Animal body electrode myelination electric control system and method thereof

Abstract

The invention provides an animal body electrode myelination electric control system and method, and the system comprises an electrode mechanism which comprises an electrode substrate, and a first electrode, a second electrode, a third electrode, a fourth electrode, a fifth electrode and a sixth electrode which are disposed on the electrode substrate. The electrode mechanism, the resistor and the waveform function generator are sequentially connected in series to form an electric control system. When the electrode mechanism is used, the electrode mechanism is implanted into a to-be-operated object and then fixed on soft tissue of an injured area through an absorbable suture, and the two ends of a series electric control circuit are connected with an external power supply to form an electric control system. The first electrode, the second electrode, the third electrode, the fourth electrode, the fifth electrode and the sixth electrode are externally connected with copper wires for electrode stimulation selection, multi-path and multi-direction electrical stimulation scheme adjustment is achieved, growth of nerve axons can be promoted, remyelination of the axons can be promoted, recovery of nerve functions is accelerated, and popularization and application are facilitated.

Description

Technical Field

[0001] The invention belongs to the technical field of electrical stimulation, and in particular relates to an animal body electrode myelination electrical control system and a method thereof. Background Art

[0002] With the continuous deepening of research on spinal cord injury, existing methods mainly focus on two key points: reconstruction of structure and restoration of function. Remyelination of nerve axons promotes functional recovery and is an important basis for nerves to effectively perform conduction functions. There are currently many methods to try to promote axonal remyelination, such as the introduction of neurotrophic factors and cell transplantation. Although they can promote the formation of myelin to a certain extent, the recovery of myelination function is not ideal, and its efficacy needs to be improved. Studies have shown that spontaneous myelin regeneration of endogenous Schwann cells is a potential therapeutic target for SCI repair, but there is a lack of effective intervention treatments. Therefore, it is urgent to find a suitable treatment method to mobilize endogenous Schwann cells, strengthen and accelerate spontaneous myelin regeneration, and promote functional recovery after SCI. Studies have found that after the central nervous system of rodents, non-human primates and humans is damaged, Schwann cells (Schwanncells) have a significant remyelination effect on central nervous system axons, which is most significant in the dorsal spinal cord. In this case, myelin regeneration mediated by endogenous Schwann cells dominates the SCI injury area. Compared with oligodendrocytes, Schwann cell-mediated spinal axon myelin regeneration is rapid, efficient and stable, which can effectively promote the recovery of motor function after SCI. In addition, the myelin sheath formed by Schwann cells in central nervous system axons has persistence and normal conduction function. This spontaneous endogenous Schwann cell myelin repair also faces the problem of insufficient intrinsic driving force. Therefore, mobilizing endogenous Schwann cells and strengthening and accelerating spontaneous myelin regeneration will hopefully significantly improve the prognosis of SCI.

[0003] Based on this, an animal body electrode myelination electrical control system and method are proposed. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide an animal body electrode myelination electrical control system and method thereof in view of the deficiencies of the above-mentioned prior art, so as to solve the problems raised in the above-mentioned background technology.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is: In a first aspect, an animal body electrode myelination electrical control system includes an electrode mechanism, wherein the electrode mechanism includes an electrode substrate and a first electrode, a second electrode, a third electrode, a fourth electrode, a fifth electrode, and a sixth electrode disposed on the electrode substrate; Among them, a first electrode layout area and a second electrode layout area are provided on the electrode substrate. The first electrode layout area and the second electrode layout area are symmetrically arranged. The first electrode, the second electrode, and the third electrode are arranged in parallel at equal intervals in the first electrode layout area. The fourth electrode, the fifth electrode, and the sixth electrode are arranged in parallel at equal intervals in the second electrode layout area; The electrode mechanism, the resistor, and the waveform function generator are connected in series in sequence to form a series electric control circuit, and an external power supply is connected to both ends of the series electric control circuit to form an electric control system.

[0006] As a further description of the present invention, the electrode substrate is made of polydimethylsiloxane, and the first electrode, the second electrode, the third electrode, the fourth electrode, the fifth electrode, and the sixth electrode are made of titanium alloy.

[0007] As a further description of the present invention, copper wires are externally connected to the first electrode, the second electrode, the third electrode, the fourth electrode, the fifth electrode, and the sixth electrode, and the on-off operation control of the first electrode, the second electrode, the third electrode, the fourth electrode, the fifth electrode, or the sixth electrode can be completed through the copper wires.

[0008] As a further description of the present invention, the second electrode and the fifth electrode have the same length, and for the first electrode, the third electrode, the fourth electrode, and the sixth electrode, the length of the second electrode is greater than the length of the first electrode.

[0009] As a further description of the present invention, the resistance value of the resistor is 1Ω, and an oscilloscope is also connected to both ends of the resistor, and the voltage across the resistor is measured through the oscilloscope.

[0010] In a second aspect, a control method for an electrocontrol system of animal body electrode myelination includes the following steps: After implanting the electrode mechanism into the object to be operated, fix the electrode mechanism on the soft tissue in the damaged area with absorbable sutures, and then externally connect the resistor and the waveform function generator in series in sequence to form a series electric control circuit. Connect an external power supply to both ends of the series electric control circuit to form an electric control system. Use the copper wires externally connected to the first electrode, the second electrode, the third electrode, the fourth electrode, the fifth electrode, and the sixth electrode to perform electrode stimulation selection, with parameters: voltage 2.5V, frequency 100Hz, 1h / d, to complete the electrostimulation operation.

[0011] The present invention has the following advantages compared with the prior art: The electronic control system in the present invention includes an electrode mechanism. The electrode mechanism includes an electrode substrate and a first electrode, a second electrode, a third electrode, a fourth electrode, a fifth electrode, and a sixth electrode disposed on the electrode substrate. The first electrode, the second electrode, and the third electrode are arranged in parallel at equal intervals in the first electrode layout area. The fourth electrode, the fifth electrode, and the sixth electrode are arranged in parallel at equal intervals in the second electrode layout area. During use, after the electrode mechanism is implanted into the object to be operated, the electrode mechanism is fixed to the soft tissue in the damaged area with absorbable sutures. Then, an external resistor and a waveform function generator are connected in series in turn to form a series electronic control circuit, and an external power supply is connected to both ends of the series electronic control circuit to form an electronic control system. The stimulation selection of the electrodes is realized by connecting copper wires to the first electrode, the second electrode, the third electrode, the fourth electrode, the fifth electrode, and the sixth electrode, so as to adjust the multi-path and multi-directional electrical stimulation scheme, which can not only promote the growth of nerve axons, but also promote the remyelination of axons, thus accelerating the recovery of nerve function and facilitating popularization and use. Description of the Drawings

[0012] Figure 1 is a schematic structural diagram of the electrode substrate of the present invention; Figure 2 is a schematic diagram of the results in Experimental Example 1 of the present invention; Figure 3 is a schematic diagram of the results in Experimental Example 2 of the present invention; Figure 4 is a schematic diagram of the results in Experimental Example 3 of the present invention; Figure 5 is a schematic diagram of the results in Experimental Example 4 of the present invention; Figure 6 is a schematic diagram of the results in Experimental Example 5 of the present invention; Figure 7 is a schematic diagram of the results in Experimental Example 6 of the present invention.

[0013] Description of the Reference Numerals: 1 - Electrode substrate; 11 - First electrode layout area; 12 - Second electrode layout area; 2 - First electrode; 3 - Second electrode; 4 - Third electrode; 5 - Fourth electrode; 6 - Fifth electrode; 7 - Sixth electrode. Detailed Embodiments

[0014] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0015] Such as Figure 1As shown in the figure, the present invention provides a technical solution: an electro-control system for myelinization of an animal body electrode, including an electrode mechanism. The electrode mechanism includes an electrode substrate 1 and a first electrode 2, a second electrode 3, a third electrode 4, a fourth electrode 5, a fifth electrode 6, and a sixth electrode 7 disposed on the electrode substrate 1; Among them, a first electrode layout area 11 and a second electrode layout area 12 are provided on the electrode substrate 1. The first electrode layout area 11 and the second electrode layout area 12 are symmetrically arranged. The first electrode 2, the second electrode 3, and the third electrode 4 are arranged in parallel at equal intervals in the first electrode layout area 11, and the fourth electrode 5, the fifth electrode 6, and the sixth electrode 7 are arranged in parallel at equal intervals in the second electrode layout area 12; The first electrode 2, the second electrode 3, the third electrode 4, the fourth electrode 5, the fifth electrode 6, and the sixth electrode 7 are all externally connected with copper wires, and the on-off operation control of the first electrode 2, the second electrode 3, the third electrode 4, the fourth electrode 5, the fifth electrode 6, or the sixth electrode 7 can be completed through the copper wires.

[0016] For convenient cooperation, the second electrode 3 and the fifth electrode 6 have the same length. The first electrode 2, the third electrode 4, the fourth electrode 5, and the sixth electrode 7, and the length of the second electrode 3 is greater than the length of the first electrode 2.

[0017] The electrode substrate 1 is made of polydimethylsiloxane, and the first electrode 2, the second electrode 3, the third electrode 4, the fourth electrode 5, the fifth electrode 6, and the sixth electrode 7 are made of titanium alloy, which can significantly improve the electric energy transmission efficiency and reduce crosstalk pollution, and realize the adjustment of multi-path and multi-directional electrical stimulation schemes by controlling the discharge process of the electrodes in the first electrode layout area 11 and the second electrode layout area 12.

[0018] The electrode mechanism, the resistor, and the waveform function generator are sequentially connected in series to form a series electro-control circuit, and an external power supply is connected to both ends of the series electro-control circuit to form an electro-control system.

[0019] The resistance value of the resistor is 1Ω, and an oscilloscope is also connected to both ends of the resistor, and the voltage across the resistor is measured through the oscilloscope.

[0020] The control method of the above-mentioned electro-control system for myelinization of an animal body electrode includes the following steps: After implanting the electrode mechanism into the subject to be operated on, fix the electrode mechanism on the soft tissue in the damaged area with absorbable sutures. Then, externally connect a resistor and a waveform function generator in series to form a series electric control circuit, and connect an external power supply across the series electric control circuit to form an electric control system. Use the copper wires externally connected to the first electrode 2, the second electrode 3, the third electrode 4, the fourth electrode 5, the fifth electrode 6, and the sixth electrode 7 to select the stimulation of the electrodes. The parameters are: voltage 2.5V, frequency 100Hz, 1h / d, and complete the electrical stimulation operation.

[0021] Experimental example 1, the myelinization electric control system has good biological safety. To test the safety of the myelinization electric control system, electrodes were implanted into the exposed spinal cord at the T8-10 level, and then the electrodes were implanted above the spinal cord. After 4 weeks of electrical stimulation, HE staining was performed to observe the pathological characteristics. Compared with the Control group, HE staining of the main organs, heart, liver, spleen, lung, and kidney, as well as the spinal cord, in the ES group of rats did not show any pathological abnormalities. As Figure 2 shown; Figure 2 Among them, (A) HE staining of the main organs; (Bar = 200μm) (B) HE staining of the spinal cord; (Bar = 100μm) (C), (D) ELISA method to detect the secretion of inflammatory factors TNF-α and IL-1α in spinal cord tissue; (E), (F) detect the secretion of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). n = 5, ns means no statistical difference; To verify whether there is an inflammatory reaction in the spinal cord tissue at the stimulation site after electrical stimulation, by detecting the secretion of spinal cord inflammatory factors TNF-α and IL-1α, it shows that the secretion levels of TNF-α and IL-1α in the spinal cord in the ES group have no obvious difference from those in the Control group, which indicates that ES does not cause cell inflammation and is a safe physical stimulation method. In addition, the systemic toxicity after electrode implantation was tested. The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the ES group of rats have no difference compared with the Control group, indicating that the implanted electrodes do not cause systemic toxicity.

[0022] Experimental example 2, the myelinization electric control system can promote the functional recovery of animals with spinal cord injury. The BBB scale and footprint were used to comprehensively evaluate the effect of ES on the motor function recovery after spinal cord injury. The results of the BBB score are as Figure 3 shown; Figure 3 Among them, (A) Motor function of each group using the BBB score; (B) Representative footprint images of each group; (C) Average step length statistics. n = 5, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, The results showed that starting from the 7th day, the motor functions of all SCI groups recovered to varying degrees; on the 14th day, the scores of the SCI-ES group and the SCI-ES-SCs group were higher than those of the SCI group, and the differences among the three were statistically significant. Moreover, the score of the SCI-ES-SCs group was the highest, which might be related to the neuroprotection and inflammatory regulation of nutritional factors in the subacute phase after cell transplantation; on the 28th day, the score of the SCI-ES-SCs group was the highest, followed by the SCI-ES group, and the SCI group was the lowest, with statistically significant differences. The above results indicate that ES can significantly promote the improvement of motor function, while ES-SCs cell transplantation can better promote the recovery of motor function in rats. The footprint showed similar results: on the 28th day, the scores of the SCI-ES-SCs group and the SCI-ES group were higher than those of the SCI group, and the difference between the SCI-ES-SCs group and the SCI group was statistically significant.

[0023] Experimental Example 3: The myelinization electric control system can promote the remyelination of axons after spinal cord injury. The spinal cord tissues at 7, 14, and 28 days after spinal cord injury were taken for MBP / neurofilament NF200 immunofluorescence staining. MBP is a specific biomarker for myelin differentiation, and NF200 is a specific biomarker for neurofilaments, which are important indicators for evaluating myelin regeneration and axon regeneration. Thus, the axonal remyelination situation after SCI-ES-SCs treatment was observed. Myelin is the membrane that wraps axons and can act as an insulator to prevent electrical signals from passing from one axon to another. The formation of myelin at the nerve injury site provides a protective barrier for axon regeneration. The results are as Figure 4 shown Figure 4 in (A) Immunofluorescence staining of MBP (red, myelin basic protein marker) and NF200 (green, neurofilament marker) in spinal cord tissues at 7, 14, and 28 days after spinal cord injury (Bar = 500μm); (B) Statistics of the MBP+ immunofluorescence intensity in spinal cord tissues; (C) Statistics of the NF200+ immunofluorescence intensity in spinal cord tissues. n = 5, ns indicates no statistical difference, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, The results showed that on the 7th day after injury, the fluorescence staining intensity of MBP+ / NF200+ in the SCI-ES-SCs group was significantly higher than that in the SCI and SCI-ES groups, while there was no obvious difference between the SCI and SCI-ES groups. On the 14th day after injury, the fluorescence staining intensity of NF200+ in the SCI-ES-SCs group was significantly higher than that in the SCI and SCI-ES groups, and the fluorescence staining intensity of the SCI-ES group was also higher than that of the SCI group. On the 28th day after injury, more myelin regeneration was observed in the SCI-ES-SCs group compared with the SCI group, and the differences were all statistically significant.

[0024] Experimental Example 4. The myelinization electro-control system can promote nerve regeneration after spinal cord injury. Spinal cord tissues at 7, 14, and 28 days after spinal cord injury were taken for neuron Tuj1 immunofluorescence staining to observe the growth of neurons after SCI-ES-SCs treatment. The results are as Figure 5 shown, Figure 5 in (A) immunofluorescence staining of Tuj1 (red, neuron marker) at 7, 14, and 28 days after spinal cord injury; (Bar = 1000μm) (B) statistical analysis of the average fluorescence intensity of Tuj1+ cells. n = 5, ns indicates no statistical difference, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, The results showed that a small number of neuron regenerations could be observed in the injury area of the SCI-ES-SCs group at 7 days after injury. At 14 days after injury, the expression of Tuj1+ in the SCI-ES-SCs group was significantly higher than that in the SCI and SCI-ES groups, and the fluorescence staining intensity of Tuj1+ in both ES groups was enhanced compared with that in the SCI group, indicating that ES has an obvious effect on nerve regeneration. By 28 days, the neurons in the injury area became denser. Compared with the SCI group, the expression of neurons in the SCI-ES and SCI-ES-SCs groups was significantly increased, and the difference was statistically significant.

[0025] Experimental Example 5. The myelinization electro-control system inhibits the activation of astrocytes and reduces the formation of scars. The distribution and activation of microglia / macrophages and astrocytes in spinal cord tissues at 7, 14, and 28 days after spinal cord injury were detected, as Figure 6 shown, Figure 6 in (A) immunofluorescence staining of astrocytes GFAP (green, astrocyte marker) and microglia Iba1 (red, microglia marker) in spinal cord tissues at 7, 14, and 28 days after spinal cord injury (Bar = 400μm); (B) statistical analysis of the immunofluorescence intensity of GFAP+ in spinal cord tissues; (C) statistical analysis of the immunofluorescence intensity of Iba1+ in spinal cord tissues. n = 5, ns indicates no statistical difference, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0026] The results showed that after spinal cord injury, the normal and intact tissue structure was damaged, and microglia / macrophages began to activate and proliferate. The number of microglia reached its peak at 7 days. When the inflammation entered the subacute phase, astrocytes began to take over microglia / macrophages, and the expression of GFAP was significantly up-regulated. The two types of glial cells jointly participated in the formation of scars. By 28 days, a syrinx had formed and the scar structure had become denser. Compared with the SCI group, both SCI-ES and SCI-ES-SCs could down-regulate the expression levels of IBA1 and GFAP after spinal cord injury and reduce scar formation.

[0027] Experimental Example 6. The myelinization electric control system can inhibit the secretion of inflammatory factors. Neurotoxic astrocytes are induced by C1q, IL-1α, and TNF-α secreted by activated microglia. To explore whether SCI-ES-SCs inhibit astrocyte activation in the subacute phase by reducing the cytokines secreted by microglia or by directly acting on astrocytes themselves. The protein expression levels in the spinal cord tissue at 7, 14, and 28 days after SCI were detected by ELISA, as Figure 7 shown, Figure 7 in which, n = 5, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; The results showed that the levels of C1q, IL-1α, and TNF-α in the SCI group were significantly higher than those in the Sham group, while those in the SCI-ES and SCI-ES-SCs groups were lower than those in the SCI group, indicating that SCI-ES and SCI-ES-SCs can reduce the up-regulation of inflammatory factors to a certain extent.

[0028] It should be noted that in this article, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the term "comprising", "including" or any other variant thereof is intended to cover non-exclusive inclusion, so that a process, method, article or device comprising a series of elements not only includes those elements, but also includes other elements not expressly listed, or also includes elements inherent to such process, method, article or device.

[0029] Although the embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made in these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. An animal body electrode myelination electric control system, characterized in that: The invention comprises an electrode structure, wherein the electrode structure comprises an electrode substrate (1) and a first electrode (2), a second electrode (3), a third electrode (4), a fourth electrode (5), a fifth electrode (6) and a sixth electrode (7) arranged on the electrode substrate (1); The electrode substrate (1) is provided with a first electrode layout area (11) and a second electrode layout area (12); the first electrode layout area (11) and the second electrode layout area (12) are symmetrically arranged; the first electrode (2), the second electrode (3), and the third electrode (4) are arranged in parallel at equal intervals in the first electrode layout area (11); and the fourth electrode (5), the fifth electrode (6), and the sixth electrode (7) are arranged in parallel at equal intervals in the second electrode layout area (12); The electrode mechanism, the resistor and the waveform function generator are sequentially connected in series to form a series electric control circuit, and an external power supply is connected to both ends of the series electric control circuit to form an electric control system.

2. The animal body electrode myelination electric control system according to claim 1, characterized in that: The electrode substrate (1) is made of polydimethylsiloxane, and the first electrode (2), the second electrode (3), the third electrode (4), the fourth electrode (5), the fifth electrode (6) and the sixth electrode (7) are made of titanium alloy.

3. The animal body electrode myelination electric control system according to claim 1, characterized in that: The first electrode (2), the second electrode (3), the third electrode (4), the fourth electrode (5), the fifth electrode (6) and the sixth electrode (7) are all externally connected to copper wires, and the on-off operation control of the first electrode (2), the second electrode (3), the third electrode (4), the fourth electrode (5), the fifth electrode (6) or the sixth electrode (7) can be completed via the copper wires.

4. The animal body electrode myelination electric control system according to claim 1, characterized in that: The second electrode (3) and the fifth electrode (6) have the same length as the first electrode (2), the third electrode (4), the fourth electrode (5) and the sixth electrode (7), and the length of the second electrode (3) is greater than the length of the first electrode (2).

5. The animal body electrode myelination electric control system according to claim 1, characterized in that: The resistance value of the resistor is 1Ω, and an oscilloscope is connected to both ends of the resistor to measure the voltage across the resistor.

6. A control method for an animal body electrode myelination electric control system according to any one of claims 1 to 5, characterized in that: The following steps are involved: After the electrode mechanism is implanted in the object to be operated, the electrode mechanism is fixed to the soft tissue of the damaged area with an absorbable suture, and then an external resistor and a waveform function generator are connected in series in sequence to form a series electric control circuit. An external power supply is connected to both ends of the series electric control circuit to form an electric control system. The first electrode (2), the second electrode (3), the third electrode (4), the fourth electrode (5), the fifth electrode (6) and the sixth electrode (7) are connected with an external copper wire to select the electrode stimulation. The parameters are: voltage 2.5V, frequency 100Hz, 1h / d, to complete the electric stimulation operation.

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