TWI622382B - Evaluating method for skin texture - Google Patents

Abstract

The subject of the present invention is to provide a novel measurement or evaluation method using a biochemical index that can easily and reliably measure or evaluate the texture state, instead of a method of removing the skin surgically and irritating the skin. The evaluation method of the skin texture of the present invention is based on the expression level of the interleukin-1 receptor antagonist in the cuticle of the skin as an index.

Description

Evaluation method of texture

The present invention relates to a method for evaluating texture.

There are fine lines called "texture" on the surface of human skin. Specifically, it is a mesh-like groove, and a part of the groove is called a "sulcus cutis". In addition, a portion surrounded by a "skin ditch" is called a "crista cutis".

It is said that skin with fine texture is beautiful skin. In addition, various foundations have been developed for skin that looks fine. In order to look beautiful and healthy skin, it is important to make the texture small and inconspicuous.

As a technique for evaluating texture, a device has been developed that takes pictures by emphasizing the distribution of light and dark on the skin surface, and objectively quantifies the visual roughness of wrinkles and textures to evaluate wrinkles and textures, and then evaluates the application of cosmetics Effect of hiding wrinkles and texture (Patent Document 1: Japanese Patent Application Laid-Open No. 6-189942).

In addition, a cosmetic test piece such as a texture test piece (patent document 2: Japanese Patent Application Laid-Open No. 9-262135) for visual selection for selecting a foundation based on the texture of the user has been developed.

Since everyone has different skins, Based on the above viewpoints, we have also developed the following technology: comparing the skin condition of other people with relative evaluation and evaluating the relative quality of the skin, using a microscope to visually observe the reproduction of the texture, and reducing muscle fatigue The texture of the "mouth" is compared with the texture of the "cheek" having high muscle fatigue (Patent Document 3: Japanese Patent Application Laid-Open No. 2005-58756).

In addition, focusing on the small texture, the skin has less brightness and gloss, and developed a technology that evaluates the texture fineness by the replication method and is effective for the evaluation of skin brightness and gloss (Patent Document 4: Japanese Patent (2010-273736).

Generally, it is common to make a skin replica model that divides the texture of the skin into 4 stages (fraction of texture) and use it as a reference, or directly look at the skin and evaluate the texture by experts.

Due to the large individual differences in texture, as useful information for selecting foundations, and as an indicator of skin beauty, the evaluation of texture is particularly important, but it is difficult to observe the system directly with the naked eye, using replicas to observe enlarged images in detail Technology is required and the operation is cumbersome. In addition, the visual evaluation of the texture may be biased by the evaluator. Therefore, a simple objective evaluation method of texture is desired.

[Prior technical literature] [Patent Literature]

[Patent Document 1] Japanese Unexamined Patent Publication No. 6-189942

[Patent Document 2] Japanese Unexamined Patent Publication No. 9-262135

[Patent Document 3] Japanese Patent Laid-Open No. 2005-58756

[Patent Document 4] Japanese Patent Laid-Open No. 2010-273736

As described above, a method for objectively evaluating texture is desired. The subject of the present invention is to provide a novel evaluation method that can easily and reliably evaluate the texture, instead of a method of removing the skin surgically and irritating the skin.

The present invention has the following configuration.

(1) A method for evaluating skin texture, which uses an expression amount of an interleukin-1 receptor antagonist in the stratum corneum of the skin as an index.

(2) The evaluation method according to (1), wherein the skin stratum corneum is collected by a tape peeling method.

(3) An evaluation method of skin texture, which is composed of the following steps: the step of collecting the stratum corneum, and the measurement of the expression of the interleukin-1 receptor antagonist in the stratum corneum collected in the foregoing step of collecting. Step, and comparing the expression level of the interleukin-1 receptor antagonist measured in the aforementioned measurement step with the texture score in the collective cuticle and the expression level distribution of the interleukin-1 receptor antagonist measured in advance Comparison steps.

According to the present invention, the texture of the skin can be objectively evaluated by measuring the expression level of the interleukin-1 receptor antagonist of the skin stratum corneum, thereby replacing the visual evaluation method performed by an expert.

In addition, since the evaluation method of the present invention collects evaluation samples by a simple operation method such as a tape peeling method, the burden on the subject is small, regardless of Anyone can easily evaluate. In addition, since it is a biochemical test method, the same result can be obtained regardless of who measures it, so it is not necessary to make a visual judgment by an expert as in the past.

In addition, since the method according to the present invention can objectively quantify and easily evaluate the effect of adjusting texture with cosmetics, the effects of cosmetics and the like can be further objectively evaluated.

FIG. 1 is a graph comparing a skin replica model used for texture evaluation and a texture score.

Fig. 2 is an image taken of skin texture states classified by texture scores from 1 to 4.

Fig. 3 is a box diagram showing the measurement results of the amount of interleukin-1 receptor antagonist in the stratum corneum collected from skin classified by texture score.

Fig. 4 is a scattergram showing the horizontal axis as the texture score and the vertical axis as the amount of interleukin-1 receptor antagonist in the stratum corneum.

Figure 5 is a graph showing the number distribution of texture scores for women aged 30 to 39 years.

The present invention is an evaluation method of skin texture using the expression amount of an interleukin-1 receptor antagonist in the cuticle of the skin as an index.

Hereinafter, the present invention will be described in detail.

The texture evaluation method of the present invention is characterized by taking the expression level of an interleukin-1 receptor antagonist (hereinafter, referred to as IL-1Ra) in the stratum corneum of the skin as a characteristic index.

The texture in the present invention refers to fine lines on the surface of human skin, specifically, mesh-like grooves. The part of the ditch is called "skin ditch". In addition, the part surrounded by the "skin ditch" is called "skin ridge". It is said that the skin with fine texture, that is, the skin with fine mesh, regular, clear skin is beautiful.

The individual shape of the texture varies greatly, and it can be said that each person is born with a skin shape. With the increase of age, the texture tends to become rough and become irregular and unclear. However, due to the large individual differences in shape, even young people have unclear texture, so the aging and the roughness of the texture may not be directly correspond.

The fineness of the texture can be determined by preparing a standard texture replica image shown in FIG. 1 and comparing the standard image with an image of the skin surface to visually determine the fineness of the texture. It was found that the fineness of the texture determined in this way was correlated with the expression level of IL-1Ra in the stratum corneum of the skin.

IL-1Ra is an anti-inflammatory interleukin produced by monocytes, macrophages, and the like, and increases during inflammation and normalizes the skin. IL-1Ra antagonizes the function of interleukin-1. As cell level IL-1Ra increases, matrix metalloproteinase-1, which is an enzyme that breaks down and fragments collagen, decreases.

The stratum corneum is the tissue located on the top layer of the skin and has the function of protecting the skin from foreign bodies and irritation from the outside.

The site to be evaluated in the present invention may include any site as long as the stratum corneum can be obtained, and the main site can be cited as The face, neck, and upper arm of the evaluation target area. The special line evaluation is particularly important for the evaluation of the texture of the stratum corneum of the face. The stratum corneum derived from the skin of the above region can be obtained by a conventional method. However, as described above, since a method such as surgically exfoliating the skin places a burden on the user, a method such as a tape peeling method and a scraping method that can easily obtain the stratum corneum is preferred. Extra good tape peeling method.

The expression amount of IL-1Ra in each sample thus prepared can be measured by a conventionally known method. For example, a method such as an enzyme immunoassay, a radioimmunoassay, a Western blotting, or the like based on a reaction with an antibody against IL-1Ra can be used. Such a measurement method is commercially available. As a typical IL-1Ra enzyme immunoassay kit, human IL-1Ra / IL-1F3 Quantikine ELISA Kit manufactured by R & D Systems, and IL-1Ra manufactured by Life Technologies can be exemplified. Human ELISA Kit.

IL-1Ra is extracted from each sample as a soluble component by a biochemical method known per se, such as a freeze-thaw method, an ultrasonic disruption method, a homogenization method, and the like. Measurement was performed quickly after extraction.

Since the expression of IL-1Ra in the stratum corneum of a person with a small texture is significantly higher than the expression of IL-1Ra in a stratum corneum of a person with a rough texture, the amount of IL-1Ra expression in the stratum corneum is used instead of the texture score Indicators of texture.

The stratum corneum is collected by a tape stripping method or the simple method of collecting only the superficial part of the stratum corneum of the skin with a stratum corneum tape.

Can be easily collected because the tape peeling method does not burden the subject It's very good to collect samples.

The evaluation method of the present invention includes the following steps: a collection step of collecting stratum corneum, a measurement step of measuring the expression level of IL-1Ra in the stratum corneum collected in the foregoing collection step, and an IL-1Ra measured in the foregoing measurement step. A comparison step in which the expression level is compared with the IL-1Ra expression level distribution in the collective cuticle measured in advance. The IL-1Ra expression level of the subject was compared with the IL-1Ra expression level distribution in the collective cuticle measured in advance. When the expression level was large, the texture was judged to be small, and when the expression level was small, the texture was judged to be rough.

Hereinafter, specific examples will be described, but the present invention is not limited to the following examples.

(Example) Test example Confirmation test on the relationship between the collective evaluation of the subject's texture and the expression of IL-1Ra <Subject>

172 healthy women (18 to 65 years old, average age 40 years) were taken as subjects.

<Evaluation of cheek texture>

A magnified photograph of the subject's cheek was taken using an iPhone (brand name: Apple Inc.) equipped with Pocket Micro (Scalar Co., Ltd.), and the texture was visually judged by a professional beauty researcher, and the score was recorded according to the following criteria There are 4 stages.

The criteria for stage 4 are 1: the sulcus and ridges are not clear; 2: the sulcus and ridges are flowing and rough; 3: the sulcus and ridges are present, but they are messy; 4: exist Neatly in the furrows and skin ridges.

Photographs of skin replicas for each standard are shown in Figure 1.

In addition, a representative enlarged cheek photograph evaluated as each score is shown in FIG. 2.

<Cut layer sample extraction method, protein quantification>

A cuticle test sheet (manufactured by Asahibodimed Co., Ltd.) was attached to the cheek of the subject to collect the cuticle. From the stratum corneum test piece, a bead mill method (a method of adding a test substance, glass beads with a diameter of about 2 mm, and an extraction solution to a container and performing shake extraction) was performed using 500 μl of T-PER extraction buffer (Tissue Protein Extraction Reagent Thermo Scientific (Product # 78510) to extract cuticle protein. The protein mass of each sample was measured using Pierce BCA protein Assay Kit (Thermo Scientific # 23225). During the measurement, 10 μl of the cuticle protein extract was added. 200 μl of the Pierce BCA protein Assay Kit reaction reagent solution was incubated at 60 ° C. for 30 minutes, and then the absorbance at 562 nm was measured. At the same time, a standard curve was prepared with BSA, and the protein mass was calculated from the absorbance value.

<Measurement of IL-1Ra in the stratum corneum>

Similarly, the cuticle protein extract solution extracted from the tape was used to measure the expression amount of IL-1Ra using an ELISA kit [Human IL-1Ra / IL-1F3 Quantikine ELISA Kit] manufactured by R & D systems. First, in a 96-well plate (# 3590) made by costar, a solid-phase antibody that becomes a primary antibody was solid-phase coated at a concentration specified in a kit at 20 ° C. overnight. Next, the plate was blocked with 1% BSA at 37 ° C for 1 hour, and the plate was washed with 0.01% Tween-PBS to form a standard. 100 μl of standard samples and 100 μl of samples were sampled and incubated at 25 ° C. for 2 hours. After the culture plate was washed with 0.01% Tween-PBS, the secondary antibody was cultured at a concentration specified in the kit at 25 ° C for 2 hours. Furthermore, after the culture plate was washed with 0.01% Tween-PBS, Streptavidin-HRP was cultured at a concentration specified in the kit at 25 ° C for 30 minutes. Finally, the plate was washed with 0.01% Tween-PBS, and TMB solution (Promega, G7431) was added as a coloring matrix at 100 μl / well. After the time specified in the kit, 100 μl of stop solution (0.5M sulfuric acid) was added. ), Measure the absorbance at 450 nm, and calculate the expression from the standard curve.

The obtained IL-1Ra values were classified by texture scores, and boxplots and scatter plots were made for analysis.

<Result>

Fig. 3 shows the distribution using a box plot obtained from the texture score values of the four stages on the horizontal axis and the IL-1Ra value in the stratum corneum of the subject on the vertical axis.

Figure 3 shows that the subjects with high IL-1Ra expression were fine in texture (texture score 4), and those with low IL-1Ra expression were rough in texture (texture score 1).

In addition, a scatter plot showing the texture score on the horizontal axis and the expression of IL-1Ra on the vertical axis is shown in FIG. 4. The scatter plot shows that the expression of IL-1Ra is positively correlated with the increase in texture score, and can be used as an indicator instead of texture score.

<Reference example>

Females aged 30 to 39 years were selected from the subjects, and the distribution of texture scores (30) was confirmed.

As shown in Figure 5, it can be seen that even for the same age group, the texture score cannot be represented by a specific score, but is widely distributed.

It can be confirmed from the photograph shown in FIG. 2 that the evaluation of the skin texture with the naked eye is very unclear and difficult to judge compared with the replica. It is clear from the third figure that the IL-1Ra value of the stratum corneum of the skin is highly consistent with the texture score evaluated by experts. Therefore, by measuring IL-1Ra of the stratum corneum of the skin, it is possible to evaluate the texture of the skin even if it is not an expert.

Claims (3)

A method for evaluating skin texture, which uses the expression of interleukin-1 receptor antagonist in the stratum corneum of the skin collected by tape peeling or scraping as an index, and uses the interleukin of the subject The expression level of the -1 receptor antagonist is compared with the expression level distribution of the interleukin-1 receptor antagonist in the collective cuticle measured in advance. When the expression level is large, it is judged to be small in texture and when the expression level is small. , Judged as rough texture. The evaluation method according to item 1 of the scope of patent application, wherein the cuticle of the skin is collected by a tape peeling method. A method for evaluating skin texture, which is composed of the following steps: a step of collecting stratum corneum by tape peeling or scraping, and measuring interleukin-1 receptor antagonism in the stratum corneum collected in the foregoing step of collecting A step of measuring the expression amount of the agent, and the expression amount of the interleukin-1 receptor antagonist measured in the aforementioned measurement step and the texture score and interleukin-1 receptor antagonist in the collective cuticle measured in advance A comparison step for comparing the performance distributions.