Tissue preservation solution for autologous tooth transplantation as well as preparation method and application of tissue preservation solution
By using a preservation solution composed of high-glucose DMEM culture medium, antioxidant L-ascorbic acid-2-phosphate, and anti-inflammatory dexamethasone, the problems of cell viability and short preservation time in autologous tooth transplantation have been solved, achieving high viability and long-term preservation of periodontal ligament cells, thus improving the success rate of autologous tooth transplantation.
Patent Information
- Application Number
- CN202510785011.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-12
- Publication Date
- 2025-11-14
AI Technical Summary
Current autologous tooth transplant preservation solutions have a single composition, which cannot effectively maintain the activity of periodontal ligament cells. They lack antioxidants and anti-inflammatory factors, resulting in cell damage and short preservation time, making them unsuitable for long-term preservation.
A preservation solution composed of high-glucose DMEM culture medium, ITS, dexamethasone, L-ascorbic acid-2-phosphate, and penicillin/streptomycin was used to extend the preservation time and promote cell repair by providing energy support, anti-inflammation, and antioxidant effects.
It significantly improves the activity and anti-inflammatory capacity of periodontal ligament cells, extends the preservation time to more than 6 hours, and improves the success rate and clinical effect of autologous tooth transplantation.
Smart Images
Figure CN120944797A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of biotechnology, and more specifically, to a tissue preservation solution for autologous tooth transplantation, its preparation method, and its application. Background Technology
[0002] In autologous tooth transplantation, the preservation solution serves as a crucial link between tooth preservation and transplantation, and is essential for maintaining the viability of periodontal ligament cells. Existing preservation solutions, such as sodium chloride injection (NS) or Hofbart balanced salt solution (HBSS), are widely used. These solutions primarily maintain ion balance and osmotic pressure regulation; however, their compositions are too singular, failing to provide the necessary support components for periodontal ligament cells. This leads to decreased metabolic capacity of cells during preservation, a lack of antioxidant protection, and the accumulation of free radicals causing cell damage, thus affecting periodontal ligament cell survival and post-transplantation survival rates. Furthermore, existing preservation solutions lack anti-inflammatory or growth factors, failing to effectively suppress inflammatory responses during preservation and lacking the function of promoting tissue repair. As a result, preservation times are generally short, typically not exceeding 0.5 hours, making it difficult to meet the needs of long-term preservation, especially in long-distance transportation or complex surgical scenarios, and failing to guarantee high activity and repair capacity of tooth tissue.
[0003] Therefore, there is an urgent need for a tissue preservation solution for autologous tooth transplantation, its preparation method, and its application to solve the above problems. Summary of the Invention
[0004] The purpose of this invention is to provide a tissue preservation solution for autologous tooth transplantation, its preparation method, and its application, in order to improve the aforementioned problems. To achieve the above objective, the technical solution adopted by this invention is as follows:
[0005] In one aspect, this application provides a tissue preservation solution for autologous tooth transplantation, comprising the following components: high-glucose DMEM culture medium, ITS (insulin-transferrin-selenium), dexamethasone, L-ascorbic acid-2-phosphate, and penicillin / streptomycin.
[0006] Preferably, the concentrations of each component are: 1% ITS (insulin-transferrin-selenium), 0.1 μM dexamethasone, 0.2 mL ascorbic acid-2-phosphate and 1% penicillin / streptomycin.
[0007] Secondly, this application also provides a method for preparing the above-mentioned tissue preservation solution for autologous tooth transplantation, comprising the following steps:
[0008] Dexamethasone was dissolved in anhydrous dimethyl sulfoxide according to the preset concentration requirements to prepare a dexamethasone inhibitor stock solution.
[0009] Dissolve a pre-concentrated sodium L-ascorbic acid-2-phosphate in sterile water and adjust the pH to neutral to obtain an antioxidant stock solution.
[0010] Under a sterile operating table, using high-glucose DMEM as the base solution, 1% ITS (insulin-transferrin-selenium), 0.1 μM dexamethasone, 0.2 mL ascorbic acid-2-phosphate and 1% penicillin-streptomycin antibiotic were added in sequence according to the ratio. After mixing evenly, the pH value was adjusted with sterile NaOH or HCl solution to obtain the tissue preservation solution.
[0011] The tissue preservation solution was aseptically filtered using a 0.22 μm filter membrane and then dispensed into sterile centrifuge tubes to obtain the final tissue preservation solution product.
[0012] Preferably, the pH value of the antioxidant stock solution is adjusted to be in the range of 7.2 to 7.4.
[0013] Preferably, the pH value of the sterile NaOH or HCl solution is adjusted to be in the range of 7.2 to 7.4.
[0014] Thirdly, this application also provides the application of the above-mentioned tissue preservation solution for autologous tooth transplantation in the tissue preservation of autologous tooth transplantation.
[0015] The specific steps for tissue preservation using autologous tooth transplant tissue preservation solution provided by this invention are as follows:
[0016] The extracted target tooth is immediately placed in a tissue preservation solution that has met the assessment criteria;
[0017] The tissue preservation solution placed in the target tooth is kept at a temperature of 4°C and transported to the transplant site for tooth transplantation.
[0018] Preferably, the root surface of the target tooth is rinsed before the target tooth is transplanted, and the tooth transplantation operation is performed after rinsing.
[0019] Preferably, the transportation time to the transplant site does not exceed 6 hours.
[0020] Preferably, when the extracted target tooth is immediately placed into the tissue preservation solution, the target tooth must be completely immersed in the tissue preservation solution.
[0021] The beneficial effects of this invention are as follows:
[0022] This invention effectively enhances the activity of periodontal ligament cells, strengthens their anti-inflammatory and tissue repair capabilities, and prolongs preservation time during the preservation process. To this end, this invention uses high-glucose DMEM culture medium as the base medium, combined with antioxidants such as L-ascorbic acid-2-phosphate to effectively scavenge free radicals and reduce oxidative damage. Furthermore, the addition of dexamethasone and ITS (insulin, transferrin, selenium) enhances anti-inflammatory effects and promotes cell proliferation, optimizing tissue repair. This preservation solution provides sufficient metabolic substrates, improves cellular energy supply, and ensures high periodontal ligament cell activity for preservation periods exceeding 6 hours. It is suitable for long-term transport and complex clinical surgical environments, thereby significantly improving the success rate and clinical outcomes of autologous tooth transplantation. This tissue preservation solution not only improves cell viability protection but also enhances anti-inflammatory and repair capabilities, filling a gap in existing technologies.
[0023] Other features and advantages of the invention will be set forth in the following description, and will be apparent in part from the description, or may be learned by practicing embodiments of the invention. The objects and other advantages of the invention may be realized and obtained by means of the structures particularly pointed out in the written description, claims, and drawings. Attached Figure Description
[0024] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.
[0025] Figure 1 This is a comparison chart of the Western Blot results of tissue preservation solution and sodium chloride injection solution preserved for 1 hour in the embodiments of the present invention.
[0026] Figure 2 This is a comparison chart of the Western Blot results of tissue preservation solution and sodium chloride injection solution after 3 hours in the embodiments of the present invention.
[0027] Figure 3 This is a comparison chart of the Western Blot results of tissue preservation solution and sodium chloride injection solution preserved for 6 hours in the embodiments of the present invention. Detailed Implementation
[0028] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. The components of the embodiments of the present invention described and shown in the accompanying drawings can generally be arranged and designed in various different configurations. Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.
[0029] It should be noted that similar reference numerals and letters in the following figures indicate similar items; therefore, once an item is defined in one figure, it does not need to be further defined and explained in subsequent figures. Furthermore, in the description of this invention, terms such as "first," "second," etc., are used only to distinguish descriptions and should not be construed as indicating or implying relative importance.
[0030] Example 1:
[0031] The present invention provides a tissue preservation solution for autologous tooth transplantation, characterized in that it comprises the following components: high glucose DMEM culture medium, ITS (insulin-transferrin-selenium), dexamethasone, L-ascorbic acid-2-phosphate and penicillin / streptomycin.
[0032] As one implementation method, the concentrations of each component are: 1% ITS (insulin-transferrin-selenium), 0.1 μM dexamethasone, 0.2 mL ascorbic acid-2-phosphate and 1% penicillin / streptomycin.
[0033] The tissue preservation solution described in this invention uses high-glucose DMEM as the base solution, supplemented with a precise ratio of 1% ITS, 0.1 μM dexamethasone, 0.2 mM L-ascorbic acid-2-phosphate, and 1% penicillin / streptomycin. This achieves multiple synergistic effects in maintaining cell metabolism, anti-inflammatory and antioxidant properties, and antibacterial activity, effectively enhancing the activity and stability of periodontal ligament cells during in vitro preservation. High-glucose DMEM provides energy substrates to support basic metabolism, ITS promotes cell proliferation and repair, dexamethasone inhibits inflammatory responses, L-ascorbic acid-2-phosphate scavengees free radicals, and penicillin / streptomycin maintains a sterile preservation environment, significantly extending preservation time to over 6 hours, far superior to traditional preservation solutions, effectively improving the success rate of autologous tooth transplantation and the flexibility of clinical procedures.
[0034] This invention provides a method for preparing the tissue preservation solution described in the above technical solution, comprising the following steps:
[0035] Dexamethasone was dissolved in anhydrous dimethyl sulfoxide according to the preset concentration requirements to prepare a dexamethasone inhibitor stock solution.
[0036] Dissolve a pre-concentrated sodium L-ascorbic acid-2-phosphate in sterile water and adjust the pH to neutral to obtain an antioxidant stock solution.
[0037] Under a sterile operating table, using high-glucose DMEM as the base solution, 1% ITS (insulin-transferrin-selenium), 0.1 μM dexamethasone, 0.2 mL ascorbic acid-2-phosphate and 1% penicillin-streptomycin antibiotic were added in sequence according to the ratio. After mixing evenly, the pH value was adjusted with sterile NaOH or HCl solution to obtain the tissue preservation solution.
[0038] The tissue preservation solution was aseptically filtered using a 0.22 μm filter membrane and then dispensed into sterile centrifuge tubes to obtain the final tissue preservation solution product.
[0039] As one implementation, the pH value of the antioxidant stock solution is adjusted to a range of 7.2 to 7.4.
[0040] As one embodiment, the sterile NaOH or HCl solution is used to adjust the pH value to a range of 7.2 to 7.4.
[0041] This invention achieves sterility, safety, and component homogeneity and stability in the final preservation solution product. This method not only ensures the activity and compatibility of the components but also expands the regulatory mechanism of the preservation solution through the use of substitutable small molecules, providing more options for individualized preservation needs and significantly improving the ability to maintain the bioactivity of tooth tissue during autologous tooth transplantation.
[0042] Based on the above advantages, the present invention provides the application of the tissue preservation solution described in the above technical solution or the tissue preservation solution prepared by the preparation method described in the above technical solution in the tissue preservation of autologous tooth transplantation.
[0043] The specific steps involved in tissue preservation include: immediately placing the extracted target tooth into a tissue preservation solution that has met the assessment criteria;
[0044] The tissue preservation solution placed in the target tooth is kept at a temperature of 4°C and transported to the transplant site for tooth transplantation.
[0045] As one implementation method, the root surface of the target tooth is rinsed before the target tooth is transplanted, and the tooth transplantation operation is performed after rinsing is completed.
[0046] As one implementation method, the transportation time to the transplant site does not exceed 6 hours.
[0047] As one implementation, when the extracted target tooth is immediately placed into the tissue preservation solution, the target tooth must be completely immersed in the tissue preservation solution.
[0048] This invention proposes a specific application method for this preservation solution in autologous tooth transplantation, fully leveraging its advantages in maintaining periodontal ligament cell activity, inhibiting inflammation, and prolonging preservation time. In the specific application process, the extracted target tooth must be immediately placed in a qualified tissue preservation solution to ensure that the tooth tissue is in a favorable biological environment from the moment it is removed from the oral cavity, thereby maximizing the preservation of cell function. The preservation solution significantly slows metabolism and reduces oxidative damage at a low temperature of 4°C, while ensuring its bioactivity is maintained evenly within 6 hours of transport to the transplantation site, providing a greater time buffer for clinical procedures. Rinsing the root surface before transplantation further removes potential sources of contamination, helping to improve the postoperative healing environment. During preservation, the tooth must be completely immersed in the preservation solution to ensure uniform hydration and prevent localized drying that could lead to cell death. In summary, the application steps proposed in this invention are not only scientifically sound but also fully match the pharmacological properties of the preservation solution, providing comprehensive assurance for improving the success rate of autologous tooth transplantation and the quality of tissue healing.
[0049] Additionally, such as Figure 1 , Figure 2 and Figure 3 The preservation solution of the present invention can significantly improve the activity and survival rate of periodontal ligament stem cells extracted after 1-6 hours of storage at room temperature, and has a more obvious promoting effect compared with sodium chloride injection solution.
[0050] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.
[0051] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.
Claims
1. A tissue preservation solution for autologous tooth transplantation, characterized in that, It includes the following components: high glucose DMEM culture medium, ITS (insulin-transferrin-selenium), dexamethasone, L-ascorbic acid-2-phosphate and penicillin / streptomycin.
2. The tissue preservation solution for autologous tooth transplantation according to claim 1, characterized in that... The concentrations of each component are: 1% ITS (insulin-transferrin-selenium), 0.1 μM dexamethasone, 0.2 mL ascorbic acid-2-phosphate and 1% penicillin / streptomycin.
3. The method for preparing the tissue preservation solution for autologous tooth transplantation according to any one of claims 1-2, characterized in that... This includes the following steps: Dexamethasone was dissolved in anhydrous dimethyl sulfoxide according to the preset concentration requirements to prepare a dexamethasone inhibitor stock solution. Dissolve a pre-concentrated sodium L-ascorbic acid-2-phosphate in sterile water and adjust the pH to neutral to obtain an antioxidant stock solution. Under a sterile operating table, using high-glucose DMEM as the base solution, 1% ITS (insulin-transferrin-selenium), 0.1 μM dexamethasone, 0.2 mL ascorbic acid-2-phosphate and 1% penicillin-streptomycin antibiotic were added in sequence according to the ratio. After mixing evenly, the pH value was adjusted with sterile NaOH or HCl solution to obtain the tissue preservation solution. The tissue preservation solution was aseptically filtered using a 0.22 μm filter membrane and then dispensed into sterile centrifuge tubes to obtain the final tissue preservation solution product.
4. The method for preparing the tissue preservation solution for autologous tooth transplantation according to claim 3, characterized in that... ,include: The pH range of the antioxidant stock solution is 7.2 to 7.
4.
5. The method for preparing the tissue preservation solution for autologous tooth transplantation according to claim 3, characterized in that... ,include: The pH value of the sterile NaOH or HCl solution is adjusted to a range of 7.2 to 7.
4.
6. The tissue preservation solution for autologous tooth transplantation according to any one of claims 1-2 is used in the tissue preservation of autologous tooth transplantation.
7. The application according to claim 4, characterized in that, Specifically, the following steps are included: The extracted target tooth is immediately placed in a tissue preservation solution that has met the assessment criteria; The tissue preservation solution placed in the target tooth is kept at a temperature of 4°C and transported to the transplant site for tooth transplantation.
8. The application according to claim 6, characterized in that, include: Before transplanting the target tooth, the root surface of the target tooth is rinsed, and the tooth transplantation procedure is performed after rinsing.
9. The application according to claim 4, characterized in that, include: The transportation time to the transplant site shall not exceed 6 hours.
10. The application according to claim 4, characterized in that, include: When the extracted target tooth is immediately placed in the tissue preservation solution, the target tooth must be completely immersed in the tissue preservation solution.