System for carrying out intracellular electric treatment based on ultrahigh-frequency ultrashort pulse and application of system
The ultra-high frequency and ultra-short pulse electric field system enables precise regulation of the cell membrane structure, solving the problems of insufficient target selectivity and regulation ability in existing technologies, and is suitable for precision medicine of tumor cells.
Patent Information
- Application Number
- CN202511278460.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-08
- Publication Date
- 2025-12-12
AI Technical Summary
Existing electrical pulse technology has shortcomings in target selectivity, energy utilization efficiency, and real-time control capabilities, and cannot precisely control different cell death modes, thus limiting its application in precision medicine.
The system employs an ultra-high frequency ultra-short pulse electric field system, which outputs unipolar or bipolar pulse electric fields of 10-100ns and 0.5-10MHz through a high-voltage ultra-short pulse generation module. Combined with a control module and an adherent cell processing module, it achieves targeted regulation of the intracellular membrane structure and precisely induces modes such as ferroptosis, apoptosis, and immunogenic cell death.
It achieves precise regulation of specific organelles, is suitable for different treatment needs, and has the advantages of being drug-free and reversible. It has low temperature rise and avoids systemic toxicity, making it suitable for precision medicine of tumor cells.
Smart Images
Figure CN121109129A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of bioelectric medicine and cell therapy technology, specifically relating to a system for intracellular electrical processing based on ultra-high frequency and ultra-short pulses and its application. Background Technology
[0002] Cellular electromodulation technology is a class of biotechnologies that intervene in cellular behavior by applying external electric fields or currents. Its core lies in using electrical signals to influence the physiological state of cells. This type of technology has broad application prospects in basic research, disease treatment, and bioengineering.
[0003] Traditional irreversible electroporation (IRE) and nanosecond pulsed electric field (nsPEF) techniques are widely used for cell electroregulation. Traditional IRE primarily uses microsecond-level pulses of 100-500 μs combined with a moderate field strength of 0.5-3 kV / cm to induce cell death by disrupting cell membrane integrity. While traditional IRE has been successfully applied in clinical settings such as tumor ablation, its effect is limited to the cell membrane level, failing to achieve targeted regulation of organelles such as mitochondria and endoplasmic reticulum. The recently developed nanosecond pulsed electric field (nsPEF) technique shortens the pulse width to 200-500 ns and increases the field strength to 5-100 kV / cm, theoretically enabling it to penetrate the cell membrane and affect the endometrial system; studies have reported its ability to induce apoptosis. However, in practical applications, nanosecond pulsed electric field technology still suffers from insufficient precision in regulating specific cell death pathways (such as ferroptosis and immunogenic cell death), and systematic parameter-effect relationship studies are lacking. Further technological exploration has led to the development of high-frequency pulse techniques (such as H-FIRE) that attempt to reduce muscle stimulation using bipolar pulse sequences of 10-100 kHz. However, these techniques still rely on microsecond-level pulse widths, resulting in limited intracellular effects. Meanwhile, while chemical induction methods can achieve specific cell death patterns (such as Erastin-induced ferroptosis), they suffer from inherent drawbacks such as low drug delivery efficiency and high off-target toxicity.
[0004] In summary, existing electrical pulse technology has significant shortcomings in terms of target selectivity, energy utilization efficiency, and real-time control capabilities. In particular, it lacks an intelligent system that can precisely control different cell death modes according to treatment needs, which severely limits its application prospects in precision medicine. Summary of the Invention
[0005] In view of this, to overcome the shortcomings of existing technologies such as single target and uncontrollable cell death modes, this invention proposes a system and its application based on ultra-high frequency and ultra-short pulse intracellular electrical treatment. This invention utilizes ultra-high frequency and ultra-short pulse electric fields to target and regulate the intracellular membrane structure, precisely controlling different cell death modes, such as ferroptosis, apoptosis, and immunogenic cell death, according to therapeutic needs, which is beneficial for precision medicine.
[0006] One of the objectives of this invention is to provide a system for intracellular electrical processing based on ultra-high frequency and ultra-short pulses.
[0007] To achieve the above objectives, the present invention adopts the following technical solution:
[0008] A system for intracellular electrical processing based on ultra-high frequency ultrashort pulses includes a high-voltage ultrashort pulse generation module, a control module, and an adherent cell processing module. The high-voltage ultrashort pulse generation module is connected to both the control module and the adherent cell processing module. The control module presets multiple pulse parameter combinations, and the high-voltage ultrashort pulse generation module outputs a pulsed electric field with preset parameters according to the instructions issued by the control module, which is then applied to organelles through the adherent cell processing module.
[0009] Preferably, the high-voltage ultrashort pulse generation module is used to output a unipolar or bipolar pulse electric field with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and an amplitude of 0-±5kV, and a waveform rise / fall time of <5ns.
[0010] Preferably, the control module presets pulse electric field parameters according to different cell death modes.
[0011] As preferred, the different cell death modes and their pulsed electric field parameters are as follows:
[0012] The ferrodeath induced mode pulse electric field parameter combination includes: field strength of 20kV / cm (i.e., amplitude of 2kV), pulse width of 50ns, unipolarity, pulse sequence frequency of 10MHz, and number of pulse sequences of 50.
[0013] The necrosis-induced apoptosis mode uses pulsed electric field parameters including: field strength of 30 kV / cm (i.e., amplitude of 3 kV), pulse width of 100 ns, unipolarity, pulse sequence frequency of 0.5 MHz, and 20 pulse sequences.
[0014] The pyroptosis-inducing mode uses pulsed electric field parameters including: field strength of 10 kV / cm (i.e., amplitude of 1 kV), pulse width of 15 ns, bipolarity, pulse sequence frequency of 2 MHz, and number of pulse sequences of 100.
[0015] The apoptosis-inducing mode uses pulsed electric field parameters including: field strength of 50 kV / cm (i.e., amplitude of 5 kV), pulse width of 80 ns, bipolarity, pulse sequence frequency of 1 MHz, and 100 pulse sequences.
[0016] Preferably, the adherent cell processing module is a microelectrode array, which uses an FR-4 epoxy resin substrate as an insulating carrier and has a metal conductive layer on its surface to form an electrode structure.
[0017] Preferably, the electrodes in the microelectrode array are metal electrodes or poly(3,4-ethylenedioxythiophene) / polystyrene sulfonate (PEDOT:PSS) conductive polymer patches.
[0018] Preferably, the electrodes adopt a finger-shaped cross design, with a single electrode width of 130-180 micrometers and an electrode spacing of 200-300 micrometers.
[0019] Preferably, the width of a single electrode is 150 micrometers and the spacing between electrodes is 250 micrometers.
[0020] Preferably, the high-voltage ultrashort pulse generation module is connected to the adherent cell processing module via a wire.
[0021] Preferably, the cell is a cancer cell, including one or more of glioma cells, colon cancer cells, and pancreatic cancer cells; the organelles include mitochondria and endoplasmic reticulum.
[0022] A second objective of this invention is to provide the application of the aforementioned system and / or pulsed electric field based on ultra-high frequency and ultra-short pulses for intracellular electrical treatment in the preparation of products for inducing ferroptosis, necroptosis, pyroptosis and / or apoptosis in tumor cells.
[0023] To achieve the above objectives, the present invention adopts the following technical solution:
[0024] The aforementioned system and / or pulsed electric field are used in the preparation of products for inducing ferroptosis, necroptosis, pyroptosis and / or apoptosis in tumor cells, wherein the tumor includes any one or more of glioma, colon cancer, and pancreatic cancer.
[0025] Preferably, the pulsed electric field is a unipolar or bipolar pulsed electric field with a pulse width of 10-100ns, a frequency of 0.5-10MHz, an amplitude of 0-±5kV, and a waveform rise / fall time of <5ns.
[0026] As a preferred embodiment, the pulsed electric field parameters induced by ferroptosis are: field strength of 20kV / cm (i.e., amplitude of 2kV), pulse width of 50ns, unipolarity, pulse sequence frequency of 10MHz, and number of pulse sequences of 50.
[0027] The pulsed electric field parameters induced by necrosis and apoptosis were: field strength of 30 kV / cm (i.e., amplitude of 3 kV), pulse width of 100 ns, unipolarity, pulse sequence frequency of 0.5 MHz, and number of pulse sequences of 20.
[0028] The pulsed electric field parameters induced by pyroptosis were: field strength of 10 kV / cm (i.e., amplitude of 1 kV), pulse width of 15 ns, bipolarity, pulse sequence frequency of 2 MHz, and number of pulse sequences of 100.
[0029] The parameters of the apoptosis-induced pulsed electric field were: field strength of 50 kV / cm (i.e., amplitude of 5 kV), pulse width of 80 ns, bipolarity, pulse sequence frequency of 1 MHz, and number of pulse sequences of 100.
[0030] Preferably, the products include, but are not limited to, medical devices / devices, pharmaceutical compositions, and fields.
[0031] Preferably, the system is used in the preparation of devices for inducing tumor cell ferroptosis, necrosis-apoptosis, pyroptosis, and / or apoptosis; or in the preparation of pulsed electric fields for inducing tumor cell ferroptosis, necrosis-apoptosis, pyroptosis, and / or apoptosis.
[0032] A third objective of this invention is to provide another application of the aforementioned system for intracellular electrical processing based on ultra-high frequency and ultra-short pulses and / or pulsed electric fields.
[0033] To achieve the above objectives, the present invention adopts the following technical solution:
[0034] The aforementioned system and / or pulsed electric field are used in the preparation of products for promoting lipid ROS levels, inhibiting GPX4 protein expression and / or inducing iron-dependent lipid peroxidation; or the aforementioned system and / or pulsed electric field are used in the preparation of products for promoting IL-1β and IL-18 expression; or the aforementioned system and / or pulsed electric field are used in the preparation of products for promoting mitochondrial membrane potential collapse and / or activating caspase.
[0035] Preferably, the products include, but are not limited to, medical devices / devices, pharmaceutical compositions, and fields.
[0036] Preferably, the pulsed electric field is a unipolar or bipolar pulsed electric field with a pulse width of 10-100ns, a frequency of 0.5-10MHz, an amplitude of 0-±5kV, and a waveform rise / fall time of <5ns.
[0037] Preferably, the system and / or pulsed electric field induce ferroptosis in tumor cells by increasing lipid ROS levels and decreasing glutathione peroxidase 4 (GPX4) activity, thereby inducing iron-dependent lipid peroxidation. The system and / or pulsed electric field induce necroptosis and apoptosis in tumor cells by activating the RIPK1 / RIPK3 / MLKL signaling pathway, leading to progressive loss of cell membrane integrity. The system and / or pulsed electric field induces pyroptosis in tumor cells by activating proteins and inducing the massive release of IL-1β and IL-18. The system and / or pulsed electric field induces apoptosis in tumor cells by triggering mitochondrial membrane potential collapse and activating caspases.
[0038] The beneficial effects of this invention are as follows:
[0039] 1. This invention uses high-frequency (0.5–10MHz) ultrashort pulses (10–100ns), whose unique electric field distribution characteristics can penetrate the cell membrane without causing permanent damage and directly act on specific organelles such as mitochondria, endoplasmic reticulum, and lysosomes.
[0040] 2. This invention can flexibly induce four major programmed cell death modes by adjusting the combination of pulse parameters, which is suitable for different treatment needs and contributes to precision medicine.
[0041] 3. Traditional chemotherapy or targeted drugs are often accompanied by problems such as systemic toxicity and drug resistance, while this invention uses pure physical energy regulation, which has the advantages of no drug dependence and reversible regulation;
[0042] 4. Traditional microsecond pulses cause a temperature rise of >2℃, while the present invention uses a high-frequency ultrashort pulse electric field with a temperature rise of <0.5℃. Attached Figure Description
[0043] Figure 1 This is a structural diagram of the ultra-high frequency ultra-short pulse intracellular electrochemical treatment system according to an embodiment of the present invention, wherein 100 is a control module, 200 is a high voltage short pulse generation module, and 300 is an adherent cell treatment module;
[0044] Figure 2 A diagram showing the integration of a microelectrode array into an adherent cell processing module; where, Figure 2 -A is a front view of the integrated microelectrode array. The interdigitated electrodes have a gold plating to reduce cytotoxicity. The width of a single finger electrode is 150 micrometers and the interdigital spacing is 250 micrometers. Figure 2 -B is a schematic diagram of the back of the integrated microelectrode array. The pulse generator is connected to the back through the circular solder joints to enable the front interdigitated electrode array to apply an ultra-high frequency and ultra-short pulse electric field to the cell.
[0045] Figure 3 The image shows the results of human glioma cells U87-MG after treatment with a high-frequency ultrashort pulse electric field (20kV / cm field strength, 50ns pulse width, unipolar, 10MHz pulse sequence frequency, 0-50 pulse sequences) and the cell viability as assessed by PI / CalceinAM.
[0046] Figure 4 The image shows the results of detecting the expression of GPX4 protein, a marker of ferroptosis, in residual cells of human glioma cells U87-MG after treatment with a high-frequency ultrashort pulse electric field (20kV / cm field strength, 50ns pulse width, unipolarity, 10MHz pulse sequence frequency, 0-50 pulse sequences).
[0047] Figure 5The image shows the results of HCT116 colon cancer cells after treatment with a high-frequency ultrashort pulse electric field (30kV / cm field strength, 100ns pulse width, unipolar, 0.5MHz pulse sequence frequency, 0-20 pulse sequences), and the cell viability and mortality status as assessed by PI / CalceinAM.
[0048] Figure 6 The figure shows the detection results of RIPK1 protein content in residual cell protein precipitate after HCT116 colon cancer cells were treated with a high-frequency ultrashort pulse electric field (30kV / cm field strength, 100ns pulse width, unipolarity, 0.5MHz pulse sequence frequency, 0-20 pulse sequences).
[0049] Figure 7 The image shows the results of pancreatic cancer cells Panc-1 after treatment with a high-frequency ultrashort pulse electric field (10kV / cm field strength, 15ns pulse width, bipolar, 2MHz pulse sequence frequency, 0-100 pulse sequences) and the cell viability as assessed by PI / CalceinAM.
[0050] Figure 8 The figure shows the results of detecting the expression level of Gasdermin-D protein, a pyroptosis marker, in residual cells of pancreatic cancer cells Panc-1 after treatment with a high-frequency ultrashort pulse electric field (10kV / cm field strength, 15ns pulse width, bipolar, 2MHz pulse sequence frequency, 0-100 pulse sequences). Detailed Implementation
[0051] The technical solution of the present invention will be described more clearly and completely below with reference to specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Therefore, based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the protection scope of the present invention.
[0052] In the description of this invention, unless otherwise stated, "a plurality of" means two or more; the terms "upper," "lower," "left," "right," "inner," "outer," "front end," "rear end," "head," "tail," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing the invention and simplifying the description, and do not indicate or imply that the system, device, or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of the invention. Furthermore, the terms "first," "second," "third," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance.
[0053] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "connected" and "linked" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.
[0054] The system and principles of the present invention will now be further explained with reference to the accompanying drawings.
[0055] In some embodiments, such as Figure 1 As shown, the system for intracellular electrochemical treatment based on ultra-high frequency ultrashort pulses of the present invention includes a high-voltage ultrashort pulse generation module 200, a control module 100, and an adherent cell treatment module 300; the high-voltage ultrashort pulse generation module 200 is connected to the control module 100 and the adherent cell treatment module 300 respectively; the control module 100 presets multiple pulse parameter combinations, and the high-voltage ultrashort pulse generation module 200 outputs a pulse electric field with preset parameters according to the instructions issued by the control module 100, and acts on organelles through the adherent cell treatment module 300.
[0056] In some embodiments, such as Figure 2 As shown, the adherent cell processing module 300 integrates a microelectrode array using printed circuit board technology, with its structure designed using specialized circuit design software. The array uses a 1 mm thick FR-4 epoxy resin substrate as the insulating carrier, with a 40 μm thick conductive metal layer covering the surface to form the electrode structure. The electrodes employ a finger-like, interlaced design, with a single electrode width of 150 μm and an inter-electrode spacing of 250 μm. This design ensures a uniform distribution of the electric field. The conductive layer undergoes a special gold plating treatment, ensuring good biocompatibility while effectively avoiding the potential toxicity of copper ions.
[0057] In some embodiments, the high-voltage ultrashort pulse generation module 200 is connected to the adherent cell processing module 300 via a wire.
[0058] In some embodiments, the high-voltage ultrashort pulse generator module 200 is used to output a unipolar or bipolar pulsed electric field with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and an amplitude of 0-±5kV, and a waveform rise / fall time of <5ns.
[0059] In some embodiments, the control module 100 presets pulse electric field parameters according to different cell death modes.
[0060] In some embodiments, the pulsed electric field parameters induced by ferroptosis are: field strength of 20 kV / cm (i.e., amplitude of 2 kV), pulse width of 50 ns, unipolarity, pulse sequence frequency of 10 MHz, and number of pulse sequences of 50.
[0061] In some embodiments, the pulsed electric field parameters induced by necrosis and apoptosis are: field strength of 30 kV / cm (i.e., amplitude of 3 kV), pulse width of 100 ns, unipolarity, pulse sequence frequency of 0.5 MHz, and number of pulse sequences of 20.
[0062] In some embodiments, the pulsed electric field parameters induced by pyroptosis are: field strength of 10 kV / cm (i.e., amplitude of 1 kV), pulse width of 15 ns, bipolarity, pulse sequence frequency of 2 MHz, and number of pulse sequences of 100.
[0063] In some embodiments, the apoptosis-induced pulsed electric field parameters are: field strength of 50 kV / cm (i.e., amplitude of 5 kV), pulse width of 80 ns, bipolarity, pulse sequence frequency of 1 MHz, and number of pulse sequences of 100.
[0064] Example 1. Ultra-high frequency and ultra-short pulse intracellular electrical processing system
[0065] (1) System Composition
[0066] High voltage ultrashort pulse generator: outputs nanosecond-level (10-100ns), high-frequency (0.5-10MHz) electrical pulses, supports unipolar (0-5kV) or bipolar (0-±5kV) waveforms, and waveform rise / fall time <5ns.
[0067] Control terminal: Customizable pulse parameters (pulse width, frequency, field strength, polarity), supports preset treatment protocols (such as ferroptosis mode, apoptosis mode).
[0068] Adherent cell processing module: integrates a microelectrode array, adapted for adherent cell processing, ensuring uniform electric field coverage of target cells.
[0069] (2) Working principle
[0070] The working principle of this system is based on the specific regulatory effect of ultra-high frequency and ultra-short pulse electric fields on the intracellular membrane system. The system first generates high-precision electrical pulses with a pulse width of 10-100ns and a frequency of 0.5-10MHz through a high-voltage pulse generator. These pulses have extremely fast rise times (<5ns) and adjustable amplitudes (0-±5kV), which can penetrate the cell membrane without causing permanent perforation and directly act on key organelles such as mitochondria and endoplasmic reticulum within the cell.
[0071] When a pulsed electric field acts on target cells, its unique high-frequency component creates a non-uniform electric field distribution within the cell, producing selective effects at the subcellular scale. Depending on different treatment needs, the system can intelligently adjust the combination of pulse parameters: ① Ferraphobia induction mode: significantly increases lipid ROS levels, reduces glutathione peroxidase 4 (GPX4) activity, and induces iron-dependent lipid peroxidation (parameter example: 20kV / cm (amplitude 2kV), 50ns, unipolarity, pulse sequence frequency 10MHz, number of pulse sequences 50); ② Necrosis and apoptosis induction mode: activates the RIPK1 / RIPK3 / MLKL signaling pathway, causing progressive loss of cell membrane integrity (parameter example: 30kV / cm (amplitude 3kV), 10ns, 5 ... ① 0 ns, unipolar, pulse sequence frequency 0.5 MHz, number of pulse sequences 20); ② Pyroptosis induction mode: significantly activates proteins, inducing massive release of IL-1β and IL-18 (parameter example: 10 kV / cm (i.e., amplitude 1 kV), 15 ns, bipolar, pulse sequence frequency 2 MHz, number of pulse sequences 100); ③ Apoptosis induction mode: triggers mitochondrial membrane potential collapse and caspase activation (parameter example: 50 kV / cm (i.e., amplitude 5 kV), 80 ns, bipolar, pulse sequence frequency 1 MHz, number of pulse sequences 100).
[0072] Example 2
[0073] (1) In vitro tumor cell ferroptosis induction experiment
[0074] Results of in vitro tumor cell ferroptosis induction experiment as follows Figure 3 and Figure 4 As shown. Human glioma cells U87-MG were injected at 1×10⁻⁶. 5 Cells were seeded at a density of 1000 cells / mL in 6-well plates and treated with a high-frequency ultrashort pulse electric field of 20 kV / cm (i.e., 2 kV pulse amplitude, 1 mm electrode spacing), 50 ns pulse width, unipolarity, 10 MHz pulse sequence frequency, and 50 pulse sequences. 24 hours after treatment, C11-BODIPY fluorescent probe detection showed a significant increase in lipid ROS levels compared to the control group (fluorescence intensity increased 3.5-fold). Western blot analysis showed a 52% decrease in GPX4 protein expression, and electron microscopy revealed characteristic mitochondrial shrinkage and increased membrane density. These results confirm that this parameter combination can effectively induce iron-dependent lipid peroxidation, achieving specific induction of ferroptosis.
[0075] (2) Activation experiment of necrosis-apoptosis signaling pathway
[0076] Results of the necroptosis signaling pathway activation experiment are as follows Figure 5 and Figure 6As shown in the figure, HCT116 colon cancer cells were seeded in confocal culture dishes and treated with parameters of 30 kV / cm field strength (i.e., 3 kV pulse amplitude, 1 mm electrode spacing), 100 ns pulse width, unipolarity, 0.5 MHz pulse sequence frequency, and 20 pulse sequences. Immunofluorescence staining showed that 6 hours after treatment, the formation rate of the RIPK1 / RIPK3 complex increased by 4.2-fold, and the phosphorylation level of MLKL protein increased by 3.8-fold. Live-cell imaging dynamic observation revealed a progressive loss of cell membrane integrity (PI-positive cell rate reached 65% at 12 hours), but the intracellular ATP level remained at 55% of that in the control group. These characteristic changes are consistent with the typical biological manifestations of necrosis and apoptosis.
[0077] (3) Verification experiment on the pyroptosis effect of pancreatic cancer cells Panc-1
[0078] The results of the validation of the pyroptosis effect in pancreatic cancer cells Panc-1 are as follows: Figure 7 and Figure 8 As shown, the parameters used were 10 kV / cm field strength (i.e., 1 kV pulse amplitude, 1 mm electrode spacing), 15 ns pulse width, bipolarity, 2 MHz pulse sequence frequency, and 100 pulse sequences. ELISA analysis showed that IL-1β and IL-18 secretion in the culture supernatant increased by 15-fold and 12-fold, respectively, after 24 hours of treatment. Confocal microscopy revealed that the gasdermin D protein pore formation rate was as high as 83%, and the cells exhibited obvious swelling and rupture morphology. This experiment confirms that ultrashort pulses can efficiently induce immunogenic pyroptosis by activating the inflammasome pathway.
[0079] Example 3
[0080] Replacing traditional metal electrodes with a poly(3,4-ethylenedioxythiophene) / polystyrene sulfonate (PEDOT:PSS) conductive polymer patch reduced the surface impedance from 50Ω to 15Ω (1kHz test). Conductivity decreased by less than 5% after 100 bending cycles, significantly improving wearability and athletic performance.
[0081] Example 4
[0082] Reducing the number of pulse sequences from 100 to 50 can shorten the time between treatments, and at the same time shorten the time between blood-brain barrier openings without affecting drug delivery efficiency, thus reducing the risk of complications.
[0083] Example 5
[0084] The original scheme was reduced from 3kV to 1kV, but the model's penetration rate of 3kDa dextran was still 2.1±0.3 times that of the control group (p<0.01), and live cell imaging (PI staining) confirmed that the cell death rate decreased from 6% to 1%. This parameter combination improved the cross-barrier transport efficiency of doxorubicin by 2.7 times (HPLC detection).
Claims
1. A system for intracellular electrical processing based on ultra-high frequency and ultra-short pulses, characterized in that, The system includes a high-voltage ultrashort pulse generation module, a control module, and an adherent cell processing module; the high-voltage ultrashort pulse generation module is connected to the control module and the adherent cell processing module respectively; the control module presets multiple pulse parameter combinations, and the high-voltage ultrashort pulse generation module outputs a pulse electric field with preset parameters according to the instructions issued by the control module, and acts on the organelles through the adherent cell processing module.
2. The system according to claim 1, characterized in that, The high-voltage ultrashort pulse generation module is used to output a unipolar or bipolar pulse electric field with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and an amplitude of 0-±5kV, with a waveform rise / fall time of <5ns.
3. The system according to claim 1, characterized in that, The control module presets pulse electric field parameters according to different cell death modes.
4. The system according to claim 3, characterized in that, Different cell death modes and their pulsed electric field parameters are as follows: Ferrocyte-induced mode, pulsed electric field parameter combination includes: field strength of 20kV / cm, pulse width of 50ns, unipolarity, pulse sequence frequency of 10MHz, and number of pulse sequences of 50. The necroptosis-inducing mode uses pulsed electric field parameters including: field strength of 30 kV / cm, pulse width of 100 ns, unipolarity, pulse sequence frequency of 0.5 MHz, and 20 pulse sequences. The pyroptosis-inducing mode uses pulsed electric field parameters including: field strength of 10 kV / cm, pulse width of 15 ns, bipolarity, pulse sequence frequency of 2 MHz, and 100 pulse sequences. The apoptosis-inducing mode uses pulsed electric field parameters including: field strength of 50 kV / cm, pulse width of 80 ns, bipolarity, pulse sequence frequency of 1 MHz, and 100 pulse sequences.
5. The system according to claim 1, characterized in that, The adherent cell processing module is a microelectrode array, which uses an FR-4 epoxy resin substrate as an insulating carrier and has a metal conductive layer on its surface to form an electrode structure.
6. The system according to claim 5, characterized in that, The electrodes adopt a finger-like cross design, with a single electrode width of 130-180 micrometers and an electrode spacing of 200-300 micrometers.
7. The system according to claim 1, characterized in that, The high-voltage ultrashort pulse generation module is connected to the adherent cell processing module via wires.
8. The system according to claim 1, characterized in that, The cells are cancer cells, including any one or more of glioma cells, colon cancer cells, and pancreatic cancer cells; the organelles include mitochondria and endoplasmic reticulum.
9. The use of the system and / or pulsed electric field according to any one of claims 1-8 in the preparation of products for inducing ferroptosis, necroptosis, pyroptosis, and / or apoptosis in tumor cells, characterized in that, The tumor includes any one or more of the following: glioma, colon cancer, and pancreatic cancer.
10. The use of the system and / or pulsed electric field according to any one of claims 1-8 in the preparation of products for promoting lipid ROS levels, inhibiting GPX4 protein expression and / or inducing iron-dependent lipid peroxidation; Or the application of the system and / or pulsed electric field in the preparation of products for promoting the expression of IL-1β and IL-18; Or the application of the system and / or pulsed electric field in the preparation of products for promoting mitochondrial membrane potential collapse and / or activating caspase.