Wearable oral diagnostic and treatment device and manufacturing method thereof

Abstract

The present invention is applicable to the technical field of oral diagnosis and treatment equipment, and provides a wearable oral diagnosis and treatment device and a manufacturing method thereof. The device integrates a flexible electrochemical biosensor through a dental brace, which can realize continuous perception of oral microenvironment biomarkers and provide a data basis for early diagnosis of diseases. The electroresponsive drug release structure it carries can explore the controlled release of antibacterial drugs. The flexible double-sided structure is adopted to meet the dual disease management while optimizing the electrode space and improving wearing comfort. By monitoring and analyzing oral health data, it is expected to help reduce the irrational use of antibiotics. At the same time, based on the collaborative model of "wearable device + terminal device", a preliminary framework for oral health data management has been constructed, providing a new technical path for the comprehensive prevention and treatment of chronic oral diseases.

Description

Technical Field

[0001] The present invention belongs to the technical field of oral diagnosis and treatment equipment, and in particular relates to a wearable oral diagnosis and treatment equipment and a manufacturing method thereof. Background Art

[0002] Chronic oral diseases (such as dental caries and periodontitis) are a persistent challenge in global public health. Their impact goes beyond the local oral cavity and is closely related to the occurrence and development of systemic diseases such as sepsis, Alzheimer's disease, and rheumatoid arthritis. However, traditional diagnostic methods rely on visual inspection or imaging assessment, which can only be detected in the middle and late stages of the disease, when irreversible tissue damage has already occurred. On the other hand, the main treatment strategy is still centered on "post-intervention", and there are significant technical shortcomings in early identification, precise treatment, and dynamic management. Overall, these challenges highlight the urgent need for innovative solutions, such as wearable platforms that combine continuous biomarker monitoring with on-demand treatment capabilities to achieve personalized treatment and closed-loop management of chronic oral diseases.

[0003] At present, the dilemma in the diagnosis and treatment of chronic oral diseases lies in the fragmentation of the diagnosis and treatment system: at the diagnostic level, early diagnosis is delayed and dynamic monitoring capabilities are insufficient; in terms of treatment, the model is passive and single, and the problem of drug abuse is prominent; in equipment application, monitoring and treatment functions are independent of each other, and a closed-loop management system cannot be formed, making it difficult to meet the needs of precision medicine. Summary of the Invention

[0004] The purpose of the embodiments of the present invention is to provide a wearable oral diagnostic and treatment device and a manufacturing method thereof, aiming to solve the problems raised in the above-mentioned background technology.

[0005] The embodiment of the present invention is implemented as follows: an oral wearable diagnostic and treatment device includes: a flexible double-sided electrode array and a near-field communication module;

[0006] The flexible double-sided electrode array includes a base layer, an upper serpentine interconnection wire layer is provided on the upper surface of the base layer, one end of the upper serpentine interconnection wire layer is connected to a via electrode interface provided on the lower surface of the base layer through an electrode via provided on the base layer, an upper insulating layer is provided at the end of the upper serpentine interconnection wire layer, an upper conductive ink layer is provided on the upper insulating layer, and the upper conductive ink layer includes a working electrode W pH , working electrode W cT , reference electrode R pH and reference electrode R cT , and the working electrode W pH , working electrode W cT , reference electrode R pH and reference electrode R cTThe lower surface of the base layer is provided with a lower serpentine interconnection wire layer, the end of the lower serpentine interconnection wire layer is provided with a lower insulating layer, the lower insulating layer is provided with a lower conductive ink layer, and the lower conductive ink layer includes a counter electrode C RNS , working electrode W RNS , reference electrode R RNS and working electrode W pT , and the counter electrode C RNS , working electrode W RNS , reference electrode R RNS and working electrode W pT are respectively connected to a serpentine wire in the lower serpentine interconnection wire layer;

[0007] The front of the near-field communication module is provided with a near-field communication chip and a single-chip microcomputer chip, and the near-field communication chip is connected to the near-field communication coil. The back of the near-field communication module is provided with an electrode interface for matching the upper serpentine interconnection wire layer and the lower serpentine interconnection wire layer.

[0008] A further technical solution is that the working electrode W pH The reference electrode R pH The working electrode W is modified with a PVB solid composite electrolyte layer; cT It is modified with a Ppy / pTs drug-loaded electrode layer.

[0009] A further technical solution is that the working electrode W RNS The working electrode W pT It is also modified with a Ppy / pTs drug-loaded electrode layer.

[0010] Another object of an embodiment of the present invention is to provide a method for preparing a wearable oral diagnostic and therapeutic device, based on the above-mentioned wearable oral diagnostic and therapeutic device, comprising the following steps:

[0011] Step 1: Design and fabrication of flexible double-sided electrode arrays:

[0012] The flexible double-sided electrode array is fabricated using computer-aided design and flexible printed circuit board (FPCB) technology. The upper and lower conductive ink layers are formed by screen-printing conductive carbon and Ag / AgCl inks. Using high-precision FPCB technology, the upper and lower serpentine interconnect layers are formed on the substrate by laser processing. The upper serpentine interconnect layer is then connected to a via-electrode interface on one side of the lower serpentine interconnect layer.

[0013] Step 2: Electrode functionalization modification:

[0014] At the working electrode WpH Iridium oxide nanotubes (IrOx NTs) layer was deposited on the surface; pH The surface is coated with a polyvinyl butyral (PVB) solid composite electrolyte layer; RNS The gold nanoparticles (Au NPs) layer was deposited on the interface by chronoamperometry; cT and working electrode W pT Surface modified Ppy / pTs drug-loaded electrode layer;

[0015] Step 3: System Integration:

[0016] The pins of the upper serpentine interconnect wire layer and the lower serpentine interconnect wire layer are respectively connected to the electrode interface, packaged, and integrated with the customized transparent braces to complete the preparation.

[0017] In a further technical solution, the IrOx NTs layer is prepared by an optimized electrospinning method:

[0018] 0.25 g hydrated iridium trichloride, 2 mL N, N-dimethylformamide solution, and 0.4 g polyvinyl pyrrolidone were heated in a water bath to prepare a precursor solution. After electrospinning, the solution was annealed at 500 °C for 3 h. The solution was evenly dispersed in deionized water to a concentration of 5 mg / mL and 5 μL was accurately drop-coated on the pretreated working electrode W. pH The surface is then shaped by infrared-assisted drying.

[0019] A further technical solution is to prepare the PVB solid composite electrolyte layer as follows:

[0020] 78 mg of PVB and 50 mg of NaCl were dissolved in 1 mL of methanol solution and ultrasonically treated to form a solid composite electrolyte layer. Then 3 μL of the PVB mixture was added dropwise to the reference electrode R pH and dried at room temperature overnight.

[0021] A further technical solution is to prepare the Au nanoparticle layer as follows:

[0022] The pretreated working electrode W RNS The samples were immersed in a 2 mM HAuCl4 solution in 0.1 M phosphate buffer (PBS) and a constant potential of -0.2 V vs. Ag / AgCl was applied for 120 s.

[0023] Further technical solutions, the preparation of the Ppy / pTs drug-loaded electrode layer is as follows:

[0024] First, in a mixed solution of 0.2 M pyrrole and 0.05 M pTs, 2 mA / cm -2The constant current was applied for 10 min to achieve electrochemical deposition of Ppy thin films;

[0025] The electrodes were then immersed in the drug solution, and a potential of -0.6 V relative to Ag / AgCl was applied thereto for 0-40 minutes.

[0026] An embodiment of the present invention provides an oral wearable diagnostic and treatment device and a manufacturing method thereof. The device integrates a flexible electrochemical biosensor through a dental brace, which can achieve continuous perception of oral microenvironment biomarkers, provide a data basis for early diagnosis of diseases, and to a certain extent make up for the shortcomings of traditional detection methods in capturing early lesion signals. The electrically responsive drug release structure it carries can explore the controlled release of antibacterial drugs. In terms of structural design, a flexible double-sided structure is adopted to meet the dual disease management while optimizing the electrode space and improving wearing comfort. By monitoring and analyzing oral health data, it is expected to help reduce the irrational use of antibiotics, and thus help control the risk of bacterial resistance. At the same time, based on the collaborative model of "wearable device + terminal device", a preliminary framework for oral health data management has been constructed, providing a new technical path for the comprehensive prevention and treatment of chronic oral diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 A schematic diagram of the structure of a flexible double-sided electrode array in a wearable oral diagnostic and treatment device provided by an embodiment of the present invention;

[0028] Figure 2 A schematic diagram of the structure of a near-field communication module in a wearable oral diagnostic and treatment device provided by an embodiment of the present invention (where a is the front side and b is the back side);

[0029] Figure 3 A diagram showing the wearing effect of a wearable oral diagnostic and treatment device provided by an embodiment of the present invention;

[0030] Figure 4 The potential response and fitting curve of the flexible double-sided electrode array for pH detection (a is the potential response and b is the fitting curve);

[0031] Figure 5 This is a test diagram of pH detection selectivity of flexible double-sided electrode array;

[0032] Figure 6 This is a graph showing the reproducibility of pH detection using a flexible double-sided electrode array (a is the potential response graph of multiple electrodes, and b is the corresponding potential-pH fitting curve).

[0033] Figure 7 This is a test diagram of the functional modification time of the flexible double-sided electrode array RNS electrode;

[0034] Figure 8 The test diagram for the optimal working voltage of the flexible double-sided electrode array RNS (where a is The current response diagram between 50-650 μM, b is the corresponding concentration and current response fitting curve);

[0035] Figure 9 The amperometric current response and fitting curve of the flexible double-sided electrode array RNS (where a is The current response diagram between 0-30 mM, b is the corresponding concentration and current response fitting curve);

[0036] Figure 10 Selective test chart for RNS detection;

[0037] Figure 11 This is a data graph of pH value monitored in an in vitro simulated caries environment;

[0038] Figure 12 Simulating periodontitis microenvironment monitoring for RAW 264.7 macrophages Data graphs from sensors;

[0039] Figure 13 This is a diagram showing the drug release effect in response to electrical stimulation;

[0040] Figure 14 Figure 2 is the statistical result of the antibacterial CFU of S. mutans and P. gingivalis (where a is the CFU count result of S. mutans biofilm formation under different treatment conditions, and b is the CFU count result of P. gingivalis biofilm formation under different treatment conditions);

[0041] Figure 15 SEM images of bacteria impact;

[0042] Figure 16 CLSM images of bacterial effects;

[0043] Figure 17 These are images of rat histological sections.

[0044] In the figure: upper conductive ink layer 1; working electrode W cT 1-1; Reference electrode R cT 1-2; Reference electrode R pH 1-3; working electrode W pH 1-4; upper insulating layer 2; upper serpentine interconnection wire layer 3; base layer 4; electrode via 4-1; lower serpentine interconnection wire layer 5; via electrode interface 5-1; lower insulating layer 6; lower conductive ink layer 7; counter electrode C RNS 7-1; working electrode WpT 7-2; Reference electrode R RNS 7-3; working electrode W RNS 7-4; near-field communication chip 8; single-chip microcomputer chip 9; near-field communication coil 10; electrode interface 11. DETAILED DESCRIPTION

[0045] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention will be further described in detail below with reference to 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.

[0046] The specific implementation of the present invention is described in detail below with reference to specific embodiments.

[0047] like Figure 1 and Figure 2 (in Figure 2 a is the front view, Figure 2 b is the reverse side), which shows an oral wearable diagnostic and treatment device provided by one embodiment of the present invention, comprising: a flexible double-sided electrode array and a near-field communication module;

[0048] The flexible double-sided electrode array includes a base layer 4, an upper serpentine interconnection wire layer 3 is provided on the upper surface of the base layer 4, one end of the upper serpentine interconnection wire layer 3 is connected to a via electrode interface 5-1 provided on the lower surface of the base layer 4 through an electrode via 4-1 provided on the base layer 4, an upper insulating layer 2 is provided at the end of the upper serpentine interconnection wire layer 3, an upper conductive ink layer 1 is provided on the upper insulating layer 2, and the upper conductive ink layer 1 includes a working electrode W cT 1-1. Working electrode W pH 1-4. Reference electrode R cT 1-2 and reference electrode R pH 1-3, and the working electrode W cT 1-1. Working electrode W pH 1-4. Reference electrode R cT 1-2 and reference electrode R pH 1-3 are respectively connected to a serpentine wire in the upper serpentine interconnection wire layer 3; the lower surface of the base layer 4 is provided with a lower serpentine interconnection wire layer 5, the end of the lower serpentine interconnection wire layer 5 is provided with a lower insulating layer 6, and a lower conductive ink layer 7 is provided on the lower insulating layer 6, and the lower conductive ink layer 7 includes a counter electrode C RNS 7-1. Working electrode W pT 7-2. Reference electrode R RNS 7-3 and working electrode W RNS 7-4, and the counter electrode C RNS 7-1. Working electrode W pT 7-2. Reference electrode RRNS 7-3 and working electrode W RNS 7-4 is respectively connected to a serpentine wire in the lower serpentine interconnect wire layer 5;

[0049] The front of the near-field communication module is provided with a near-field communication chip 8 and a single-chip microcomputer chip 9. The near-field communication chip 8 is connected to the near-field communication coil 10. The back of the near-field communication module is provided with an electrode interface 11 for matching the upper serpentine interconnection wire layer 3 and the lower serpentine interconnection wire layer 5.

[0050] As a preferred embodiment of the present invention, the working electrode W pH 1-4 is modified with an IrOx NTs layer; the reference electrode R pH 1-3 is modified with a PVB solid composite electrolyte layer; the working electrode W cT 1-1 is modified with a Ppy / pTs drug-loaded electrode layer.

[0051] As a preferred embodiment of the present invention, the working electrode W RNS 7-4 is modified with an Au nanoparticle layer; the working electrode W pT 7-2 is also modified with a Ppy / pTs drug-loaded electrode layer.

[0052] An embodiment of the present invention provides a method for preparing a wearable oral diagnostic and therapeutic device, based on the above-mentioned wearable oral diagnostic and therapeutic device, comprising the following steps:

[0053] Step 1: Design and fabrication of flexible double-sided electrode arrays:

[0054] The flexible double-sided electrode array was fabricated using flexible printed circuit (FPCB) technology after computer-aided design using 3ds Max 2020 software (Autodesk, USA). The upper conductive ink layer 1 and the lower conductive ink layer 7 are created by screen-printing conductive carbon and Ag / AgCl inks. The double-sided design conforms to oral anatomy and facilitates dual disease management. Using high-precision FPCB technology, the upper serpentine interconnect layer 3 and the lower serpentine interconnect layer 5 are formed on the base layer 4 by laser processing. The upper serpentine interconnect layer 3 is connected to the via electrode interface 5-1 on one side of the lower serpentine interconnect layer 5. This geometrically optimized routing design significantly enhances the mechanical flexibility of the electrode. The optimized wiring layout concentrates the pin connections on one side of the base layer 4, ensuring a stable connection to the near-field communication module (i.e., the control circuit).

[0055] Step 2: Electrode functionalization modification:

[0056] By using the working electrode W pH1-4 Surface-deposited iridium oxide nanotubes (IrOx NTs) were prepared as the H⁺ sensitive layer. IrOx NTs were prepared using an optimized electrospinning method to enhance their structural stability and electrochemical performance. Reference electrode R pH 1-3 was prepared by coating a polyvinyl butyral (PVB) solid composite electrolyte layer on the electrode surface to stabilize the Cl - Concentration. Working electrode W RNS The 7-4 interface was constructed by chronoamperometric electrodeposition of gold nanoparticles (Au NPs), thereby improving the electrocatalytic activity and sensing sensitivity of the electrode to nitrite. cT 1-1 and working electrode W pT 7-2 is prepared using a two-step process. First, a polypyrrole / p-toluenesulfonic acid (Ppy / pTs) thin film is deposited on the electrode surface via constant current electropolymerization. Subsequently, charged drugs are electrochemically introduced into the polymer network in a drug solution.

[0057] Step 3: System Integration:

[0058] The pins of the upper serpentine interconnection wire layer 3 and the lower serpentine interconnection wire layer 5 are respectively connected to the electrode interface 11, and after packaging, they are integrated with the customized transparent braces to complete the preparation. The assembled device is as follows Figure 3 shown.

[0059] As a preferred embodiment of the present invention, the IrOx NTs layer was prepared using an optimized electrospinning method: 0.25 g of hydrated iridium trichloride, 2 mL of N,N-dimethylformamide solution, and 0.4 g of polyvinyl pyrrolidone were heated in a water bath to prepare a precursor solution. After electrospinning, the solution was annealed at 500°C for 3 hours. The solution was evenly dispersed in deionized water to a concentration of 5 mg / mL and 5 μL was accurately drop-coated on the pretreated working electrode W. pH 1-4 surfaces, followed by infrared-assisted drying to ensure uniform film formation and improve its stability and adhesion.

[0060] As a preferred embodiment of the present invention, the preparation of the PVB solid composite electrolyte layer is as follows:

[0061] 78 mg of PVB and 50 mg of NaCl were dissolved in 1 mL of methanol solution and ultrasonicated to form a solid composite electrolyte layer. The PVB mixture (3 μL) was then added dropwise to the reference electrode R pH 1-3 and dried at room temperature overnight to ensure that the electrolyte is uniformly formed and has good stability.

[0062] As a preferred embodiment of the present invention, the preparation of the Au nanoparticle layer is as follows:

[0063] The pretreated working electrode W RNS 7-4 was immersed in a 2 mM chloroauric acid (HAuCl4) solution prepared in 0.1 M phosphate buffer (PBS) and a constant potential of -0.2 V (vs. Ag / AgCl) was applied for 120 s to achieve controllable reduction and uniform deposition of Au NPs, thereby improving the electrocatalytic activity and sensing sensitivity of the electrode to nitrite.

[0064] As a preferred embodiment of the present invention, the preparation of the Ppy / pTs drug-loaded electrode layer is as follows:

[0065] First, in a mixed solution of 0.2 M pyrrole and 0.05 M pTs, 2 mA / cm -2 A constant current was applied for 10 minutes to achieve electrochemical deposition of the Ppy film. The electrodes were then immersed in the drug solution and a potential of -0.6 V (vs. Ag / AgCl) was applied thereto for 0-40 minutes.

[0066] As a preferred embodiment of the present invention, a near-field communication module utilizes the principle of inductive coupling to enable bidirectional energy and data exchange between the biological microenvironment and electronic systems, supporting wireless energy capture, wireless data transmission, electrochemical signal processing, and electroresponsive drug release. The resonant frequency is approximately 13.56 MHz, and the wireless communication range is 0-15 mm.

[0067] As a preferred embodiment of the present invention, transparent braces are customized based on volunteers. First, the tooth morphology is accurately captured using a 3Shape scanning device. The data is exported in STL format and then a 3D printer is used to create a solid model. Then, a flexible double-sided electrode array and a near-field communication module are installed at the designated location. Then, a "tooth model after wearing" containing the module installation state is prepared again using 3Shape scanning and 3D printing technology. The brace body is then thermoformed with TPU material and trimmed to form the brace body. Finally, the brace and electronic module are integrated.

[0068] In the embodiment of the present invention, the finally prepared wearable oral diagnostic and treatment device can be used for the detection of the oral microenvironment and can also be used for the antibacterial treatment of caries and periodontitis.

[0069] To systematically verify the device's ability to monitor the oral microenvironment, its pH and RNS ( ), which are key biomarkers for the early detection of dental caries and periodontitis, respectively. After calibration with a Sartorius pH meter, 0.1 M HCl / NaOH buffer solutions with a pH range of 3-8 were prepared to simulate the dynamic acidification processes of tooth enamel during demineralization (pH < 5.5) and healthy conditions (pH ~ 7.0). The open-circuit potential (OCP) method was used to record the potential response of the pH sensor. Figure 4 (Where a is the potential response and b is the fitting curve) The open circuit potential response as the pH value changes is recorded. During the acidification process (pH=8-3), as the pH continues to decrease, the potential shows a step response, while during the alkalization process (pH=3-8), it shows a completely reversible and stable potential change. Figure 4 The calibration curve showed nearly ideal linearity (R²=0.9992), reflecting near-Nernstian performance. This may be attributed to the IrOx NTs modified on the working electrode surface, which can act as an ion-selective membrane in selective sensing on the one hand, and its high conductivity and high specific surface area on the other hand contribute to efficient signal extraction. Considering that other electrolytes and metabolites present in the oral microenvironment may have a negative impact on the sensor output, the selectivity of the sensor was tested. Specifically, an artificial saliva solution was prepared to simulate the interference environment under physiological conditions, which contained K + (40 mM), Na + (20 mM), Ca 2+ (3 mM), Mg 2+ (0.5 mM) and bovine serum albumin (BSA, 3 mg / mL). Figure 5 As shown in the results, the addition of non-target electrolytes and metabolites did not substantially affect the sensor performance. The sensor showed high specificity, which may be attributed to the specific hydrogen ion selection of IrOx, thereby minimizing the potential response of common interferents. To explore the reproducibility of the developed pH sensor, multiple measurements were performed using multiple independently prepared electrodes in the same buffer solution with a pH range of 3-8, showing similar step responses and slopes ( Figure 6 , where a is the potential response diagram of multiple electrodes and b is the corresponding potential-pH fitting curve diagram), which shows that the electrodes obtained from different batches have similar response trends and sensitivities.

[0070] RNS ) sensing electrode was subjected to a constant potential of -0.2 V vs. Ag / AgCl in a 2 mM HAuCl4 solution for 120 seconds by chronoamperometry to reduce and deposit gold nanoparticles. Cyclic voltammetry (CV) was used to investigate the relationship between the deposition time of gold nanoparticles (AuNPs) and RNS ( ) Structure-activity relationship of sensing performance. Figure 7 It shows that when the deposition time is 120 seconds, The peak current of oxidation reaches the highest, so the deposition time of 120 seconds is the optimal condition for preparing RNS-sensitive electrodes.

[0071] To determine RNS ( ) monitoring, and the amperometric response characteristics in the potential range of 0.75–0.95 V (vs. Ag / AgCl) were systematically studied in phosphate buffer saline (PBS, pH=7.4). Figure 8 (where a is The current response diagram between 50-650 μM, b is the corresponding The concentration and current response fitting curves show that at different potentials of 0.75-0.95V, the The chronoamperometric response and the corresponding calibration curve in the concentration range of 0-650 μM. In contrast, as the voltage increases, the current response value increases and the sensitivity increases, but the waveform gradually becomes distorted, and R 2 The lower the voltage, the poorer the measurement accuracy. Therefore, considering the sensitivity and stability, 0.85 V was finally selected as the operating voltage for all subsequent experiments to obtain better test performance.

[0072] Gold nanoparticle-based flexibility The sensor was systematically evaluated by chronoamperometry in artificial saliva. Figure 9 (where a is The current response diagram between 0-30 mM, b is the corresponding The concentration and current response fitting curve shows that the measured current intensity increases with The concentration increases, which shows that The sensor has a significant current response as low as 1 μM, indicating that it has extremely high sensitivity. The amperometric response of the electrode has two linear ranges, the first of which is consistent with the range of 1 μM-5 mM. The range of the correlation coefficient is 0.993. This range covers physiological and pathological concentrations and can monitor the progression of inflammation in situ. Similarly, the selectivity of the electrode was tested. Figure 10 As shown, the interfering substances (Na + , K + , Ca 2+ Mg 2+ The interference of the electrodes with the ions (e.g., Uresa, Suc, Glu, and BSA) was negligible, indicating that the sensor had good anti-interference ability. Therefore, the developed device has the ability to monitor RNS in the oral microenvironment in real time.

[0073] To verify the clinical applicability of the developed device, an in vitro model was constructed to simulate the development of dental caries. The cells were then inoculated into BHI brain heart infusion broth containing 2% (w / v) sucrose and cultured anaerobically. The temperature was maintained at 37°C and the gas environment was 10% H2, 5% CO2, and 85% N2. The prepared microelectrode was used to record the pH value at 30-minute intervals during the 24-hour culture period to simulate the actual situation in vivo. Before detection, the electrode was calibrated with a standard buffer solution (pH = 7.0), and all experiments were performed with three biological replicates (n = 3). Figure 11 As shown, with prolonged incubation, the acid-generating activity of Streptococcus mutans increased, and the pH gradually decreased from an initial level of around 7 to below the critical threshold for enamel demineralization (pH 5.5). This is consistent with the expected mechanism of caries development. These results demonstrate that this device can achieve real-time pH monitoring in a simulated caries environment, offering potential clinical applications for early caries detection and management.

[0074] In this study, RAW 264.7 macrophages were used as an in vitro inflammatory model to simulate the periodontal microenvironment and evaluate the detection performance of the developed device. Specifically, RAW 264.7 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% (v / v) fetal bovine serum (FBS). The cells were incubated at 37°C in a humidified atmosphere with 5% CO2. When the cells reached 80%–90% confluence, they were harvested using a cell scraper and centrifuged at 1000 rpm for 5 minutes. Subsequently, the cells were repeatedly washed with 0.01 M phosphate-buffered saline (PBS) to remove residual culture medium components. To induce macrophage activation and stimulate reactive nitrogen species (RNS) production, the cells were pretreated with lipopolysaccharide (LPS, 1 µg / mL) to induce M1 polarization. L-arginine (L-Arg, 1 mM) was used as a substrate for nitric oxide synthesis. In addition, Nω-nitro-L-arginine methyl ester (L-NAME, 0.5 mM) was added to inhibit RNS generation. Real-time amperometric detection (working potential 0.85 V vs. Ag / AgCl) was performed using the device (previously activated in 0.1 M PBS for 30 minutes) and real-time recording was performed. The current response of the release. Figure 12 As shown, a significant increase in current was observed immediately after the addition of L-Arg. However, the simultaneous addition of LNAME (a NO inhibitor) and L-Arg completely suppressed the current response, indicating that the current signal was indeed generated by the release of RNS by macrophages. Furthermore, no signal was generated when there were no cells in the system, eliminating the interference of L-Arg. These data confirm that the device is capable of monitoring the dynamic changes of RNS in the periodontitis microenvironment in real time. Therefore, it shows significant potential in the clinical monitoring of periodontal oxidative stress and the evaluation of personalized therapeutic effects.

[0075] To study the controlled release of drugs under electrical stimulation, drug-loaded electrodes were immersed in deionized water and periodically electrically stimulated to simulate an on-off control mode. Subsequently, the released drug concentration was measured periodically using a UV-visible spectrophotometer to analyze the electrical response control performance. Figure 13 An “on-off” drug release pattern was displayed, indicating that this system has the potential for controlled drug delivery.

[0076] To systematically validate the therapeutic efficacy against dental caries and periodontitis, a multimodal antimicrobial evaluation was conducted against the key pathogens Streptococcus mutans and Porphyromonas gingivalis. Biofilm inhibition was systematically assessed using a triple analysis system using colony-forming unit (CFU) counting, scanning electron microscopy (SEM), and live / dead fluorescence staining (CLSM).

[0077] The CFU count results of S. mutans and P. gingivalis biofilm formation under different treatment conditions are shown as follows: Figure 14 As shown in the figures (a shows the CFU counts of S. mutans biofilms formed under different treatment conditions, and b shows the CFU counts of P. gingivalis biofilms formed under different treatment conditions), the electrically stimulated drug-loaded electrode group demonstrated excellent antibacterial efficacy against bacteria, with a CFU reduction of over 3 log. The stark contrast in antibacterial efficacy between the electrically stimulated drug-loaded group and the non-electrical stimulation drug-loaded group demonstrates the device's ability to provide controlled, on-demand drug release, inhibiting pathogenic biofilm formation and demonstrating its potential for treating dental caries and periodontitis. In the figure, Ctrl, Ppy / E, Ppy+ES, Drug-ES, and Drug+ES represent the blank control group, electrode group, electrical stimulation alone group, non-electrical stimulation drug-loaded group, and electrical stimulation drug-loaded group, respectively.

[0078] SEM microstructure analysis ( Figure 15 ) Direct observation of cell membrane damage in the electrically stimulated drug-loaded group revealed that >90% of S. mutans and P. gingivalis cells exhibited membrane collapse and structural breakdown. Three-dimensional confocal microscopy enabled visualization of changes in biofilm thickness and bacterial activity after different treatments. Figure 16 As shown, the electrostimulation-drug-loaded group exhibited the highest percentages of dead S. mutans and P. gingivalis biofilms, at 87.92% and 89.71%, respectively (quantified by SYTO 9 / PI), and the thinnest biofilm thicknesses, at 38.6% and 42.48%, respectively, compared to the control group (Ctrl). These results are consistent with the SEM and CFU data, demonstrating the robust closed-loop management capabilities of the device, enabling on-demand drug release and effective biofilm inhibition. This approach offers a promising strategy for the treatment of caries and periodontitis.

[0079] To ensure the long-term biosafety of the device, an in vivo toxicity assessment was performed. According to the protocol approved by the Animal Research Committee of Jilin University, the animals treated in different groups were sacrificed, and the main organs, heart, liver, lung, spleen, and kidney, were collected, fixed with formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (H&E). Figure 17 It was shown that rats treated with the flexible wearable electrode array of the present invention did not experience morphological changes or other adverse reactions, indicating that the present invention has good biosafety.

[0080] 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 scope of protection of the present invention.

Claims

1. A wearable oral diagnostic and treatment device, characterized in that: include: Flexible double-sided electrode array and near-field communication module; The flexible double-sided electrode array includes a base layer, an upper serpentine interconnection wire layer is provided on the upper surface of the base layer, one end of the upper serpentine interconnection wire layer is connected to a via electrode interface provided on the lower surface of the base layer through an electrode via provided on the base layer, an upper insulating layer is provided at the end of the upper serpentine interconnection wire layer, an upper conductive ink layer is provided on the upper insulating layer, and the upper conductive ink layer includes a working electrode W pH , working electrode W cT , reference electrode R pH and reference electrode R cT , and the working electrode W pH , working electrode W cT , reference electrode R pH and reference electrode R cT The lower surface of the base layer is provided with a lower serpentine interconnection wire layer, the end of the lower serpentine interconnection wire layer is provided with a lower insulating layer, the lower insulating layer is provided with a lower conductive ink layer, and the lower conductive ink layer includes a counter electrode C RNS , working electrode W RNS , reference electrode R RNS and working electrode W pT , and the counter electrode C RNS , working electrode W RNS , reference electrode R RNS and working electrode W pT are respectively connected to a serpentine wire in the lower serpentine interconnection wire layer; The front side of the near-field communication module is provided with a near-field communication chip and a single-chip microcomputer chip, the near-field communication chip is connected to the near-field communication coil, and the back side of the near-field communication module is provided with an electrode interface for matching the upper serpentine interconnection wire layer and the lower serpentine interconnection wire layer; The working electrode W pH The reference electrode R pH The working electrode W is modified with a PVB solid composite electrolyte layer; cT It is modified with a Ppy / pTs drug-loaded electrode layer; The working electrode W RNS The working electrode W pT It is also modified with a Ppy / pTs drug-loaded electrode layer; The preparation method of the wearable oral diagnostic and treatment device comprises the following steps: Step 1: Design and fabrication of flexible double-sided electrode arrays: The flexible double-sided electrode array is fabricated using computer-aided design (CAD) and FPCB technology. The upper and lower conductive ink layers are formed by screen-printing conductive carbon and Ag / AgCl inks. Based on the FPCB process, the upper and lower serpentine interconnect layers are formed on the substrate by laser processing. The upper serpentine interconnect layer is then connected to a via-electrode interface on one side of the lower serpentine interconnect layer. Step 2: Electrode functionalization modification: At the working electrode W pH IrOx NTs layer is deposited on the surface; at the reference electrode R pH The surface is coated with a PVB solid composite electrolyte layer; the working electrode W RNS The gold nanoparticle layer was deposited on the interface by chronoamperometry; cT and working electrode W pT Surface modified Ppy / pTs drug-loaded electrode layer; Step 3: System Integration: The pins of the upper serpentine interconnect wire layer and the lower serpentine interconnect wire layer are respectively connected to the electrode interface, packaged, and integrated with the customized transparent braces to complete the preparation; The IrOx NTs layer was prepared using an optimized electrospinning method: A precursor solution was prepared by heating 0.25 g of hydrated iridium trichloride, 2 mL of N,N-dimethylformamide solution, and 0.4 g of polyvinyl pyrrolidone in a water bath. After electrospinning, the solution was annealed at 500 °C for 3 h. Disperse it evenly in deionized water to a concentration of 5 mg / mL, and accurately apply 5 μL to the pretreated working electrode W. pH The surface is then shaped by infrared-assisted drying.

2. The wearable oral diagnostic and treatment device according to claim 1, characterized in that: The preparation of the PVB solid composite electrolyte layer is as follows: 78 mg of PVB and 50 mg of NaCl were dissolved in 1 mL of methanol solution and ultrasonically treated to form a solid composite electrolyte layer. Then 3 μL of the PVB mixture was added dropwise to the reference electrode R pH and dried at room temperature overnight.

3. The wearable oral diagnostic and treatment device according to claim 1, characterized in that: The Au nanoparticle layer was prepared as follows: The pretreated working electrode W RNS The sample was immersed in a 2 mM chloroauric acid solution prepared in 0.1 M phosphate buffer and a constant potential of -0.2 V vs. Ag / AgCl was applied for 120 s.

4. The wearable oral diagnostic and treatment device according to claim 1, characterized in that: The preparation of the Ppy / pTs drug-loaded electrode layer is as follows: First, in a mixed solution of 0.2 M pyrrole and 0.05 M pTs, apply 2 mA / cm -2 The constant current was applied for 10 min to achieve electrochemical deposition of Ppy thin films; The electrodes were then immersed in the drug solution, and a potential of -0.6 V relative to Ag / AgCl was applied thereto for 0-40 minutes.

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