Medical kit combined with reversible electroporation and used for organ transplantation and transferring and application of medical kit

Abstract

The invention belongs to the technical field of medical equipment, and particularly discloses a reversible electroporation combined medical kit for organ transplantation and transfer and application of the reversible electroporation combined medical kit. According to the medical kit for organ transplantation, a novel electric pulse device is additionally designed on the basis of original design, a large amount of emergency response generated after organ transplantation can be remarkably reduced, and the survival rate of organs is increased. The novel electric pulse device is designed to be attached to the inner wall of the box body, the designed electric pulse electrode plate is attached to the buffer pad, the buffer pad is made of an insulating non-toxic heat preservation material, and organ preservation liquid is contained in the organ preservation bag. According to the medical kit for organ transplantation, the additional electrodes can be used for applying electric pulses to organs in the organ storage bag, medicine in the organ storage liquid is promoted to enter the organs, meanwhile, the optimal applied pulse intensity is confirmed in combination with cell experiments, the potential distribution condition is simulated and confirmed in combination with simulation software, and emergency reaction generated after perfusion can be reduced; therefore, normal organ functions are ensured, and the success rate of transplantation is improved.

Description

Technical Field

[0001] This invention belongs to the field of medical device technology, and relates to a medical kit for organ transplantation combined with reversible electroporation and its application. Specifically, it relates to a medical kit for organ transplantation based on reversible electroporation that can reduce the emergency of organ reperfusion. Background Technology

[0002] Since the inception of organ transplantation, organ transplantation equipment and technology have advanced rapidly, but progress in organ preservation has been limited. In early kidney transplant surgeries, the donor and recipient were in adjacent beds, so the preservation status of the kidney transplant was almost unaffected by organ transport. However, with the decreasing availability of organ donors and the increasing collaboration in organ transplantation nationwide and globally, there is an urgent need for better organ preservation technologies to facilitate long-distance transplant transport. Because organ transplants inevitably suffer ischemia-reperfusion injury, their condition declines rapidly, resulting in a short preservation time after leaving the donor. Therefore, kidney transplants may be discarded after long-distance transport due to poor condition. Glutathione (GSH) is often added to the perfusion solution used for organ preservation during renal ischemia-reperfusion as an antioxidant to inhibit oxidative stress. Promoting drug delivery through GSH may further inhibit oxidative stress caused by ischemia-reperfusion injury. Reversible electroporation can temporarily disrupt cell membrane integrity, promoting drug molecules to enter cells and enhancing the therapeutic effect of drugs. Applying reversible electroporation to organ transplantation can reduce intracellular oxidative stress damage in transplanted cells, thereby improving the success rate of organ transplantation. Summary of the Invention

[0003] To address the aforementioned technical problems, this invention provides a medical kit for organ transplantation that incorporates reversible electroporation.

[0004] The specific technical solution is as follows:

[0005] A medical kit for organ transplantation transfer combined with reversible electroporation includes a kit body and a cover connected to the kit body by a hinge. An insulating and heat-preserving cushion is provided on the inner wall of the kit body. A first electrode plate and a second electrode plate are provided on the inner side of the cushion. A control system connection port is provided on the outer side of the kit body for connecting to an external control system. An organ preservation bag is provided inside the kit body, and the organ preservation bag contains organ preservation fluid. The electrode plates can apply reversible electrical pulses according to the control system.

[0006] The electrode sheet is a solid cuboid shape, made of conductive material, and consists of two electrodes. The electrode sheet is 14cm long, 6cm wide, and 0.5cm thick.

[0007] The electrode plate can be subjected to reversible electrical pulses according to the control system.

[0008] The organ preservation bag contains an organ preservation solution, which is primarily used for the storage and transportation of donated organs such as kidneys, hearts, and lungs. Its main components are calcium chloride (dihydrate), sodium hydroxide, hydroxyethylpiperazine ethanesulfonic acid (free acid), potassium phosphate (monobasic base), mannitol, glucose, sodium gluconate, magnesium gluconate, ribose, hydroxyethyl starch, glutathione (reduced state), adenine (free base), and water for injection. These components all meet the basic regulatory requirements of reversible electrical pulses.

[0009] Furthermore, the external control system of the transplant box enables the electrode plates to generate electrical pulses with a voltage of 0-500V, a pulse width of 0-50μs, and a pulse frequency of 0-250Hz.

[0010] Furthermore, the electrode sheet is made of conductive metal or other stable, non-toxic organic or inorganic conductive materials, such as CuMOF and CoMOF materials.

[0011] Furthermore, the conductive metals are silver, copper, aluminum, or conductive polymers, and carbon materials with good conductivity such as graphene and multi-walled carbon nanotubes.

[0012] Furthermore, the organ preservation solution inside the organ preservation bag is continuously infused and maintained at 4-10°C.

[0013] Furthermore, the enclosure is made of non-toxic insulating material.

[0014] Furthermore, non-toxic insulating materials include polyethylene, ultrafine glass wool, and ceramic wool.

[0015] A medical case for organ transplantation incorporating reversible electroporation is disclosed. The device comprises a main body, a lid, a cushioning pad, electrode pads, and an organ preservation bag. The lid is hinged to the main body, and the organ preservation bag is fitted within the main body. An insulating, non-toxic, and heat-insulating cushioning pad is adhered to the inner wall of the case. The organ preservation bag contains organ preservation fluid, and the electrode pads are attached to the cushioning pad. An external control system is also included.

[0016] In this design, the electrode plates can apply reversible electrical pulses according to the control system;

[0017] The organ preservation bag in this scheme contains organ preservation fluid;

[0018] This invention also provides a method for organ transplant transfer using a medical box combined with reversible electroporation, as described above. First, suitable pulse width, pulse number, pulse frequency, and voltage are selected through cell experiments. The control system is then connected to the medical box's interface 7, and the parameters selected through cell experiments are set on the control system. The optimal placement orientation of the organ preservation bag is confirmed using the simulation software COMSOL Multiphysics. The control system releases pulses into the organ preservation bag, continuously irrigating it with organ preservation fluid.

[0019] The organ preservation solution used in this method is mainly used for the storage and transportation of donated kidneys. Its main components are calcium chloride (dihydrate), sodium hydroxide, hydroxyethylpiperazine ethanesulfonic acid (free acid), potassium phosphate (monobasic base), mannitol, glucose, sodium gluconate, magnesium gluconate, ribose, hydroxyethyl starch, glutathione (reduced form), adenine (free base), and water for injection.

[0020] In this method, the external control system of the transplant box enables the electrode plates to generate electrical pulses with a voltage of 0-300V, a pulse width of 0-50μs, and a pulse frequency of 0-250Hz.

[0021] In this scheme, the electrode sheet is a conductive metal, preferably silver, copper, or aluminum, but it can also be a stable organic or inorganic conductive material.

[0022] In this scheme, the organ preservation solution inside the organ preservation bag is continuously infused and maintained at 4-10℃.

[0023] The enclosure used in this scheme is made of non-toxic insulating materials such as polytetrafluoroethylene and polyethylene.

[0024] The beneficial effects of this invention are:

[0025] 1. The present invention provides a medical box for organ transplantation that incorporates reversible electroporation, which has the advantage of reducing intracellular oxidative stress damage in transplanted cells, thereby improving the success rate of organ transplantation and meeting the needs of organ transplantation transportation.

[0026] 2. The box structure of the present invention is simple and has the advantages of simple manufacturing and low manufacturing cost.

[0027] 3. The insulating material used in this invention can avoid the shielding effect of the metal parts inside the organ transplant transport box on the pulse electric field, the electric field distribution in the organ tissue is uniform, and no toxic substances will be dissolved under the action of electric field and temperature field. Attached Figure Description

[0028] Figure 1 This is a front view of an organ transplant medical kit.

[0029] Figure 2This is a rear view of the organ transplant medical kit.

[0030] Figure 3 This is a cross-sectional view of the inside of an organ transplant medical kit.

[0031] Figure 4 The following are simulation diagrams of the electric field: (A) Schematic diagram of the electric potential distribution when an electric field acts on a kidney transplant; (B) Cross-sectional view of the electric potential distribution; (C) Schematic diagram of the electric field distribution when an electric field acts on a kidney transplant.

[0032] Figure 5 The effect of pulsed electric fields of 0–600V on the survival rate of HEK293 cells.

[0033] Figure 6 The survival rate of HEK293 cells under different pulse durations.

[0034] Figure 7 The survival rate of HEK293 cells with different reversible pulse parameters is (A) width; (B) frequency. Figure 8 This is a structural diagram of an organ transplant medical kit.

[0035] The components include: 1. First electrode plate; 2. Second electrode plate; 3. Buffer pad; 4. Organ preservation bag; 5. Box body; 6. Cover; 7. Hinge; 8. Control system connection port. Detailed Implementation

[0036] The present invention will now be described in further detail with reference to the accompanying drawings and embodiments. It should be noted that the described embodiments are preferred embodiments of the present invention, and the actual structure is not limited thereto. Based on the embodiments described in this invention, if those skilled in the art obtain similar embodiments without departing from the spirit of the present invention, they should all fall within the protection scope of this invention.

[0037] See Figure 1 This invention provides a medical box for organ transplantation incorporating reversible electroporation, mainly comprising electrode pads, a box body, a cushioning pad, and an organ preservation bag. 1 and 2 are electrode pads, 3 is the cushioning pad, 4 is the organ preservation bag, 5 is the box body, 6 is the hinge, and 7 is the control system connection port. The electrode pads are made of conductive metals such as silver, copper, and aluminum. An external control system is connected, enabling the release of electrical pulses with a voltage of 0-500V, a pulse width of 0-50μs, and a pulse frequency of 0-250Hz. The organ preservation perfusion solution within the organ preservation bag is continuously infused and maintained at 4-10℃. This invention obtains the input pulse parameters to the control system through cell experiments. Then, the control system transmits specific parameter pulses to the electrode pads via interface 7, which then release them onto the organ. The potential distribution is confirmed through software simulation.

[0038] A method for organ transplantation transfer using a medical box combining reversible electroporation involves first screening suitable pulse width, pulse number, pulse frequency, and voltage through cell experiments, connecting the control system to the medical box interface 7, and setting the parameters screened by cell experiments on the control system; confirming the optimal placement orientation of the organ preservation bag using the simulation software COMSOL Multiphysics; and releasing pulses into the organ preservation bag, continuously irrigating the organ preservation fluid inside the bag.

[0039] The organ preservation solution mainly consists of calcium chloride (dihydrate), sodium hydroxide, hydroxyethylpiperazine ethanesulfonic acid (free acid), potassium phosphate (monobasic base), mannitol, glucose, sodium gluconate, magnesium gluconate, ribose, hydroxyethyl starch, glutathione (reduced state), adenine (free base), and water for injection. The external control system of the transplant box enables the electrode pads to generate electrical pulses with a voltage of 0-300V, a pulse width of 0-50μs, and a pulse frequency of 0-250Hz. The organ preservation solution inside the organ preservation bag is continuously infused and maintained at 4-10℃.

[0040] Example 1: Determining the optimal parameters of the control system through cell experiments.

[0041] 1. Reversible electrical pulse stimulation of cells: Electrical pulses were applied to human embryonic kidney cells (HEK293 cells). The pulse voltage ranged from 0 to 600 V, the pulse width was 30 μs, the pulse frequency was 1 Hz, and the number of pulses was 60. Cell viability was determined using the CCK-8 assay. All experiments were repeated three times under the same conditions. Figure 5 The graph shows the survival rate of HEK293 cells within a pulse voltage range of 0-600V. When the pulse voltage reaches 400V, the survival rate of HEK293 cells drops sharply to 58.47%. When the pulse voltage is less than 300V, the cell survival rate decreases slightly to 93.76%, still exceeding 90%.

[0042] The cell viability graph shows that when the pulse voltage exceeds 300V, further increases in pulse voltage lead to a continuous decrease in cell viability. To determine the optimal experimental conditions, a pulse width of 30μs, a pulse frequency of 1Hz, and a pulse count of 60 were selected. The reversible pulse voltage range for HEK293 cells was found to be 0–300V.

[0043] 2. Optimization of reversible pulse parameters.

[0044] HEK293 cells were selected as the experimental subject. Figure 6The figure shows the change in the survival rate of HEK293 cells with the number of pulses under pulse parameters of 150V, 30μs, and 1Hz. It is known that increasing the number of reversible pulsed electric fields applied to HEK293 cells increases the total duration of the pulse voltage. (Observation...) Figure 6 It was found that as the total duration of the reversible pulse voltage increased, the cell survival rate decreased. The cell survival rate reached its maximum at 30 pulses, at which point it was 97.95%. When the number of pulses was 30, i.e., the total duration of the reversible pulse electric field was 900 μs, the electric field had the best effect on cell proliferation, resulting in the highest cell survival rate. Therefore, the optimized reversible pulse conditions for subsequent experiments were 150 V, 1 Hz, 30 μs, and 30 pulses.

[0045] like Figure 7 As shown in (A), with fixed reversible parameters of a total pulse duration of 900 μs and a pulse width of 1 Hz, the specific combinations of pulse width and number of pulses (pulse width, number of pulses) are: (10 μs, 90), (20 μs, 45), (30 μs, 30), (36 μs, 25), (50 μs, 18), (90 μs, 10), and (100 μs, 9). The cell viability is highest at 50 μs, at 102.96%. At 10 μs and 20 μs, the cell viability is approximately 100%. Therefore, with a total pulse duration of 900 μs, the optimal combination of pulse width and number of pulses is 50 μs and 18 pulses. Figure 7 (B) shows the cell viability of HEK293 cells under different pulse frequencies. Except for the pulse frequency, the other pulse parameters are the optimized pulse parameters: 150V, 50μs, and 18 cells. As shown in the figure, under these pulse conditions, cell viability initially increases and then decreases. The highest and lowest cell viability values ​​are observed at reversible pulse frequencies of 1Hz and 50Hz, respectively. Therefore, 1Hz was selected as the optimized pulse frequency. After multiple optimizations, the parameters for reversible electroporation of HEK293 cells are 150V, 50μs, 18 cells, and 1Hz.

[0046] Example 2: Determining the potential distribution through software simulation.

[0047] In engineering applications, the organ subjected to reversible pulse stimulation is the kidney transplant, a biological tissue composed of tens of millions of cells with unique structure and properties. In addition, the cells and blood vessels within the kidney transplant contain large amounts of plasma, water, organ preservation fluid, and other fluids. When an electric field is applied to the kidney transplant, the non-uniformity of the conductive medium within the transplant causes energy loss and scattering, resulting in a voltage on the transplant being lower than the actual applied voltage. Furthermore, the kidney transplant has a thickness of 3-4 cm and non-uniform tissue conductivity, which also affects the penetration of the applied electric field. Therefore, the finite element method coupled with multiphysics can be used to simulate the reversible pulse stimulation of the kidney transplant in an organ transplant transport box, thereby obtaining the distribution of the electric and temperature fields within the kidney transplant.

[0048] The Comsol software simulation model uses a three-dimensional axisymmetric spatial dimension and a current interface as the physics field. A flattened ellipsoid is constructed, 120mm long, 40mm wide, and 60mm thick. This model contains two quarter-circles on the left side of the kidney model, and these two arc-shaped cylinders are symmetrical. These cylinders represent the ureter and renal vessels, respectively. Figure 4 As shown, when the control system applies a voltage of 300V, the minimum voltage in the kidney transplant is 142V and the maximum voltage is 162V.

[0049] The above-described embodiments are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the concept of the present invention, and these improvements and modifications are also within the protection scope of the present invention.

Claims

1. A medical case for organ transplantation transfer combined with reversible electroporation, comprising a case body and a lid connected to the case body via a hinge, characterized in that, An insulating and heat-preserving cushioning pad is provided on the inner wall of the box. A first electrode plate and a second electrode plate are provided on the inner side of the cushioning pad. A control system connection port is provided on the outer side of the box body for connecting to an external control system. An organ preservation bag is provided inside the box, and the organ preservation bag contains organ preservation fluid. The electrode plates can apply reversible electrical pulses according to the control system.

2. The organ transplant transfer kit combined with reversible electroporation according to claim 1, characterized in that, The electrode sheet is made of conductive metal or other stable, non-toxic organic or inorganic conductive materials.

3. The organ transplant transfer kit combined with reversible electroporation according to claim 2, characterized in that, The non-toxic organic and inorganic conductive materials are CuMOF materials and CoMOF materials, etc.

4. The organ transplant transfer kit combined with reversible electroporation according to claim 2, characterized in that, The conductive metals are silver, copper, aluminum, or conductive polymers, or carbon with good conductivity.

5. The organ transplant transfer kit combined with reversible electroporation according to claim 4, characterized in that, The carbon with good electrical conductivity is a material such as graphene or multi-walled carbon nanotubes.

6. The organ transplant transfer kit combined with reversible electroporation according to claim 1, characterized in that, The enclosure is made of non-toxic insulating material.

7. The organ transplant transfer kit combined with reversible electroporation according to claim 6, characterized in that, Non-toxic insulating materials include polyethylene, ultrafine glass wool, and ceramic wool.

8. A method for using a medical kit for organ transplantation combined with reversible electroporation as described in any one of claims 1-7, characterized in that, First, suitable pulse width, pulse number, pulse frequency, and voltage are screened through cell experiments. The control system is then connected to the medical box interface 7, and the parameters screened through cell experiments are set on the control system. The optimal placement direction of the organ preservation bag is confirmed through the simulation software COMSOL Multiphysics. The control system releases pulses to the organ preservation bag, and the organ preservation fluid inside the organ preservation bag is continuously irrigated. The organ preservation solution is mainly composed of calcium chloride (dihydrate), sodium hydroxide, hydroxyethylpiperazine ethanesulfonic acid (free acid), potassium phosphate (monobasic base), mannitol, glucose, sodium gluconate, magnesium gluconate, ribose, hydroxyethyl starch, glutathione (reduced state), adenine (free base), and water for injection.

9. The method for organ transplant transfer using a medical kit combined with reversible electroporation as described in claim 8, characterized in that, The external control system of the transplant box enables the electrode plates to generate electrical pulses with a voltage of 0-300V, a pulse width of 0-50μs, and a pulse frequency of 0-250Hz.

10. The method for organ transplant transfer using a medical kit combined with reversible electroporation according to claim 8, characterized in that, The organ preservation solution inside the organ preservation bag is continuously infused and maintained at 4-10℃.

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