Tooth protector assembled with liquid collecting module and application of tooth protector in early warning of oral cancer

Abstract

The invention relates to a tooth protector assembled with a liquid collection module and application of the tooth protector in early warning of oral cancer, and belongs to the field of biomedical treatment instruments. The invention is inspired by a common nepenthes lip water collection structure, and discloses an early oral cancer early-warning tooth protection device with a liquid collection module, which comprises a tooth socket and the liquid collection module, the liquid collecting module is provided with a common nepenthes bionic microstructure, the liquid collecting module is in an arch bridge shape, the two ends of the arch bridge structure are a supply side and a receiving side respectively, and the arch bridge structure comprises evenly-arranged protrusions; the bottom of the inner side of the tooth socket is connected with a groove; the supply side of the liquid collection module faces downwards, and the receiving side of the liquid collection module faces upwards and adheres into the groove. The device is used as an intraoral system and is combined with a wearable tooth protector, so that tumor necrosis factor alpha (TNF-alpha) in saliva can be permanently, specifically and sensitively detected in situ, and early-warning of early oral cancer can be carried out in time.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical treatment instruments, and more specifically, relates to a mouthguard equipped with a liquid collection module and its application in oral cancer early warning. Background Art

[0002] Oral cancer, including malignancies of the lips, tongue, gums, palate, floor of mouth, and other oral structures, is a major challenge to global public health. It is estimated that the global incidence of oral cancer exceeds 70,000 cases per year, placing a heavy burden on healthcare systems and individuals. Early oral cancer is divided into stages I and II, which are generally confined to the primary site of the oral cavity and have a good prognosis. In contrast, advanced oral cancer (including stages III and IV) is characterized by the spread of cancer cells to deeper tissues, lymph nodes, or distant organs, which greatly increases the mortality rate. This emphasizes the need for early detection and intervention, which is crucial to curb disease progression and improve overall survival. However, early oral cancer is easily overlooked due to its hidden location (such as the root of the tongue, floor of the mouth, and buccal mucosa) and atypical symptoms (such as mild pain, difficulty chewing, and foreign body sensation), which poses a great challenge to early diagnosis. Therefore, current research efforts are dedicated to enhancing the understanding of the etiology of oral cancer, identifying biomarkers for initial screening, and developing innovative treatment strategies to combat this terrible malignancy.

[0003] Despite the availability of multiple oral cancer diagnostic strategies (e.g., regular intraoral examinations, imaging, histopathological evaluation, and biomarker testing), there are still major clinical challenges that deserve further attention: (1) the insensitivity and nonspecificity of early detection of oral cancer (e.g., collection and detection of biomarkers); (2) the high cost of advanced imaging tests such as computed tomography and magnetic resonance imaging may limit patient acceptance; (3) intraoral tissue biopsies may result in adverse congenital lesions and carry multiple risks of bleeding, infection, and even cancer spread; (4) the reliance on professional knowledge and limited home use further limit the accessibility of diagnosis. Encouragingly, saliva-based diagnostic methods have attracted great interest as a promising method for diagnosing oral diseases, mainly because the direct and persistent contact of saliva with intraoral lesions ensures the high sensitivity and convenience of saliva. In addition, compared with traditional methods, this method also has significant advantages such as noninvasive sample collection, large sample quantity, and low risk of infection. In particular, more than 100 biomarkers (e.g., proteins, nucleic acids, metabolites, and microorganisms) have been found in saliva, which provides a huge opportunity for diversified diagnosis of oral diseases. Attractively, case-control exploratory studies have highlighted that the salivary cytokine TNF-α is significantly dysregulated (approximately 30-200 pg / mL) in patients with oral cancer compared with healthy controls (generally less than 9 pg / mL), reflecting the inflammatory and immune disorders associated with tumorigenesis. However, current methods for detecting TNF-α (e.g., enzyme-linked immunosorbent assay, immunofluorescence, flow cytometry, and mass spectrometry) still face many challenges. Therefore, it is necessary to explore convenient, sensitive, economical, and reliable strategies for detecting oral tumor necrosis factor α (TNF-α) to improve the survival rate and prognosis of patients with oral cancer, reduce medical expenses, and improve patients' medical experience. Summary of the invention

[0004] In order to overcome the problems of insensitivity, nonspecificity, high cost, professional operation and poor patient compliance of the existing oral cancer diagnosis strategies (such as regular intraoral examination, imaging examination, histopathological evaluation and biomarker detection, etc.), the present invention provides a mouthguard equipped with a liquid collection module and its application in oral cancer early warning, including a liquid collection module with a pitcher plant bionic microstructure, which can collect saliva, and dopamine is coated on the groove of the mouthguard for ultraviolet irradiation to form a polydopamine layer, and then the polydopamine layer is used to adsorb the capture antibody to capture TNF-α in the saliva of oral cancer patients, and then the chitosan modified aggregation-induced luminescent nano dye modified with the detection antibody is combined with the captured TNF-α, and if it shows fluorescence, it indicates that the person to be tested has a risk of oral cancer. It is gratifying that the minimum detection limit of TNF-α detected by the present invention is 19.5pg / mL, which well covers the concentration range of TNF-α in the saliva of oral cancer patients, and improves its clinical application potential in oral cancer early warning.

[0005] According to a first aspect of the present invention, there is provided a mouthguard equipped with a liquid collection module, comprising a mouth guard and a liquid collection module; the liquid collection module is in an arch bridge shape, the two ends of the arch bridge structure of the liquid collection module are a supply side and a receiving side respectively, and the surface of the liquid collection module is a uniformly arranged protrusion;

[0006] The bottom of the inner side of the dental brace is connected with a groove; the liquid collection module is bonded into the groove with the supply side facing downward and the receiving side facing upward.

[0007] Preferably, the groove has a polydopamine layer, and the polydopamine layer is used to adsorb a capture antibody capable of capturing tumor necrosis factor TNF-α.

[0008] Preferably, the number of the liquid collection modules is 1-10.

[0009] Preferably, when there are a plurality of liquid collection modules, each of the liquid collection modules is bonded side by side into the groove.

[0010] According to another aspect of the present invention, there is provided a method for preparing a mouthguard equipped with a liquid collection module as described in any one of the preceding claims, comprising the following steps:

[0011] S1: Designing an arch-bridge-shaped liquid collection module (2) having a pitcher plant bionic microstructure, and printing it on a 3D printer using a washable resin; the two ends of the arch-bridge structure of the liquid collection module (2) are a supply side (3) and a receiving side (4), respectively, and the liquid collection module (2) includes evenly arranged protrusions (5);

[0012] S2: Design and print the dental brace resin master mold, make and assemble the inner and outer molds of the polyvinyl chloride dental brace mold to form a cavity to accommodate the model, and then fill the cavity with polydimethylsiloxane, which is cross-linked to make a polydimethylsiloxane dental brace with grooves;

[0013] S3: The liquid collection module obtained in step S1 is bonded into the groove of the polydimethylsiloxane brace prepared in step S2 with the supply side facing downward and the receiving side facing upward, so as to obtain the mouthguard equipped with the liquid collection module.

[0014] Preferably, after step S3, the method further includes coating dopamine on the groove, and then subjecting the groove to ultraviolet irradiation so that the dopamine self-assembles to form a polydopamine layer; and then utilizing the polydopamine layer to adsorb a capture antibody capable of capturing tumor necrosis factor TNF-α.

[0015] According to another aspect of the present invention, there is provided a use of any one of the above-mentioned mouthguards equipped with a liquid collection module for preparing an oral cancer early warning device.

[0016] Preferably, the application is specifically as follows: dopamine is coated on the groove of the mouthguard and subjected to ultraviolet irradiation to form a polydopamine layer; then, a capture antibody capable of capturing tumor necrosis factor TNF-α is adsorbed using the formed polydopamine layer or directly using the polydopamine layer in the groove to capture the tumor necrosis factor TNF-α in saliva; then, an aggregation-induced luminescence nano-dye modified with chitosan modified with a detection antibody is combined with the captured tumor necrosis factor TNF-α, and if fluorescence is presented, it indicates that the subject to be tested suffers from oral cancer; if no fluorescence is presented, it indicates that the subject to be tested is not an oral cancer patient.

[0017] Preferably, the preparation of the chitosan-modified aggregation-induced luminescence nano dye modified with the detection antibody is specifically as follows:

[0018] (1) dissolving chitosan in an acetic acid solution, dissolving sodium tripolyphosphate in distilled water, and dissolving 1,2-di(4-carboxyphenyl)-1,2-diphenylethylene in a dimethyl sulfoxide solution to form a stable yellow solution;

[0019] (2) adding catalysts 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide to the yellow solution, then adding the solution to the chitosan solution, and then adding sodium tripolyphosphate dropwise to prepare chitosan-modified aggregation-induced luminescence nanoparticle dye; adding a detection antibody to the chitosan-modified aggregation-induced luminescence nanoparticle dye, wherein the detection antibody is an antibody capable of detecting tumor necrosis factor TNF-α, and vortexing and incubating the mixture with the 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide solution to obtain a chitosan-modified aggregation-induced luminescence nanoparticle dye labeled with the detection antibody.

[0020] Preferably, in step (2), the concentration of the chitosan solution is 5-20 mg / mL; and the concentration of the detection antibody is 10-100 mg / mL.

[0021] In general, the above technical solution conceived by the present invention has the following technical advantages compared with the prior art:

[0022] (1) Inspired by the special water collection structure of the lip of Nepenthes, the present invention designs an innovative early oral cancer warning mouthguard device that can automatically collect saliva. As an intraoral system, it is combined with a wearable mouthguard to detect TNF-α in saliva in a persistent, in situ, specific and sensitive manner to timely warn of early oral cancer. The present invention adopts an efficient liquid collection module that can collect saliva into the groove of the mouthguard in a complex oral environment. Furthermore, TNF-α in saliva is adsorbed by a specific capture antibody fixed on the groove of the mouthguard. Subsequently, the chitosan-modified aggregation-induced luminescence nanodye modified with the detection antibody binds to the captured TNF-α. After the reaction is completed, the liquid collection module can be removed in order to more conveniently observe the detection results of the mouthguard. Finally, under the irradiation of ultraviolet light, oral cancer is predicted by whether it fluoresces or not. The invention has demonstrated excellent capabilities in detecting target biomarkers and protecting dental health in a simulated intraoral environment, and has been proven to be an ideal candidate product for oral cancer warning, with the potential for clinical transformation and improved patient care experience.

[0023] (2) The present invention has a simple design and is easy to operate, making it particularly suitable for home use. Compared with traditional oral cancer early warning methods (such as oral tissue biopsy and magnetic resonance imaging), the present invention does not rely on complex laboratory equipment or expensive medical instruments, which makes the entire detection process more efficient and easier to implement. Patients only need to wear the mouthguard to automatically collect saliva samples, eliminating the need for cumbersome sampling and processing steps. At the same time, the test results only require a household ultraviolet lamp to activate, reducing the economic burden and improving the accessibility of testing. This is especially important for patients living in remote areas or lacking medical resources, allowing them to more easily monitor their oral health.

[0024] (3) The present invention has high specificity and strong sensitivity. The present invention adopts a highly targeted biomarker detection method, which can still accurately identify specific molecules (TNF-α) related to oral cancer under the interference of other biological factors, demonstrating strong detection potential in the complex saliva environment of the oral cavity. The present invention can effectively detect the target biomarker (TNF-α) at ​​extremely low concentrations (minimum detection limit = 19.5pg / mL), showing its excellent sensitivity. This enables the present invention to capture early lesions of oral cancer in a timely manner, thereby providing patients with timely early warning information and improving the success rate of treatment and patient survival rate. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 .Schematic diagram of the structure of the present invention, wherein: 1-braces, 2-liquid collection module, 3-supply side, 4-receiving side, 5-protrusion, 6-groove.

[0026] Figure 2 .Photo of the directional drainage of liquid by the lip structure of the pitcher plant (a), transmission electron microscope photo of the lip structure of the pitcher plant (b), schematic diagram of the production of the liquid collection module (c), actual picture of the liquid collection module (bionic Nepenthes lip structure) printed with polylactic acid (PLA) as the material (d), schematic diagram of the liquid collection module for directional collection of liquid (e).

[0027] Figure 3 .Screenshot of the video showing the liquid collection module directing the drainage of liquid.

[0028] Figure 4 .A real picture of a mouthguard with grooves made of PDMS (a), a schematic diagram of a side cut (b), and a real picture of a side cut (c). In (b, c), the blue dotted line represents the groove and the red dotted line represents the outline of the mouthguard.

[0029] Figure 5 .The overall physical picture of the product of the present invention (i), the partial enlarged physical picture from the front view (ii) and the partial enlarged physical picture from the rear view (iii).

[0030] Figure 6 . Schematic diagram of the process of forming polydopamine in the groove of the product of the present invention to adsorb the capture antibody (a) and a scanning electron microscope photo of the polydopamine layer adsorbed in the groove (b).

[0031] Figure 7 Transmission electron microscopy photograph (a) and particle size analysis (b) of the chitosan-modified aggregation-induced luminescence nanoparticles of the present invention, as well as fluorescence photograph (c) and fluorescence intensity statistics (d) of the solvent under ultraviolet light irradiation in water / dimethyl sulfoxide solvent.

[0032] Figure 8. Schematic diagram of the process of detecting TNF-α derived from oral tongue squamous cell carcinoma cells by the product of the present invention (a) and the fluorescent photograph (b) and fluorescence intensity data statistics (c) of the final detection result; (d) The statistical picture of the fluorescence intensity of the standard samples bovine serum albumin (125pg / mL) and TNF-α (125pg / mL) detected by the product of the present invention; (e) The fluorescence intensity statistics of the TNF-α standard samples of different concentrations (0pg / mL, 5pg / mL, 25pg / mL, 50pg / mL, 250pg / mL and 500pg / mL) detected by the product of the present invention, and the minimum detection limit of 19.5pg / mL (the minimum detection limit is defined as the mean value of the blank sample + 3 standard deviations); (f) A photo of a healthy adult volunteer wearing the product of the present invention (i) and the fluorescent photograph (ii) and statistical data (iii) of the TNF-α standard samples (0pg / mL, 25pg / mL and 250pg / mL) dropped from the outside into the mouth of a healthy adult volunteer by the product of the present invention. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0034] Inspired by the water collection structure of the lip of Nepenthes, the present invention designs an early oral cancer warning mouthguard device equipped with a liquid collection module; as an intraoral system, it can automatically collect saliva, and detect tumor necrosis factor α (TNF-α) in saliva persistently, in situ, specifically and sensitively to timely warn of early oral cancer. The mouthguard of the present invention includes a brace 1 and a liquid collection module 2; the liquid collection module 2 has a bionic microstructure of Nepenthes, the liquid collection module 2 is in the shape of an arch bridge, and the liquid collection module 2 contains a supply side 3 and a receiving side 4, that is, the liquid collection module 2 includes evenly arranged protrusions 5; the bottom of the inner side of the brace 1 is connected to a groove 6; the liquid collection module 2 is bonded to the groove 6 with the supply side 3 facing downward and the receiving side 4 facing upward ( Figure 1 ).

[0035] The method for preparing a mouthguard device for early oral cancer warning of the present invention comprises the following steps:

[0036] (1) Preparation of liquid collection module: A bridge-shaped liquid collection module with a pitcher plant bionic microstructure was designed using Solidworks software and printed on a 3D printer using a washable resin (ultraviolet wavelength 365-405 nm) ( Figure 2 The module's liquid collection function is mainly due to the protrusions evenly distributed on the surface (imitating the tiny protrusions on the lip structure of the pitcher plant) ( Figure 2 These protrusions together form a synergistic functional interface that divides the liquid into tiny droplets and, by changing the balance between the liquid surface tension and gravity, causes the liquid to move in a directional manner for a specific distance (h), draining from the supply side to the receiving side ( Figure 3 ). Specifically, the equilibrium state of the liquid can be described by the following formula: 2γπrcosβ=ρπr2hg. Further, the height of the liquid rise is inferred: h=2γcosβ / ρgr. Among them, γ, ρ, β, g, h and r represent the surface tension coefficient, liquid density, liquid-solid interface contact angle, gravity constant, liquid rise height and liquid contact radius on the solid surface respectively.

[0037] (2) Preparation of the mouthguard: Based on the mouthguard resin master mold designed by Solidworks and 3D printed, two polyvinyl chloride (PVC) mouthguard molds (inner mold and outer mold) were made and assembled to form a cavity to accommodate the model. Then, polydimethylsiloxane (PDMS) was filled in the cavity and cross-linked to form a PDMS mouthguard with a semi-cylindrical groove in the middle ( Figure 4 ). In order to achieve continuous interaction with saliva in situ, the liquid collection module is integrated into the semi-cylindrical groove of the intraoral system (homemade mouthguard) to achieve persistent accumulation of saliva and subsequent biochemical reactions. This design maximizes the drainage function of the liquid collection module by closing the lower gum where saliva spontaneously accumulates, thereby collecting liquid conveniently and efficiently. In addition, the mouthguard conforms to the shape of the teeth and has minimal incision, which has been proven to be highly operable and feasible in the in situ detection of oral cancer.

[0038] (3) Integrating the liquid collection module into the mouthguard: In order to achieve continuous interaction between the liquid collection module and saliva, multiple liquid collection modules are arranged in parallel to form a large liquid collection device, and bonded to the groove of the PDMS mouthguard in a specific direction (supply side facing down, receiving side facing up) (using 706 Kraft glue as adhesive) to form an early oral cancer warning intelligent mouthguard system ( Figure 5 ).

[0039] (4) Using a smart mouthguard system to warn of early oral cancer: dopamine is coated on the groove of the mouthguard and irradiated with ultraviolet light to form a polydopamine layer ( Figure 6 ); then the polydopamine layer is used to adsorb the capture antibody to capture TNF-α in the saliva of oral cancer patients; then, the chitosan modified aggregation-induced luminescence nanodye ( Figure 7) binds to the captured TNF-α; after the reaction is completed, in order to more conveniently and clearly observe the test results of the mouthguard, the liquid collection module is removed; finally, under ultraviolet light, observe whether the mouthguard exhibits fluorescence ( Figure 8 ).

[0040] In some embodiments, in the design of the liquid collection module in step (1), the liquid collection module has a width of 1-5 mm, a length of 1-9 mm, and a bending radius of 1-10 mm.

[0041] In some embodiments, the liquid collection module in step (1) has 10-100 protrusions evenly distributed on a surface adapted to the curvature, and each protrusion has a width of about 100-1000 microns, a length of about 100-800 microns, and a height of about 400-800 microns.

[0042] In some embodiments, the number of liquid collection modules in the liquid collection module design in step (3) is 1-10.

[0043] In some embodiments, the mouthguard in step (2) is designed to have a height of 1-10 cm, a width of 1-10 cm, and a thickness of 1-8 mm.

[0044] In some embodiments, in the design of the mouth guard described in step (2), the length of the semi-cylindrical groove on the mouth guard is 1-5 cm and the radius is 1-8 mm.

[0045] In some embodiments, the dopamine concentration in step (4) is 10-50 mg / ml.

[0046] In some embodiments, the ultraviolet irradiation time of dopamine in step (4) is 10-30 minutes.

[0047] In some embodiments, the concentration of chitosan in step (4) is 5-20 mg / ml.

[0048] In some embodiments, the concentration of the aggregation-induced emission dye in step (4) is 2-20 mg / ml.

[0049] In some embodiments, the concentration of the detection antibody in step (4) is 10-100 μg / ml.

[0050] In some embodiments, the concentration of the capture antibody in step (4) is 10-100 μg / ml.

[0051] In some embodiments, the chitosan-modified aggregation-induced luminescence nano dye in step (4) has an average particle size of 10-300 nanometers.

[0052] In some embodiments, the monitoring of TNF-α in step (4) can be achieved by a simple household ultraviolet lamp.

[0053] The early oral cancer warning mouthguard device capable of automatically collecting saliva is used in monitoring oral health, especially in monitoring the development of early oral cancer.

[0054] The following are specific embodiments

[0055] Example 1: Preparation of an early oral cancer warning intelligent mouthguard

[0056] The liquid collection module and the resin mouthguard master mold with grooves were designed using Solidworks software and printed out with white resin on a 3D printer. The PVC film covering the resin master mold was reshaped using a vacuum heating system consisting of a hollow iron frame, a hot solvent gun, and a circulating water vacuum pump to obtain two PVC mouthguard molds (inner mold and outer mold). PDMS was filled into the cavity formed after the two PVC mouthguard molds were assembled, and then peeled off after curing to obtain the PDMS mouthguard. The PDMS solution consists of two parts: vinyl (Part A) and hydroxyl (Part B). Subsequently, the liquid collection module was integrated into the PDMS mouthguard using an adhesive to obtain an early oral cancer warning intelligent mouthguard system that can automatically collect liquid.

[0057] Example 2: Synthesis of chitosan-modified aggregation-induced luminescence nanoparticle dyes

[0058] Chitosan was dissolved in an acetic acid solution to adjust the pH; sodium tripolyphosphate was dissolved in distilled water to adjust the pH. 1,2-di(4-carboxyphenyl)-1,2-diphenylethylene (DCPD) was dissolved in a DMSO solution to form a stable yellow solution. Then, EDC / NHS was added to the DCPD solution. The DCPD solution was added to the chitosan solution and stirred, and sodium tripolyphosphate was added dropwise and stirred to prepare a chitosan-modified aggregation-induced luminescence nanoparticle dye. Then, centrifuged in a low-temperature centrifuge and the supernatant was removed. Finally, it was freeze-dried in a vacuum freeze dryer, weighed and stored. Then, the detection antibody was incorporated into the aggregation-induced luminescence nanoparticle dye and vortexed and incubated with EDC / NHS for a period of time. Then, the excess EDC / NHS was removed by centrifugation in a low-temperature centrifuge, and the mixture was resuspended in PBS buffer to obtain the aggregation-induced luminescence nanoparticle dye labeled with the detection antibody.

[0059] Example 3: Early warning of oral cancer using a smart mouthguard system

[0060] The specificity of the system was verified by using the smart mouthguard system to detect saliva containing standard bovine serum albumin and TNF-α. In addition, the detection limit of the system was determined by detecting a series of TNF-α standards with different concentrations using the smart mouthguard system. Further, CAL-27 cells were inoculated into a cell culture dish to construct an in vitro oral cancer cell model. Dopamine was coated on the groove of the mouthguard and irradiated with ultraviolet light to form a polydopamine layer. Then, the polydopamine layer was used to adsorb specific antibodies that captured TNF-α using the characteristics of adsorbing biomacromolecules. It was sealed with ELISPOT diulent and then rinsed with PBS buffer. Then, the mouthguard was placed in a cell culture dish and incubated for a period of time at different culture times to collect TNF-α in the supernatant. Then, the groove part of the mouthguard was immersed in the aggregation-induced luminescence nanoparticle dye labeled with the detection antibody and then rinsed with PBS buffer. After that, the liquid collection module was separated from the mouthguard for easy observation. Finally, use ultraviolet light to illuminate the mouth guard to observe whether there is fluorescence in the groove. If the result is fluorescence, it indicates that there is a certain amount of TNF-α in the mouth, which indicates the risk of oral cancer.

[0061] It will be easily understood by those skilled in the art that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A mouthguard equipped with a liquid collection module, characterized in that: It comprises a dental brace (1) and a liquid collection module (2); the liquid collection module (2) is in the shape of an arch bridge, the two ends of the arch bridge structure of the liquid collection module (2) are respectively a supply side (3) and a receiving side (4), and the surface of the liquid collection module (2) is provided with evenly arranged protrusions (5); The bottom of the inner side of the dental brace (1) is connected with a groove (6); the supply side (3) of the liquid collection module (2) is bonded into the groove (6) with the supply side (3) facing downward and the receiving side (4) facing upward.

2. The mouthguard equipped with a liquid collection module according to claim 1, characterized in that: The groove (6) has a polydopamine layer therein, and the polydopamine layer is used to adsorb a capture antibody capable of capturing tumor necrosis factor TNF-α.

3. The mouthguard equipped with a liquid collection module according to claim 1, characterized in that: The number of the liquid collection modules (2) is 1-10.

4. The mouthguard equipped with a liquid collection module according to claim 3, characterized in that: When there are multiple liquid collection modules (2), each of the liquid collection modules (2) is bonded side by side into the groove (6).

5. The method for preparing a mouthguard equipped with a liquid collection module according to any one of claims 1 to 4, characterized in that: The following steps are involved: S1: Designing an arch-bridge-shaped liquid collection module (2) having a pitcher plant bionic microstructure, and printing it on a 3D printer using a washable resin; the two ends of the arch-bridge structure of the liquid collection module (2) are a supply side (3) and a receiving side (4), respectively, and the liquid collection module (2) includes evenly arranged protrusions (5); S2: designing and printing a dental brace resin master mold, making and assembling an inner mold and an outer mold of a polyvinyl chloride dental brace mold to form a cavity for accommodating the model, and then filling the cavity with polydimethylsiloxane, which is cross-linked to form a polydimethylsiloxane dental brace with a groove (1); S3: The liquid collection module (2) obtained in step S1 is bonded into the groove of the polydimethylsiloxane brace prepared in step S2 with the supply side (3) facing downward and the receiving side (4) facing upward, thereby obtaining the mouthguard equipped with the liquid collection module.

6. The method for preparing a mouthguard equipped with a liquid collection module according to claim 5, characterized in that: After step S3, the method further includes coating dopamine on the groove, and then irradiating with ultraviolet light to allow the dopamine to self-assemble into a polydopamine layer; and then using the polydopamine layer to adsorb a capture antibody capable of capturing tumor necrosis factor TNF-α.

7. Use of the mouthguard equipped with a liquid collection module as described in any one of claims 1 to 4 for preparing an oral cancer early warning device.

8. The use according to claim 7, characterized in that The application is specifically as follows: dopamine is coated on the groove of the mouthguard and subjected to ultraviolet irradiation to form a polydopamine layer; then, a capture antibody capable of capturing tumor necrosis factor TNF-α is adsorbed using the formed polydopamine layer or directly using the polydopamine layer in the groove to capture the tumor necrosis factor TNF-α in saliva; then, an aggregation-induced luminescent nano-dye modified with chitosan modified with a detection antibody is combined with the captured tumor necrosis factor TNF-α, and if fluorescence is presented, it indicates that the subject to be tested suffers from oral cancer; if no fluorescence is presented, it indicates that the subject to be tested is not an oral cancer patient.

9. The use according to claim 8, characterized in that The preparation of the chitosan-modified aggregation-induced luminescence nano dye modified with the detection antibody is specifically as follows: (1) dissolving chitosan in an acetic acid solution, dissolving sodium tripolyphosphate in distilled water, and dissolving 1,2-di(4-carboxyphenyl)-1,2-diphenylethylene in a dimethyl sulfoxide solution to form a stable yellow solution; (2) adding catalysts 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide to the yellow solution, then adding the solution to the chitosan solution, and then adding sodium tripolyphosphate dropwise to prepare chitosan-modified aggregation-induced luminescence nanoparticle dye; adding a detection antibody to the chitosan-modified aggregation-induced luminescence nanoparticle dye, wherein the detection antibody is an antibody capable of detecting tumor necrosis factor TNF-α, and vortexing and incubating the mixture with the 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide solution to obtain a chitosan-modified aggregation-induced luminescence nanoparticle dye labeled with the detection antibody.

10. The use according to claim 9, characterized in that In step (2), the concentration of the chitosan solution is 5-20 mg / mL; the concentration of the detection antibody is 10-100 mg / mL.

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