching treatment on the enamel of the first molar and the enamel of the third molar by adopting an acid etching agent, and coating an adhesive on the surface of the enamel; then placing flowing resin on the tooth surface subjected to acid etching, embedding a ligature wire into the flowing resin, and carrying out illumination curing; and finally, performing infection prevention management to obtain the rat molar transplantation experiment model. The construction method has the characteristics of short experimental period and high operation repeatability, and is suitable for subsequent periodontal membrane healing mechanism research, dental pulp revascularization process observation and screening evaluation of dental biological materials.

Description

TECHNICAL FIELD

[0001] The application belongs to the experimental technical field of oral basic medicine, and particularly relates to a rat molar transplantation experimental model for autologous tooth transplantation research, a construction method and application. BACKGROUND

[0002] Autologous tooth transplantation, as a biological repair method to replace missing teeth, has shown high success rate and long-term stability in clinical practice. However, due to the ethical restrictions and individual variable control of human clinical research, it is extremely challenging to directly observe the tissue healing mechanism (such as periodontal membrane regeneration and pulp revascularization) after transplantation. Therefore, developing stable and repeatable in vitro experimental models and animal models has become a key tool to promote the basic research of autologous tooth transplantation.

[0003] At present, the commonly used animal models in this field mainly rely on large animals such as dogs, rabbits and monkeys, and the research objects are mostly concentrated on the transplantation of single anterior teeth. However, these models have problems such as high cost, long experimental period and complex operation, which limit the feasibility of large-scale research. In contrast, molar transplantation (such as transplantation of the third molar to replace the first or second molar) has important application value in clinical practice, but due to the complex anatomical structure of the molar (multiple roots, narrow operation space), related research is still relatively scarce.

[0004] In recent years, rat models have been gradually introduced into the field of tooth transplantation research due to their high economy, clear genetic background and strong experimental repeatability. However, standardized experimental methods for rat molar transplantation are still in the exploratory stage, especially in the precise positioning after tooth extraction and stable fixation after transplantation, which lack systematic optimization. This technical gap seriously restricts the subsequent molecular mechanism research and biological material screening.

[0005] Therefore, there is an urgent need in the field to establish a highly efficient and standardized rat molar transplantation model construction method to support the basic scientific research needs of periodontal tissue repair mechanism research and new dental materials evaluation, thereby providing a theoretical basis for future clinical translation. SUMMARY

[0006] The purpose of the present application is to establish a standardized, repeatable, economical and efficient rat molar transplantation animal model that can better simulate the periodontal membrane healing and pulp revascularization process after clinical autologous tooth transplantation. Specifically, a rat molar transplantation experimental model for autologous tooth transplantation research, a construction method and application are provided.

[0007] The specific technical solutions adopted by the present application are as follows: In a first aspect, the present application provides a construction method of a rat molar transplantation experimental model for autologous tooth transplantation research, the specific steps of which are as follows: S1: obtaining a to-be-treated transplantation rat model in which a third molar is inserted into a socket of a second molar in advance; the distal root, the mesial buccal root and the palatal root of the third molar are respectively inserted into the mesial palatal root, the distal buccal root and the mesial buccal root socket of the second molar, and suturing is completed; S2: performing acid-etching treatment on the enamel of the first molar in the to-be-treated transplantation rat model and the third molar in the socket of the second molar by using an acid-etching agent; S3: after the acid-etching treatment is completed, the excess acid-etching agent is wiped off, and an adhesive is coated on the surface of the acid-etched enamel; then, the flowing resin is placed on the acid-etched tooth surface by using a probe, and a ligation wire is embedded in the flowing resin, and light curing is performed; then, preventive infection management is performed, and a rat molar transplantation experimental model is obtained.

[0008] Preferably, the acid-etching agent in step S2 is a phosphoric acid solution with a concentration of 30% to 50%.

[0009] Preferably, the acid-etching treatment in step S2 is performed for 15 to 25 seconds.

[0010] Preferably, the length of the ligation wire in step S3 is 0.9 to 1.1 mm.

[0011] Preferably, the light curing in step S3 is performed for 15 to 25 seconds.

[0012] Preferably, the preventive infection management in step S3 includes preventive antibiotic treatment for 3 to 5 days, and the daily dosage is 20,000 IU of penicillin sodium.

[0013] In a second aspect, the present application provides a rat molar transplantation experimental model obtained by using the construction method in the first aspect.

[0014] In a third aspect, the present application provides an application of a rat molar transplantation experimental model, and the rat molar transplantation experimental model in the second aspect is used for studying the mechanism of periodontal ligament healing and pulp revascularization after autologous tooth transplantation.

[0015] In a fourth aspect, the present application provides an application of a rat molar transplantation experimental model, and the rat molar transplantation experimental model in the third aspect is used for evaluating the application effect of dental biomaterials in tooth transplantation.

[0016] Compared with the prior art, the present application has the following beneficial effects: (1) The molar transplantation model construction method provided by the present application has a short experimental period, can quickly complete the construction of the model and related experiments, greatly saves research time, improves scientific research efficiency, and has low cost.

[0017] (2) The method provided by the application has a standardized operation process, strict step design, and ensures the accuracy and consistency of experimental operation. The standardized operation process makes the experimental results have good repeatability.

[0018] (3) The model constructed by the application is suitable for various detection methods, including histology (HE staining), imaging (Micro-CT), and molecular biology (gene expression analysis), and can comprehensively analyze the biological behavior after transplantation.

[0019] (4) The model provided by the application can also be used to study the key mechanisms of periodontal ligament healing and pulp revascularization, and provides an important tool for basic research in the field of dentistry. Periodontal ligament healing and pulp revascularization are key links for successful tooth transplantation, and through the model, the regulatory factors and influencing factors of these mechanisms can be further explored, providing a theoretical basis for the improvement and optimization of clinical tooth transplantation technology.

[0020] (5) The model provided by the application provides an effective platform for material intervention. For example, the model can be used to study the intervention of hydrogel, bioactive carrier and other materials, and observe the influence of these materials on the healing process after molar transplantation, so as to screen out materials more conducive to tooth repair and regeneration, and provide strong support for the research and application of new dental materials. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 The construction method flowchart of the molar transplantation experimental model provided by the application is shown in the figure. Figure 2 The cross-sectional view of the third molar transplantation in the maxilla to the second molar alveolar socket is shown in the figure. Figure 3 The Micro-CT image of the third molar transplantation in the example is shown in the figure. Figure 4 The comparison chart of periodontal tissue healing HE staining of the third molar transplantation in the example after 4 weeks (b) and the natural tooth (a) is shown in the figure. Figure 5 The picture of the third molar transplantation in the example after 4 weeks is shown in the figure. DETAILED DESCRIPTION

[0022] The application will be further described and explained in conjunction with the accompanying drawings and specific embodiments. The technical features of each embodiment in the application can be combined accordingly without conflict.

[0023] Example 1 The construction method of a rat molar transplantation experimental model for autologous tooth transplantation mechanism research is provided in this example. The construction method flowchart of this example is shown in the figure. Figure 1The specific steps are as follows: (1) Obtain a to-be-treated transplanted rat model in which the third molar is inserted into the alveolar crypt of the second molar in advance. It should be noted that the present application does not specifically limit the obtaining method of the to-be-treated transplanted rat model. Those skilled in the art can directly purchase a commercially available rat model or perform self-transplantation. The self-transplantation method is adopted in this embodiment: Select 20 male Sprague-Dawley rats (SD rats) aged 8-9 weeks and weighing 230-280 g, and adaptively feed them for one week, feed them with soft food, and let them drink water freely.

[0024] After anesthesia treatment, the third molar and the second molar are pulled out by using the Taiji tooth extraction method: first, the gum around the neck of the third molar is gently separated by using a probe. Then, the probe is inserted into the mesial surface, distal surface, buccal side and palatal side of the third molar in turn by using the “Taiji method” minimally invasive extraction technique, and a stable force is continuously applied in the opposite direction for 10-15 seconds, and then the force is released until the tooth is loose. Then, the tooth crown is pulled out by using a hemostat to clamp the tooth crown, and then the tooth is soaked in physiological saline. The second molar is completely pulled out by using the same method.

[0025] As shown in Figure 2 , the third molar of the SD rat is appropriately rotated and transplanted into the alveolar crypt of the second molar in the original position, so that the distal root, mesial buccal root and palatal root of the third molar are respectively inserted into the mesial palatal root, distal buccal root and mesial buccal root of the second molar. The gum tissue is sutured in place by using 5-0 absorbable suture, and it is ensured that the soft tissue is well covered.

[0026] (2) Then, a phosphoric acid solution with a concentration of 37% is used as an etching agent to etch the enamel of the transplanted third molar and the first molar, and the etching treatment time is 20 seconds. After the etching treatment is completed, the excess etching agent is wiped off, and an adhesive is applied to the surface of the etched enamel and is subjected to light treatment for 20 seconds. Then, a small amount of flowing resin is placed on the etched tooth surface by using a probe, and a length of about 1 mm of the ligation wire is embedded in the flowing resin, and after light curing, the fixing is completed.

[0027] (3) Preventive infection management: 20,000 IU of penicillin sodium is injected intramuscularly every day for 3 days, and soft food is fed to reduce mechanical stress, and finally the rat molar transplantation experimental model is obtained.

[0028] (4) Model evaluation At 4 weeks after the operation, the stability of the transplanted tooth, the healing of the alveolar bone and the biological response of the dental pulp / periodontal membrane are evaluated by using multiple modalities, such as direct observation in the mouth, Micro-CT imaging, Hematoxylin and Eosin staining (HE) and the like, to verify the feasibility and stability of the model. The results are as follows:Figure 3 and Figure 4 as shown.

[0029] Figure 3 is the Micro-CT image of the third molar immediately after transplantation. The Micro-CT image immediately after transplantation shows that the third molar has been stably implanted in the original alveolar crypt of the second molar, and the tooth axial direction and the root embedding state are good, indicating that the operation method described in the application has feasibility and repeatability in achieving the initial stability of the molar transplantation of the rat.

[0030] Figure 4 is the comparison of HE staining of the periodontal tissue healing of the third molar 4 weeks after transplantation and the natural tooth in this embodiment. Figure 4 (a) shows that the periodontal membrane structure is regular, the fiber arrangement is clear, and the cementum and bone tissue structure are complete. Figure 4 (b) shows that the periodontal membrane in the region of the transplanted tooth is visible but the fiber arrangement is disordered, and the root surface cementum layer is discontinuous and local bone tissue reconstruction is visible, indicating that the model has appeared periodontal tissue reconstruction and vascularization physiological process at 4 weeks after transplantation, and is a feasible experimental platform for studying the repair mechanism of periodontal membrane and bone tissue after transplantation.

[0031] This embodiment also takes a photograph of the third molar 4 weeks after transplantation, and the result is shown in Figure 5 According to the figure, the transplanted tooth position is stable, and no shedding or obvious displacement is observed, and the gingival tissue morphology is good, indicating that the model can maintain stable occlusion and local soft tissue healing state after operation. Among the 20 initial SD rats, 16 rats were successfully modeled, and the modeling success rate of this embodiment was 80%.

[0032] The above-described embodiments are only a preferred scheme of the application, and are not intended to limit the application. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the application. Therefore, any technical solution obtained by equivalent replacement or equivalent transformation falls within the protection scope of the application.

Claims

1. A method for constructing a rat molar transplantation experimental model for autologous tooth transplantation research, characterized by, The specific steps are as follows: S1: obtaining a to-be-treated transplantation rat model in which a third molar is inserted into a second molar alveolar crypt in advance; the distal root, mesial buccal root and palatal root of the third molar of the to-be-treated transplantation rat model are respectively inserted into the mesial palatal root, distal buccal and mesial buccal root alveolar crypts of the second molar, and suturing is completed; S2: using an acid etching agent to perform acid etching treatment on the enamel of the first molar in the to-be-treated transplantation rat model and the third molar in the second molar alveolar crypt; S3: after the acid etching treatment is completed, the excess acid etching agent is wiped off, and an adhesive is coated on the surface of the acid-etched enamel; then, a probe is used to place flowing resin on the acid-etched tooth surface, and a ligation wire is embedded in the flowing resin, and light curing is performed; then, preventive infection management is performed, and a rat molar transplantation experimental model is obtained.

2. The method of constructing a rat molar transplantation test model according to claim 1, wherein The acid etching agent in step S2 is a phosphoric acid solution with a concentration of 30%-50%.

3. The method of claim 1, wherein the method is characterized by: The acid etching treatment in step S2 is performed for 15-25 seconds.

4. The method of claim 1, wherein the method is characterized by, The length of the ligation wire in step S3 is 0.9-1.1 mm.

5. The method of claim 1, wherein the method is characterized by: The light curing in step S3 is performed for 15-25 seconds.

6. The method of constructing a rat molar transplantation experimental model according to claim 1, wherein The preventive infection management in step S3 includes preventive antibiotic treatment for 3-5 days, and the daily dosage is 20,000 IU of penicillin sodium.

7. A rat molar transplantation experimental model obtained by using the construction method of any one of claims 1-6.

8. Use of a rat molar transplantation model, characterized in that The rat molar transplantation experimental model of claim 7 is used for studying the mechanism of periodontal membrane healing and pulp revascularization after autologous tooth transplantation.

9. Use of a rat molar transplantation model, characterized in that The rat molar transplantation experimental model of claim 7 is used for evaluating the application effect of dental biomaterials in tooth transplantation.

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