JP2002062294A - Method and device for measuring skin surface condition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method and a device for measuring a skin surface condition so as to measure and digitize a human sense given by a stimulation to the skin surface and to provide a method and a device for measuring effect of a protective drug against the sense if the sense is mentally minus. SOLUTION: A skin section is floated on the surface of a culture solution stored in a vessel with the skin section surface side directed upward, and a potential difference between the skin section skin surface and the culture solution is measured. By giving stimulation to the skin surface or adding a stimulant to the culture solution, the potential difference is measured for measuring the degree of stimulation to the skin. By applying the drug to the skin section after giving the stimulation to the skin surface or adding the stimulant to the culture solution, the potential difference is measured for measuring the degree of reduction of skin stimulation due to the drug. By adding the drug to the culture solution after giving the stimulation to the skin surface or adding the stimulant to the culture solution, the potential difference is measured for measuring the degree of reduction of skin stimulation due to the drug.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for measuring the condition of the skin surface and an apparatus for measuring the same. More specifically, a method of creating a skin model in which a skin slice is floated with the surface facing up on the surface of the culture solution and measuring the state of the skin surface from the potential difference between the skin surface and the culture solution, A method for measuring the degree of irritation from a potential difference that changes by giving a stimulating factor to a culture solution, a method for measuring the degree of inhibition of skin irritation from a change in the potential difference when a drug is added to these irritating skin, and these methods A device suitable for implementation.

[0002]

2. Description of the Related Art The surface of human skin is subjected to various stimuli from nature. When irritated, it often appears as a symptom on the skin, and at the same time, there are many things that are expressed by human sensations, such as itching, itching, and sticking, and there are also many things that do not show any symptoms but only sensation . Some of these stimuli are comfortable, but many are unpleasant, and stimuli such as itching are unpleasant for humans.
These stimuli are in the category of human sensation and can be awkward to others.

[0003] However, there has been no method for measuring the surface condition of the skin in the past, so that the degree of sensation had to rely on human complaints. Therefore, if such a stimulus can be replaced with an objective numerical value or the like, it is possible to share a subjective sensation, and further, if the above numerical value or the like is used, the development speed of an unpleasant stimulus suppressing drug is expected to rapidly change. You.

[0004]

SUMMARY OF THE INVENTION The present invention has been made in view of the above circumstances, and it is an object of the present invention to provide a method and an apparatus for measuring the surface condition of the skin. The present invention provides a method for measuring and quantifying a human sensation caused by a human being, and a method for measuring the effect of a drug that protects the sensation when the sensation is mentally negative for a human, and an apparatus for use in the method. It is in.

[0005]

Means for Solving the Problems The present inventors have attempted to solve the above problems by focusing on the potential of the skin surface. Then, a simple skin model was created in which a skin slice was floated with the surface facing up on the liquid surface of the culture solution, and a method capable of easily measuring the potential difference between the skin surface and the culture solution was found. Was solved.

That is, according to the method for measuring the condition of the skin surface according to the present invention, a skin slice is floated on the surface of a culture solution contained in a container with the surface facing upward, and the potential difference between the skin surface of the skin slice and the culture solution is determined. It is characterized by measuring.

[0007] Further, a skin slice is floated on the surface of the culture solution contained in the container with the surface facing upward, and when the potential difference between the skin surface of the skin slice and the culture solution is measured, the skin surface is stimulated. And a potential difference between the measured value and the potential difference when no stimulus is given.

[0008] In the measurement of the degree of skin irritation of the present invention, the stimulus may be a stimulus substance or a physical stimulus.

[0009] Further, a skin slice is floated on the surface of the culture solution contained in the container with its surface facing upward, and in measuring the potential difference between the skin surface of the skin slice and the culture solution, an irritant is added to the culture solution. It is characterized in that the potential difference is measured after addition, and the degree of skin irritation is measured from the difference between the value and the potential difference when no stimulant is added.

The most typical irritants used for measuring the degree of irritation of the skin are itch-generating substances.

In addition, a stimulated skin slice is floated on the surface of the culture solution contained in the container with its surface facing upward, and the potential difference between the skin surface of the skin slice and the culture solution is measured. Before or after applying a stimulus to the skin surface, apply a drug to the skin surface and measure the potential difference.
The method is characterized in that the degree of inhibition of skin irritation by a drug is measured from the difference from the potential difference when the drug is not applied.

Further, a stimulated skin slice is floated on the surface of the culture solution contained in the container with its surface facing upward, and the potential difference between the skin surface of the skin slice and the culture solution is measured. Before or after stimulating the skin, a drug is added to the culture solution to measure the potential difference, and the degree of inhibition of skin irritation by the drug is measured from the difference between the value and the potential difference when no drug is added. And

In the measurement of the degree of inhibition of skin irritation according to the present invention,
The stimulus can be a stimulus or a physical stimulus.

Further, a skin slice is floated on the surface of the culture solution containing the stimulant contained in the container with the surface facing upward, and the potential difference between the skin surface of the skin slice and the culture solution is measured. Before or after adding the stimulant to the solution, apply a drug to the skin surface and measure the potential difference,
From the difference between that value and the potential difference when the drug is not applied,
It is characterized in that the degree of inhibition of skin irritation by a drug is measured.

Further, the skin slice is floated on the surface of the culture solution containing the stimulant contained in the container, with the surface facing upward.
In measuring the potential difference between the skin surface of the skin slice and the culture solution, before or after adding the stimulant to the culture solution, a drug is added to the culture solution to measure the potential difference, and the value and the stimulus substance are measured. From the difference from the potential difference when not added,
It is characterized in that the degree of inhibition of skin irritation by a drug is measured.

The most typical irritants used for measuring the degree of inhibition of the skin irritation are itching substances.

An apparatus for carrying out such a skin surface condition measuring method comprises a container containing a culture solution, a skin section floating on the surface of the culture solution, and a standard device connected to the culture solution. An electrode unit, a measurement electrode unit connected to the skin surface of the skin cut, and a potential difference detection unit that detects a potential difference between the standard electrode unit and the measurement electrode unit.

A preferred standard electrode portion used in the skin surface condition measuring apparatus of the present invention is an inner tube containing an internal liquid in which the standard electrode is immersed, and a double tube formed by containing the inner tube therein. An outer tube containing an inner liquid, wherein a liquid junction is attached to a wall of the inner tube.

In this case, the communication between the culture solution and the standard electrode section is made by a salt bridge connected between the culture solution and the internal solution in the outer tube.

The measuring electrode section used in the skin surface condition measuring apparatus of the present invention includes an inner tube containing an internal liquid in which the measuring electrode is immersed, and a double tube containing the inner tube therein. An outer tube containing an internal liquid to be formed is provided, and a liquid junction is attached to the inner tube wall.

In this case, communication between the skin surface of the skin slice and the measuring electrode unit is performed by a salt bridge connected between the skin surface of the skin slice and the internal liquid in the outer tube.

The culture solution used in each of the above methods and apparatuses is preferably a physiological saline solution or a culture solution containing amino acids.

According to the method and apparatus for measuring the surface condition of the skin according to the present invention as described above, the potential difference between the skin surface of the skin section and the culture solution on which the skin section floats is measured by using a suitable apparatus, whereby the skin is measured. The state of the surface can be easily measured. In addition, by applying a stimulus factor to the skin surface or the culture solution and measuring the change in the potential difference, the stimulus can be objectively expressed without using a human directly and beyond the human subjectivity. Further, the change in the potential difference due to the addition of a drug can be used for searching for a drug effective against skin irritation, and a great effect can be achieved for searching for a new drug, which is very useful.

In the present invention, the skin surface condition is a concept including various state changes occurring on the skin surface, and includes, for example, a state change due to irritation, suppression of irritation and the like, and a degree thereof. It is.

[0025]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The method of the present invention and the device of the present invention will be described below in this order.

In the first method of the present invention, a skin slice is floated on the surface of a culture solution contained in a container with its surface facing upward, and the potential difference (mV) between the skin surface of the skin slice and the culture solution is determined. Measure. Thereby, the surface condition of the skin is measured.

In the second method of the present invention, in measuring the skin surface condition in the first method, a stimulus is further applied to a skin model comprising a skin slice and a culture solution to measure the potential difference.
This potential difference is changed by applying a stimulus. This change means that a flow of ions has occurred in the epidermis. The direction of ion flow differs depending on the stimulus. If there is a change compared to the value of the untreated skin, it is interpreted that a stimulus that causes the movement of ions to the epidermis was given. The degree of skin irritation is measured from the change value. In applying the stimulus, the location may be either the skin surface or the culture solution. In the case of the skin surface, the stimulus may be applied by applying an irritant to the skin, by physical stimulation, or the like. Also,
In the case of a culture solution, a method of adding a stimulant is used. These stimulants include neuropeptides, histamine, surfactants, organic acids, capsaicin and the like. Among these, there are substance P and the like as neuropeptides. In the present invention, it is particularly useful that the stimulant is an itch-generating substance. Examples of the physical stimulus include an example of barrier destruction by Cellotape (registered trademark) and the like.

The third method of the present invention is a method for measuring the degree of inhibition of the skin stimulated by the second method due to the addition of a drug. In the present invention, after applying a stimulus to the skin model system, a drug is added, the potential difference is measured, and the degree of suppression of skin irritation of the drug is measured from the value of the potential difference changed by the addition.

This method can take various methods depending on the method of adding the drug. (1); Float the skin section with the skin surface stimulated on the culture solution with the surface facing up, apply a drug to the skin surface before or after applying the skin irritation, measure the potential difference, and change with the drug application The degree of inhibition of skin irritation of the drug is measured from the value of the potential difference. (2); instead of applying the drug to the skin surface of (1), the drug is added to the culture solution to measure the potential difference, and the degree of inhibition of skin irritation of the drug is determined from the value of the potential difference that changes with the addition of the drug to the culture solution. Is measured. (3); Float the skin slice on the culture solution to which the stimulant has been added, apply a drug to the skin surface before or after adding the stimulus, measure the potential difference, and determine the potential difference that changes due to application of the drug. The degree of inhibition of skin irritation of the drug is measured from the value. (4);
Instead of applying the drug to the skin surface in (3), the drug is added to the culture solution and the potential difference is measured, and the degree of inhibition of the skin irritation of the drug is measured from the value of the potential difference changed by the addition of the drug to the culture solution.

The application of the drug may be performed before or after applying the stimulus. If the application of the drug is after applying the stimulus, it can be measured as an effect to relieve the stimulus, and before applying the stimulus, it can be measured as an effect to prevent the stimulus.

As the culture solution used in the first to third measurement methods, physiological saline, an aqueous solution containing amino acids and the like are used. The skin slice used for the measurement may be cultured skin, human skin obtained in plastic surgery or the like, animal-derived skin such as a hairless mouse, or the like.

When the skin of a person is stimulated, it often appears as a symptom on the skin, and at the same time, a thing expressed by human sensations such as itching, itching, and tingling occurs. There are many things that are only sensations. According to the first to third methods of the present invention, since such a subjective sensation can be expressed as an objective value, for example, a search for a drug that relieves or suppresses a stimulus without using humans It can greatly contribute to the development of new drugs.

Next, an apparatus preferably used in the method of the present invention will be described with reference to the drawings.

FIG. 1 shows an example of a skin surface condition measuring apparatus according to the present invention. As shown in FIG. 1, the skin surface condition measuring device comprises a container 3 containing a culture solution 1, a skin section 2 floating on the surface of the culture solution 1, and a standard electrode section connected to the culture solution 1. 4, a measuring electrode unit 5 connected to the skin surface of the skin section 2, and a potential difference detecting unit 6 for detecting a potential difference between the standard electrode unit 4 and the measuring electrode unit 5.
And

The standard electrode section 4 has a double outer tube 41 and an inner tube 42 made of plastic, and a porous wall having a function of allowing the solution to pass through the tube wall of the inner tube 42 but not allowing the passage of electrons or ions. And a liquid junction 43 made of high quality ceramic. This double outer tube 4
1 and the inside of the inner tube 42 are filled with internal solutions 44 and 45 each consisting of a KCl solution of a predetermined concentration, and a standard electrode 46 made of silver-silver chloride is immersed in the internal solution 45 of the inner tube 42. I have. This standard electrode 46
The inner tube 4 is attached to an electrode holder 47 also serving as a cap for closing the upper ends of the double outer tube 41 and the inner tube 42, and suspended therefrom via a conductor.
2 is immersed in the internal liquid 45. On the peripheral surface near the upper end of the outer tube 41, the inner liquid 4
A liquid replenishing port 48 for replenishing the liquid 4 is provided.

In the standard electrode section 4 shown in FIG. 1, both the double outer tube 41 and the inner tube 42 may be made of glass. The liquid junction 43 may be a known one such as a solution having a pseudocrystal structure made of gelatin, agar, or the like, or a liquid junction used as a salt bridge such as porous Vycor glass. In addition, the standard electrode 4
6 may use an electrode made of calomel. Also,
In addition to the KCl solution, a chloride solution such as sodium chloride and lithium chloride can be used as the internal solution 45. The internal liquid 44 in the outer tube 41 may be a physiological saline in addition to the chloride used for the internal liquid 45 in the inner tube 42.

The measuring electrode section 5 has the same structure as the standard electrode section 4. That is, it has a double outer tube 51 and an inner tube 52 made of plastic, and has a liquid junction 53 made of porous ceramic on the tube wall of the inner tube 52. The insides of the double outer tube 51 and the inner tube 52 are filled with internal solutions 54 and 55 each composed of a KCl solution having a predetermined concentration, and the internal solution 55 of the inner tube 52 is composed of silver-silver chloride. Electrode 56
Is immersed. The measurement electrode 56 is attached to an electrode holder 57 which also serves as a cap for closing the upper ends of the double outer tube 51 and the inner tube 52, and the inner electrode 52 in the inner tube 52 is suspended therefrom via a conductor. 55. A liquid replenishing port 58 for replenishing the inner liquid 54 into the outer tube 51 is provided on a peripheral surface near the upper end of the outer tube 51.

The measuring electrode unit 5 shown in FIG. 1 can have the same embodiment as the standard electrode unit 4.
That is, both of the double outer tube 51 and the inner tube 52 can be made of glass. Also, the liquid junction 53 may be made of porous Vycor glass. Further, the measurement electrode 56 may be an electrode made of calomel. In addition to the KCl solution, a solution of a chloride such as sodium chloride or lithium chloride can be used as the internal solution 55. The internal solution 54 in the outer tube 51 may be a physiological saline solution other than the chloride used for the internal solution 55.

Further, the standard electrode section 4 in FIG. 1 is connected to a flexible plastic tube immersed in the internal solution 44 in the outer tube 41 via a salt bridge 7 filled with agarose containing physiological saline, and then the DMEM is provided. (Dulbecco's mo
(dified Eagle's medium). This DMEM is a known culture solution containing amino acids, and is available as a commercial product. In addition, known substances such as those containing physiological saline can be used. Also,
The salt bridge 7 may be a known one, for example, a rubber tube made of silicone or the like can be used.

The measuring electrode section 5 is provided with an outer tube 5
The flexible plastic tube immersed in the internal liquid 54 in the container 1 is connected to the skin surface of the skin section 2 via a salt bridge 8 filled with agarose containing physiological saline and a filter paper 9 placed on the surface. ing. This filter paper 9 is for uniform contact of the skin section 2 of the salt bridge 8 with the skin surface. The salt bridge 8 may be a known one, for example, a rubber tube made of silicone or the like can be used.

The standard electrode 46 of the standard electrode unit 4 and the measuring electrode 56 of the measuring electrode unit 5 are connected to the potential difference detecting unit 6,
The potential difference between them is measured.

As the standard electrode section in the skin surface condition measuring apparatus, a standard electrode section having a liquid junction formed on the tube wall of the outer tube 41 of the standard electrode section 4 in FIG. 1 can be used.
The outer tube may be provided with a liquid replenishing port for replenishing the internal liquid as in FIG.

This standard electrode unit has the same function as the standard electrode unit 4 of the skin surface condition measuring apparatus in FIG. 1 and can be used in place of the standard electrode unit 4 in FIG. However, communication with the culture solution is performed through the liquid electrode provided on the outer tube wall, with the standard electrode portion immersed in the culture solution. The embodiment in the standard electrode section is the same as that in FIG.

Further, the inner tube 42 of the standard electrode unit 4 shown in FIG.
, Ie, only the inner tube 42, the liquid junction 43, the inner liquid 45, and the standard electrode 46 can be used.

This standard electrode unit also has the same function as the standard electrode unit 4 of the skin surface condition measuring apparatus in FIG. 1 and can be used in place of the standard electrode unit 4 in FIG. However, communication with the culture solution is performed through the liquid electrode provided on the tube wall by immersing the standard electrode portion in the culture solution. The embodiment in the standard electrode section is the same as that in the case of FIG.

[0046]

Next, more specific embodiments of the present invention will be described.

Example 1 Using the skin surface condition measuring device shown in FIG. 1, the potential difference (m
V) was measured. The skin sample used for the measurement was a skin section of a hairless mouse, and (1) a sample without any treatment on the skin surface, and (2) a substance P which is an irritant substance
(SP) added to a culture solution, and (3) cultured together with SP, an SP antagonist ([D-Arg ¹ , D-Trp ^7,9 , Leu ¹¹ ] -Substance P) known as a stimulus protective substance Three types, one added to the liquid, were used. The results are shown in FIG.

In FIG. 2, the abscissa indicates the elapsed time after the addition of SP. When an SP antagonist is added, the time at which the SP antagonist is added is set to zero. The vertical axis indicates the potential difference. The measured values are represented by an average of five measurements, and their variations are also shown.

As shown in FIG. 2, when SP was added to the culture solution with no treatment on the skin surface (control), a sharp change in the potential difference was observed. One hour after the addition, the state of a large change in the potential difference continues with a peak, and returns to a value close to the control 2 hours later. On the other hand, when the SP antagonist was added to the culture solution before the addition of SP, it was found that the potential difference was almost the same as that of the control until 2 hours after the addition, without a sharp change in the potential difference. From this, S
It can be seen that when an SP antagonist is added together with P, the SP antagonist suppresses SP stimulation and suppresses a potential difference change.

Example 2 Using the skin surface condition measuring device shown in FIG. 1, the potential difference (m
V) was measured. The skin sample used for the measurement is a skin section of a hairless mouse, and it can act as a stimulus inhibitor after (1) skin whose cell surface is barrier-destructed (tape stripping) with cellotape, and (2) tape stripping. Two types of skin, verapamil, which is a known calcium channel blocker, was added to a culture solution. The results are shown in FIG.

In FIG. 3, the horizontal axis indicates the elapsed time after tape stripping. The measured value of the elapsed time of zero is a value of the skin before tape stripping.
When verapamil was added to the culture solution, the time at which it was added was regarded as zero, and the measured value at the time of zero indicates the value of the skin before tape stripping. The vertical axis indicates the potential difference. The measured values are represented by an average of five measurements, and their variations are also shown.

As shown in FIG. 3, it can be seen that the stimulation by the tape stripping causes a sharp change in the potential difference compared to before. In contrast, when verapamil is added to the culture solution after tape stripping, the change in potential difference is suppressed. This indicates that verapamil suppresses the tape stripping stimulus, thereby reducing the change in potential difference.

Example 3 Using the skin surface condition measuring device shown in FIG. 1, the potential difference (m
V) was measured. The skin sample used for the measurement was a skin section of a hairless mouse, and (1) skin in which the skin surface was subjected to barrier destruction (tape stripping) with cellotape, (2) potassium tape blocker after tape stripping, Aminopyridine (AP) known to exhibit inhibitory effects
When tetrodotoxin (TTX), which is a sodium channel blocker and shows no stimulus-inhibiting effect after tape stripping, is added to the culture solution. , Three types of skin. FIG. 4 shows the results.

In FIG. 4, the horizontal axis indicates the elapsed time after tape stripping. The measured value of the elapsed time of zero is a value of the skin before tape stripping.
When AP or TTX is attached to the culture solution,
The time of attachment is defined as zero, and the measured value at the time of zero indicates the value of the skin before tape stripping. The vertical axis indicates the potential difference. The measured values are represented by an average of five measurements, and their variations are also shown.

As shown in FIG. 4, it can be seen that the stimulation by the tape stripping causes a sharp change in the potential difference compared to before. In contrast, when AP is added to the culture solution after tape stripping, the change in potential difference is suppressed. Further, when TTX is added to the culture solution, the change in the potential difference is not suppressed. From these results, it can be seen that the AP suppresses the tape stripping stimulus, resulting in a decrease in the potential difference change, and the TTX does not suppress the tape stripping stimulus, and thus does not reduce the potential difference change.

Example 4 Using the skin surface condition measuring device shown in FIG. 1, the potential difference (m
V) was measured. The skin sample used for the measurement was a skin section of a hairless mouse, and (1) a skin surface without any treatment, (2) a capsaicin, which is a stimulant applied to the skin surface, and (3) capsaicin. Was applied to the skin surface, and then applied with capsazepine, which is known as a protective substance against irritation. FIG. 5 shows the results.

In FIG. 5, the abscissa indicates the elapsed time since the application of capsaicin. When capsazepine is applied, the time when the capsazepine is applied is set to zero. The vertical axis indicates the potential difference. The measured values are represented by an average of five measurements, and their variations are also shown.

As shown in FIG. 5, when capsaicin was applied to the skin surface in contrast to the case where no treatment was applied to the skin surface (control), a sharp change in the potential difference was observed. The change of the potential difference is maximum in 5 hours. On the other hand, if capsaicin is applied after applying capsaicin to the skin surface, the range of change in the potential difference decreases,
It can be seen that as a result of the applied capsazepine suppressing irritation of the skin surface by capsaicin, the change in potential difference is reduced.

In addition to the above examples, similar results were obtained when histamine, dodecylbenzene sulfate (surfactant) and lactic acid (organic acid) were used as stimulants.

[0060]

As described above, according to the skin surface condition measuring apparatus of the present invention, a skin model in which a skin slice is floated with the surface facing upward on the liquid surface of the culture solution is prepared, and the skin surface and the culture solution are formed. Can be easily measured.
In addition, by applying a stimulating factor to the skin surface or the culture solution, the potential difference between the skin surface and the culture solution increases, and the degree of irritation applied to the skin can be confirmed. Furthermore, by applying or adding a stimulus-protecting substance to the skin before and after giving a stimulating factor to the skin, the degree of change in the potential difference is reduced, and a substance that inhibits skin irritation can be searched for.

[Brief description of the drawings]

FIG. 1 is a diagram showing an example of a skin surface state measuring device according to an embodiment of the present invention.

FIG. 2 is a graph showing an example of a result of measuring a skin surface potential difference with respect to a culture solution using the skin surface condition measuring device according to the present invention.

FIG. 3 is a graph showing another example of a result of measuring a skin surface potential difference with respect to a culture solution using the skin surface condition measuring device according to the present invention.

FIG. 4 is a graph showing another example of a result of measuring a skin surface potential difference with respect to a culture solution using the skin surface condition measuring apparatus according to the present invention.

FIG. 5 is a graph showing another example of a result of measuring a skin surface potential difference with respect to a culture solution using the skin surface condition measuring apparatus according to the present invention.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 ... Culture liquid 2 ... Skin slice 3 ... Container 4 ... Standard electrode part 5 ... Measurement electrode part 6 ... Potential difference detecting part 7,8 ... Shiohashi 9 ... Filter paper 41,51 ... Outer tube 42,52 ... Inner tube 43,53 ... liquid junction 44, 45, 54, 55 ... internal liquid 46 ... standard electrode 47, 57 ... electrode holder 48, 58 ... liquid replenishing port 56 ... measuring electrode

 ────────────────────────────────────────────────── ─── Continued on the front page (72) Inventor Kaori Inoue 2-12-1 Fukuura, Kanazawa-ku, Yokohama-shi, Kanagawa F-term in Shiseido Second Research Center Co., Ltd. (Reference) 2G045 AA40 CB09 FB05 GC18

Claims (22)

[Claims] 1. A skin surface condition wherein a skin slice is floated on a liquid surface of a culture solution contained in a container with its surface facing upward, and a potential difference between the skin surface of the skin slice and the culture solution is measured. Measurement method. 2. The method according to claim 1, wherein the culture solution is a culture solution containing physiological saline or amino acids. 3. A skin slice is floated on the surface of a culture solution contained in a container with its surface facing upward, and when measuring a potential difference between the skin surface of the skin slice and the culture solution, stimulation is applied to the skin surface. A method of measuring the skin potential from the difference between the measured potential difference and the potential difference when no stimulation is applied. 4. The method according to claim 3, wherein the stimulus is a stimulus substance. 5. The stimulus according to claim 3, wherein the stimulus is a physical stimulus.
The method for measuring the degree of skin irritation described in the above. 6. A skin slice is floated on the surface of a culture solution contained in a container with its surface facing upward, and when measuring the potential difference between the skin surface of the skin slice and the culture solution, an irritant is added to the culture solution. A method in which the potential difference is measured after addition, and the degree of skin irritation is measured from the difference between the value and the potential difference when no stimulant is added. 7. The method for measuring the degree of skin irritation according to claim 4, wherein the irritant is an itch-producing substance for the skin. 8. The method for measuring the degree of skin irritation according to claim 3, wherein the culture solution is a culture solution containing physiological saline or amino acids. 9. A stimulated skin slice is floated on the surface of a culture solution contained in a container with its surface facing upward, and the skin surface is measured when measuring the potential difference between the skin surface of the skin slice and the culture solution. A method of measuring the potential difference by applying a drug to the skin surface before or after applying a stimulus to the skin, and measuring the degree of inhibition of skin irritation by the drug from the difference between the value and the potential difference when the drug is not applied. 10. A stimulated skin slice is floated on the surface of a culture solution contained in a container with its surface facing upward, and the skin surface is measured when measuring the potential difference between the skin surface of the skin slice and the culture solution. A method in which a potential difference is measured by adding a drug to a culture solution before or after stimulating the skin, and the degree of inhibition of skin irritation by the drug is measured from the difference between the value and the potential difference when the drug is not added. 11. The method according to claim 9, wherein the stimulus is a stimulus substance. 12. The method according to claim 9, wherein the stimulus is a physical stimulus. 13. A skin slice is floated on a liquid surface of a culture solution to which a stimulant substance contained in a container is added, with the surface thereof facing upward, and in measuring the potential difference between the skin surface of the skin slice and the culture solution, the culture is performed. Before or after adding the stimulant to the solution, apply the drug to the skin surface and measure the potential difference, and measure the degree of inhibition of skin irritation by the drug from the difference between the value and the potential difference when the drug is not applied how to. 14. A skin slice is floated on a liquid surface of a culture solution to which a stimulant substance contained in a container is added, with the surface thereof facing upward, and when measuring a potential difference between the skin surface of the skin slice and the culture solution, culture is performed. Before or after adding the stimulant to the solution, add a drug to the culture solution and measure the potential difference, and measure the degree of inhibition of skin irritation by the drug from the difference between that value and the potential difference when no stimulant is added how to. 15. The method for measuring the degree of inhibition of skin irritation according to claim 11, wherein the stimulant is an itch-producing substance for the skin. 16. The method for measuring the degree of suppression of skin irritation according to any one of claims 9 to 15, wherein the culture solution is a culture solution containing physiological saline or amino acids. 17. A container containing a culture solution, a skin slice floating above the liquid surface, a standard electrode portion connected to the culture solution, and a skin surface of the skin slice. A skin surface condition measuring device comprising: a measuring electrode unit; and a potential difference detecting unit that detects a potential difference between the standard electrode unit and the measuring electrode unit. 18. The standard electrode part comprises: an inner tube containing an internal liquid in which a standard electrode is immersed; and an outer tube containing an internal liquid, which contains the inner tube and forms a double tube. 2. A standard electrode part, wherein a liquid junction is attached to the inner tube wall.
A skin surface condition measuring device according to claim 7. 19. The communication between the culture solution and a standard electrode unit,
19. The skin surface condition measuring device according to claim 18, wherein a salt bridge is connected between the culture solution and the internal solution in the outer tube. 20. The measurement electrode section comprising: an inner tube containing an internal liquid in which the measurement electrode is immersed; and an outer tube containing the internal liquid, which contains the inner tube to form a double tube. 2. A measurement electrode unit, wherein a liquid junction is provided on the inner tube wall.
The skin surface condition measuring device according to any one of claims 7 to 19. 21. The communication between the skin surface of the skin slice and the measurement electrode unit is by a salt bridge connected between the skin surface of the skin slice and the internal liquid in the outer tube. The skin surface condition measuring device according to claim 20. 22. The skin surface condition measuring device according to claim 17, wherein the culture solution is a culture solution containing physiological saline or amino acids.