JP2013252117A - Method and apparatus for proliferation and activation of fibroblast - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an electric treatment method and an electric apparatus which are based on discovery by experiments and clinical trials, have extremely high safety and can easily proliferate and activate a fibroblast since a fibroblast existing in dermis or the like under the skin is a basic cell that can secrete an extracellular matrix in a supporting tissue (a connective tissue) in the body, a conventional method for proliferating/activating a fibroblast artificially comprises applying a relatively high voltage/electrical field and this method may cause the deterioration in safety or the damage to the skin.SOLUTION: As a result of repeated experiments and clinical trials, it is found that a fibroblast can be physiologically activated and can self-propagate by applying a far lower electric field than those employed in conventional methods. Consequently, it is confirmed that electrical field intensity of 0.001 to 0.3 V/cm is a proper electrical field region to be applied to a subepidermal region. On the basis of this confirmation, a method and an apparatus both for proliferating/activating a fibroblast by applying such an electrical field are invented. The electrical apparatus is so adapted that a battery composed of a metal and a semiconductor material having different electrical affinities from each other is used as a skin attachment tool.

Description

  The present invention relates to a method and apparatus for self-proliferating and bioactivating fibroblasts.

  Fibroblasts are basic cells that secrete extracellular matrices such as proteins, polysaccharides and glycoproteins in the body's supporting tissue (connective tissue). In the subcutaneous fat region, fibroblasts change into adipocytes and also into chondrocytes. Fibroblasts also produce cells on the inner wall of blood vessels. In particular, in the skin, it is distributed in the dermis and subcutaneous tissue. In the dermis, proteins such as collagen, hyaluronic acid, elastin, and proteoglycan, mucopolysaccharides, and glycoproteins are secreted to form a fibrous connective tissue to support the epidermis.

  When the skin is damaged by trauma, the fibroblasts migrate to the damaged site and are activated to generate and supply an extracellular matrix to the damaged site. This is the process of natural healing of skin wounds, but the migration and physiological activation of fibroblasts is due to electrical properties, and focusing on the negative electrode orientation promotes healing by energizing the wound Medical practice to be performed is mainly performed in the United States (Non-patent Document 1). There has also been a clinical report that wounds are healed at a rate 1.5 to 2.5 times higher than in the case of non-energization (Non-Patent Document 2). In order to examine how treatment of such wounds is actually related to the action of fibroblasts, the applied electric field dependence of skin fibroblasts was measured in vitro (Non-patent Document 3). According to FIG. 6 showing the experimental results of applying an electric field by immersing undifferentiated fibroblasts of human skin seeded on a porous filter and culturing in a culture solution in a glass container, the physiological activity of fibroblasts is shown. Showed a clear electric field dependence, and the self-propagation (DNA proliferation) showed almost the same electric field dependence.

  FIG. 6 (A) is data showing the electric field dependence of fibroblasts placed at an intermediate distance from the positive and negative electrodes when a pulsed electric field is applied as a relative ratio with no electric field applied as a standard. B) is shown in comparison with the data of fibroblasts placed in the vicinity of the negative electrode and the positive electrode in the same experiment. The horizontal axis of each figure shows the voltage applied between the electrodes. The distance between the positive and negative electrodes in each experiment is 7.5 cm. As shown by the arrows in the graph of FIG. 6, the protein secreted by fibroblasts reaches a peak at 17.8 V / cm when the applied voltage between the electrodes is changed to the electric field strength, and DNA synthesis (self-replication) It can be seen that the same calculation method has a generation peak at about 20 V / cm, and fibroblasts are physiologically activated and self-proliferate in the range of an applied electric field of about 13 to 30 V / cm. FIG. 6 (B) shows that activation appears strongly in the vicinity of the negative electrode, and it can be seen that fibroblasts exhibit a clear negative electrode orientation in this activated region. As can be seen from FIG. 6A, when the applied voltage exceeds about 260 V (approx. 35 V / cm in terms of applied electric field strength), the physiological activity and self-growth of fibroblasts are conversely suppressed.

L.C.Kloth and J.M.McCullock; Advances in Wound Care, vol.9, No.5 (1996) pp.42-45 P.J.Carley and S.F.Wainaipel; Arch. Phys. Med, Rehabil., Vol. 66 (1985) pp.443-445 G.B.Bourguignon and L.Y.W.Bourguignon; FASEB J. (1987) pp.398-402 S. Tokumoto et al.; Int. J. Pharm. 3 / (2006) pp.13-19

The above-described physiological activation and self-proliferation of fibroblasts by application of an electric field are practically important techniques, but are dangerous when applied to a living body. That is, the above-mentioned wound treatment in the United States is carried out for severe patients in the second to fourth stages in which the epidermis has been damaged and disappeared, and is accompanied by the risk of infection, so it is performed by a medical professional such as a doctor. Yes. On the other hand, if the technique is applied in a state where the epidermis is not damaged and the fibroblasts are non-invasively physiologically activated and self-propagated, a voltage is applied between the electrodes arranged on the epidermis. Will be applied. There is a stratum corneum layer having a thickness of about 20 μm at the top of the epidermis, and its electric resistance is several orders of magnitude higher than the skin layer below it (100 kΩ at the upper arm). Therefore, most of the applied voltage is applied to the epidermal stratum corneum. The stratum corneum has a very dense structure, but it has been found that the tissue order can be easily destroyed by simply applying an electric pulse of 100 V or more several times, and macromolecules can easily enter the skin. (For example, Non-Patent Document 4). For this reason, when a voltage is applied to the skin from the electrode on the epidermis, it is practically difficult to give an electric field strength of 13 to 30 V / cm to the dermis region (the stratum corneum is temporarily destroyed in the process) Faced with the risk of causing viral infection and allergen invasion). Damaged stratum corneum order is not easily recovered when the voltage reaches several hundred volts or higher, or when a voltage of 100 volts or higher is applied continuously.
An object of the present invention is to provide a method and apparatus for proliferating and activating fibroblasts that enables the activation and proliferation of fibroblasts without adversely affecting the stratum corneum and the epidermis.

  The present invention continuously applies an electric field having an intensity of 0.001 to 0.3 V / cm to fibroblasts in a culture environment, thereby allowing fibroblasts in the energized region of the electric field application to self- A method for proliferating and activating fibroblasts that is proliferated and physiologically activated is disclosed.

Furthermore, the present invention is a fibroblast proliferation / activation device for growing and activating fibroblasts in a culture environment,
A power source, and a first electrode and a second electrode connected to the power source by a conductive wire and disposed in or near the culture environment. The power source is 0.001 to 0.001 in the culture environment. Disclosed is a fibroblast proliferation / activation device for increasing the capacity to generate an electric field of 0.3 V / cm, and for proliferating and physiologically activating fibroblasts in this energized region.

  Furthermore, the present invention is a fibroblast proliferation / activation device for proliferating and activating fibroblasts in a culture environment, wherein the metal layer is connected to the metal layer by a conductive wire, and the metal layer and the electron are connected to each other. It has a semiconductor layer with different affinity and a contact surface. This metal layer and the semiconductor layer are in contact with this contact surface, and the surface opposite to this contact surface can touch the skin surface in the culture environment. The metal layer and the semiconductor layer having different electron affinities in order to proliferate and physiologically activate fibroblasts in the epidermis region in the skin surface in the culture environment. Discloses a fibroblast proliferation / activation device having a capacity to generate an electric field of 0.001 to 0.3 V / cm in the culture environment.

  Further, in the present invention, the metal layer and the semiconductor layer are disposed in a strip shape with a predetermined interval width on the sheet, and the electrode is disposed so as to intersect the strip-shaped metal layer and the semiconductor layer. A fibroblast proliferation / activation device according to claim 3 is disclosed.

  According to the present invention, fibroblast activation and self-proliferation can be realized with extremely low electric field strength.

It is an Example figure of the electric apparatus for the proliferation and activation of the fibroblast of this invention. It is a data figure of the fibroblast activation of the negative electrode vicinity at the time of the weak electric field application of this invention. It is a data figure of the fibroblast activation of the center part between electrodes at the time of the weak electric field application of this invention. It is a data figure of the fibroblast activation in the positive electrode vicinity at the time of the weak electric field application of this invention. It is a healing data figure of the electric apparatus which is an Example of this invention. It is a data example figure of activation of the conventional fiber cell.

In order to solve the above-mentioned problems, the present inventors have searched for an electric field strength region suitable for fibroblasts to be physiologically activated and proliferate at a much lower applied electric field than before. The present inventors have found out that a suitable electric field region of 001 to 0.3 V / cm exists, and have achieved the present invention.
The conforming electric field region of the present invention is 3 to 4 orders lower than the conventional conforming region described above, and in the conventional conforming region, the fibroblasts showed a clear negative polarity orientation, whereas the nonpolar orientation, that is, the polarity of the electric field Regardless of the field strength, the activation is uniformly dependent only on the electric field strength, so it can be said that the electric field region is completely different from the conventional technique.

Since the electric resistance value of the dermis region is several tens of ohms, which is three orders of magnitude lower than that of the stratum corneum, when 10 V is applied between the electrodes on the epidermis, the dermis region is biased to several to 10 mV. Therefore, if the electrode spacing is maintained at 1 to 2 cm, this bias voltage provides an electric field strength suitable for nonpolar activation (activation regardless of polarity) of the dermal fibroblasts. In this case, the electric field strength applied to the stratum corneum having a thickness of 20 μm is about 5 × 10 ³ V / cm and does not immediately damage the stratum corneum order. However, when energization is performed on the same skin over a long period of time, there is no guarantee that even if 10 V is applied, the skin will not be affected.

Therefore, a skin electromotive element can be recommended as an electrical device that can apply an effective electric field of 0.001 to 0.3 V / cm to the dermis region while further improving the safety to the skin. The skin electromotive element is a small battery that uses the skin as an electrolyte instead of an external power source. When an n-type semiconductor is used for the negative electrode, electrons flow to the positive metal side through the external conductor after skin contact, and holes are generated in the negative electrode. As a result, electrons from the positive electrode metal and holes from the negative electrode semiconductor are simultaneously injected into the skin to reduce and oxidize intradermal ions to generate an ionic current in the skin (Japanese Patent No. 2797118). By adjusting the combination of the positive and negative electrode materials (electromotive force) and the distance between the positive and negative electrodes, an effective electric field of 0.001 to 0.3 V / cm can be created in the dermis region by this ion current. For example, if gold is used for the positive electrode and zinc oxide is used for the negative electrode and the distance between the positive and negative electrodes is 1 to 2 cm, an electric field of 0.005 to 0.01 V / cm is generated in the dermis region. This electric field activates fibroblasts in the dermis region nonpolarly. That is, fibroblasts in a region to which an electric field is applied regardless of the energization direction are physiologically activated and perform self-proliferation. According to the data of the examples described later, the activation rate is increased by 50 to 80% in 90 minutes as compared to when no electric field is applied. At this time, the voltage measured on the skin is about 0.8V. Even if all of these voltages are applied to the stratum corneum, the electric field strength of the stratum corneum remains at 4 × 10 ² V / cm, and there is no danger of damaging the stratum corneum even when energized for a long time (for example, 48 hours).

Intracutaneous energization by the skin electromotive element can also be performed through an electrolyte layer in close contact with the epidermis. In this case, skin electromotive force occurs in both the electrolyte layer and the intradermal electrolyte layer on the epidermis, and the skin electromotive battery is connected in parallel to the external conductor.
In addition, the suitable electric field area | region 0.001-0.3V / cm in this invention was set to the area | region where the self-growth rate of a fibroblast increases 10% or more compared with the time of no electric field application. This is based on the activation measurement result in Example 1 and is because variation in activation of individual fibroblasts was seen about ± 10% in the course of the proliferation experiment.

  As described above, in the present invention, the fibroblasts in the region are physiologically activated and self-proliferated by applying an electric field strength 2 to 4 orders of magnitude lower than that conventionally used to the culture environment where the fibroblasts are present. I can do it. Unlike the activation in the conventional electric field strength region (negative electrode orientation type activation region), this effect is a nonpolar orientation type, so that the entire current-carrying region can be activated equally, and the effect expression range can be greatly increased. . Moreover, when this technique is applied to fibroblasts existing under the epidermis, there is a very great advantage that the stratum corneum can be prevented from being broken especially when energized for a long time.

(Part 1)
A culture solution having the following composition was filled in a glass container having inner dimensions of width 7.5 cm, depth 2.5 cm, and height 1.5 cm. In this, undifferentiated human skin fibroblasts seeded at a density of 5 × 10 ³ cells / filter are immersed on a Millipore filter (25 mm, GSWP02400; Millipore). Human skin fibroblasts were collected from healthy skin specimens by the Expand method, and were cultured in a medium for 48 hours at 37 ° C. and 5% CO ₂ .

  As the medium, Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum was used. It was confirmed that the fibroblasts before immersing in the culture medium in the glass container after the culture were in the process of growth.

  The culture solution in the glass container is a DMEM containing 10% fetal bovine serum with HEPES (4- (2-hydroxythyl) -1-piperazineethanesulfonic acid) buffer, 1% penicillin and 1% streptomycin added to pH 7.0. It is.

A stainless steel electrode having a size of 2.4 × 2.0 cm ² was arranged on both side walls in the glass container and connected to a pulse power source. The cultured fibroblast-mounted filter was cut to a size of 1.0 × 1.0 cm ² and immersed in the bottom of the container, one sheet at a position 5 mm away from each electrode and one sheet at the center of the glass container. . In this state, a triangular wave unidirectional pulse voltage having a duration of 100 μs and a pulse density of 100 / sec was applied between the stainless steel electrodes to change the peak voltage (the horizontal axis in FIGS. 2 to 4 is represented by the electric field strength). The application experiment was continued for 90 minutes at room temperature.

Thereafter, the fibroblast-loaded filter pulled up from the glass container was again placed in the medium and cultured at 37 ° C. in a 5% CO ₂ atmosphere for 2 hours. Thereafter, the amount of protein and DNA produced by fibroblasts was examined.

Protein production is measured with an activation label. That is, when the activated amino acid [ ³ H] proline 1Ci (Curie) / ml is mixed with the cultured fibroblast group at 37 ° C. for 20 minutes, it binds to the synthesized protein. After 20 minutes, the fibroblast-loaded filter was immersed in an ice-cooled 5% trichloroacetic acid solution to stop the amino acid-protein binding reaction. Thereafter, the filter was further washed three times with 10 ml of 5% ice-cold trichloroacetic acid solution, washed with water and dried. Thereafter, measurement was performed using a normal liquid scintillation counter.

The amount of DNA produced is maintained by maintaining the cultured fibroblasts in a culture solution having the same composition as that in the glass container and adding 0.1 Ci / ml of nucleoside [ ³ H] thymidine at 37 ° C. for 12 hours. Measured with different specimens. After holding for 12 hours, the sample-loaded filter is immersed in an ice-cooled 5% trichloroacetic acid solution to stop the binding reaction between DNA and nucleosides, washed and dried, and the amount of production is measured in the same manner as for protein. It was.

  For comparison with an electric field applied sample, a specimen immersed for 90 minutes without applying an electric field to a culture solution in the same glass container was prepared, and the amount of protein and DNA produced was measured by the same method as described above to obtain a reference value ( Control). FIG. 2 to FIG. 4 show the data of the electric field application sample as a percentage ratio with respect to the control data.

2 to 4 show the state of physiological activation and self-proliferation of human skin fibroblasts obtained in vitro using a weakly applied electric field as a parameter. 2 shows the data obtained in the vicinity of the negative electrode, FIG. 3 shows the data obtained in the intermediate position, and FIG. 4 shows the data obtained in the vicinity of the positive electrode. These figures are
(1) The amount of protein and DNA produced is 10% or more higher than the case where no electric field is applied in the applied electric field range of 0.001 to 0.3 V / cm (production rate). (2) Weak electric field applied At times, the amount of protein and DNA produced by fibroblasts shows a peak value at almost the same electric field strength (about 0.01 V / cm) regardless of the distance from the electrode of the specimen, that is, the physiological activation of fibroblasts and It shows that self-propagation is nonpolar and depends only on the electric field strength.

  This fibroblast activation region is in an applied electric field region that is two orders of magnitude lower than the conventional experimental data shown in FIG. 6, and is completely different from the conventional physiological activation showing a clear negative electrode orientation. Since it shows different electric field dependence, it can be said that it is an applied electric field region newly discovered by the present inventors.

  Such application of a low electric field activates skin fibroblasts and not only efficiently produces proteins that are the extracellular matrix of skin and subcutaneous tissue, but also proliferates the active fibers by proliferating the fibroblast's own DNA. It is extremely important for practical use that blasts are generated compoundively. In this example, only protein production was measured. However, considering the action of fibroblasts, production of other secretions such as hyaluronic acid, elastin and adipocytes can be easily activated as well. It is done.

  The knowledge obtained in this example can be applied to regenerative medicine. That is, a large amount of product can be obtained by holding fibroblasts collected from a patient in a incubator under a weak electric field. Unlike the prior art, it is possible to produce the entire electric field application region evenly by applying an extremely low electric field. If products such as proteins and fibroblasts that have been efficiently proliferated in this way and the fibroblasts themselves are transplanted into a patient by injection, there is no rejection reaction and treatment is possible in a short period of time.

  Furthermore, if this method is applied to living skin, the intradermal connective tissue can be proliferated very effectively in a non-invasive manner, ensuring the beauty (rejuvenation) of elderly and sick people with impaired proliferation function and ensuring flexibility of peripheral blood vessels. Expected to be effective in (reducing arteriosclerosis).

(Part 2)
In the in vitro examples described above, the electric field application to the fibroblasts was performed by pulse energization with an external power source. However, when the technique of the present invention is applied to living skin and subcutaneous tissue, which can be regarded as a culture environment, it is necessary to continuously perform physiological activation of fibroblasts on a daily basis. Is inappropriate. In addition, it is necessary to confirm that the same effect can be obtained not by pulse energization but by DC energization.

FIG. 1 shows an embodiment of an electrical apparatus for non-invasively applying a DC weak electric field to living body skin. This apparatus includes electrodes 1 and 2, a lead wire 3, a base film 4, and an adhesive tape 5. The base film 4 is made of PET having a thickness of 12 μm, and strip-like electrodes 1 and 2 are alternately stuck on the film 4 with an interval a. The front side of the paper is the skin attachment surface. The electrode 1 is a positive electrode, for example, gold, and the electrode 2 is a negative electrode, for example, zinc oxide. The electrodes 1 and 2 are each formed by vacuum deposition on a PET film having a thickness of 6 μm, and the electrode width is, for example, 15 mm. The electrodes 1 and 2 are attached to the PET base film 4 with the electrode surface facing up. The base film 4 accurately maintains the distance a between the electrodes 1 and 2 and serves to reinforce the strength of the thin electrodes 1 and 2. On these strip electrode groups 1 and 2, lead wires 3 are affixed at intervals b perpendicularly. The lead wire 3 is, for example, copper having a thickness of 3 μm and a width of 3 mm, and is adhered to the electrodes 1 and 2 and the PET base film 4 from the front side of the paper surface by a waterproof single-sided adhesive tape 5. The width of the tape 5 is wider than the lead wire 3, and the lead wire 3 is completely cut off from the external atmosphere. Moreover, although the waterproof single-sided tape 5 contacts the skin, this tape 5 serves as a shield, and the lead wire 3 does not directly contact the skin. The electric device of FIG. 1 has a thickness of about 30 μm at the thickest portion and is extremely flexible and light overall, so that it is very convenient without hindering daily life. The size of this electrical device is, for example, 50 × 100 mm ² . Thus, if the electrodes 1 and 2 are alternately arranged with an interval a and connected by a common lead wire 3, an electric field having a constant strength can be applied to the subcutaneous region regardless of the affected skin area. It is also suitable for mass production.

  When this electric device is affixed to living body skin, the skin can be electrogenerated as an electrolyte, and direct current can be applied to the skin directly under the affixed site. The electromotive force is theoretically determined by the material combination of the electrodes 1 and 2 having different electric affinities, but in the case of the gold / zinc oxide electrode shown in FIG. When used, it was a gold / zinc oxide electrode with an open circuit voltage of 1.24V). Therefore, the electric field strength applied to the skin can be changed by changing the combination of the materials of the electrodes 1 and 2 and the distance a.

  The potential drop (internal loss of the battery) in the skin is about 0.4 V in the case of the gold / zinc oxide, but most of it occurs in the epidermal stratum corneum, and the effective voltage applied to the epidermis is 0. It is about .01V. Therefore, if a is 1 cm, an electric field of about 0.01 V / cm is applied to the epidermis, and if a is 2 cm, an electric field of about 0.005 V / cm is applied to the epidermis. According to the data of FIGS. 2 to 4, this electric field strength is a value capable of physiologically activating epidermal fibroblasts and efficiently performing self-proliferation. If b is about 8 to 10 cm, a sufficiently uniform electric field can be applied to the entire surface of the sheet.

  When a gauze impregnated with an electrolyte layer, for example, 0.5% physiological saline is adhered between the electrode sheet and the epidermis shown in FIG. 1 and the electrode surface of the electrode sheet is attached to the gauze, Electricity is generated both inside the skin. From the gauze electrolyte layer under the negatively charged gold electrode, chlorine ions are rapidly introduced into the skin together with the electrons, and from the gauze electrolyte layer under the positively charged zinc oxide electrode, sodium ions are rapidly injected into the skin. Therefore, the power generation amount of the skin battery increases with time, and conversely, the power generation amount of the skin battery including the gauze electrolyte layer increases the resistance of the electrolyte layer (as a result of electrolyte ions permeating into the skin, the central part between the electrodes). Therefore, the ion concentration decreases and the electrolyte resistance increases). For this reason, in the case of continuous energization, it is preferable to use this method because the combination selection range of the electrodes 1 and 2 for securing the necessary electric field in the skin is widened.

  If the resistance of the stratum corneum decreases due to the rapid penetration of the stratum corneum by the iontophoresis effect of monoatomic ions (chlorine ions and sodium ions) that are also present in the skin, the amount of current flowing into the skin Increases rapidly, and the voltage applied to the subcutaneous region (subcutaneous current × subcutaneous resistance), that is, the intradermal electric field increases, and as a result, the physiological activation of fibroblasts can be favorably affected.

  FIG. 1 shows an embodiment of the electric device used in the present invention. Of course, various shapes are possible depending on the mounting site, and various electrode material combinations and various lead wire material selections are possible. . Moreover, although it was set as the gauze electrolyte layer, there is also an example of a gel electrolyte layer.

In the apparatus shown in FIG. 1, gold is used as the positive electrode 1 and zinc oxide is used as the negative electrode 2 as it is, and the lead wire 3 is zinc and the distance between the electrodes 1 and 2 is 10 mm, which is applied to pressure ulcer treatment. The thighs of Wistar rats weighing 400 to 450 g were anesthetized after shaving, and a pressure of 1 kg / cm ² was applied to the shaved part with a cylindrical weight. Pressure application was continued for 6 hours a day for 5 days to form a pseudo-decubitus with remaining epidermis. The wound condition was observed over time for each of the control group (control), the drug administration group, and the sheet (electrical device) wearing group of the present invention. The drug used was acetylsalicylic acid cream (concentration 1%), which was pointed out to be effective for pressure ulcers (Japanese Patent Application No. 8-345103). Acetylsalicylic acid was administered at 1 mg / wound site / day. In addition, a hydrogel sheet (electrolyte layer) containing 0.9% NaCl was interposed between the energizing sheet of the present invention and the rat skin surface. For comparison, data were also collected for two mice per group without the hydrogel sheet (when electrodes 1 and 2 were directly in contact with the rat epidermis). FIG. 5 shows the wound area of each group with respect to the wound area (major axis × minor axis) measured immediately after the pressure sore formation, and the ratio of the area was shown over time.

  As is clear from the figure, the pressure ulcer spreads over time in the control group and the drug administration group, whereas the method of the present example shows that the wound is shrinking and is healing. This is presumed to be the result of fibroblasts in the subcutaneous region self-proliferating and physiologically activated to significantly increase the regeneration rate of the subcutaneous tissue at the site. Even when the electrolyte layer (hydrogel sheet) is not used on the epidermis, the effect is clearly higher than that of the drug administration group. However, due to stratum corneum resistance and contact resistance, the strength of the electric field applied in the skin is smaller than that in the case of using a gel sheet, and as a result, it can be seen that the physiological activation effect of fibroblasts is relatively small.

  The data of FIG. 5 using the apparatus of FIG. 1 confirms that continuous DC electric field application has an excellent effect on physiological activation of fibroblasts in the skin as well as pulse electric field application of the previous example. Yes. Considering that the applied electric field region imparts nonpolar activation to the fibroblasts, it is obvious that the same activation effect can be expected not only by the DC electric field but also by applying the AC electric field.

  The present invention is expected to be used in the beauty field and the medical field such as regeneration of living skin and treatment of arteriosclerosis.

DESCRIPTION OF SYMBOLS 1 Positive electrode 2 Negative electrode semiconductor 3 Lead wire 4 Base film 5 Waterproof single-sided adhesive tape

Claims (4)

  By continuously applying an electric field having a strength of 0.001 to 0.3 V / cm to fibroblasts in a culture environment, the fibroblasts in the energized region to which the electric field is applied self-proliferates, Fibroblast proliferation / activation method to be activated. A fibroblast proliferation / activation device for growing and activating fibroblasts in a culture environment,
A power source, and a first electrode and a second electrode connected to the power source by a conductive wire and disposed in or near the culture environment. The power source is 0.001 to 0.001 in the culture environment. A fibroblast proliferation / activation device that has a capacity that generates an electric field of 0.3 V / cm, and that causes the fibroblasts in this energized region to proliferate and physiologically activate.   A fibroblast proliferation / activation device for proliferating and activating fibroblasts in a culture environment, wherein the metal layer is connected to the metal layer by a conductive wire and has an electric affinity different from that of the metal layer. An electrolyte layer having a semiconductor layer and a contact surface, the metal layer and the semiconductor layer are in contact with the contact surface, and the surface opposite to the contact surface is capable of skin contact with the skin surface in a culture environment; The metal layer and the semiconductor layer having different electron affinities are used to grow and physiologically activate fibroblasts in the epidermal region in the skin surface in the culture environment. A fibroblast proliferation / activation device having a capacity to generate an electric field of 0.001 to 0.3 V / cm within the device.   The metal layer and the semiconductor layer are arranged in a strip shape with a predetermined interval width on the sheet, and the electrode is arranged so as to cross the strip-like metal layer and the semiconductor layer. The fibroblast proliferation / activation device according to claim 3.

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