Oral wearable diagnosis and treatment equipment and manufacturing method thereof

Abstract

The invention is suitable for the technical field of oral cavity diagnosis and treatment equipment, and provides oral cavity wearable diagnosis and treatment equipment and a manufacturing method thereof, the equipment integrates a flexible electrochemical biosensor through a tooth socket, continuous perception of oral cavity microenvironment biomarkers can be realized, and a data basis is provided for early diagnosis of diseases. And the carried electric response drug release structure can exploratively realize the controllable release of the antibacterial drug. And a flexible double-sided structure is adopted, so that the electrode space is optimized while double-disease management is met, and the wearing comfort is improved. By monitoring and analyzing oral health data, help is expected to be provided for reducing unreasonable use of antibiotics. Meanwhile, a preliminary framework of oral health data management is constructed based on a collaborative mode of wearable equipment and terminal equipment, and a new technical path is provided for 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 an oral wearable diagnosis and treatment equipment and a manufacturing method thereof. Background Art

[0002] Chronic oral diseases (such as dental caries, periodontitis) are persistent problems in the field of global public health. Their impact has transcended the local oral scope and is closely related to the occurrence and development of systemic diseases such as sepsis, Alzheimer's disease, and rheumatoid arthritis. According to statistics, about 3.5 billion people worldwide suffer from dental caries, and more than half of adults are troubled by periodontitis, which has brought a heavy burden to global public health, especially the economic cost is particularly significant among low-income groups. However, traditional diagnostic methods rely on visual inspection or imaging assessment and can only detect diseases in the middle and late stages, when irreversible damage has occurred to tissues. On the other hand, the main treatment strategies still focus on "post-intervention", and there are significant technical shortcomings in early identification, precise treatment, and dynamic management. Generally speaking, 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] Currently, 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 lags behind and the ability of dynamic monitoring is insufficient; in terms of treatment, the mode is passive and single, and the problem of drug abuse is prominent; in the application of equipment, the 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 an oral wearable diagnosis and treatment equipment and a manufacturing method thereof, aiming to solve the problems raised in the above background art.

[0005] The embodiments of the present invention are implemented as follows. An oral wearable diagnosis and treatment equipment includes: a flexible double-sided electrode array and a 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. The upper conductive ink layer includes 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 pHand the reference electrode R cT are respectively connected to one serpentine wire in the upper serpentine interconnection wire layer; a lower serpentine interconnection wire layer is provided on the lower surface of the base layer, a lower insulating layer is provided at the end of the lower serpentine interconnection wire layer, and a lower conductive ink layer is provided on the lower insulating layer. The lower conductive ink layer includes the counter electrode C RNS , the working electrode W RNS , the reference electrode R RNS and the working electrode W pT , and the counter electrode C RNS , the working electrode W RNS , the reference electrode R RNS and the working electrode W pT are respectively connected to one serpentine wire in the lower serpentine interconnection wire layer; On the front surface of the near-field communication module, a near-field communication chip and a microcontroller chip are provided. The near-field communication chip is connected to the near-field communication coil. On the back surface of the near-field communication module, electrode interfaces for matching the upper serpentine interconnection wire layer and the lower serpentine interconnection wire layer are provided.

[0006] In a further technical solution, an IrOx NTs layer is modified on the working electrode W pH ; a PVB solid composite electrolyte layer is modified on the reference electrode R pH ; a Ppy / pTs drug-loaded electrode layer is modified on the working electrode W cT .

[0007] In a further technical solution, an Au nanoparticle layer is modified on the working electrode W RNS ; a Ppy / pTs drug-loaded electrode layer is also modified on the working electrode W pT .

[0008] Another object of the embodiments of the present invention is a preparation method of an oral wearable diagnosis and treatment device. Based on the above oral wearable diagnosis and treatment device, the method includes the following steps: Step 1: Design and fabrication of a flexible double-sided electrode array: The flexible double-sided electrode array is fabricated by computer-aided design and then by using flexible printed circuit board (FPCB) technology; the upper conductive ink layer and the lower conductive ink layer are made by screen printing conductive carbon and Ag / AgCl ink. Based on the high-precision FPCB process, the upper serpentine interconnection wire layer and the lower serpentine interconnection wire layer are formed on the base layer by laser processing, and the upper serpentine interconnection wire layer is connected to the via electrode interface on one side of the lower serpentine interconnection wire layer; Step 2: Electrode functionalization modification: Deposit an iridium oxide nanotube (IrOx NTs) layer on the surface of the working electrode W pH ; deposit a layer of PVB solid composite electrolyte on the reference electrode RpH The surface is coated with a polyvinyl butyral (PVB) solid composite electrolyte layer; on the working electrode W RNS At the interface, a layer of gold nanoparticles (Au NPs) is electrochemically deposited by chronoamperometry; on the working electrode W cT and the working electrode W pT The surface is modified with a Ppy / pTs drug-loaded electrode layer; Step 3: System integration: The pins of the upper serpentine interconnection wire layer and the lower serpentine interconnection wire layer are respectively docked with the electrode interfaces. After encapsulation, they are integrated with a customized transparent dental appliance to complete the preparation.

[0009] A further technical solution is that the IrOx NTs layer is prepared by an optimized electrospinning method: 0.25 g of iridium(III) chloride hydrate, 2 mL of N,N-dimethylformamide solution, and 0.4 g of polyvinylpyrrolidone are used to prepare a precursor solution by water bath heating. After electrospinning, it is annealed at 500 °C for 3 hours; it is uniformly dispersed in deionized water, formulated into a concentration of 5 mg / mL, and 5 μL is precisely dropped onto the pretreated working electrode W pH The surface, and then formed by infrared-assisted drying.

[0010] A further technical solution is that the PVB solid composite electrolyte layer is prepared as follows: 78 mg of PVB and 50 mg of NaCl are dissolved in 1 mL of methanol solution, and a solid composite electrolyte layer is formed by ultrasonic treatment; then 3 μL of the PVB mixture is dropped onto the reference electrode R pH and dried overnight at room temperature.

[0011] A further technical solution is that the Au nanoparticle layer is prepared as follows: The pretreated working electrode W RNS is immersed in a 2 mM chloroauric acid (HAuCl4) solution prepared with 0.1 M phosphate buffer solution (PBS), and a constant potential of -0.2 V vs. Ag / AgCl is applied for 120 s.

[0012] A further technical solution is that the Ppy / pTs drug-loaded electrode layer is prepared as follows: First, in a mixed solution of 0.2 M pyrrole and 0.05 M pTs, a constant current of 2 mA / cm -2 is applied for 10 minutes to achieve the electrochemical deposition of the Ppy film; Then the electrode is immersed in the drug solution respectively, and a potential of -0.6 V vs. Ag / AgCl is applied thereto for 0 - 40 minutes.

[0013] An oral wearable diagnosis and treatment device and its manufacturing method provided by an embodiment of the present invention. The device integrates a flexible electrochemistry biosensor through a dental appliance, enabling continuous perception of biomarkers in the oral microenvironment, providing a data basis for early disease diagnosis, and making up for the deficiencies of traditional detection means in capturing early lesion signals to a certain extent. Its carried electro-responsive drug release structure can explore and achieve controllable release of antibacterial drugs. In terms of structural design, a flexible double-sided structure is adopted, which meets the dual disease management, optimizes the electrode space, and improves the wearing comfort. By monitoring and analyzing oral health data, it is expected to help reduce the unreasonable use of antibiotics, thereby assisting in controlling the risk of bacterial resistance. At the same time, based on the collaborative mode of "wearable device + terminal device", a preliminary framework for oral health data management is constructed, providing a new technical path for the comprehensive prevention and treatment of chronic oral diseases. Description of the Drawings

[0014] Figure 1 Schematic diagram of the flexible double-sided electrode array structure in an oral wearable diagnosis and treatment device provided by an embodiment of the present invention; Figure 2 Schematic diagram of the structure of the near-field communication module in an oral wearable diagnosis and treatment device provided by an embodiment of the present invention (where a is the front side and b is the back side); Figure 3 Wearing effect diagram of an oral wearable diagnosis and treatment device provided by an embodiment of the present invention; Figure 4 pH detection potential response and fitting curve graph of the flexible double-sided electrode array (where a is the potential response and b is the fitting curve); Figure 5 pH detection selectivity test graph of the flexible double-sided electrode array; Figure 6 pH detection reproducibility test graph of the flexible double-sided electrode array (where a is the potential response graph of multiple electrodes and b is the corresponding potential-pH fitting curve graph); Figure 7 RNS electrode functionalization modification time test graph of the flexible double-sided electrode array; Figure 8 RNS detection optimal working voltage test graph of the flexible double-sided electrode array (where a is the current response graph between 50 - 650 μM, and b is the corresponding concentration and current response fitting curve); Figure 9 RNS detection amperometric current response and fitting curve graph of the flexible double-sided electrode array (where a is the current response graph between 0 - 30 mM, and b is the corresponding concentration and current response fitting curve graph); Figure 10 It is a selective test diagram for RNS detection; Figure 11 It is a data diagram for monitoring pH value in an in vitro simulated caries environment; Figure 12 It is for monitoring RAW 264.7 macrophages simulating the periodontitis microenvironment of the sensor data diagram; Figure 13 It is an effect diagram of drug release under electrical stimulation response; Figure 14 It is a statistical result diagram of the antibacterial CFU of S. mutans and P. gingivalis (where a is the CFU counting result of S. mutans biofilm formation under different treatment conditions, and b is the CFU counting result of P. gingivalis biofilm formation under different treatment conditions); Figure 15 It is an SEM diagram of the influence of bacteria; Figure 16 It is a CLSM diagram of the influence of bacteria; Figure 17 It is a histological section image of a rat.

[0015] In the attached drawings: 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 interconnecting wire layer 3; base layer 4; electrode via 4-1; lower serpentine interconnecting 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 W pT 7-2; reference electrode R RNS 7-3; working electrode W RNS 7-4; near-field communication chip 8; microcontroller chip 9; near-field communication coil 10; electrode interface 11. Detailed implementation manners

[0016] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the following further details the present invention in combination with the attached drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0017] The following describes the specific implementation of the present invention in detail in combination with specific embodiments.

[0018] Such as Figure 1 and Figure 2 wherein Figure 2a is a front view, Figure 2 as shown in b which is a rear view, a kind of oral wearable diagnosis and treatment device provided by an embodiment of the present invention includes: a flexible double-sided electrode array and a near-field communication module; The flexible double-sided electrode array includes a base layer 4, an upper serpentine interconnecting wire layer 3 is arranged on the upper surface of the base layer 4, one end of the upper serpentine interconnecting wire layer 3 is connected to a via electrode interface 5-1 arranged on the lower surface of the base layer 4 through an electrode via 4-1 arranged on the base layer 4, an upper insulating layer 2 is arranged at the end of the upper serpentine interconnecting wire layer 3, an upper conductive ink layer 1 is arranged on the upper insulating layer 2, and the upper conductive ink layer 1 includes 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 interconnecting wire layer 3; a lower serpentine interconnecting wire layer 5 is arranged on the lower surface of the base layer 4, a lower insulating layer 6 is arranged at the end of the lower serpentine interconnecting wire layer 5, a lower conductive ink layer 7 is arranged on the lower insulating layer 6, and the lower conductive ink layer 7 includes 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 R RNS 7-3 and working electrode W RNS 7-4 are respectively connected to a serpentine wire in the lower serpentine interconnecting wire layer 5; On the front of the near-field communication module, a near-field communication chip 8 and a single-chip microcomputer chip 9 are arranged, the near-field communication chip 8 is connected to a near-field communication coil 10, and on the rear of the near-field communication module, an electrode interface 11 for matching the upper serpentine interconnecting wire layer 3 and the lower serpentine interconnecting wire layer 5 is arranged.

[0019] As a preferred embodiment of the present invention, an IrOx NTs layer is modified on the working electrode W pH 1-4; a PVB solid composite electrolyte layer is modified on the reference electrode R pH 1-3; a Ppy / pTs drug-loading electrode layer is modified on the working electrode W cT 1-1.

[0020] As a preferred embodiment of the present invention, the working electrode W RNS is modified with a layer of Au nanoparticles on 7-4; the working electrode W pT is also modified with a Ppy / pTs drug-loaded electrode layer on 7-2.

[0021] A method for preparing an oral wearable diagnosis and treatment device provided by an embodiment of the present invention, based on the above-mentioned oral wearable diagnosis and treatment device, includes the following steps: Step 1: Design and fabrication of a flexible double-sided electrode array: The flexible double-sided electrode array is prepared by using flexible printed circuit board (FPCB) technology after computer-aided design based on 3ds Max 2020 software (Autodesk, Inc., USA). The upper conductive ink layer 1 and the lower conductive ink layer 7 are made by screen printing conductive carbon and Ag / AgCl inks. The double-sided design conforms to the oral anatomical shape, facilitating dual disease management. Based on the high-precision FPCB process, the upper serpentine interconnecting wire layer 3 and the lower serpentine interconnecting wire layer 5 are formed on the base layer 4 by laser processing, and the upper serpentine interconnecting wire layer 3 is connected to the via electrode interface 5-1 on one side of the lower serpentine interconnecting wire layer 5. Its geometrically optimized trace design significantly improves the mechanical flexibility of the electrode. The wire layout is optimized, and the pin connections are concentrated on one side of the base layer 4, thus achieving a stable connection with the near-field communication module (i.e., the control circuit).

[0022] Step 2: Electrode functionalization modification: It is prepared by depositing iridium oxide nanotubes (IrOx NTs) as the H⁺ sensitive layer on the surface of the working electrode W pH 1-4. The IrOx NTs are prepared by an optimized electrospinning method to enhance their structural stability and electrochemical performance. The reference electrode R pH 1-3 is prepared by coating a polyvinyl butyral (PVB) solid composite electrolyte layer on the electrode surface to stabilize the Cl - concentration at the electrode interface. The interface of the working electrode W RNS 7-4 is constructed by chronoamperometric electrodeposition of gold nanoparticles (Au NPs), thereby improving the electrocatalytic activity and sensing sensitivity of the electrode to nitrite. The working electrode W cT 1-1 and the working electrode W pT 7-2 are prepared by a two-step method. First, a polypyrrole / p-toluenesulfonic acid (Ppy / pTs) film is deposited on the electrode surface by constant current electropolymerization. Subsequently, charged drugs are introduced into the polymer network electrochemically in the drug solution respectively.

[0023] Step 3: System integration: The pins of the upper serpentine interconnecting wire layer 3 and the lower serpentine interconnecting wire layer 5 are respectively docked with the electrode interface 11. After encapsulation, it is integrated with a customized transparent dental appliance to complete the preparation. The assembled device is as shown in Figure 3 as follows.

[0024] As a preferred embodiment of the present invention, the IrOx NTs layer is prepared by an optimized electrospinning method: 0.25 g of iridium trichloride hydrate, 2 mL of N,N-dimethylformamide solution, and 0.4 g of polyvinylpyrrolidone are used to prepare a precursor solution by water bath heating. After electrospinning, it is annealed at 500 °C for 3 hours. It is uniformly dispersed in deionized water, formulated into a concentration of 5 mg / mL, and 5 μL is precisely drop-coated on the pretreated working electrode W pH 1-4 surfaces. Subsequently, infrared-assisted drying is carried out to ensure uniform film formation and improve its stability and adhesion.

[0025] As a preferred embodiment of the present invention, the preparation of the PVB solid composite electrolyte layer is as follows: 78 mg of PVB and 50 mg of NaCl are dissolved in 1 mL of methanol solution, and a solid composite electrolyte layer is formed by ultrasonic treatment. Then the PVB mixture (3 μL) is dropped onto the reference electrode R pH 1-3 and dried overnight at room temperature to ensure uniform formation of the electrolyte and good stability.

[0026] As a preferred embodiment of the present invention, the preparation of the Au nanoparticle layer is as follows: The pretreated working electrode W RNS 7-4 is immersed in a 2 mM chloroauric acid (HAuCl4) solution prepared with 0.1 M phosphate buffer solution (PBS), and a constant potential of -0.2 V (vs. Ag / AgCl) is 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.

[0027] As a preferred embodiment of the present invention, 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, a constant current of 2 mA / cm -2 is applied for 10 minutes to achieve electrochemical deposition of the Ppy film. Then the electrode is immersed in the drug solution respectively, and a potential of -0.6 V (vs. Ag / AgCl) is applied thereon for 0-40 minutes.

[0028] As a preferred embodiment of the present invention, the near-field communication module realizes the bidirectional interaction of energy and data between the biological microenvironment and the electronic system by using the inductive coupling principle, and supports wireless energy capture, wireless data transmission, electrochemical signal processing, and electro-responsive drug release. The resonance frequency is about 13.56 MHz, and the wireless communication distance is 0 - 15 mm.

[0029] As a preferred embodiment of the present invention, the transparent dental appliance is customized according to the volunteers. First, the tooth morphology is accurately captured by a 3Shape scanning device, and the data is exported in STL format and used to manufacture a physical model with a 3D printer. Then, a flexible double-sided electrode array and a near-field communication module are installed at the designated positions. Subsequently, the 3Shape scanning and 3D printing technologies are used again to prepare a "tooth model after wearing" including the module installation state, which is thermoformed with TPU material and trimmed into the main body of the dental appliance. Finally, the dental appliance and the electronic module are integrated.

[0030] In the embodiment of the present invention, the finally prepared oral wearable diagnosis and treatment device can be used for the detection of the oral microenvironment and also for the antibacterial treatment of dental caries and periodontitis.

[0031] To systematically verify the monitoring ability of this device for the oral microenvironment, its sensing abilities for pH and RNS ( ) were quantitatively evaluated, which are key biomarkers for the early detection of dental caries and periodontitis respectively. After calibration with a Sartorius pH meter, a buffer solution with pH = 3 - 8 was prepared using 0.1 M HCl / NaOH to simulate the dynamic acidification process of enamel demineralization (pH < 5.5) and healthy state (pH ~ 7.0), and the potential response of the pH sensor was recorded using the open circuit potential (OCP) method. Figure 4 (where a is the potential response and b is the fitting curve) recorded the open circuit potential response varying with the pH value. During the acidification process (pH = 8 - 3), as the pH continuously decreased, the potential showed a step response, while during the alkalization process (pH = 3 - 8), it showed a completely reversible and stable potential change. Figure 4 The calibration curve showed nearly ideal linearity (R² = 0.9992), demonstrating near-Nernst performance. This may be attributed to the IrOx NTs modified on the surface of the working electrode, which can serve as an ion-selective membrane in selective sensing on the one hand, and its high conductivity and high specific surface area contribute to efficient signal extraction on the other hand. Considering that other electrolytes and metabolites present in the oral microenvironment may have a negative impact on the sensor output, the selectivity of this 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), Mg2+ (0.5 mM) and bovine serum albumin (BSA, 3 mg / mL). As Figure 5 shown, the addition of non-target electrolytes and metabolites does not substantially affect the sensor performance, and the sensor shows high specificity, which may be attributed to the specific hydrogen ion selectivity of IrOx, thus minimizing the potential response of common interferents. To explore the reproducibility of the developed pH sensor, multiple independently prepared electrodes were used to perform multiple measurements in buffer solutions with the same pH = 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), indicating that the electrodes obtained in different batches have similar response trends and sensitivities.

[0032] RNS ( ) sensing electrodes were applied with a constant potential of -0.2 V vs. Ag / AgCl for 120 s in 2 mM HAuCl4 solution by chronoamperometry to reduce and deposit gold nanoparticles. Cyclic voltammetry (CV) was used to explore the structure-activity relationship between the deposition time of gold nanoparticles (AuNPs) and the sensing performance of RNS ( ). Figure 7 It shows that when the deposition time is 120 s, the peak current of oxidation reaches the highest, so the deposition time of 120 s is the optimal condition for preparing electrodes sensitive to RNS.

[0033] To determine the optimal working parameters for RNS ( ) monitoring, the amperometric response characteristics in the potential range of 0.75 - 0.95 V (vs. Ag / AgCl) were systematically studied in phosphate buffer solution (PBS, pH = 7.4). Figure 8 (where a is the current response diagram between 50 - 650 μM, and b is the corresponding concentration and current response fitting curve) shows the chronoamperometric response and the corresponding calibration curve with the concentration in the range of 0 - 650 μM at different potentials of 0.75 - 0.95 V. In contrast, as the voltage increases, the current response value becomes larger and the sensitivity is higher, but the waveform gradually distorts and the R 2 is lower and the measurement accuracy is poor. Therefore, considering both sensitivity and stability, 0.85 V was finally selected as the working voltage for all subsequent experiments to obtain better test performance.

[0034] The flexible sensor based on gold nanoparticles was systematically evaluated by chronoamperometry in artificial saliva. Figure 9 (where a is the The current response graph between 0 - 30 mM, and b is the corresponding concentration and current response fitting curve graph) shows that the measured current intensity increases with the increase in concentration, which exhibits a representative amperometric signal for . And the sensor has an obvious current response even at as low as 1 μM, indicating that the sensor has extremely high sensitivity. The amperometric response of the electrode has two linear ranges, where the first linear range conforms to the range of 1 μM - 5 mM, and the correlation coefficient is 0.993. This range covers physiological and pathological concentrations and can in-situ monitor the progress of inflammation. Similarly, selectivity tests were performed on the electrode. As Figure 10 shown, the interference of interferents (Na + , K + , Ca 2+ , Mg 2+ , Uresa, Suc, Glu, BSA) on the electrode is negligible, which indicates that the sensor has good anti-interference ability. Therefore, the developed device has the ability to real-time monitor RNS in the oral microenvironment.

[0035] To verify the clinical applicability of the developed device, an in vitro model was constructed to simulate the occurrence process of dental caries. The concentration of Streptococcus mutans was standardized to and then inoculated in BHI brain heart infusion broth medium containing 2% (w / v) sucrose for anaerobic culture at 37°C with a gas environment of 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 in vivo situation. 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). As Figure 11 shown, with the extension of the culture time, the acid production activity of Streptococcus mutans increases, and the pH gradually decreases from about 7 initially to below the critical threshold of enamel demineralization (pH 5.5). This is in line with the expected mechanism of dental caries occurrence. The results show that the device can achieve real-time pH monitoring in a simulated dental caries environment, providing potential clinical application value for early dental caries detection and management.

[0036] In this study, RAW 264.7 macrophages were selected as an in vitro inflammation model to simulate the periodontitis microenvironment and evaluate the detection performance of the developed device. Specifically, RAW 264.7 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% (v / v) fetal bovine serum (FBS). The cells were cultured in a humidified environment at 37 °C and 5% CO2. When the cells reached 80%–90% confluence, the cells were collected using a cell scraper and separated by centrifugation at 1000 rpm for 5 minutes. Subsequently, they were washed repeatedly with 0.01 M phosphate buffer saline (PBS) to remove residual components of the culture medium. To induce macrophage activation and stimulate the production of reactive nitrogen species (RNS), the cells were pretreated with lipopolysaccharide (LPS, 1 µg / mL) to induce M1 polarization, and 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 production. Real-time amperometric detection (working potential 0.85 V vs. Ag / AgCl) was performed using the device (pre-activated in 0.1 M PBS for 30 minutes), and the released current response was recorded in real time. As Figure 12 shown, a significant increase in the current signal was immediately observed after the addition of L-Arg, while the current response was completely inhibited when L-NAME (a NOs inhibitor) and L-Arg were added simultaneously, indicating that the current signal was indeed generated by the release of RNS from macrophages. In addition, no signal was generated when there were no cells in the system, ruling out the interference of L-arg. These data confirmed that the device was able to real-time monitor the dynamic changes of RNS in the periodontitis microenvironment. Therefore, it showed significant potential in the clinical monitoring of periodontal oxidative stress and individualized efficacy evaluation.

[0037] To study the controlled release behavior of drugs under electrical stimulation, the drug-loaded electrodes were immersed in deionized water and periodic electrical stimulation was applied to simulate the on-off control mode. Subsequently, the concentration of the released drug was determined regularly using a UV-visible spectrophotometer to analyze the electro-responsive control performance. Figure 13 showed an "on-off" drug release pattern, indicating the potential of the system for controllable drug delivery.

[0038] To systematically verify the therapeutic effects on caries and periodontitis, multimodal antibacterial evaluations were carried out against the key pathogenic bacteria Streptococcus mutans and Porphyromonas gingivalis. The biofilm inhibition effect was systematically evaluated through a triple analysis system of colony-forming unit (CFU) counting, scanning electron microscopy (SEM), and live / dead fluorescence staining (CLSM).

[0039] The CFU counting results of S. mutans and P. gingivalis biofilm formation under different treatment conditions are as follows Figure 14 (where a is the CFU counting result of S. mutans biofilm formation under different treatment conditions, and b is the CFU counting result of P. gingivalis biofilm formation under different treatment conditions). The electrostimulated drug-loaded electrode group showed excellent antibacterial efficacy against bacteria, with a CFU reduction of more than 3 logs. The distinct contrast in antibacterial effects between the electrostimulated drug-loaded group and the non-electrostimulated drug-loaded group confirmed that the device can provide controllable drug release on demand, inhibit the formation of pathogenic biofilms, and show potential for the treatment of dental caries and periodontitis. In the figure, Ctrl, Ppy / E, Ppy+ES, Drug-ES, and Drug+ES represent the blank control group, the electrode group, the simple electrostimulation group, the non-electrostimulated drug-loaded group, and the electrostimulated drug-loaded group, respectively.

[0040] SEM microstructure analysis ( Figure 15 ) directly observed cell membrane damage in the electrostimulated drug-loaded group, with >90% of S. mutans and P. gingivalis showing envelope collapse and structural breakdown. Confocal microscopy three-dimensional imaging was able to visualize changes in biofilm thickness and bacterial viability after different treatments. Figure 16 As shown, the electrostimulated drug-loaded group had the highest proportion of dead bacteria in the biofilms of S. mutans and P. gingivalis, 87.92% and 89.71% respectively (SYTO 9 / PI quantification), and the thinnest biofilm thickness, which was reduced by 38.6% and 42.48% respectively compared to the control group (Ctrl). These results were consistent with the SEM and CFU results, indicating that the device has strong closed-loop management capabilities, can release drugs on demand, and effectively inhibit biofilms. This method provides a promising strategy for the treatment of dental caries and periodontitis.

[0041] To ensure the long-term biosafety of the device, in vivo toxicity assessments were performed. According to the protocol approved by the Animal Research Committee of Jilin University, animals treated in different groups were sacrificed, and the main organs, heart, liver, lung, spleen, and kidney, were collected, fixed in formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (H&E). Figure 17 The results showed that no morphological changes or other adverse reactions occurred in the rats treated with the flexible wearable electrode array of the present invention, indicating good biosafety of the present invention.

[0042] The above are only the preferred embodiments of the present invention and are 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 within the protection scope of the present invention.

Claims

1. An oral wearable diagnosis and treatment device, characterized in that, Comprising: A flexible double-sided electrode array and a near-field communication module; The flexible double-sided electrode array includes a base layer, on the upper surface of which an upper serpentine interconnection wire layer is provided. 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, and an upper conductive ink layer is provided on the upper insulating layer. The upper conductive ink layer includes working electrode W pH and 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 are respectively connected to one serpentine wire in the upper serpentine interconnection wire layer. A lower serpentine interconnection wire layer is provided on the lower surface of the base layer. A lower insulating layer is provided at the end of the lower serpentine interconnection wire layer, and a lower conductive ink layer is provided on the lower insulating layer. The lower conductive ink layer includes 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 one serpentine wire in the lower serpentine interconnection wire layer; On the front side of the near-field communication module, a near-field communication chip and a microcontroller chip are provided. The near-field communication chip is connected to a near-field communication coil. On the back side of the near-field communication module, an electrode interface for matching the upper serpentine interconnection wire layer and the lower serpentine interconnection wire layer is provided.

2. The oral wearable diagnosis and treatment device according to claim 1, characterized in that, The working electrode W pH is modified with an IrOx NTs layer; the reference electrode R pH is modified with a PVB solid composite electrolyte layer; the working electrode W cT is modified with a Ppy / pTs drug-loaded electrode layer.

3. The oral wearable diagnosis and treatment device according to claim 2, wherein The working electrode W RNS is modified with a layer of Au nanoparticles; the working electrode W pT is also modified with a Ppy / pTs drug-loaded electrode layer.

4. A preparation method of an oral wearable diagnosis and treatment device, based on the oral wearable diagnosis and treatment device according to any one of the above claims 1-3, characterized in that, Including the following steps: Step 1: Design and fabrication of the flexible double-sided electrode array: The flexible double-sided electrode array is prepared by computer-aided design and then by using the FPCB technology; the upper conductive ink layer and the lower conductive ink layer are made by screen-printing conductive carbon and Ag / AgCl inks. Based on the FPCB process, the upper serpentine interconnection wire layer and the lower serpentine interconnection wire layer are formed by laser processing on the base layer, and the upper serpentine interconnection wire layer is connected to the via electrode interface on one side of the lower serpentine interconnection wire layer; Step 2: Electrode functionalization modification: On the working electrode W pH surface, deposit an IrOx NTs layer; on the reference electrode R pH surface, coat a PVB solid composite electrolyte layer; at the interface of the working electrode W RNS electrodeposit a gold nanoparticle layer by chronoamperometry; on the working electrode W cT and the working electrode W pT surface is modified with a Ppy / pTs drug-loaded electrode layer; Step 3: System integration: The pins of the upper serpentine interconnection wire layer and the lower serpentine interconnection wire layer are respectively docked with the electrode interface. After encapsulation, it is integrated with a customized transparent dental brace to complete the preparation.

5. The preparation method of the oral wearable diagnosis and treatment device according to claim 4, characterized in that The IrOx NTs layer is prepared by an optimized electrospinning method: 0.25 g of iridium(III) chloride hydrate, 2 mL of N,N-dimethylformamide solution, and 0.4 g of polyvinylpyrrolidone are used to prepare a precursor solution by water bath heating. After electrospinning, it is annealed at 500 °C for 3 hours; Disperse it evenly in deionized water to prepare a solution with a concentration of 5 mg / mL, and accurately drop 5 μL onto the pretreated working electrode W pH surface, and then form it by infrared-assisted drying.

6. The preparation method of the oral wearable diagnosis and treatment device according to claim 4, wherein The preparation of the PVB solid composite electrolyte layer is as follows: Dissolve 78 mg of PVB and 50 mg of NaCl in 1 mL of methanol solution, and form a solid composite electrolyte layer by ultrasonic treatment; then drop 3 μL of the PVB mixture onto the reference electrode R pH and dry it overnight at room temperature.

7. The preparation method of the oral wearable diagnosis and treatment device according to claim 4, characterized in that, The preparation of the Au nanoparticle layer is as follows: Immerse the pre-treated working electrode W RNS in a 2 mM chloroauric acid solution prepared with 0.1 M phosphate buffer and apply a constant potential of -0.2 V vs. Ag / AgCl for 120 s.

8. The preparation method of the oral wearable diagnosis and treatment device according to claim 4, wherein 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 a constant current of 2 mA / cm -2 for 10 minutes to achieve the electrochemical deposition of the Ppy film; Then the electrodes are respectively immersed in the drug solution, and a potential of -0.6 V vs. Ag / AgCl is applied thereto for 0 - 40 minutes.

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