Application of ultrashort waves in the fabrication of devices for modulating brain neural activity

Abstract

This application relates to the fields of biomedical engineering and neuromodulation, disclosing the application of ultrashortwave in the fabrication of devices for modulating brain neural activity. The modulation of brain neural activity is achieved through the non-thermal effects of ultrashortwave, which refers to ultrashortwave with a frequency of 27.12MHz ± 0.6%, a wavelength of 11.06 nm, and a power of 1.6 ± 0.72 W. This application breaks through the traditional technical prejudice that high-frequency electric fields are unsuitable for the central nervous system, proving that non-thermal ultrashortwave can be safely used for brain neural modulation, opening up a completely new technical direction for neural modulation. With "ultrashortwave + application scenario" as its core protection element, it provides fundamental patent support for research on the treatment of various central nervous system diseases such as Parkinson's disease, stroke, depression, and Alzheimer's disease, possessing extremely high core patent value and broad clinical translation prospects.

Description

Technical Field

[0001] This application relates to the fields of biomedical engineering and neuromodulation technology, and more specifically, to the application of an ultrashortwave in the fabrication of a device for modulating brain neural activity. Background Technology

[0002] Neuromodulation technology has become an important means of treating central nervous system diseases such as Parkinson's disease, post-stroke sequelae, and depression. However, existing neuromodulation techniques have various drawbacks: deep brain stimulation (DBS) is invasive; transcranial magnetic stimulation (TMS) and transcranial direct current stimulation (tDCS) are superficial; and time-domain interferometric stimulation (TI) and transcranial focused ultrasound (tFUS) are technically difficult and costly to perform. Therefore, there is an urgent need in this field for a novel neuromodulation technology that can achieve deep, uniform, safe, non-invasive, and cost-effective treatment.

[0003] Shortwave diathermy is traditionally used in clinical practice to treat inflammation of the musculoskeletal and respiratory systems, such as arthritis and pneumonia, primarily utilizing its thermal effects to achieve anti-inflammatory, analgesic, and improved blood circulation effects. However, the thermal effects pose significant risks when applied to the brain, severely limiting the application of high-frequency electric field technology, which has better penetration effects, in the treatment of central nervous system diseases.

[0004] Therefore, exploring the non-thermal biological effects and mechanisms of action of ultrashort waves in the central nervous system is crucial to breaking through the limitations of traditional thermal effects and expanding their application in neural regulation. Summary of the Invention

[0005] The purpose of this invention is to verify that ultra-short waves can be safely applied to the brain under specific conditions, and to verify that ultra-short waves have a regulatory effect on brain neural activity under specific conditions.

[0006] This application discovers that, under specific stimulation parameters, the non-thermal effects of ultrashortwave can achieve safe and effective neuromodulation, providing fundamental data support for the application of ultrashortwave in central nervous system diseases.

[0007] To achieve the above-mentioned objectives, this application adopts the following technical solution:

[0008] Firstly, the application of the ultrashortwave provided in this application in the preparation of devices for modulating brain neural activity.

[0009] Furthermore, the regulation of brain neural activity is achieved through the non-thermal effects of ultrashort waves.

[0010] Furthermore, the ultra-short wave refers to an ultra-short wave with a frequency of 27.12MHz±0.6%, a wavelength of 11.06, and a power of 1.6±0.72W.

[0011] Furthermore, the device is configured to apply the ultrashort wave to the subject's head in a non-invasive manner.

[0012] Furthermore, the device is configured to apply ultra-short waves according to a preset stimulation protocol.

[0013] Furthermore, the preset stimulation protocol includes each stimulation lasting 10 minutes, with stimulation occurring once every 12 hours, for a cumulative total of 10 stimulations. Furthermore, the regulation of brain neural activity includes at least one of the following: a) Reduce the average power in the Alpha and Beta bands; b) Regulates neuronal calcium activity; c) Activate neurons in the striatum region; d) Regulate neurotransmitter expression.

[0014] Secondly, this application provides a device for regulating brain neural activity, the device comprising: Ultra-shortwave generation unit, used to generate ultra-shortwave waves; and An ultra-shortwave application unit is used to apply the ultra-shortwave to the subject's head.

[0015] Furthermore, the ultrasonic generating unit generates an ultrashort wave with a frequency of 27.12MHz±0.6%, a wavelength of 11.06, and a power of 1.6±0.72W.

[0016] In summary, this application has the following beneficial effects: This invention breaks through the traditional technical prejudice that high-frequency electric fields are unsuitable for use in the central nervous system, proving that calorific ultrashortwave can be safely used for brain neuromodulation, thus opening up a completely new technical direction for neuromodulation. With "ultrashortwave + application scenario" as its core protective elements, it provides fundamental patent support for research on the treatment of various central nervous system diseases such as Parkinson's disease, stroke, depression, and Alzheimer's disease, possessing extremely high core patent value and broad prospects for clinical translation. Attached Figure Description

[0017] Figure 1 : Schematic diagram of the intervention method. A. Diagram of the ultra-shortwave intervention pattern; B. Top view of the mouse during intervention; C. Front view of the mouse during intervention.

[0018] Figure 2The surface temperature of the mouse brain remained stable. A. Representative infrared thermographic images of each group during the intervention period; B. Quantitative statistical graph of the area under the curve (AUC) of the temperature-time curve; C. Real-time dynamic change curve of mouse brain surface temperature during the intervention period (0-600s). Data are expressed as mean ± standard error (Mean ± SEM) (n=6). Differences between groups were analyzed using independent samples t-test. ns, p > 0.05 indicated no statistically significant difference.

[0019] Figure 3 The mice exhibited normal behavioral characteristics. A. Time spent moving on the rotundus; B. Time to turn head and total time in the Pole test; C. Time to fall and overall score in the Hanging wire test; D. Open-field movement trajectory, movement speed, and anxiety index; E. Y-maze movement trajectory and spontaneous alternation rate; F. Cold and heat pain response time. Data are expressed as mean ± SEM, and statistical analysis was performed using the Student T-test (n=6).

[0020] Figure 4 Evans blue did not leak from the blood-brain barrier. Evans blue leakage amount, top view of the mouse brain, and coronal view of a brain slice including the striatum. Data are expressed as mean ± SEM, and statistical analysis was performed using the Student T-test (n=6).

[0021] Figure 5 Shortwave brain stimulation is non-cytotoxic. A. Nissell staining of cortical neurons, scale bar: 50 μm; B. Immunofluorescence staining and density statistics of cortical neurons (NeuN, TUNEL, DAPI, blue), scale bar: 50 μm; C. Myelin staining, scale bar: 100 μm; D. Western blotting and statistical analysis of synaptic protein expression; E. Immunofluorescence staining and density statistics of hippocampal astrocytes (GFAP, green), microglia (IBA1, red), DAPI, blue), scale bar: 200 μm; Data are expressed as mean ± SEM, and statistical analysis was performed using the Student T-test, n=6.

[0022] Figure 6Shortwave brain stimulation can modulate brain electrical activity. A. Schematic diagram of the experimental setup for mouse EEG recording; B. Timeline of the EEG recording protocol, showing the surgical procedure and the recording periods before stimulation, after the first stimulation (1X recording), and after the tenth stimulation (10X recording); C. Representative time-frequency graphs of EEG signals after single (1X) and repeated (10X) stimulation in the sham surgery group (Sham) and the shortwave stimulation (SW) group. The time scale represents a 10-minute window; D. Quantitative analysis of the average power in the Alpha and Beta bands; E. Quantification of the power spectral density (PSD) curves and area under the curve (AUC) of the SW group before, during, and after stimulation; F. Corresponding PSD curves and AUC quantification of the Sham group; G. Schematic diagram of cortical injection of rAAV-hSyn-GCaMP7f and representative immunofluorescence images confirming viral expression and fiber optic implantation sites; H. Timeline of calcium signal recording protocol; I. Temporal dynamics of calcium signal (ΔF / F) recorded at baseline (SW-Pre), first stimulation (SW-1X), and tenth stimulation (SW-10X). Dashed lines represent the stimulation window period, dark solid lines represent the mean, and light shaded areas represent standard errors (n = 12); J. Quantitative analysis of the area under the calcium signal curve (AUC) during the pre-stimulation, during-stimulation, and post-stimulation phases (10 minutes each) in the baseline (SW-Pre), first stimulation (SW-1X), and tenth stimulation (SW-10X) recording periods; K. Quantitative analysis of calcium transient frequency; L. C-Fos immunofluorescence staining and positive cell counts in the cortex, striatum, and hippocampus, scale bar: 100 μm; Data are expressed as mean ± SEM. P<0.05, P < 0.001; ns, not significant.

[0023] Figure 7 Heatmap of monoamine and amino acid neurotransmitter expression in the cortex, striatum, and hippocampus. Detailed Implementation

[0024] The technical solutions and effects of this application will be further described in detail below with reference to the embodiments and accompanying drawings. It should be understood that the specific embodiments described herein are merely for explaining the invention and are not intended to limit the invention.

[0025] The following examples provide the application of an ultrashortwave at a frequency of 27.12 MHz ± 0.6%, a wavelength of 11.06 nm, and a power of 1.6 ± 0.72 W in the fabrication of a device for modulating neural activity in the brain. The device is configured to apply the ultrashortwave to the head of a mouse in a non-invasive manner.

[0026] Example Intervention method: Eight-week-old male C57BL / 6 mice were used. A mouse restraint device was applied to ensure that the mice were awake and received ultra-shortwave stimulation, while the material did not affect the electric field distribution. Figure 1 The ultrashortwave stimulation parameters were: frequency 27.12MHz ± 0.6%, wavelength 11.06 nm, power 1.6 ± 0.72 W, electrode distance from the midline of the mouse's head 5 cm, and bilateral electrode distance kept constant at 10 cm. Each stimulation lasted 10 minutes, with a 12-hour interval between stimulations, for a total of 10 stimulations.

[0027] Security verification: (1) Cranial temperature monitoring showed no obvious thermal effect. Infrared thermal imaging was used to monitor the temperature changes of the mouse brain surface in real time during a 10-minute intervention period. The experiment showed that the temperature distribution in the mouse brain region was uniform during the intervention, and no obvious local heat accumulation was observed. AUC quantitative analysis of the temperature-time curves showed no significant difference in AUC values ​​between the intervention group (LHE) and the control group (Sham) (p>0.05). These results demonstrate that under the intervention parameters used in this experiment, no significant thermal effect was produced. Figure 2 ) (2) Behavioral assessment showed normal motor, sensory, cognitive, and emotional performance. Behavioral testing revealed that, compared with the sham stimulation control group (Ctr group), mice in the ultra-short wave stimulation group (LHE group) spent more time on the rotarod. Figure 3 A), the time for turning the head during the pole climbing experiment, and the total time spent climbing the pole ( Figure 3 .B) No significant difference; the results of the suspension test showed that the LHE group mice had a shorter fall time and a lower overall score reflecting the number of falls and the number of times they reached the platform. Figure 3 .C) showed no significant difference from the sham-stimulated control group; in the open field test, spontaneous activity and anxiety-like behavior in mice showed no significant difference from the sham-stimulated control group. Figure 3 .D); Y Maze ( Figure 3 E) and pain testing ( Figure 3 The results (.F) showed that the cognitive and sensory functions of the mice were not affected by the ultra-short wave.

[0028] (3) The blood-brain barrier function is intact. Evans blue staining showed no significant increase in Evans blue penetration in mice in the shortwave diathermy group (LHE group), and no obvious blue exudation was observed in either top-view or coronal section of the brain, demonstrating that the blood-brain barrier function remained intact. Figure 4 ).

[0029] (4) Ultrashortwave brain stimulation has no cytotoxicity Nissl staining of neurons revealed that neurons in the low-height (LHE) group were arranged in a regular manner and had abundant Nissl bodies within their cells. Figure 5 NeuN and TUNEL staining showed no decrease in cortical neuronal density and no obvious apoptosis, proving that LHE did not cause neuronal damage. Figure 5 B); Myelin staining showed that the myelin sheath structure was intact ( Figure 5 .C); no significant fluctuations in the expression of synaptic structure-related proteins were observed. Figure 5 .D); GFAP and Iba-1 staining showed no significant activation of astrocytes and microglia in the hippocampus region ( .D); Figure 5 (E). The above results demonstrate that the described paradigm of ultrashortwave brain stimulation does not cause neurocytotoxicity or damage to brain structure.

[0030] Verification of neuromodulation effects: (1) Shortwave brain stimulation can regulate brain electrical activity. Changes in electroencephalographic activity in mice at different stimulation time points after shortwave diathermy intervention (SED) Figure 6 A, B). The results showed that after the first stimulation, no changes in EEG activity were observed in the ultra-short wave stimulation group (SW group), but after 10 repeated stimulations, the average power of both the Alpha and Beta bands decreased significantly. Figure 6 (C, D) Power spectral density analysis further revealed that the area under the curve during stimulation was significantly lower than before stimulation, while no significant change was observed in the sham stimulation group. Figure 6 E, F). Fiber optic recording technology was used to monitor calcium activity in cortical neurons (E, F). Figure 6 The results showed that, compared with baseline, the first stimulation did not induce an increase in neuronal calcium signaling, while the 10th stimulation induced a significantly greater amplitude of calcium signaling than the first stimulation. Figure 6 Quantitative analysis confirmed that the area under the calcium signal curve was significantly increased in the 10-time stimulation group during the stimulation period (I). Figure 6 (J), but returned to normal levels immediately after stimulation ended. Furthermore, calcium transient frequency analysis showed that the neuronal activity frequency decreased in the 10-time stimulation group (J). Figure 6 .K).

[0031] C-Fos activation assays of neurons in the cortex, striatum, and hippocampus showed an increase in the number of C-Fos-positive cells in the striatum, indicating neuronal activation. Figure 6 The above results confirm that this ultra-shortwave brain stimulation intervention paradigm can regulate cortical brain electrical power and neuronal calcium activity, activate neurons, and has a neuromodulatory effect.

[0032] (2) Ultrashort wave brain stimulation can regulate neurotransmitter expression. Neurotransmitter detection showed that ultrashortwave brain stimulation had significant brain region specificity in regulating neurotransmitters. The striatum was mainly characterized by overall activation, the cortex by neurotransmitter balance remodeling, and the hippocampus primarily affected the 5-HT energy system. Figure 7 The above results demonstrate that shortwave brain stimulation can modulate the brain's microenvironment.

[0033] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. Application of ultra-short waves in the fabrication of devices for modulating brain neural activity.

2. The application according to claim 1, characterized in that, The regulation of brain neural activity is achieved through the non-thermal effects of ultra-short waves.

3. The application according to claim 1, characterized in that, The ultra-short wave refers to an ultra-short wave with a frequency of 27.12MHz±0.6%, a wavelength of 11.06, and a power of 1.6±0.72W.

4. The application according to claim 1, characterized in that, The device is configured to apply the ultrashort wave to the subject's head in a non-invasive manner.

5. The application according to claim 1, characterized in that, The device is configured to apply ultra-short waves according to a preset stimulation protocol.

6. The application according to claim 5, characterized in that, The preset stimulation program includes each stimulation lasting 10 minutes, with a 12-hour interval between stimulations, for a total of 10 stimulations.

7. The application according to claim 1, characterized in that, The regulation of brain neural activity includes at least one of the following: a) Reduce the average power in the Alpha and Beta bands; b) Regulates neuronal calcium activity; c) Activate neurons in the striatum region; d) Regulate neurotransmitter expression.

8. A device for regulating brain neural activity, characterized in that, The device includes: Ultra-shortwave generation unit, used to generate ultra-shortwave waves; and An ultra-shortwave application unit is used to apply the ultra-shortwave to the subject's head.

9. The device for regulating brain neural activity according to claim 8, characterized in that, The ultrasonic generating unit produces an ultrashort wave with a frequency of 27.12MHz±0.6%, a wavelength of 11.06, and a power of 1.6±0.72W.

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