WO2012073892A1

WO2012073892A1 - Method for evaluating texture using oxytalan fibers as index

Info

Publication number: WO2012073892A1
Application number: PCT/JP2011/077397
Authority: WO
Inventors: 青木　宏文; 美郷豊田; 勝田　雄治; 茂麦倉
Original assignee: 株式会社資生堂
Priority date: 2010-11-29
Filing date: 2011-11-28
Publication date: 2012-06-07
Also published as: JP2012117827A

Abstract

The purpose of the present invention is to establish a physiologically based method for evaluating the texture of skin, provide a method for screening a skin texture improvement method/improvement drug which uses a method for physiologically evaluating the texture of skin, and provide a method for improving the texture of skin on the basis of physiological findings relating to texture, obtained by the present invention. By the present invention, the total oxytalan fiber length per unit length of a skin section is measured. The total oxytalan fiber length in the skin section serves as an index for the texture of the skin.

Description

Skin texture evaluation method using oxytalan fiber as an index

The present invention relates to a method for measuring the fineness of skin texture using oxytalan fibers as an index. The present invention further relates to a method for improving skin texture by increasing oxytalan fibers, and a method for screening a skin texture improving agent using oxytalan fibers as an index.

The texture composed of skin grooves and cuticles on the skin surface causes light scattering on the skin surface and is involved in the youthfulness and beauty of the skin. In general, infants' skin has many fine textures, giving them a smooth and transparent feeling that is unique to infants. However, with the age, the small texture disappears and the texture becomes larger, and the skin groove flows and the directionality is increased. It will cause wrinkles and fine wrinkles.

Therefore, maintaining the texture of the skin is particularly useful in cosmetics, and measuring and evaluating the texture is important in cosmetics and in the research and development of products related to beauty and the provision of services. So far, the method of evaluating the skin condition by photographing the surface of the skin with a microscope and performing information processing on the skin groove and the cuticle (Patent Document 1), and the skin condition using the replica method A method for evaluating skin (Patent Document 2) and a skin texture evaluation method using an acoustic system (Patent Document 3) have been developed.

However, the skin texture evaluation methods known so far are based on external measurement of the surface condition of the skin, but the texture is evaluated histologically and physiologically from the inside of the skin. The method has not yet been developed.

JP 2006-305184 A Japanese Patent Laid-Open No. 2001-170028 JP 2008-114067 A

Therefore, the problems to be solved by the present invention are to establish a method for histologically and physiologically evaluating skin texture, and to improve and improve the skin texture by using a physiological evaluation method for skin texture. It is to provide a drug screening method and to provide a skin texture improving method based on the histological and physiological knowledge of the texture obtained by the present invention.

As a result of intensive studies aimed at solving the above-mentioned problems, the present inventors have found that many oxytalan fibers running vertically to the epidermis are present inside the cuticle part, but are present inside the skin groove part. I found that I did not. Based on this discovery, the present inventors have found that the rise of the dermis is caused by oxytalan fibers running longitudinally in the epithelium, and oxytalan fibers are involved in the formation of the texture composed of the dermis and skin groove. I hypothesized that. As a result of further verification of this hypothesis, the total oxytalan fiber length per unit length inside the dermis of the skin section was measured and compared with the dermis density grade defined as an indicator of the fineness of the dermis. It was surprisingly found that there is a high correlation between a decrease in density grade and a decrease in the fiber length. In other words, it is considered that the decrease in the total oxytalan fiber length inside the skin mound, the decrease in fineness of the texture, and the deterioration of the shape are deeply involved.

Based on this discovery, a texture evaluation method was established by measuring the total oxytalan fiber length per unit length of the skin section. Furthermore, the texture evaluation method further includes a step of comparing the measured total oxytalan fiber length with an index for the average total oxytalan length in each dendrite density grade. Furthermore, as described above, the present inventors have clarified the structure of the texture from the histological and / or physiological aspects, so that the total oxytalan fiber length per unit skin section before and after the administration of the candidate drug. The present invention provides a method for screening a skin texture-improving drug by measuring, and a method for improving skin texture by administering a drug that increases oxytalan fibers.

Conventionally, only the evaluation method of the texture by measuring the skin surface has been known, and the present invention provides a method for evaluating the texture histologically and physiologically. There is an advantage in providing a holistic approach. Furthermore, the present invention provides histological and physiological knowledge about the formation of the texture, and enables screening of drugs by focusing on the increase in oxytalan fibers in the texture improving drug and the texture improving method. There are advantages.

FIG. 1 is a diagram showing a hide density density grade. FIG. 2 is a skin section of a 35 year old adult stained with Elastica Wangison (Weigert) stain. FIG. 3 is a skin section of a 66 year old adult stained with Elastica Wangyson (Weigert) stain. FIG. 4 is a diagram in which oxytalan fibers are identified and traced in a skin section of a subject stained with Elastica Wangison (Weigert) staining. FIG. 5 is a graph plotting the total oxytalan fiber length against the skin density grade, and the Pearson correlation coefficient is 0.54.

The present invention relates to a skin texture evaluation method by measuring the total oxytalan fiber length per unit length of a skin section. More specifically, the present invention relates to a skin texture evaluation method by measuring the total oxytalan fiber length per arbitrary length (for example, 1 mm, 2 mm, 0.5 cm, 1 cm, 2 cm, 5 cm, etc.) of a skin section. . The present inventors have shown that the total oxytalan fiber length per unit length directly under the skin epidermis correlates with the dermis density grade, and by measuring the total oxytalan fiber length per unit length of the skin section, Can evaluate skin texture.

In one aspect of the present invention, the texture of the skin is evaluated by comparing the total oxytalan fiber length per unit length of the target with the total oxytalan fiber length per unit length as an index. This index can be created from a graph obtained by plotting the total oxytalan fiber length per unit length against the skin density density of the subject group. Since bark density grades 1-2, 2-3, and 3-4 are judged as poor, normal, and good, respectively, the total oxytalan fiber length corresponding to

bark density grades

2 and 3 is used as an index. It can be. The total oxytalan fiber length corresponding to an arbitrary point such as a skin density grade 1.5, 2.5, and 3.5 can also be used as an index. By applying the total oxytalan fiber length per unit length of the subject to such a graph, the texture of the subject's skin can be evaluated. In addition, any total oxytalan fiber length per unit length can be used as an index. For example, 500 μm, 1000 μm, 1500 μm, 2000 μm, 2500 μm, or 3000 μm can be selected as an index per 1 mm of skin length.

In another aspect of the present invention, the skin by comparing the total oxytalan fiber length per unit length measured with the total oxytalan fiber length per unit length in the skin of the same subject, the same or a corresponding site, measured in advance It is related with the texture evaluation method. In this case, it is possible to evaluate whether or not the use of a texture improving agent or a skin care product contributes to the improvement of the texture.

Another aspect of the present invention relates to a method for screening a skin texture improving agent by measuring the total oxytalan fiber length per unit length before and after administration of a candidate drug. By measuring the oxytalan fibers involved in the formation of the texture, it is possible to screen the skin improvement effect of the candidate drug with a sensitivity superior to the evaluation method of the texture by observing the epidermis.

In a further aspect of the present invention, the present invention also relates to a method for improving skin texture by increasing oxytalan fibers. Since the present inventors have discovered that oxytalan fibers in the skin epidermis are involved in the formation of skin texture, it has been found that increasing oxytalan fibers is important for improving the texture. Therefore, the skin texture can be improved by administration of a drug that increases oxytalan fibers in the epithelium or skin care. The improvement of the texture can be done for cosmetic, therapeutic and preventive purposes. Examples of such skin care include, but are not limited to, massage and moisturizing.

In the present invention, the term “texture” refers to a surface form formed by a cuticle and a skin groove on the surface of skin. The skin hill part constituting the texture is surrounded by a skin groove, and many take a triangular shape, but some take a polygonal shape. A well-textured state refers to a state where the number of fine triangular cuticles and crevices that are well-formed is large, and the number and density of the crevice decreases with age as the crevice and cuticles become blurred. However, the length of the crevice tends to increase, and such a state is called a bad state. Since the hide density density grade defined in this specification represents the fineness of the hide, the larger the hide density density grade, the better the texture (FIG. 1). The present inventors have discovered that many oxytalan fibers running vertically to the epidermis are present inside the dermis, but not inside the sulcus, and the swell of the dermis is raised. It was concluded that is caused by oxytalan fibers running longitudinally in the epithelium. While dense and long oxytalan fibers are present in the cuticle part, and no oxytalan fibers are present in the skin groove part, the skin groove is deepened and the texture state is excellent. By reducing the density and length of the oxytalan fibers in the cuticle, the crevice becomes shallow, the boundary between the crevice and the cuticle becomes blurred, and the texture deteriorates. Therefore, it is possible to evaluate the texture state by measuring the fiber length of oxytalan per unit length.

Any site can be used as the site of the skin for measuring the total oxytalan fiber length per unit length, but the skin of the face such as the cheeks and the forehead, the back of the hand, and the skin of the hand are particularly preferable. For example, a slice of arm skin is used. Furthermore, a surplus skin fragment obtained by diet surgery can also be used. In this case, the total oxytalan fiber length per unit length is measured for the abdominal skin.

Measurement of the total oxytalan fiber length per unit length of the skin section is performed after the skin section is stained so that the oxytalan fiber can be identified. The skin section may be stained by any method as long as oxytalan fibers can be identified. For example, Elastica Wangyson (Weigert) staining, which can stain elastic fibers, is performed, and oxytalan fibers are stained purple-black with Weigert's solution. In the skin section, the oxytalan fibers run perpendicular to the epidermis surface, and a large amount exists particularly in the cuticle (FIGS. 2 and 3).

The stained image of the skin section is captured with a CCD camera, the oxytalan fibers are traced with computer software (Win Roof), and the total trace length per 1 mm of the epidermis is summed to measure the total oxytalan fiber length per unit length of the epidermis. be able to.

“Oxytalan fiber” is a fiber component that creates mechanical strength in the connective tissue of a living body, and is one of elastic fibers. Elastic fibers are particularly abundant in organs and tissues subjected to mechanical pressure, such as skin, blood vessels, lungs, and ligaments. Elastic fibers are composed of elastin and microfibril protein, and are classified into three types according to the composition ratio of elastin: elastic fiber, elaurin fiber, and oxytalan fiber. Among these, oxytalan fibers are those in which the content of elastin fibers is low and the microfibril protein forms a dense bundle. In tissues, it has been reported that they exist as periodontal ligaments that support teeth and as fibers directly under the epidermis that run perpendicular to the epidermis surface in the skin (Non-patent Document 2).

“Dune density grade” is an indicator of the fineness of the hide. The texture is divided into four groups: bad, slightly bad, normal, and good, and representative images of each are listed in FIG. By comparing the skin texture images of the subject's skin, the hill density grade in the subject's skin can be determined.
Specifically, several experienced researchers familiar with the enlarged image of the texture compare the texture image group to be evaluated with this dendrite density grade, and have the fineness of the nearest dend. The grade number that can be felt is selected for each texture image. At this time, attributes such as age of the subject of each texture image are completely hidden by both the evaluation researcher and the test management researcher, so that a double blind test is performed. By calculating the average value of the hill density grades from a plurality of researchers obtained for each texture image, the skin density grade of the texture image is obtained.
Since the skin condition and the dermis density grade are closely related, if it is judged that it falls under the hill density grade 1-2, it is judged that the skin condition is bad and falls within the hill density grade 2-3. If it is determined that the texture state is normal, and it is determined that it corresponds to the hill density grade 3 to 4, it can be determined that the texture state is good.

The term “administration” as used herein includes, but is not limited to, transdermal administration, oral administration, intradermal administration, subcutaneous administration, intramuscular administration, intravenous administration, intraperitoneal administration, and the like. It is not a thing. Preferably, the medicament is administered by transdermal administration, subcutaneous administration, or intradermal administration.

For the surplus skin fragment of the abdomen obtained by Elastica wanggieson (Weigert) staining diet operation, a frozen section of the skin was prepared according to a conventional method, the tissue was fixed with 10% neutral buffered formalin solution, and then twice with PBS It was washed and immersed in Weiger tresol synthesize fuchsin staining solution (Mudo Chemical No. 4032) for 40 minutes or more. Next, it was immersed in 100% ethanol for a short time three times and then washed with water for 10 minutes. Tissue sections were embedded in water and observed under an optical microscope. Elastic fibers including oxytalan fibers were stained purple-black (FIGS. 2 and 3).

Measurement of Oxytalan Fiber Length The above-mentioned stained image of Elastica Wanghisson (Weigert) staining is captured with a CCD camera, and fibers (oxytalan fibers) that run perpendicular to the epidermis surface are used with computer software (Win Roof (registered trademark)). The total trace length per 1 mm of the epidermis was totaled to obtain the total oxytalan fiber length per unit length (FIG. 4).

Correlation between Oxytalan Fiber Length and Peel Density Grade The texture image in Fig. 1 is defined as skin mound density grade 1 to 4 respectively, and four skilled researchers familiar with the enlarged texture image are the texture images to be evaluated. The group was compared with the image of FIG. 1, and a grade number that was visually felt as having the fineness of the nearest skin was selected for each texture image. At this time, the attributes such as age of the subject of each texture image are hidden by both the evaluation researcher and the test management researcher to make a double blind test, and the skins from multiple researchers obtained for each texture image By calculating the average value of the density grades, it was set as the hill density grade of the texture image.

Table 1 below shows the relationship between the bark density grade thus obtained and the total oxytalan fiber length per unit length. FIG. 5 was obtained by plotting the total oxytalan fiber length per unit length against the bark density grade. Pearson's correlation coefficient was 0.54, indicating a correlation between the dermis density grade and the total oxytalan fiber length per unit length.

The skin density grade is an index of the fineness of the skin, and since it has a close relationship with the texture, the texture can be evaluated using the oxytalan fibers of the skin as an index.

Claims

A texture evaluation method performed for cosmetic purposes by measuring the total oxytalan fiber length per unit length of a skin section.
2. The texture evaluation method according to claim 1, further comprising a step of comparing the measured total oxytalan fiber length per unit length with an index of the total oxytalan fiber length per unit length representing the texture state.
A screening method for a skin texture-improving drug by measuring the total oxytalan fiber length per unit length of a skin section before and after administration of a candidate drug.
A method of improving skin texture by administering a drug that increases oxytalan fibers.