Method for estimating hair cycle, method for estimating collagen degradation and / or neogenesis

Abstract

To provide a new technology of estimating a hair cycle.SOLUTION: The present invention is a hair cycle estimation method for estimating a hair cycle of hair follicles in an evaluation target based on analysis results of collagen degradation and / or neogenesis in perifollicular tissues including hair follicles.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for estimating the hair growth cycle and a method for estimating collagen degradation and / or neogenesis. [Background technology]

[0002] Hair follicles are tissues that produce hair, such as hair. The hair matrix cells and hair stem cells that make up these follicles control the cyclical regeneration and shedding of hair. This cycle of hair regeneration and shedding is called the hair cycle, and is divided into the anagen phase, catagen phase, and telogen phase. It is known that abnormalities in the hair growth cycle can cause hair growth disorders. In recent years, research has been conducted on hair, particularly hair, to elucidate the mechanisms underlying hair thinning and hair loss.

[0003] Patent Document 1 focuses on the relationship between hair cycle abnormalities and hair loss, and discloses a method for evaluating candidate substances for hair growth agents or hair care ingredients using an alopecia model in which hair cycle abnormalities are induced.

[0004] Furthermore, accurate classification of the hair growth cycle is clinically important in order to accurately assess the symptoms of patients with alopecia, etc. Observations of human hair focusing on its histological structure have been widely conducted to provide an index for accurately assessing the hair growth cycle (Non-Patent Document 1).

[0005] Collagen is a protein secreted mainly by fibroblasts. Among collagens, fibrous type I collagen is a component that forms the skeletal structure of the dermis and is involved in maintaining the elasticity and flexibility of the skin. It is also known that the promotion of the degradation of fragmented collagen promotes the production of new collagen (e.g., Non-Patent Document 2). In recent years, methods for improving skin firmness, elasticity, etc. by promoting collagen metabolism have attracted attention. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Patent No. 3953709 [Non-patent literature]

[0007] [Non-Patent Document 1] Ji Won Oh et al., J Invest Dermatol. 2016 Jan; 136(1): 34‐44. [Non-patent document 2] Iwahashi H. et al., Photodermatol Photoimmunol Photomed. 2021 Sep 1. Summary of the Invention [Problem to be solved by the invention]

[0008] In view of the above prior art, an object of the present invention is to provide a new technique for estimating the hair growth cycle. Another object of the present invention is to provide a new technique for inferring collagen degradation and / or neogenesis. [Means for solving the problem]

[0009] The present invention, which solves the above problem, is a method for estimating the hair cycle of a hair follicle in an evaluation subject based on the analysis results of collagen degradation and / or neogenesis in the tissue surrounding the hair follicle, including the hair follicle. By using the present invention, it is possible to estimate the hair cycle of hair follicles in the skin tissue being evaluated.

[0010] The present invention was made based on the discovery by the present inventors that the amount and location of collagen degradation and neogenesis in the tissues surrounding the hair follicle change depending on the hair cycle. That is, according to the present invention, the hair growth cycle can be easily estimated simply by focusing on collagen metabolism, such as collagen degradation and / or collagen synthesis.

[0011] The present invention can also be applied to estimating the effects of drugs or cosmetic procedures that contribute to normalizing the hair cycle.

[0012] In a preferred embodiment of the present invention, the means for analyzing collagen degradation and / or neogenesis comprises analyzing the amount and / or site of collagen degradation and / or the amount and / or site of collagen neogenesis. By analyzing the amount and / or location of collagen degradation and / or neogenesis, the hair cycle can be estimated without the need for detailed analysis of the histological structure of the tissue surrounding the hair follicle.

[0013] In a preferred embodiment of the present invention, the means for analyzing collagen degradation and / or collagen synthesis is a means for detecting degraded collagen and / or procollagen.

[0014] In a preferred embodiment of the present invention, a collagen degradation detection step of detecting degraded collagen in the perifollicular tissue is provided. a collagen synthesis detection step of detecting procollagen in the perifollicular tissue; and an estimation step of estimating the hair cycle based on the amount of collagen decomposition and the collagen new synthesis site and / or collagen new synthesis amount. By using the amount of collagen degradation and the amount and / or site of collagen synthesis as indicators, the hair cycle can be classified into three stages: the resting phase, the anagen phase, and the regressing phase.

[0015] In a preferred embodiment of the present invention, the estimation step uses collagen synthesis in the lower part of the hair bulb as an indicator.

[0016] In a preferred embodiment of the present invention, a collagen metabolism information acquisition step of acquiring information on collagen degradation and / or neogenesis from the subject to be evaluated; and an estimation step of applying the information regarding collagen degradation and / or neogenesis acquired in the collagen metabolism information acquisition step to a previously prepared formula or model that indicates the correlation between the hair cycle and collagen degradation and / or neogenesis, thereby estimating the hair cycle of the subject to be evaluated.

[0017] In a preferred embodiment of the present invention, the hair growth cycle of body hair is estimated. According to the present invention, it is possible to estimate the hair growth cycle of not only head hair but also body hair other than head hair.

[0018] The present invention also provides A storage means for storing correlation data between the degradation and / or neogenesis of collagen in the perifollicular tissue and the hair cycle of the perifollicular tissue; an estimation means for applying information on collagen degradation and / or neogenesis obtained from the subject to the correlation data stored in the storage means to estimate the hair growth cycle of the subject; The hair growth cycle estimation device is provided with:

[0019] The present invention also provides a computer comprising: A storage means for storing correlation data between the degradation and / or neogenesis of collagen in the perifollicular tissue and the hair cycle of the perifollicular tissue; an estimation means for applying information on collagen degradation and / or neogenesis obtained from the subject to the correlation data stored in the storage means to estimate the hair growth cycle of the subject; This is a hair growth cycle estimation program that functions as a hair growth spurt.

[0020] The present invention also relates to a method for estimating collagen degradation and / or neogenesis, which estimates the state of collagen degradation and / or neogenesis in skin tissue to be evaluated based on the hair cycle. According to the present invention, by identifying the hair cycle of skin tissue, it is possible to estimate the degradation and / or neogenesis of collagen in the skin tissue.

[0021] Furthermore, the present invention can be applied to estimating the effects of drugs and cosmetic procedures on collagen degradation and / or neogenesis.

[0022] In a preferred embodiment of the present invention, a hair cycle determining step of determining the hair cycle of the perifollicular tissue from the evaluation subject; and an estimation step of applying the information about the hair cycle obtained in the hair cycle identification step to a previously prepared equation or model that indicates the correlation between the hair cycle and collagen degradation and / or neogenesis, and estimating the state of collagen degradation and / or neogenesis of the subject to be evaluated.

[0023] The present invention also provides A storage means for storing correlation data between the degradation and / or neogenesis of collagen in the perifollicular tissue and the hair cycle of the perifollicular tissue; an estimation means for applying information on the hair growth cycle acquired from the subject to be evaluated to the correlation data stored in the storage means, and estimating the state of collagen degradation and / or neogenesis of the subject to be evaluated; The present invention is a device for estimating collagen degradation and / or new synthesis, comprising:

[0024] The present invention also provides a computer comprising: A storage means for storing correlation data between the degradation and / or neogenesis of collagen in the perifollicular tissue and the hair cycle of the perifollicular tissue; The program for estimating collagen degradation and / or regeneration functions as an estimation means for estimating the state of collagen degradation and / or regeneration of the subject by applying information about the hair growth cycle obtained from the subject to be evaluated to the correlation data stored in the storage means. [Effects of the Invention]

[0025] According to the present invention, a new technique for estimating the hair growth cycle can be provided. Furthermore, the present invention can provide a new technique for estimating collagen degradation and / or neogenesis. [Brief explanation of the drawings]

[0026] [Figure 1] These are fluorescence images that detect collagen degradation and regeneration in the tissue surrounding the hair follicle during the telogen, anagen, and catagen phases. [Figure 2](a) HE-stained image of the perifollicular tissue in mid-catagen (b). The arrows indicate the location of the hair follicle. DETAILED DESCRIPTION OF THE INVENTION

[0027] In the present invention, the hair cycle refers to the cyclical changes of hair and hair follicles, which are composed of three stages: telogen, anagen, and catagen. The anagen stage is subdivided into the beginning of anagen, early anagen, and late anagen, and the catagen stage is subdivided into the early catagen, mid-catagen, and late catagen.

[0028] In the present invention, the term "perifollicular tissue" refers to the hair follicle and the surrounding tissues such as cells and extracellular matrix. The term "hair" refers to both scalp hair and body hair, where scalp hair refers to hair found on the scalp and body hair refers to hair found outside the scalp, including vellus hair.

[0029] <1> Estimation of hair cycle by assessing collagen degradation and / or neogenesis The present invention is a method for estimating the hair cycle in a hair follicle to be evaluated based on the results of an analysis of collagen degradation and / or neogenesis in the tissue surrounding the hair follicle, including the hair follicle.

[0030] The collagen used as an indicator in the present invention is preferably collagen constituting the dermis. Examples of such collagen include type I collagen, type III collagen, and type V collagen, and type I collagen is preferably used as an indicator in the present invention.

[0031] The evaluation subject according to the present invention is preferably skin tissue. The skin tissue to be used is not particularly limited as long as it is skin containing hair follicles, but hairy skin tissue is preferred.

[0032] The site from which the skin tissue is obtained is not particularly limited, and it can be obtained from the forehead, cheek, back of the hand, abdomen, back, arm, leg, etc.

[0033] Furthermore, the present invention can estimate the hair growth cycle of all types of hair, including scalp hair, body hair, etc. Here, the present invention preferably estimates the hair growth cycle of body hair using skin tissue obtained from the skin of the body excluding the scalp, and more preferably estimates the hair growth cycle of downy body hair.

[0034] The collagen degradation and / or neogenesis analyzed in the present invention broadly includes the site (localization) and amount of collagen degradation and / or neogenesis, as well as changes in collagen density and three-dimensional structure due to collagen degradation and / or neogenesis.

[0035] In the present invention, means for analyzing collagen degradation and / or neogenesis preferably include analysis of the amount of collagen degradation and / or neogenesis and the site of collagen degradation and / or neogenesis. Furthermore, analysis of collagen degradation and / or neogenesis is preferably carried out by analyzing at least the amount of collagen degradation, the amount of collagen newly synthesized, or the site of collagen new synthesis, more preferably by analyzing the amount of collagen degradation and the amount of collagen newly synthesized and / or the site of collagen new synthesis, and even more preferably by analyzing the amount of collagen degradation, the amount of collagen newly synthesized, and the site of collagen new synthesis.

[0036] The specific embodiment of the estimation method based on the analysis of collagen degradation and / or neogenesis is not particularly limited. For example, when estimating using the amount of collagen degradation and / or regeneration as an indicator, a standard can be set for the amount of collagen degradation and / or regeneration, and a specific hair cycle can be estimated based on that standard.

[0037] When estimating using the sites of collagen degradation and / or regeneration as an indicator, a method can be adopted in which the hair cycle is estimated by comparing the sites with predetermined standards for the sites of collagen degradation and / or regeneration for each hair cycle.

[0038] Furthermore, a specific embodiment of the present invention can include a collagen metabolism information acquisition step of acquiring information on collagen degradation and / or neogenesis from the subject to be evaluated, and an estimation step of applying the information on collagen degradation and / or neogenesis acquired in the collagen metabolism information acquisition step to a previously prepared equation or model that indicates the correlation between the hair cycle and collagen degradation and / or neogenesis, thereby estimating the hair cycle of the subject to be evaluated.

[0039] The collagen metabolism information acquisition step is a step of acquiring information on collagen degradation and / or neogenesis, i.e., information on collagen metabolism, from skin tissue acquired from the subject to be evaluated. Suitable examples of information on collagen degradation and / or neogenesis include the site (localization) and amount of collagen degradation / neogenesis, as well as changes in collagen density and three-dimensional structure due to collagen degradation and / or neogenesis. In the present invention, it is preferable to acquire information on collagen degradation and / or neogenesis, such as the site and amount of collagen degradation and / or neogenesis. Furthermore, information regarding collagen degradation and / or collagen synthesis can be obtained by staining skin tissue and skin sections prepared from the skin tissue and observing them using an appropriate microscope.

[0040] The collagen metabolism information acquisition step preferably includes a collagen degradation detection step and / or a collagen neogeneration detection step, which will be described later. The collagen degradation detection step and the collagen neogeneration detection step will be described in detail in (1-1) and (1-2) below.

[0041] The estimation step is a step of evaluating the hair cycle of the subject to be evaluated based on a previously prepared equation or the like that indicates the correlation between the degradation and / or neogenesis of collagen and the hair cycle.

[0042] In the quantitative estimation method described above, for example, a preferred embodiment is one in which an equation or model relating to the correlation between the hair growth cycle and collagen degradation and / or neogenesis is created in advance, and information relating to collagen degradation and / or neogenesis obtained from the skin tissue of the evaluation target is compared with the equation or model to estimate the hair growth cycle of the evaluation target. By adopting such an embodiment, the estimation accuracy can be improved.

[0043] To create a formula or model, multivariate analysis of the information on collagen degradation and / or neogenesis and the corresponding hair cycle is performed. The multivariate analysis is preferably one that utilizes the relationship between a target variable (dependent variable) and an explanatory variable (independent variable), and preferred examples include discriminant analysis and regression analysis (MLR, PLS, PCR, logistic). Of these, multiple regression analysis (MLR) and nonlinear regression analysis (PLS: Partial Least Squares) are particularly preferred. The formula or model is preferably a regression formula or regression model, and more preferably a multiple regression formula or a prediction formula.

[0044] In order to create an equation or model showing the correlation, it is preferable to create a database (DB) that associates information on collagen degradation and / or neogenesis with the hair cycle. Here, the number of samples in the DB is preferably 50 or more, more preferably 100 or more, and even more preferably 200 or more. The DB structure may be, for example, a matrix, with sample names entered in the rows and information on collagen degradation and / or neogenesis and the hair cycle entered in the columns.

[0045] This DB may be updated by adding information about collagen degradation and / or neogenesis and the hair cycle after estimating the newly acquired hair cycle of the subject. If necessary, the updated DB may be subjected to the above-mentioned multivariate analysis to update the formula or model. This is because such an update improves estimation accuracy.

[0046] The detection of collagen degradation and / or neogenesis in the tissue surrounding the hair follicle and the identification of the hair cycle, which are necessary for creating the formula or model or for creating the DB, can be performed by conventional methods.

[0047] More specifically, <3> By quantitatively or qualitatively evaluating the expression of Ki67 protein as described in the above, or by observing the components of the hair follicle using stained images of skin sections, it is possible to identify whether the hair cycle being evaluated is in the telogen, anagen, or catagen phase.

[0048] The above-described detection of collagen degradation and / or neogenesis and identification of the hair cycle can be performed on a statistically significant number of subjects, thereby creating the above-described formula or model and DB.

[0049] The method may also include a step of estimating the hair cycle using multiple skin tissues from the same individual, and estimating the tendency of the hair cycle in a certain range of skin tissues based on the multiple estimation results obtained. In this case, the formula or model preferably includes a formula or model that estimates the tendency of the hair cycle in a certain range of skin tissue based on the estimation results of the hair cycles for multiple evaluation targets.

[0050] Next, we will explain methods for estimating the hair growth cycle when (1-1) collagen degradation is used as an indicator, (1-2) collagen synthesis is used as an indicator, and (1-3) collagen degradation and collagen synthesis are used as indicators. In the above estimation method, it is estimated which hair cycle the hair follicle in the subject of evaluation is in, selected from the telogen, anagen, and catagen phases.

[0051] (1-1) A method for estimating the hair cycle using collagen degradation as an indicator The method for estimating the hair cycle using collagen degradation as an indicator includes a collagen degradation detection step and an estimation step of estimating the hair cycle of an object to be evaluated based on the results obtained in the collagen degradation detection step.

[0052] (Collagen degradation detection step) The collagen degradation detection step is a step of identifying the state of collagen degradation by detecting degraded collagen in the evaluation target, including the perifollicular tissue. Here, degraded collagen refers to collagen whose triple helix structure has been denatured, which occurs as part of collagen metabolism. By detecting the expression of degraded collagen, the site where collagen degradation occurs and the amount of degradation can be estimated.

[0053] The method for detecting degraded collagen is not particularly limited. For example, degraded collagen can be detected by obtaining a skin section from skin tissue and subjecting it to a staining treatment. The staining of the skin section is preferably immunostaining.

[0054] Immunostaining can be performed by conventional methods, and any of the direct, indirect, and sensitization methods can be selected. For example, immunostaining is preferably performed using a collagen-degrading probe (e.g., Collagen Hybridizing Peptide, Biotin Conjugate (3Helix)). If necessary, any blocking agent can be used.

[0055] Furthermore, the method for observing the stained skin section is not particularly limited, and the observation can be carried out using an appropriate microscope such as an optical microscope or a confocal laser microscope depending on the staining method used.

[0056] Furthermore, in order to improve detection by the collagen degradation probe, it is preferable to subject the skin section to a transparent treatment. Furthermore, the skin section is preferably a thick, three-dimensional skin section. The clearing treatment can be performed using an existing tissue clearing reagent (for example, RapiClear 1.52 (SunJin Lab)).

[0057] In addition, in the collagen decomposition detection step, it is preferable to detect the amount of decomposed collagen.

[0058] (Estimated process) The estimation step is a step of estimating the hair cycle of the subject to be evaluated by evaluating the state of collagen degradation obtained in the collagen degradation detection step. Preferably, the hair cycle is evaluated based on the amount of collagen degradation estimated from the amount of decomposed collagen as the state of collagen degradation. In the present invention, the amount of decomposed collagen detected can be estimated to be the amount of collagen decomposition.

[0059] When evaluating the amount of collagen degradation, the method is not particularly limited, and either a qualitative method or a quantitative method can be employed. Preferably, the information regarding the amount of collagen decomposition obtained in the collagen decomposition detection step is applied to a previously prepared equation or model that indicates the correlation between the hair cycle and the amount of collagen decomposition, thereby estimating the hair cycle of the subject to be evaluated.

[0060] It is also possible to estimate the hair growth cycle by determining whether the amount of collagen decomposition is equal to or exceeds a predetermined standard. Methods for determining the standard include determining a reference numerical value (reference value) based on the intensity of fluorescence obtained from a stained image acquired in advance, or determining a reference image to be used for visual comparison with the stained image to be evaluated.

[0061] As shown in the Examples, the inventors have found that the amount of collagen decomposition in the hair follicle connective tissue sheath is similar in the anagen and catagen phases, while the amount of collagen decomposition in the telogen phase is less than in the anagen and catagen phases. Therefore, the standard for collagen degradation can be determined based on the amount of collagen decomposition in the growth phase or regression phase, which has been obtained in advance, and the amount of collagen decomposition in the resting phase, which has been obtained in advance. In this specification, the term "hair follicle connective tissue sheath" refers to the region generally called the connective tissue sheath of the hair follicle.

[0062] The standard for the amount of collagen degradation is set to be equivalent to the amount of collagen degradation in the hair follicle connective tissue sheath in the anagen or regressive phase, and if the amount of collagen degradation in the hair follicle connective tissue sheath is equivalent to or exceeds the standard, it can be assumed that the hair cycle is in the anagen or regressive phase.On the other hand, if the amount of collagen degradation does not meet the standard, it can be assumed that the hair cycle is in the telogen phase.

[0063] As described above, hair follicles in the resting phase can be estimated by using the amount of collagen degradation as an indicator.

[0064] Preferably, the estimation step includes a step of estimating the hair cycle of the evaluation subject, and then estimating the tendency of the hair cycle in a certain range of skin tissue using the estimation results of the multiple evaluation subjects. This makes it possible to estimate the tendency of the hair growth cycle in the subject's skin tissue based on the detection results of collagen degradation.

[0065] (1-2) A method for estimating the hair cycle using collagen synthesis as an indicator The method for estimating the hair cycle using collagen neogenesis as an index includes a collagen neogenesis detection step and an estimation step of estimating the hair cycle of an object to be evaluated based on the results obtained in the collagen neogenesis detection step.

[0066] (Collagen synthesis detection step) The collagen neogeneration detection step is a step of identifying the state of collagen neogeneration by detecting procollagen in the evaluation target, including the perifollicular tissue. Here, procollagen refers to collagen peptide chains secreted from fibroblasts that form a triple helix structure. After both terminal peptides of procollagen are cleaved, they crosslink to form mature collagen fibers. Therefore, by detecting procollagen, it is possible to estimate the site of collagen synthesis and the amount of collagen synthesis involved in collagen metabolism.

[0067] The method for detecting collagen synthesis in the tissue surrounding the hair follicle is not particularly limited, as in the collagen degradation detection step described above, but is preferably carried out by subjecting a skin section prepared from the skin tissue to immunostaining using a procollagen antibody.

[0068] A preferred example of the procollagen antibody used is Anti-pro-Collagen I antibody (Clone: ​​M-58) (manufactured by Merck Millipore), etc. The skin section is preferably a thick, three-dimensional skin section.

[0069] In addition, in the collagen synthesis detection step, it is preferable to detect the amount and / or localization site of procollagen.

[0070] (Estimated process) The estimation step is a step of estimating the hair cycle of the subject to be evaluated by evaluating the state of collagen neogenesis obtained in the collagen neogenesis detection step. Preferably, the hair cycle is evaluated based on the amount or site of collagen neogenesis estimated from the localization of procollagen as the state of collagen neogenesis. In the present invention, the detected amount or localized site of procollagen can be used to estimate the amount or site of collagen newly produced, respectively.

[0071] The method for evaluating the site of new collagen synthesis is not particularly limited, but can be carried out, for example, by observing stained images of skin sections obtained from skin tissue.

[0072] The inventors have demonstrated that during the telogen and anagen phases, new collagen synthesis is observed only in the connective tissue sheath of the hair follicle, but during the catagen phase, new collagen synthesis is observed not only in the connective tissue sheath of the hair follicle but also in the lower part of the hair bulb. That is, when collagen is newly synthesized below the hair bulb, the hair cycle can be estimated to be in the catagen phase. On the other hand, when collagen is not newly synthesized below the hair bulb, the hair cycle can be estimated to be in the telogen phase or anagen phase.

[0073] As mentioned above, hair follicles in the regression phase can be estimated by using the site of new collagen synthesis as an indicator.

[0074] The present inventors have also found that collagen synthesis in the hair follicle connective tissue sheath increases in the order of telogen, catagen, and anagen. That is, in the present invention, the hair growth cycle can be estimated based on the amount of collagen newly produced. When evaluating the amount of collagen newly produced, the method is not particularly limited, and a qualitative method or a quantitative method can be adopted. Specifically, the amount of collagen newly produced can be evaluated using the same procedure as the estimation method described above in "(1-1) Method for estimating hair growth cycle using collagen degradation as an index."

[0075] Preferably, the estimation step includes a step of estimating the hair cycle of the evaluation subject, and then estimating the tendency of the hair cycle in a certain range of skin tissue using the estimation results of the multiple evaluation subjects. This makes it possible to estimate the tendency of the hair growth cycle in the subject's skin tissue based on the detection results of collagen neogeneration.

[0076] (1-3) A method for estimating the hair cycle using collagen degradation and collagen synthesis as indicators In a preferred embodiment of the present invention, the hair cycle of the subject is estimated based on the results of detecting collagen degradation and neogenesis in the tissue surrounding the hair follicle.

[0077] By using collagen degradation and collagen synthesis as indicators, the hair cycle can be divided into three stages: telogen, anagen, and catagen.

[0078] In this embodiment, the present invention comprises a collagen degradation detection step for detecting collagen degradation, a collagen neogeneration detection step for detecting collagen neogeneration, and an estimation step for estimating the hair cycle of the subject to be evaluated based on the results obtained in the collagen degradation detection step and the collagen neogeneration detection step.

[0079] The collagen degradation detection step and collagen neogeneration detection step according to this embodiment can be carried out by the same methods as the collagen degradation detection step and collagen neogeneration detection step described above. In the present invention, the order of the collagen degradation detection step and the collagen synthesis detection step is not limited, and the collagen degradation detection step may be performed after the collagen synthesis detection step.

[0080] The estimation step can also use the method for the estimation step described above. Specifically, the hair growth cycle can be estimated by combining the evaluation results regarding collagen degradation and the evaluation results regarding collagen synthesis, with reference to the methods described in the estimation steps (1-1) and (1-2) above.

[0081] In the present invention, the steps can be performed in any order, i.e., the method may include a single estimation step in which the collagen degradation detection step and the collagen synthesis detection step are performed separately, and then the detection results obtained in both steps are evaluated, or the method may include two or more estimation steps in which estimation is performed based on the results of each detection step.

[0082] Specifically, the following embodiments can be adopted. A form in which the estimation step is carried out after the collagen degradation detection step and collagen synthesis detection step are carried out After performing the collagen degradation detection step and the collagen synthesis detection step, a first estimation step of estimating the hair cycle based on the results of the collagen degradation detection step, and a second estimation step of estimating the hair cycle based on the results of the first estimation step and the collagen synthesis detection step are sequentially performed. After performing the collagen degradation detection step and the collagen synthesis detection step, a first estimation step of estimating the hair growth cycle based on the results of the collagen synthesis detection step, and a second estimation step of estimating the hair growth cycle based on the results of the first estimation step and the collagen degradation detection step are sequentially performed.

[0083] Preferably, the estimation step includes a step of estimating the hair cycle of the evaluation subject, and then estimating the tendency of the hair cycle in a certain range of skin tissue using the estimation results of the multiple evaluation subjects. This makes it possible to estimate the tendency of the hair growth cycle in the subject's skin tissue based on the results of detecting collagen degradation and regeneration.

[0084] The present invention also relates to an estimation device for estimating the hair growth cycle of an object to be evaluated. The estimation device of the present invention includes: a storage means for storing correlation data between collagen degradation and / or neogenesis in the tissue around the hair follicle and the hair cycle of the tissue around the hair follicle; an estimation means for applying information on collagen degradation and / or neogenesis obtained from the subject to the correlation data stored in the storage means to estimate the hair growth cycle of the subject; Equipped with.

[0085] The storage means stores data relating to the correlation between the degradation and / or regeneration of collagen in the tissue surrounding the hair follicle and the hair cycle. The correlation data includes the above-mentioned formulas or models, as well as DBs for creating them. The storage means can be realized by, for example, a ROM (Read Only Memory).

[0086] The hair cycle estimation means applies information regarding collagen degradation and / or neogenesis obtained from the subject to be evaluated to the correlation data stored in the storage means, and estimates the hair cycle of the subject to be evaluated. The estimation means can be realized by a CPU (Central Processing Unit). The above-described estimation process in the CPU can be executed in accordance with a program stored in the ROM.

[0087] Furthermore, it is preferable that the storage means additionally stores newly acquired information on the degradation and / or neogenesis of collagen to be evaluated and the hair cycle, and that the correlation data is successively updated. In this case, it is preferable to provide an update means for performing reanalysis based on the DB including the additional data stored in the storage means, calculating an updated formula or model, and overwriting and storing this in the storage means. The updating means can be realized by a CPU in accordance with a program stored in a ROM.

[0088] Furthermore, the estimation device of the present invention preferably includes a RAM (Random Access Memory) for temporarily storing an OS (Operating System) program and various application programs executed by the CPU.

[0089] The present invention also relates to a program for estimating the hair growth cycle of an object to be evaluated. The estimation program of the present invention includes a computer, a storage means for storing correlation data between collagen degradation and / or neogenesis in perifollicular tissues and the hair cycle of the perifollicular tissues, and an estimation means for applying information on collagen degradation and / or neogenesis obtained from the subject to the correlation data stored in the storage means to estimate the hair growth cycle of the subject; It is preferable to use an embodiment in which the function is as follows. Here, the present invention may be such a program recorded on a recording medium that can be read by a computer or the like.

[0090] <2> Method for estimating collagen degradation and / or neogenesis based on the hair cycle The present invention also relates to a method for estimating collagen degradation and / or neogenesis, which estimates the state of collagen degradation and / or neogenesis in skin tissue to be evaluated based on the hair cycle. According to the present invention, by identifying the hair growth cycle of the subject to be evaluated, it is possible to estimate the state of collagen degradation and / or neogenesis in the skin tissue of the subject to be evaluated.

[0091] The evaluation subject according to the present invention is preferably skin tissue. The site from which the skin tissue is obtained is not particularly limited, and it can be obtained from the forehead, cheek, back of the hand, abdomen, back, arm, leg, etc. The skin tissue used is not particularly limited as long as it is skin containing hair follicles, but hairy skin tissue is preferred.

[0092] Furthermore, the present invention can estimate the hair cycle of any type of hair, such as scalp hair, body hair, etc. Here, in a preferred embodiment of the present invention, skin tissue obtained from the skin of the body excluding the scalp is preferably used as an index to determine the hair cycle of body hair, and more preferably, the hair cycle of vellus body hair is used as an index.

[0093] The state of collagen degradation and / or neogenesis estimated in the present invention is preferably the amount and / or site of collagen degradation and / or the amount and / or site of collagen neogenesis, more preferably the amount of collagen degradation and / or the amount and / or site of collagen neogenesis, and even more preferably the amount of collagen degradation and / or neogenesis.

[0094] The method for obtaining information on the hair growth cycle to be evaluated is not particularly limited, and can be obtained by a conventional method. A preferred method for obtaining information about the hair cycle is to obtain skin tissue and subject it to a staining process to obtain information about the hair cycle.

[0095] More specifically, <3> By quantitatively or qualitatively evaluating the expression of Ki67 protein as described in the above, or by observing the components of the hair follicle using stained images of skin sections, it is possible to identify whether the hair cycle being evaluated is in the telogen, anagen, or catagen phase.

[0096] The specific embodiment of the method for estimating the histological characteristics of collagen in the present invention is not particularly limited. For example, a method may be employed in which a plurality of skin tissues to be evaluated are used to estimate the state of collagen degradation and / or regeneration in a certain range of skin tissue. Specifically, a certain area of ​​skin tissue is divided into multiple sections, and the hair cycle is identified for each section.Then, by evaluating the proportion of hair follicles in each hair cycle relative to the entire skin tissue in that area, the tendency of collagen degradation and / or neogenesis in the subject can be estimated.

[0097] The inventors have found that the amount of collagen degradation is high in certain hair cycles, for example, in the anagen and regressive phases, and that the amount of collagen neogenesis increases in the order of telogen, regressive phase, and anagen. Based on this, a standard can be set according to the ratio of hair follicles in each hair cycle to the total number of hair follicles related to multiple evaluation subjects, and an estimation method can be adopted to estimate the tendency of the amount of collagen degradation and / or neogenesis in a certain range of skin tissue according to the standard.

[0098] Alternatively, a quantitative estimation method may be employed. Specifically, a preferred embodiment is one in which an equation or model relating to the correlation between the hair growth cycle and the histological characteristics of collagen is created in advance, and the hair growth cycle obtained from the skin tissue of the evaluation target is compared with the equation or model to estimate the state of collagen degradation and / or neogenesis of the evaluation target. Such an embodiment can improve the estimation accuracy.

[0099] That is, a specific embodiment of a preferred aspect of the present invention is as follows: A hair cycle identification step of identifying the hair cycle of the tissue surrounding the hair follicle from the evaluation subject; and an estimation step of applying the information about the hair cycle obtained in the hair cycle identification step to a previously prepared equation or model that indicates the correlation between the hair cycle and collagen degradation and / or neogenesis, and estimating the state of collagen degradation and / or neogenesis of the subject to be evaluated.

[0100] Here, the method for creating the formula or model can be the same as that described above.

[0101] Furthermore, when estimating the state of collagen degradation and / or neogenesis using multiple evaluation subjects, it is preferable that the formula or model includes a formula or model that integrates hair cycle information obtained from multiple evaluation subjects, calculates the proportion of hair follicles in each hair cycle to the entire skin tissue of the evaluation subject, and estimates the tendency of collagen degradation and / or neogenesis in the skin tissue of the evaluation subject from the calculation results. This makes it possible to estimate the tendency of collagen degradation and / or regeneration based on the hair cycle for a certain range of skin tissue consisting of multiple evaluation targets.

[0102] Furthermore, the estimated results regarding the state of collagen degradation and / or synthesis can be used to evaluate the effect of promoting collagen metabolism.

[0103] Furthermore, the estimated results regarding the state of collagen degradation and / or regeneration can be used as an indicator when selecting cosmetics. Here, the cosmetics selected based on the estimated results regarding the state of collagen degradation and / or regeneration are cosmetics for improving skin conditions, such as those that promote collagen degradation and / or regeneration. More specifically, the cosmetics selected here are cosmetics for improving skin conditions that have the effect of inhibiting and / or improving wrinkles, sagging skin, etc., inhibiting dry skin, improving skin elasticity, etc.

[0104] For example, when the amount of collagen degradation and / or regeneration is used as an index, if the estimated amount of collagen degradation and / or regeneration is low, cosmetics for improving skin condition with stronger effects and properties are selected. Furthermore, the results of collagen degradation and / or regeneration determined by the present invention can also be useful indicators for counseling regarding skin care and makeup methods.

[0105] The present invention also relates to an estimation device for estimating collagen degradation and / or neogenesis in skin tissue to be evaluated. The estimation device of the present invention includes: a storage means for storing correlation data between collagen degradation and / or neogenesis in the tissue around the hair follicle and the hair cycle of the tissue around the hair follicle; an estimation means for applying information on the hair growth cycle acquired from the subject to be evaluated to the correlation data stored in the storage means, and estimating the state of collagen degradation and / or neogenesis of the subject to be evaluated; Equipped with.

[0106] Here, the above description can be applied to preferred embodiments of the estimation device of the present invention.

[0107] The present invention also relates to a prediction program for predicting collagen degradation and / or neogenesis in skin tissue to be evaluated. The estimation program of the present invention includes a computer, a storage means for storing correlation data between collagen degradation and / or neogenesis in the perifollicular tissue and the hair cycle of the perifollicular tissue, and A preferred embodiment is one in which the information regarding the hair growth cycle obtained from the subject to be evaluated is applied to the correlation data stored in the storage means, and functions as an estimation means for estimating the state of collagen degradation and / or regeneration of the subject to be evaluated.

[0108] Here, the above description can be applied to preferred embodiments of the estimation program of the present invention.

[0109] <3> Estimation of the hair cycle based on observation of cell division activity and / or components of the hair follicle Next, a method for estimating the hair follicle hair cycle in the method for estimating collagen degradation and / or neogenesis of the present invention will be described in detail. <1> and <2> This can be used to estimate the hair follicle's hair cycle.

[0110] Below, we will explain the methods for estimating the hair cycle from the perspectives of (3-1) estimating the hair cycle based on cell division activity and (3-2) estimating the hair cycle based on observation of the components of the hair follicle.

[0111] (3-1) Estimation of the hair cycle based on cell division activity In a preferred embodiment, the hair cycle of a hair follicle can be estimated by detecting cell division activity in the tissue surrounding the hair follicle, specifically by observing the expression and localization of Ki67 protein, a cell division marker.

[0112] The expression and localization of Ki67 protein can be confirmed by preparing a skin section from the skin tissue to be evaluated and using a stained image obtained by immunostaining the skin section using a Ki67 antibody.

[0113] Immunostaining can be performed by a conventional method, and any of direct, indirect, and sensitized methods can be selected. If necessary, any blocking agent can be used. Furthermore, the method for observing the stained skin section is not particularly limited, and the observation can be carried out using an appropriate microscope such as an optical microscope or a confocal laser microscope depending on the staining method used.

[0114] Next, a method for estimating the hair growth cycle using dyed images will be described in detail. In a preferred embodiment, the hair cycle is estimated using the expression level of Ki67 protein as an index. That is, when the expression level of Ki67 protein is higher than that of other samples, it is preferable to determine that the hair cycle is in the anagen phase. On the other hand, if the expression of Ki67 protein is lower than or at the same level as other samples, it is preferable to determine that the hair cycle is in the catagen or telogen phase.

[0115] In a preferred embodiment, the hair cycle can be estimated based on the location of the Ki67 protein by evaluating the expression of the Ki67 protein in the hair bulb, root sheath, and sebaceous gland.

[0116] For example, hair follicles that are in the anagen phase can be further identified as those that have just begun the anagen phase. That is, when Ki67 protein expression is observed only in the sebaceous gland of a hair follicle classified as in the anagen phase, it is preferable to determine that the hair cycle has begun in the anagen phase.

[0117] Furthermore, hair follicles in the early catagen phase can be identified from hair follicles in the catagen or telogen phase. That is, when Ki67 protein expression is observed only in the sebaceous gland of a hair follicle classified as being in the catagen or telogen phase, it is preferable to determine that the hair cycle is in the early catagen phase.

[0118] (3-2) Estimation based on observation of components of hair follicles In a preferred embodiment, the hair cycle of a hair follicle can be estimated based on the results of observing components of the hair follicle by staining a skin section obtained from skin tissue containing the hair follicle and observing the resulting stained image.

[0119] The skin sections can be stained by a conventional method using hematoxylin and eosin (HE), etc. Furthermore, the stained images can be observed using an appropriate microscope, such as an optical microscope, depending on the staining method.

[0120] The components of the hair follicle to be observed are preferably one or more, more preferably two or more, more preferably three or more, more preferably four or more, more preferably five or more, more preferably six or more, and even more preferably all, selected from the state of the dermal papilla, the presence or absence of an epithelial strand (ES), the thickness of the basement membrane, the distance from the hair bulb to the bulge relative to the distance from the bulge to the epidermis, the presence or absence of apoptotic cells in the root sheath, the presence or absence of club hair, and the presence or absence of new hair. The above seven items are indicators that fluctuate throughout the hair cycle, and by observing them, the hair cycle can be estimated.

[0121] The evaluation criteria for each of the above items will be explained below.

[0122] (i) Condition of the dermal papilla (hair papilla) When a cell population is present below the hair bulb, it is preferable to determine that the hair cycle is at the beginning of the anagen phase or in the early anagen phase. Furthermore, when the dermal papilla (hair papilla) is invaginated into the hair bulb, it is preferable to determine that the hair cycle is in the late anagen phase. Furthermore, when the dermal papilla (hair papilla) is detached from the hair bulb, it is preferable to determine that the hair cycle is in the early catagen phase. Furthermore, when a cell population is present under ES, it is preferable to determine that the hair cycle is in the mid-catagen stage.

[0123] (ii) Presence or absence of epithelial strand (ES) If there is no epithelial strand (ES), it is preferable to determine that the hair cycle is in the telogen-early catagen phase. If an epithelial strand (ES) is present, it is preferable to determine that the hair cycle is in mid-catagen.

[0124] (iii) Thickness of the basement membrane When the thickness of the basement membrane is thinner than that of other samples, it is preferable to determine that the hair cycle is in the anagen start-late anagen phase. If the thickness of the basement membrane is thicker than that of other samples, it is preferable to determine that the hair cycle is in the early catagen-mid catagen phase.

[0125] (iv) the distance from the hair bulb to the bulge relative to the distance from the bulge to the epidermis When the distance between the bulge and the epidermis relative to the distance between the hair bulb and the bulge is smaller than that of other samples, it is preferable to determine that the hair cycle is in the late catagen phase - beginning of the anagen phase. If the distance between the bulge and the epidermis relative to the distance between the hair bulb and the bulge is greater than that of other samples, it is preferable to determine that the hair cycle is in the early anagen-middle catagen stage.

[0126] (v) Presence or absence of apoptotic cells in the root sheath When apoptotic cells are present in the root sheath, it is preferable to determine that the hair cycle is in the mid-catagen to late-catagen phase. When there are no apoptotic cells in the root sheath, it is preferable to determine that the hair cycle is in the early anagen phase to late anagen phase.

[0127] (vi) Presence or absence of club hairs If club hair is present, it is preferable to determine that the hair cycle is in mid-catagen-late-catagen phase. If no club hair is present, it is preferable to determine that the hair cycle is in the beginning of the anagen phase and the beginning of the catagen phase.

[0128] (vii) Presence or absence of new hair growth When new hair is present, it is preferable to determine that the hair cycle is in the beginning of the anagen phase to the early anagen phase. If no new hair is present, it is preferable to determine that the hair cycle is in the early catagen-telogen phase.

[0129] When the thickness of the basement membrane or the distance from the bulge to the epidermis relative to the distance from the lower part of the hair bulb to the bulge is used as an index, the evaluation may be based on a predetermined reference value, or a reference stained image for comparison may be prepared in advance for each hair cycle and evaluated by comparing it with the stained image to be evaluated.

[0130] In the present invention, the hair growth cycle can be estimated using the indices (3-1) and (3-2) described above individually. Here, for accurate estimation of the hair growth cycle, it is preferable to estimate the hair growth cycle by comprehensively evaluating both indices. [Example]

[0131] The following shows various test results that support the findings that form the basis of the present invention.

[0132] Example 1: Observation of collagen degradation and regeneration in the tissue surrounding the hair follicle (1) Obtaining test materials In this example, the human skin shown in Table 1 below was used as the test material.

[0133] [Table 1]

[0134] (2) Detection of degraded collagen (2-1) Processing of test materials The skin tissues (A and B) in Table 1 (three-dimensional skin tissues approximately 3 mm long and wide, approximately 1 mm thick) were fixed with a 4% paraformaldehyde solution and washed with PBS. After washing, the skin tissues were permeabilized at room temperature in 2% PBST (PBS containing 2% Triton-X 100) with rotary shaking. The solution was then replaced with blocking buffer (PBS containing 10% goat serum and 1% Triton-X 100) and blocked with rotary shaking. Afterwards, a probe for detecting degraded collagen (Collagen Hybridizing Peptide, Cy3 Conjugate (3Helix)), TO-PRO TM Fluorescent staining was performed at 4°C using -3 Iodide (642 / 661)-1 mM Solution in DMSO (Thermo Fisher Scientific).

[0135] After fluorescent staining, the solution was replaced with washing buffer (PBS containing 3% NaCl and 0.2% Triton-X 100) and washed by rotary shaking. The solution was then replaced with PBS and washed again. After washing, the skin tissue was immersed in a 24-well plate containing RapiClear 1.52 (SunJin Lab) and cleared by shaking. After clearing, the skin tissue was mounted on a glass slide with RapiClear 1.52 using iSpacer (SunJin Lab). Images of the stained skin tissue were then captured using a confocal laser microscope (Nikon) (Figure 1).

[0136] (2-2) Estimation of hair growth cycle After observing the degraded collagen, the skin tissue was embedded in paraffin to prepare skin sections, which were then stained with HE and Ki67 by standard methods. Ki67 antibody, ki67 (MIB-1) (manufactured by DAKO), was used. Next, using the stained images obtained from the skin sections, the hair follicles in the skin tissue were classified into three cycles: anagen, catagen, and telogen, based on the classification table in Table 2. Figure 2 shows HE-stained and Ki67-stained images of a hair follicle estimated to be in the mid-catagen phase. After classifying the hair follicles according to the hair cycle, the relationship between the amount and localization of degraded collagen in the hair follicles and the hair cycle was evaluated.

[0137] [Table 2]

[0138] (2-3) Results and Discussion As shown in Figure 1, degraded collagen was detected in the connective tissue sheath of the hair follicle in all three stages: telogen, anagen, and catagen. Furthermore, it was revealed that the amount of degraded collagen was similar in anagen and catagen, but was less in telogen than in anagen and catagen.

[0139] From the above results, it is estimated that the amount of collagen decomposition is less in the resting phase than in the anagen and regressing phases. In other words, it has been shown that the hair cycle can be estimated by using the amount of collagen decomposition as an indicator. In particular, it has been found that by focusing on the amount of collagen decomposition, it is possible to estimate hair follicles in the resting phase.

[0140] (3) Detection of procollagen (3-1) Processing of test materials Paraffin sections were prepared using the skin tissues listed in Table 1 (C) by standard methods. The skin tissues were immersed in xylene and then washed sequentially with ethanol and PBS-T. Subsequently, 1% trypsin was added for antigen retrieval. After washing with PBS-T, the sections were reacted with a primary antibody (Anti-pro-Collagen I antibody (Clone. M-58) (Merck Millipore)) at 4°C. After the reaction, the sections were washed with PBS-T, and then reacted with a secondary antibody (Goat anti-rat IgG Alexa Fluor 555 (Invitrogen)) at room temperature. After the reaction, the skin tissues were washed with PBS and mounted in a slide with ProLong Gold Antifade Mountant with DAPI (Invitrogen). The stained images of the obtained skin tissue were taken using a confocal laser microscope (Nikon) (Figure 1).

[0141] (3-2) Estimation of hair growth cycle Paraffin sections obtained from the same skin tissue used to detect procollagen were subjected to standard HE staining and Ki67 immunostaining. Using the stained images obtained from the skin sections, hair follicles in the skin tissue were classified into three cycles: anagen, catagen, and telogen, based on the classification table in Table 2 above. The relationship between the amount and localization of procollagen in hair follicles and the hair cycle was then evaluated.

[0142] (3-2) Results and Discussion Procollagen is produced in the connective tissue sheath of the hair follicle during the telogen and anagen phases, but it was confirmed that it is produced in the lower part of the hair bulb (white arrowhead in Figure 1(b)) along with the connective tissue sheath of the hair follicle only during the catagen phase. The amount of procollagen in the connective tissue sheath of the hair follicle increased with the length of the hair follicle, that is, in the order of telogen, catagen, and anagen.

[0143] From these results, it is speculated that collagen synthesis occurs in the connective tissue sheath of the hair follicle during the telogen and anagen phases, and occurs only in the lower part of the hair bulb during the catagen phase. In other words, it was shown that the hair cycle can be estimated by using the collagen synthesis site as an indicator. In particular, it was found that by focusing on the collagen synthesis site, it is possible to estimate the hair follicle in the catagen phase. It was also revealed that the hair cycle can be estimated by focusing on the amount of collagen synthesis.

[0144] (4) Overall consideration From Example 1, it was revealed that when the amount of collagen degradation is focused, hair follicles in the resting phase can be estimated, and when the collagen regeneration site is focused, hair follicles in the regression phase can be estimated. It was also shown that the hair cycle can be estimated when the amount of collagen regeneration is focused. In other words, it was found that the hair cycle can be estimated by using collagen degradation and regeneration as indicators.

[0145] Furthermore, the above results showed that by using the amount of collagen degradation and the site of collagen synthesis as indicators, it is possible to estimate whether the hair cycle being evaluated belongs to the telogen, anagen, or regressor phase. [Industrial Applicability]

[0146] The technology of the present invention for estimating the hair growth cycle can be applied to the production and development of pharmaceuticals, cosmetics, etc. for improving diseases caused by the hair growth cycle and skin conditions.

Claims

1. an estimation step of estimating the hair cycle of the hair follicle in the subject based on the analysis results of collagen degradation and / or neogenesis in the tissue surrounding the hair follicle, including the hair follicle; The estimation step includes: (1) Using the presence or absence of collagen synthesis in the area below the hair bulb of the tissue surrounding the hair follicle as an indicator, it is estimated that the hair cycle of the hair follicle is in the telogen phase, the anagen phase, or the catagen phase. Including, How to estimate the hair growth cycle.

2. The estimation method of claim 1, wherein (1) of the estimation step includes estimating that the hair cycle is in the regression phase if collagen is newly synthesized in the lower part of the hair bulb, and estimating that the hair cycle is in the resting phase or the growth phase if collagen is not newly synthesized in the lower part of the hair bulb.

3. The estimation step comprises: (2) applying the amount of new collagen produced in the hair follicle connective tissue sheath of the tissue surrounding the hair follicle to a predetermined standard, and estimating whether the hair cycle of the hair follicle is in the resting phase, the anagen phase, or the regressing phase; The estimation method according to claim 1 or 2, wherein the criterion in (2) is determined based on the relationship that the amount of collagen newly produced in the hair follicle connective tissue sheath increases in the order of the telogen, the regression phase, and the anagen phase.

4. The estimation step (3) Inferring whether the hair follicle is in the anagen phase, the regressive phase, or the telogen phase based on the amount of collagen degradation in the hair follicle connective tissue sheath of the tissue surrounding the hair follicle. The estimation method according to any one of claims 1 to 3, comprising:

5. (3) of the estimation step includes estimating that the hair cycle is in the anagen or regressive phase when the amount of collagen degradation in the hair follicle connective tissue sheath is equal to or exceeds a predetermined standard, and estimating that the hair cycle is in the telogen phase when the amount of collagen degradation does not meet the standard; The criterion in (3) is determined based on the relationship that the amount of collagen degradation in the hair follicle connective tissue sheath during the telogen phase is less than the amount of collagen degradation in the hair follicle connective tissue sheath during the anagen phase and the regressive phase. The estimation method according to claim 4.

6. a collagen degradation detection step of detecting degraded collagen in the perifollicular tissue; a collagen synthesis detection step of detecting procollagen in the perifollicular tissue; Including, The estimation method according to claim 4 or 5, wherein the estimation step estimates the hair growth cycle based on the amount of collagen decomposition and the site of collagen regeneration and / or the amount of collagen regeneration.

7. a collagen metabolism information acquisition step of acquiring information on collagen degradation and / or neogenesis from the subject to be evaluated, the estimation step includes applying the information on collagen degradation and / or neogenesis acquired in the collagen metabolism information acquisition step to a previously prepared formula or model showing a correlation between the hair cycle and collagen degradation and / or neogenesis, thereby estimating the hair cycle of the subject to be evaluated; The estimation method according to any one of claims 1 to 6, wherein the formula or model is created based on at least the relationship between the presence or absence of collagen neogenesis in the lower part of the hair bulb and the hair cycle.

8. The method according to any one of claims 1 to 7, wherein the means for analyzing collagen degradation and / or neogenesis is a means for detecting degraded collagen and / or procollagen.

9. A collagen degradation detection step for detecting degraded collagen in perifollicular tissue including hair follicles; a collagen synthesis detection step of detecting procollagen in the perifollicular tissue; an estimation step of estimating the hair cycle of the hair follicle in the subject based on the analysis results of collagen degradation and / or neogenesis in the perifollicular tissue; Including, The estimation step includes: and estimating the hair cycle based on the amount of collagen decomposition and the collagen new synthesis site and / or collagen new synthesis amount, and The method includes carrying out the following steps (1) and / or (2), and (3): (1) Using the presence or absence of collagen synthesis in the area below the hair bulb of the perifollicular tissue as an indicator, it is estimated that the hair cycle of the hair follicle is in the telogen phase, the anagen phase, or the catagen phase; (2) applying the amount of new collagen produced in the hair follicle connective tissue sheath of the tissue surrounding the hair follicle to a predetermined standard, and estimating whether the hair cycle of the hair follicle is in the telogen, anagen, or catagen phase; (3) Inferring whether the hair follicle is in the anagen phase, the regressive phase, or the telogen phase based on the amount of collagen degradation in the hair follicle connective tissue sheath of the tissue surrounding the hair follicle; Here, the criterion in (2) is determined based on the relationship that the amount of collagen newly produced in the hair follicle connective tissue sheath increases in the order of the telogen, the regressive phase, and the anagen phase. How to estimate the hair growth cycle.

10. The estimation step (1) includes estimating that the hair cycle is in the catagen phase if collagen is newly synthesized in the lower part of the hair bulb, and estimating that the hair cycle is in the telogen phase or the anagen phase if collagen is not newly synthesized in the lower part of the hair bulb; The estimation step (3) includes estimating that the hair cycle is in the anagen phase or the regressive phase when the amount of collagen degradation in the hair follicle connective tissue sheath is equal to or exceeds a predetermined standard, and estimating that the hair cycle is in the telogen phase when the amount of collagen degradation does not satisfy the standard, The criterion in (3) is determined based on the fact that the amount of collagen decomposition in the hair follicle connective tissue sheath during the telogen phase is less than the amount of collagen decomposition in the hair follicle connective tissue sheath during the anagen phase and the catagen phase. The estimation method according to claim 9.

11. A collagen metabolism information acquisition step of acquiring information on collagen degradation and / or neogenesis from the subject to be evaluated, the estimation step includes applying the information on collagen degradation and / or neogenesis acquired in the collagen metabolism information acquisition step to a previously prepared formula or model showing a correlation between the hair cycle and collagen degradation and / or neogenesis, thereby estimating the hair cycle of the hair follicle in the subject to be evaluated; The formula or model comprises at least: The relationship between the presence or absence of collagen neogenesis in the lower part of the hair bulb and the hair cycle, and / or the relationship between the amount of collagen neogenesis in the connective tissue sheath of the hair follicle and the hair cycle, in which the amount of collagen neogenesis in the connective tissue sheath of the hair follicle increases in the order of the resting phase, the regression phase, and the growth phase, and the relationship between the amount of collagen degradation in the hair follicle connective tissue sheath and the hair cycle, in which the amount of collagen degradation in the hair follicle connective tissue sheath during the resting phase is less than the amount of collagen degradation in the hair follicle connective tissue sheath during the anagen and regressing phases; It is created based on The estimation method according to claim 9 or 10.

12. The estimation method according to any one of claims 1 to 11, characterized in that it estimates a hair growth cycle of body hair.

13. A storage means for storing correlation data between the degradation and / or regeneration of collagen in the tissue surrounding the hair follicle, including the hair follicle, and the hair cycle of the hair follicle; an estimation means for applying information on collagen degradation and / or neogenesis obtained from the subject to be evaluated to the correlation data stored in the storage means, and estimating the hair growth cycle of the hair follicle in the subject to be evaluated; Equipped with the estimation means uses the presence or absence of collagen synthesis in the area below the hair bulb of the perifollicular tissue as an indicator to estimate whether the hair cycle of the hair follicle is in the telogen phase, the anagen phase, or the catagen phase. Hair growth cycle estimation device.

14. A computer includes a storage means for storing data relating to the correlation between collagen degradation and / or neogenesis in the tissue surrounding the hair follicle, including the hair follicle, and the hair cycle of the hair follicle; an estimation means for applying information on collagen degradation and / or neogenesis obtained from the subject to be evaluated to the correlation data stored in the storage means, and estimating the hair growth cycle of the hair follicle in the subject to be evaluated; A hair growth cycle estimation program for functioning as a the estimation means uses the presence or absence of collagen synthesis in the area below the hair bulb of the perifollicular tissue as an indicator to estimate whether the hair cycle of the hair follicle is in the telogen phase, the anagen phase, or the catagen phase. Hair cycle estimation program.

15. A collagen degradation detection means for detecting degraded collagen in perifollicular tissue including hair follicles; a collagen synthesis detection means for detecting procollagen in the perifollicular tissue; A storage means for storing correlation data between the degradation and / or regeneration of collagen in the tissue around the hair follicle and the hair cycle of the hair follicle; an estimation means for applying information on collagen degradation and / or neogenesis obtained from the subject to be evaluated to the correlation data stored in the storage means, and estimating the hair growth cycle of the hair follicle in the subject to be evaluated; Equipped with The estimation means and estimating the hair cycle based on the amount of collagen degradation and the collagen new synthesis site and / or collagen new synthesis amount, and The method includes carrying out the following steps (1) and / or (2), and (3): (1) Using the presence or absence of collagen synthesis in the area below the hair bulb of the perifollicular tissue as an indicator, it is estimated that the hair cycle of the hair follicle is in the telogen phase, the anagen phase, or the catagen phase; (2) applying the amount of new collagen produced in the hair follicle connective tissue sheath of the tissue surrounding the hair follicle to a predetermined standard, and estimating whether the hair cycle of the hair follicle is in the telogen, anagen, or catagen phase; (3) Inferring whether the hair follicle is in the anagen phase, the regressive phase, or the telogen phase based on the amount of collagen degradation in the hair follicle connective tissue sheath of the tissue surrounding the hair follicle; Here, the standard in (2) of the hair cycle estimation device is determined based on the relationship that the amount of collagen newly produced in the hair follicle connective tissue sheath increases in the order of the resting phase, regression phase, and growth phase.

16. A computer comprising: a collagen degradation detection means for detecting degraded collagen in perifollicular tissue including hair follicles; a collagen synthesis detection means for detecting procollagen in the perifollicular tissue; a storage means for storing correlation data between the collagen degradation and / or neogenesis in the tissue around the hair follicle and the hair cycle of the hair follicle; an estimation means for applying information on collagen degradation and / or neogenesis obtained from the subject to be evaluated to the correlation data stored in the storage means, and estimating the hair growth cycle of the hair follicle in the subject to be evaluated; A hair growth cycle estimation program for functioning as a The estimation means and estimating the hair cycle based on the amount of collagen degradation and the collagen new synthesis site and / or collagen new synthesis amount, and The method includes carrying out the following steps (1) and / or (2), and (3): (1) Using the presence or absence of collagen synthesis in the area below the hair bulb of the perifollicular tissue as an indicator, it is estimated that the hair cycle of the hair follicle is in the telogen phase, the anagen phase, or the catagen phase; (2) applying the amount of new collagen produced in the hair follicle connective tissue sheath of the tissue surrounding the hair follicle to a predetermined standard, and estimating whether the hair cycle of the hair follicle is in the telogen, anagen, or catagen phase; (3) Inferring whether the hair follicle is in the anagen phase, the regressive phase, or the telogen phase based on the amount of collagen degradation in the hair follicle connective tissue sheath of the tissue surrounding the hair follicle; Here, the criterion in (2) is a hair cycle estimation program that is determined based on the relationship that the amount of collagen newly produced in the hair follicle connective tissue sheath increases in the order of the resting phase, regression phase, and growth phase.

17. The method includes a step of estimating the state of collagen degradation and / or neogenesis in the tissue surrounding the hair follicle in the skin tissue to be evaluated based on the hair cycle, The estimation step includes: applying information about the hair cycle of the hair follicle to a previously prepared equation or model that indicates a correlation between the hair cycle of the hair follicle and collagen degradation and / or neogenesis in the tissue surrounding the hair follicle, and estimating the state of collagen degradation and / or neogenesis in the tissue surrounding the hair follicle to be evaluated; The formula or model is created based on at least the relationship between the presence or absence of collagen neogenesis in the lower part of the hair bulb of the perifollicular tissue and the hair cycle. A method for estimating collagen degradation and / or neogenesis.

18. A storage means for storing correlation data between the degradation and / or regeneration of collagen in the tissue around the hair follicle and the hair cycle of the hair follicle; an estimation means for applying information on the hair growth cycle of hair follicles acquired from the subject to the correlation data stored in the storage means to estimate the state of collagen degradation and / or neogenesis in the tissue surrounding the hair follicles of the subject to be evaluated; Equipped with The correlation data is created based on at least the relationship between the presence or absence of collagen neogenesis in the lower part of the hair bulb of the perifollicular tissue and the hair cycle. A device for estimating collagen degradation and / or neoplasia.

19. A computer is provided with a storage means for storing data relating to the correlation between collagen degradation and / or regeneration in the tissue surrounding the hair follicle and the hair cycle of the hair follicle; a program for estimating collagen degradation and / or neogenesis, which functions as an estimation means for estimating a state of collagen degradation and / or neogenesis in the tissue surrounding the hair follicle of the evaluation subject by applying information on the hair growth cycle of the hair follicle acquired from the evaluation subject to the correlation data stored in the storage means, The correlation data is created based on at least the relationship between the presence or absence of collagen neogenesis in the lower part of the hair bulb of the perifollicular tissue and the hair cycle. A putative program of collagen degradation and / or neoplasia.

20. A method for evaluating a hair follicle comprising: an estimation step of estimating the state of collagen degradation and regeneration in the tissue surrounding the hair follicle in the skin tissue to be evaluated based on the hair growth cycle; the estimation step includes applying information about the hair cycle of the hair follicle to a previously prepared equation or model that indicates a correlation between the hair cycle and collagen degradation and neogenesis in the tissue surrounding the hair follicle, and estimating the state of collagen degradation and neogenesis of the subject to be evaluated; The formula or model is: The relationship between the presence or absence of collagen neogenesis in the lower part of the hair bulb of the perifollicular tissue and the hair cycle, and / or the relationship between the amount of collagen neogenesis in the follicular connective tissue sheath of the perifollicular tissue and the hair cycle, in which the amount of collagen neogenesis in the follicular connective tissue sheath increases in the order of telogen, regressive phase, and anagen, and the relationship between the amount of collagen degradation in the hair follicle connective tissue sheath and the hair cycle, in which the amount of collagen degradation in the hair follicle connective tissue sheath during the resting phase is less than the amount of collagen degradation in the hair follicle connective tissue sheath during the anagen and regressing phases; It is created based on A method for estimating collagen degradation and synthesis.

21. A storage means for storing correlation data between the degradation and regeneration of collagen in the tissue around the hair follicle and the hair cycle of the hair follicle; an estimation means for applying information on the hair growth cycle of hair follicles acquired from the subject to the correlation data stored in the storage means to estimate the state of collagen degradation and neogenesis in the tissue surrounding the hair follicles of the subject; Equipped with The correlation data is The relationship between the presence or absence of collagen neogenesis in the lower part of the hair bulb of the perifollicular tissue and the hair cycle, and / or the relationship between the amount of collagen neogenesis in the follicular connective tissue sheath of the perifollicular tissue and the hair cycle, in which the amount of collagen neogenesis in the follicular connective tissue sheath increases in the order of telogen, regressive phase, and anagen, and the relationship between the amount of collagen degradation in the hair follicle connective tissue sheath and the hair cycle, in which the amount of collagen degradation in the hair follicle connective tissue sheath during the resting phase is less than the amount of collagen degradation in the hair follicle connective tissue sheath during the anagen and regressing phases; It is created based on A hypothesis for collagen degradation and regeneration.

22. A computer, a storage means for storing data relating to the correlation between the degradation and regeneration of collagen in the tissue surrounding the hair follicle and the hair cycle of the hair follicle; a program for estimating collagen degradation and neogenesis, which functions as an estimation means for estimating the state of collagen degradation and neogenesis in the tissue surrounding the hair follicle of the evaluation subject by applying information on the hair cycle of the hair follicle acquired from the evaluation subject to the correlation data stored in the storage means, The correlation data is The relationship between the presence or absence of collagen neogenesis in the lower part of the hair bulb of the perifollicular tissue and the hair cycle, and / or the relationship between the amount of collagen neogenesis in the follicular connective tissue sheath of the perifollicular tissue and the hair cycle, in which the amount of collagen neogenesis in the follicular connective tissue sheath increases in the order of telogen, regressive phase, and anagen, and the relationship between the amount of collagen degradation in the hair follicle connective tissue sheath and the hair cycle, in which the amount of collagen degradation in the hair follicle connective tissue sheath during the resting phase is less than the amount of collagen degradation in the hair follicle connective tissue sheath during the anagen and regressing phases; It is created based on Putative program of collagen degradation and synthesis.

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