TW201526873A - Evaluating method for skin texture - Google Patents

Abstract

An objective of this invention is to provide a novel measuring or evaluating method capable of easily and accurately measuring or evaluation skin texture state by using an biochemical index, rather than surgically removing skin to thereby stimulate skin. An evaluating method for skin texture of this invention uses expression amount of interleukin-1 receptor antagonist in skin horny layer as an index.

Description

Texture evaluation method

The present invention relates to a method of evaluating texture.

There is a fine grain called "texture" on the surface of human skin. Specifically, in the case of a mesh-like groove, the portion of the groove is referred to as "sulcus cutis". In addition, the portion surrounded by the "skin groove" is called "crista cutis".

It is said that the skin with fine texture is beautiful skin. In addition, various foundations have been developed to look like a skin with a fine texture. In order to look beautiful and healthy skin, it is important to make the texture small and not obvious.

As a technique for evaluating texture, a device has been developed in which the film is imaged by emphasizing the light and dark distribution on the surface of the skin, and the visual roughness of the wrinkles and texture is objectively quantified to evaluate wrinkles and texture, and then the cosmetics are coated. The wrinkles and the concealing effect of the texture (Patent Document 1: JP-A-6-189942).

In addition, a cosmetic device having a simple comparative sample such as a texture test piece for visually selecting a foundation according to the texture of the user (Patent Document 2: JP-A-9-262135) has been developed.

Because everyone has different skin types, even with Based on the above-mentioned viewpoints, the following techniques have also been developed: the reproduction of the texture is visually observed using a microscope, and the muscle fatigue is low, and the skin condition of the other person is compared and evaluated. The texture of the "mouth" is compared with the texture of the "cheek" with high muscle fatigue (Patent Document 3: JP-A-2005-58756).

In addition, the technique of evaluating the fineness of the texture by the copying method and evaluating the brightness and gloss of the skin is developed by focusing on the fineness of the skin and the glossiness of the skin (Patent Document 4: Japanese Special) Japanese Patent Publication No. 2010-273736).

Usually, a skin replica model in which the skin texture fraction is divided into four stages (texture fractionation) is prepared, and this is used as a reference, or the skin is directly visually observed, and it is common for an expert to evaluate the texture.

Because the individual differences in texture are large, as a useful information for selecting a foundation, and as an indicator of skin beauty, the evaluation of texture is particularly important, but it is difficult to directly observe with the naked eye, and a replica is used to observe the enlarged image in detail. It requires technology and is cumbersome to operate. In addition, regarding the visual evaluation of the texture, there is a tendency for the evaluator to vary. Therefore, it is desirable to have a simple evaluation method of texture.

[Previous Technical Literature] [Patent Literature]

[Patent Document 1] Japanese Patent Laid-Open No. 6-189942

[Patent Document 2] Japanese Patent Laid-Open No. Hei 9-262135

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

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

As described above, it is desirable to have a method of objectively evaluating texture. An object of the present invention is to provide a novel evaluation method that can easily and reliably evaluate texture without providing a method of surgically removing skin and causing irritation to the skin.

The present invention has the following constitution.

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

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

(3) A method for evaluating skin texture, which comprises the steps of: collecting a stratum corneum, and measuring the expression amount of an interleukin-1 receptor antagonist in the stratum corneum collected in the above-mentioned collection step; And comparing the expression amount of the interleukin-1 receptor antagonist measured in the aforementioned measuring step with the pre-determined distribution of the texture fraction in the stratum corneum and the expression distribution of the interleukin-1 receptor antagonist Comparison step.

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

Further, since the evaluation method of the present invention collects an evaluation sample by a simple operation method such as a tape peeling method, the burden on the subject is small, regardless of Anyone can simply evaluate. Further, since it is a biochemical test method, the same result can be obtained regardless of the measurement, so that it is not necessary to perform visual judgment by an expert as in the past.

Further, since the method according to the present invention can objectively numerically and easily evaluate the effect of adjusting the texture with cosmetics, the effect of cosmetics or the like can be further objectively evaluated.

Figure 1 is a graph comparing a skin replica model for texture evaluation with a texture score.

Fig. 2 is an image taken of a skin texture state classified as 1 to 4 by a texture score.

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

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

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

The present invention is a method for evaluating skin texture using an expression level of an interleukin-1 receptor antagonist in the stratum corneum 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 the expression amount of an interleukin-1 receptor antagonist (hereinafter referred to as IL-1Ra) in the stratum corneum of the skin. index.

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

The individual differences in texture shape can be said to be the skin shape that each person is born with. Along with the increase in age, the texture becomes rough and becomes irregular and unclear. However, due to the large individual differences in morphology, even young people have unclear textures, so the roughness of texture and texture is not necessarily straight. 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 the image on the skin surface to visually determine the fineness of the texture. It was found that the fineness of the texture thus determined was related to the amount of IL-1Ra expression in the stratum corneum of the skin.

IL-1Ra is an anti-inflammatory interleukin produced by monocytes, macrophages, etc., and increases in inflammation and normalizes the skin. IL-1Ra antagonally blocks the function of interleukin-1. When the cell level IL-1Ra is increased, matrix metalloproteinase-1, which is an enzyme that decomposes and fragments collagen, is reduced.

The stratum corneum is a tissue located at the uppermost layer of the skin, and has a function of protecting the skin from foreign matter and irritation from the outside of the body.

The portion to be evaluated in the present invention may include any portion as long as it is a portion in which the stratum corneum layer can be obtained, and the main portion may be exemplified as The face, neck, and upper arm of the target site were evaluated. The Tejia Department evaluates the stratum corneum of the face, which is particularly important for the evaluation of texture. The stratum corneum from the skin at the above site can be obtained according to a conventional method. However, as described above, since the method of surgically removing the skin or the like imposes a burden on the user, a method of easily obtaining a stratum corneum by a tape peeling method or a scratching method is preferable. Excellent tape stripping method.

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

IL-1Ra is extracted as a soluble component from each sample by a biochemical method known per se, such as a freeze-thaw method, an ultrasonication method, a homogenization method, or the like. The measurement was carried out rapidly after the extraction.

Since the expression of IL-1Ra in the stratum corneum of a person with a fine texture is significantly higher than that of the stratum corneum of a rough-skinned person, the amount of IL-1Ra in the stratum corneum is substituted for the texture score. The index of texture.

The stratum corneum is collected by a tape stripping method or the same simple method of collecting only the surface layer of the stratum corneum of the skin with a stratum corneum tape.

Since the tape stripping method does not burden the subject, it can be easily taken. Set the sample, so it is especially good.

The evaluation method of the present invention comprises the steps of: collecting an extraction step of the stratum corneum, measuring a measurement amount of IL-1Ra in the angle and the quality layer collected in the acquisition step, and measuring the IL measured in the aforementioned measurement step. A comparison step of comparing the expression amount of -1Ra with the IL-1Ra expression amount distribution in the collective stratum corneum measured in advance. The IL-1Ra expression amount of the subject was compared with the IL-1Ra expression amount distribution in the collective stratum corneum measured in advance, and when the amount of expression was large, it was judged that the texture was fine, and when the amount of expression was small, it was judged that the texture was rough.

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

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

172 healthy women (aged 18 to 65 years, mean age 40 years) were enrolled.

<Evaluation of cheek texture>

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

The standard of 4 stages is 1: the furrow and the ridge are not clear; 2: the furrow and the ridge are flowing and rough; 3: there are furrows and ridges, but messy; 4: In the furrow, the ridge, neat.

Photographs of each standard skin replica are shown in Figure 1.

Further, a representative cheek magnified photograph evaluated as each score is shown in Fig. 2 .

<Skin extraction method, protein quantification>

A stratum corneum test piece (manufactured by ASAHIBIOMED Co., Ltd.) was attached to the cheek of the subject to collect the stratum corneum. From the stratum corneum test piece, using a bead mill method (a method of adding a test object, a glass bead having a diameter of about 2 mm and an extract and performing an oscillating extraction method) using 500 μl of T-PER extraction buffer (Tissue Protein Extraction Reagent Thermo Scientific) The product of the stratum corneum protein was extracted (product #78510). The protein amount of each sample was determined by Pierce BCA protein Assay Kit (Thermo Scientific #23225). When measuring, 10 μl of the aforementioned stratum corneum protein extract was added. 200 μl of the reagent solution of the Pierce BCA protein Assay Kit was incubated at 60 ° C for 30 minutes, and the absorbance at 562 nm was measured. At the same time, a standard curve was prepared using BSA, and the amount of protein was calculated from the value of the absorbance.

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

Similarly, the amount of IL-1Ra expressed was measured using a ELISA kit [Human IL-1Ra/IL-1F3 Quantikine ELISA Kit] manufactured by R&D Systems Co., Ltd. from the stratum corneum protein extract extracted from the tape. First, a solid phase antibody which became a primary antibody was subjected to solidification at a concentration specified by the kit at 20 ° C overnight in a 96-well plate (#3590) manufactured by Costar. Next, the plate was blocked with 1% BSA at 37 ° C for 1 hour, and the plate was washed with 0.01% Tween-PBS, and each 100 μl was placed. Standards and samples that became standard curves were loaded and incubated at 25 ° C for 2 hours. After washing the plate with 0.01% Tween-PBS, the antibody was incubated for 2 hours at 25 ° C at the specified concentration of the kit. Further, after washing the plate with 0.01% Tween-PBS, Streptavidin-HRP was incubated at a specified concentration of the kit at 25 ° C for 30 minutes. Finally, after washing the plate with 0.01% Tween-PBS, TMB solution (Promega, G7431) was added as a chromogenic substrate at 100 μl/well. After the time specified in the kit, 100 μl of stop solution (0.5 M sulfuric acid) was added. The absorbance at 450 nm was measured and the amount of expression was calculated from the standard curve.

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

<Result>

Fig. 3 is a view showing the distribution of the box plot using the four-stage texture score value from the horizontal axis and the IL-1Ra value in the stratum corneum on the vertical axis.

Fig. 3 shows that the subjects with high expression of IL-1Ra had fine texture (texture score 4) and those with less IL-1Ra expression (texture score 1).

Further, a scattergram in which the horizontal axis represents the texture fraction and the vertical axis represents the IL-1Ra expression amount is shown in Fig. 4. The scattergram shows that the expression level of IL-1Ra is positively correlated with the increase of the texture score, and can be used as an indicator instead of the texture score.

<Reference example>

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

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

From the photograph shown in Fig. 2, it was confirmed that the skin texture was evaluated with the naked eye in comparison with the replica, which was very unclear and difficult to judge. From Fig. 3, it was found that the IL-1Ra value of the stratum corneum of the skin was highly consistent with the texture score evaluated by the expert. Therefore, by measuring the IL-1Ra of the stratum corneum of the skin, the texture of the skin can be evaluated even if it is not an expert.

Claims (3)

A method for evaluating skin texture, which is based on the expression amount of an interleukin-1 receptor antagonist in the stratum corneum of the skin. The evaluation method according to claim 1, wherein the skin cuticle is collected by a tape peeling method. A method for evaluating skin texture, comprising the steps of: collecting a stratum corneum, measuring a performance amount of an interleukin-1 receptor antagonist in the stratum corneum collected in the collecting step, and Comparison step of comparing the expression amount of the interleukin-1 receptor antagonist measured in the aforementioned measuring step with the texture fraction of the muscle layer in the collective stratum corneum and the expression distribution of the interleukin-1 receptor antagonist .