TW201517930A - Use of starfish extract for manufacture of composition for enhancing secretion of collagen - Google Patents

Abstract

The present invention provides a starfish extract for manufacture of composition for enhancing the secretion of collagen, which comprises starfish extract. The starfish extract comprises active ingredients that are able to enhance the ability of collagen secretion of skin fibroblast cells and the preparation thereof is able to promote the economic value of starfish and is environmental friendly; thus, the present invention is suitable for applications regarding skin care and cosmetic products.

Description

Use of a starfish extract for the preparation of a composition for promoting collagen proliferation

The present invention relates to a use for promoting collagen hyperplasia, and more particularly to the use of a starfish extract for the preparation of a composition for promoting collagen proliferation.

Starfish is the name of an animal under the echinoderms. It has an amazing ability to regenerate. It not only regenerates the broken brachiopod, but some starfish can even regenerate a complete new individual from the truncated wrist. Starfish are mostly distributed in coastal areas of Asia, North America, and Australia, and the habitat is usually sandy bottom. Taiwan is an island with abundant aquatic products. There are also many kinds of starfish, such as Archaster typicus , which is common in the intertidal zone of Penghu. Starfish are widely used. In addition to being used as a food or processed food, some starfish are also a commonly used Chinese herbal medicine. Starfish are also processed into powdered feed because of its high nutritional value. In terms of medical use, the starfish can be used to make substitute plasma that is harmless to the human body after refining; in addition, studies have indicated that the components of the starfish, such as eicosapentaenoic acid, are useful in the treatment of hyperlipidemia and cardiovascular diseases. Certain effect.

Collagen is a biological polymer substance that is the most abundant protein in vertebrates and accounts for one-third of total protein in humans. It is mainly found in the extracellular matrix of connective tissue and is a scaffold between cells and cells. Human skin, Cartilage, cornea, blood vessel wall, internal organs and so on all contain collagen, especially the skin. Collagen provides the strength to support skin cells, keeping the skin firm and elastic. However, with the increase of age and the accumulation of external unfavorable factors, the ability of skin fibroblasts to produce collagen will decrease. Once the rate of collagen loss is greater than the rate of its formation, the skin will produce wrinkles and sagging.

Therefore, supplementing collagen or enhancing the collagen production ability of cells is an important issue in skin care. There are many products that supplement collagen in the market, including external application, injection or consumption. However, the molecular size of collagen is too large to penetrate the stratum corneum of the skin. The application method can hardly make the collagen reach the dermal layer of the fibroblasts. If the collagen is directly added by injection, the skin can be immediately reduced. The fine lines, but the collagen injected into the body will be decomposed by the enzymes in the body, requiring regular application, the process is complicated and costly, and the injection of collagen may cause immune rejection and increase the risk of use. If collagen is added to eat collagen-rich foods, and the collagen enters the stomach, the structure of the triple helix is destroyed to form a multi-peptide, which is then cleaved into small molecules through the digestive juice and finally broken down into Amino acid. Although the body uses these amino acids to synthesize proteins, but not limited to collagen, the actual effect of collagen supplementation is limited.

In view of this, the ability to increase collagen production in human skin fibroblasts is more effective than collagen supplementation to ensure efficient production and utilization of collagen. However, there is currently no safe and effective product on the market that promotes collagen production. .

Accordingly, it is an object of the present invention to provide a use of a starfish extract for preparing a composition for promoting collagen proliferation, wherein the starfish extract comprises a water-soluble polysaccharide, and The water soluble polysaccharide system is a starfish polysaccharide. The starfish extract can increase the collagen proliferation rate of a cell which is a skin fibroblast and is derived from a human; the starfish extract of the invention can promote the collagen fibroblast proliferation rate of the skin fibroblast of 80% or more. .

In one embodiment of the present invention, the starfish extract is obtained by extracting a starfish in vivo, and the starfish living body is prepared by freezing, milling, high temperature extraction, and ethanol extraction.

Further, the composition prepared by the starfish extract of the present invention may be an external rubbing agent, or a topical pharmaceutical product, or an external skin care product, or a cosmetic. It should be noted that the external rubbing or external pharmaceutical products include, but are not limited to, an ointment, or a cream, or a patch or a patch, or an artificial tissue containing an active ingredient. Further, the external skin care product or cosmetic may include, but is not limited to, a lotion, or a cosmetic cream, or an emulsion, or a foundation, or a foundation, or a sunscreen lotion, or a body wash.

Another object of the present invention is to provide an agent for promoting cell proliferation collagen, comprising a starfish extract, wherein the cell line is a skin fibroblast, and the starfish extract contains 89% (w/w) or more. a water soluble polysaccharide.

The starfish extract of the present invention contains an active ingredient and can remarkably promote collagen production by skin fibroblasts. The starfish extract of the invention has the natural source of the starfish extract, and the preparation method is simple and pollution-free, so it can be effectively applied to the skin care products and cosmetics.

The embodiments of the present invention will be further described in conjunction with the drawings, which are set forth to illustrate the features and applications of the present invention, and are not intended to limit the scope of the present invention. In the spirit and scope of the present invention, the scope of the present invention is defined by the scope of the appended claims.

The first figure is a flow chart for extracting the starfish extract of the present invention.

The second figure is the ability of the starfish extract of the present invention to promote collagen proliferation.

The third figure is a fluorescent staining micrograph of the starfish extract of the present invention for promoting collagen production of skin fibroblasts, wherein (a) the control group; (b) treated with 5% (v/v) starfish extract; ) treated with epidermal growth factor (EGF, 5 ng/mL).

In order to confirm that the starfish extract of the present invention can promote collagen proliferation, the present invention prepares a starfish extract by a gentle and easy-to-use method, and analyzes the active ingredient in the starfish extract. Further, the starfish extract of the present invention was also co-cultured with skin fibroblasts, and the starfish extract of the present invention was evaluated to enhance the collagen-producing ability of the skin fibroblasts.

definition

The starfish extract of the present invention is a collagen production rate which can increase the skin fibroblasts, and the content which is sufficient for the purpose is defined as an "effective dose". The amount at which this can be achieved will be determined according to a number of factors known to those skilled in the art, such as the consumer's weight, general condition, age, sex, and the effect of the composition matrix.

Example 1: Preparation of Starfish Extract

The archery typicus used in the present invention is collected from the Wuhu region of Taiwan. For the preparation of starfish extract, please refer to the first figure. In the preparation of the starfish extract of the present invention, first, the living starfish is collected and immediately stored at -4 ° C ~ -20 ° C during storage, and the frozen starfish is ground in vivo (as in step S1); The starfish is dissolved in water at a ratio of 1:3 to 1:20 (w:v) and heated to 90 ° C to 100 ° C for 1 to 5 hours (as in step S2); after high temperature extraction, it is filtered by centrifugation (4,300 g). And taking the supernatant (such as step S3); then, adding 3 volumes of ethanol at 4 ° C to precipitate the polysaccharide for 16-24 hours (as in step S4); after ethanol precipitation, by centrifugation (4,300g) Rpm) filter and take the precipitate (as in step S5); then, the precipitate is dissolved in 10-15 volumes of water (as in step S6); finally, it is filtered by centrifugation (4,300 g rpm) and taken up The clear liquid (as in step S7), the starfish extract of the present invention is obtained.

Example 2: Analysis of water-soluble polysaccharides of starfish extract

The total polysaccharide content of the starfish extract of the present invention is determined by the phenol-sulfuric acid method. 100 μL of starfish extract was mixed with different concentrations of glucose standard solutions (0, 25, 50, 75, 100, and 125 mg/mL) in test tubes. Next, 0.5 mL of a phenol solution was added and reacted with 2.5 mL of sulfuric acid. The tube was measured for its absorbance at 750 nm as a spectrophotometer.

The analysis results show that 2.2% (w/v) of the water-soluble polysaccharide is contained in the 2.5% (w/v) starfish extract solution, so that the seawater extract of the present invention contains a water-soluble polysaccharide content of about 89% (w/w).

Example 3: Analysis of the ability of starfish extract to promote collagen proliferation

Human skin fibroblasts CCD-966SK (Human skin fibroblast cells, purchased from Food Industry Development Institute) in 24-well plates, human skin fibrils, in minimal essential medium (MEM) containing 10% fetal bovine serum (FBS) The concentration of the cells was 4 x 10 ⁴ cells/well. The medium was updated after 24 hours of culture. In the experimental group, the starfish extracts (concentrations of 0.2, 0.4, 0.6, 0.8, and 1.0 mg/mL, respectively) were diluted with phosphate buffer saline (PBS) and added to each well. In the control group, the starfish extract was not added, and the supernatant was cultured at 37 ° C under 5% carbon dioxide for 48 hours. It was found that the starfish extract promoted collagen production and increased linearly with time within 48 hours. .

The content of the water-soluble type I collagen was analyzed using a Sircol® Collagen Assay Kit (Bicolor Life Science Assays, Northern Ireland, UK). At room temperature, the supernatant obtained in the above method of 100μL with 1mL Sircol ^® dyes mixed for 30 minutes. This was followed by centrifugation at 15,000 g for 10 minutes to form a collagen precipitate. The supernatant after centrifugation was removed and the collagen precipitate was dissolved in 1 mL of an alkaline solvent. The absorbance at a wavelength of 540 nm was measured with a spectral signal sensor. The amount of collagen was calculated as a water-soluble collagen standard curve (standard disease-free bovine skin type I collagen, USA).

Please refer to the second figure, which is an analysis of the collagen production of the present invention to promote collagen production, and found that the experimental group produced 45.25 ρg of collagen per unit of human skin fibroblasts after treatment with 1 mg/mL of starfish extract; It is much higher than the amount of 24.48 ρg of collagen produced by the control group. Therefore, the starfish extract of the present invention clearly demonstrates that it can effectively promote the collagen proliferation rate of at least 80% or more, preferably 84.8%.

Example 4: Fluorescent immunoassay of starfish extract promoting collagen proliferation

The human skin fibroblast CCD-966SK was fixed in phosphate buffered saline (PBS) containing 1% paraformaldehyde (Sigma, France) for 30 minutes and washed in a phosphate buffer solution. The cells were treated with the starfish extract (5% (v/v)), epidermal growth factor (5 ng/mL) of the present invention for 24 hours, and the control cells were treated with physiological saline (Phosphate Buffered Saline, PBS). The cells treated with the starfish extract or the epidermal growth factor were co-cultured with the water-soluble type I collagen rat monoclonal antibody (1/2000 dilution, abcam, UK) at 1% (w/v). a phosphate buffer solution of bovine serum albumin (PBS). Next, with phosphorus The acid salt buffer solution was thoroughly washed and co-cultured with fluorescein-labeled rat IgG antibody (1/200 dilution, KPL, USA) in a light-free environment for 60 minutes. After washing with a phosphate buffer solution, it was observed with a fluorescent conjugated focus microscope (Leica TSC SP2, Germany).

Please refer to the third figure, which is a fluorescence staining micrograph of collagen produced by skin fibroblasts, where (a) represents the control group cells, and (b) represents cells treated with 5% (v/v) starfish extract. (c) represents cells treated with epidermal growth factor (5 ng/mL). The arrows in the figure indicate collagen. As is clear from the third figure, the cells treated by the starfish extract of the present invention substantially increase the amount of collagen produced by the control group, and produce more collagen than the cells treated with the epidermal growth factor. Therefore, the present invention The starfish extract was further confirmed to be effective and significantly promote collagen proliferation.

At the same time, the starfish extract of the present invention also contains various amino acids and trace elements. For the analysis results, please refer to the following table.

Example 5: Production of cosmetics and/or pharmaceuticals for promoting collagen proliferation by starfish extract

Starfish extract can produce water-in-oil (O/W) neutral creams and gels and other cosmetics and/or pharmaceuticals. O/W neutral cream oil phase is about 30~50%, so it doesn't have too greasy feeling. It can add starfish extract to the formula, which has the effect of promoting skin collagen proliferation. The starfish extract obtained by the invention is applied to The formula of neutral cream is shown in Table 3:

The oil phase component is slowly added at a high speed, and after the emulsification is completed, a cream containing a starfish extract can be obtained, which can be stored at room temperature for 6 months.

The starfish extract obtained by the invention can also be used for producing collagen gel, and the gel can also achieve the effect of promoting skin collagen proliferation. The formula of the gel is shown in Table 4:

Example 6: Producing an essence of collagen proliferation by using starfish extract

The starfish extract of the invention can be used for producing an essence for promoting collagen proliferation, comprising a starfish extract component, a moisturizing functional component, a whitening functional component and a bacteriostatic agent. The essence for promoting collagen hyperplasia is a deep ocean water as a matrix, and the content thereof is substantially about 40% by weight, wherein the total weight of the collagen-promoting essence is increased, and the content of the starfish extract component is 15 wt%. % to 20% by weight; the moisturizing functional ingredient comprises, but is not limited to, selected from the group consisting of Glygoprotein, Hyaluronic Acid, 1,3-Butanediol, or 1,3-propanediol, the content of which is Between 10% and 20% by weight; the whitening functional component includes, but is not limited to, selected from Arbutin, Kojic Acid, Sodium Ascorbyl Phosphate, or Vitamin C. The group consisting of magnesium nitrate (Magnesium Ascorbyl Phosphate) is contained in an amount of from 10% by weight to 20% by weight; and the bacteriostatic agent is contained in an amount of from 0.1% by weight to 0.2% by weight.

The effect of the starfish extract of the present invention for producing an essence is that the starfish extract component in the essence contains a water-soluble polysaccharide, that is, a starfish polysaccharide, which can promote skin fibroblasts. The rate of collagen proliferation, and the starfish extract also contains a variety of amino acids and trace elements, which can activate skin cells, and with the moisturizing functional components and whitening functional components in the essence, can further enhance the skin's maintenance effect on the skin. .

In summary, the starfish extract of the present invention has various active ingredients and is rich in water-soluble polysaccharides, which can promote collagen proliferation of skin fibroblasts. In addition, the method for producing starfish extract of the present invention is simple and non-polluting. Therefore, it can not only effectively use natural resources without harming the environment, but also provide a highly active and safe skin care product.

However, the above description is only for the preferred embodiment of the present invention, and those skilled in the art can make various other modifications according to the above description, but the changes are still within the spirit of the present invention and as defined below. In the scope of patents.

Claims (12)

A use of a starfish extract for the preparation of a composition for promoting collagen proliferation, wherein the starfish extract comprises a water soluble polysaccharide. The use of the first aspect of the invention, wherein the starfish extract can increase the collagen proliferation rate of a cell. The use of claim 2, wherein the cell line is a dermal fibroblast. The use of claim 3, wherein the dermal fibroblast is derived from a human. The use according to claim 3, wherein the starfish extract promotes a collagen proliferation rate of the skin fibroblast of 80% or more. The use according to claim 1, wherein the water-soluble polysaccharide system is a starfish polysaccharide. The use of claim 1, wherein the starfish extract is obtained by in vivo extraction of starfish. The use according to claim 1, wherein the starfish extract is produced by freezing, milling, high temperature extraction and ethanol extraction. The use according to claim 1, wherein the composition is an external rubbing agent, a topical pharmaceutical product, a topical skin care product, or a cosmetic product. An accelerator for promoting cell proliferation of collagen, comprising a starfish extract. The accelerator of claim 10, wherein the cell line is a dermal fibroblast. The accelerator according to claim 10, wherein the starfish extract contains 89% (w/w) or more of a water-soluble polysaccharide.