KR101223599B1 - Hydrogel type fibroin films and methods of manufacture thereof - Google Patents

Abstract

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel fibroin film made of a protein based protein having excellent moisturizing ability, biocompatibility, flexibility, and skin adhesion required for a cosmetic mask pack.

In addition, the present invention is to produce a protein film having a high water content by adding natural materials such as agar, cellulose and alginic acid to fibroin.

And the hydrogel fibroin film provided in the present invention can be used as a material of the mask pack.

Silk Fibroin, Mask Pack, Protein Film, Hydrogel

Description

Hydrogel fibroin film and its manufacturing method {Hydrogel type fibroin films and methods of manufacture etc}

1 is a photograph of a mask pack fibroin film molded by the method provided in Preparation.

2 is a sample of the tensile strength and the specimen film used for the measurement.

The present invention relates to a novel protein film and a method for manufacturing the same, and more particularly, to isolate and solubilize fibroin in the proteins constituting silk silk, and to mix it with cellulose, alginic acid and agar to form a desired shape in a high moisture content state The present invention relates to a hydrogel fibroin film having high skin adhesion, flexibility, and biocompatibility.

In general, cosmetic packs are made by adding water or lotion to the nonwoven fabric or adding extracts known to be functional. Currently known to this kind include a non-woven fabric pack for improving acne (Korean Patent Publication No. 2004-0024213). However, non-woven fabrics are mainly made of synthetic polymers, and the addition of lotion or functional extracts makes them stiff or poorly used. Recently, a cosmetic mask pack using a hydrogel made of synthetic resin has been developed, but much evaluation and review are required in terms of biocompatibility and safety.

Cosmetic mask packs using collagen, a natural protein, are also on the market, but collagen is an animal-derived material and its strength is weak, making it difficult to handle.

Accordingly, the present inventors have examined the natural materials that can be used as a cosmetic mask pack, and as a result, the silk fibroin has excellent biocompatibility and is used as a surgical suture because it does not affect living tissues and treats surgical wounds. From the fact that it is being developed as a wound coating, it was studied to use it as a cosmetic mask pack.

At present, known as wound dressings include silk fibroin protein membrane for wound dressing (Korean Patent Publication No. 2000-0051938), wound dressing mainly composed of fibroin and sericin (Korean Patent Publication No. 2000-0068154), crystallinity Fibroin film wound dressings (Korean Patent Laid-Open Publication No. 1998-703243) and the like have been demonstrated to be helpful for wound healing as well as skin affinity.

However, while the wound coating is a dry film that does not contain moisture and can absorb pus on the wound area, the film for masks to be developed in this study contains moisture and has good adhesion to normal skin. There is strength and should be good. The reason is that naturally hardened fibroin gel is weak in strength and brittle, so it cannot be processed into a film-type product, so most fibroin films are dried, freeze-dried, and alcohol-treated to remove moisture. Was developed.

Thus, the present inventors have repeatedly studied that the silk fibroin protein forms a gel at room temperature or forms a film when dried, and that natural polysaccharides can contain water for a long time and improve the physical properties of fibroin gel and film. The present invention has been completed by discovering that fibroin films having high skin adhesion and good strength can be produced even in a state containing a lot of moisture.

In order to achieve the above object, the production method in the present invention is characterized by having a desired shape by mixing various types of natural polysaccharides in an aqueous solution of silk fibroin protein in a flat plate so that the thickness of the film is 1-2mm do.

Hereinafter, the present invention will be described more specifically.

The silk fibroin used in the present invention can be obtained using weaving and weaving by-products or silkworm cocoons of undyed silk cloth and silk fabrics available on the market.

The method for obtaining the silk fibroin aqueous solution is not particularly limited, and known methods, for example, natural silk and silkworm cocoon in 0.3% (w / v) aqueous sodium carbonate solution at 100 ° C. for 30 minutes and washed twice with distilled water. Dry at 90 ° C. for 24 hours. In this process, the sericin constituting the outer skin of the silk fiber is removed and a dried product of fibroin corresponding to the central strand of the silk fiber is obtained. The dried silk fibers were dissolved in small portions in a heated 37.5% (w / v) calcium chloride solution to obtain a dissolved silk fibroin salt solution at a concentration of about 11-12% (w / v).

The dissolved fibroin is desalted with distilled water through dialysis, a known method, to remove calcium chloride, thereby obtaining a pure silk fibroin solution. In the desalination process to remove high concentration of calcium chloride, first, the fibroin salt solution is sealed in the inside of the cellulose dialysis membrane of 6,000 to 8,000 Dalton permeation limit, which was boiled and washed twice in distilled water, and then refrigerated. Put in a 2L beaker and stir the magnetic bar. In this process, the salt exits the dialysis membrane and only fibroin remains inside the dialysis membrane. When the salt concentration value of the external distilled water reached the equilibrium by using a refractometer and a conductivity meter, dialysis was terminated when the electrical conductivity became lower than a predetermined value while changing the distilled water.

On the other hand, agar, alginic acid or cellulose is added and mixed to process the obtained pure fibroin solution into a hydrogel-type film containing water. The strength is continuously increased depending on the concentration of fibroin to be added, but the amount of fibroin and natural polysaccharides should be appropriately adjusted to obtain a white fibroin film that is excellent in appearance. Meanwhile, in order to use the prepared liquid mixture of silk fibroin and polysaccharide as a cosmetic mask pack, the thickness of the film should be 1 to 2 mm. For example, it is evenly arranged on a flat plate with a partition of about 1 mm around the rim. After application, the silk fibroin film may be prepared by immersing in a refrigerated container, dried or an aqueous solution containing a salt solution and alcohol.

The silk fibroin film produced according to the manufacturing method of the present invention may have a shape formed in a mold frame that is shaped in a wide plate shape, a mold having a shape or a part of a human body.

The silk fibroin film prepared according to the manufacturing method of the present invention contains several tens of times the moisture of the fibroin used, and exhibits considerable strength and excellent skin adhesion. Moreover, the silk fibroin film of the present invention has the advantage of activating the production of fibroblasts in the skin tissue, promoting the production of collagen and regenerating the skin at the same time is very effective as a mask pack.

Although an Example of this invention is described in more detail below, this invention is not limited only to these examples.

Preparation Example 1 Preparation of Silk Fibroin Film Containing Agar

1) Wait until the agar is mixed evenly with 3% aqueous solution of fibroin at a rate of 0.5% of the weight of fibroin, slowly stirring, and then heated for several tens of seconds before boiling in a microwave oven, and then centrifuged at 4000 rpm for 10 minutes. Separation removes bubbles and small amount of supernatant.

2) The mixed solution obtained in 1) was filled with a predetermined amount in a 10 x 10 cm frame with a 0.2 mm high partition on the frame produced by using an acrylic plate, and then evenly rolled out using a plastic rod.

3) The mold prepared in 2) is dried at 40 ° C. for 24 hours to obtain a translucent film. Prepare a container that fits into this frame, immerse in sterilized distilled water for about 5 minutes, and carefully separate the film using tweezers.

Preparation Example 2 Preparation of Silk Fibroin Film Containing Cellulose

1) Add cellulose powder in 5% aqueous solution of fibroin at a rate of 0.3% of the weight of fibroin, stir and mix quickly and evenly, and then stir continuously for 24 hours, and then centrifuge at 4000 rpm for 10 minutes in a centrifuge tube. The bubbles were removed to remove a small amount of supernatant which was layered at the same time.

2) The mixed solution obtained in 1) was filled with a predetermined amount into a 10 x 10 cm mold with a 0.1 mm high partition on the rim produced by using an acrylic plate, and then rolled out evenly using a plastic rod.

3) Wait 24 hours or more until the fibroin solution gels in the mold prepared in 2) above. Then carefully remove the film from the frame.

4) It is possible to shorten the time to form a gel by immersing the mold in 50% alcohol solution in 3).

Preparation Example 3 Preparation of Silk Fibroin Film Containing Alginic Acid

1) After agitation of alginic acid in a 2% aqueous solution of fibroin at a rate of 0.8% of the weight of fibroin, the mixture was slowly mixed with agitation, and then placed in a centrifuge tube and centrifuged at 4000 rpm for 10 minutes to remove bubbles and a small amount of supernatant.

2) The mixed solution obtained in 1) was filled with a predetermined amount in a 10 x 10 cm mold with a 0.1 mm high partition on the rim produced by using an acrylic plate, and then evenly rolled out using a flat plastic rod.

3) Prepare the container to fit the frame prepared in 2) and filled with 3% (w / v) of calcium chloride aqueous solution soaked and reacted for about 5 minutes. When the film began to be made, it was carefully turned over with tweezers to make contact with the opposite shaving solution.

Test Example 1 Strength Measurement

To evaluate whether it can be used as a cosmetic mask pack using the protein film of the silk fibroin prepared in Preparation Examples 1, 2 and 3. Tensile strength, water evaporation and usability were investigated.

1) Moisturizing

Moisturizing power measured the weight loss due to the evaporation of moisture at room temperature and normal pressure, but the weight loss during 2 hours, which is a time sufficient to use a cosmetic mask pack, was less than 10%, and the water content continued to decrease afterwards. .

2) Strength measurement and improvement

Dumbbell-shaped specimens were prepared to measure the mechanical strength of the hydrogel fibroin film. For the production of the specimen was used a mold frame patterned according to the shape of the specimen on the acrylic plate. Tensile strength was measured using a universal testing machine for the specimen prepared using the mold by the method provided in Preparation Example 1, Preparation Example 2 and Preparation Example 3.

The tensile strength of the silk fibroin film formed by the method provided in Preparation Example 1 was 38 KPa, and the tensile strength was continuously increased with the extension of the drying time. However, skin adhesion and appearance gradually decreased.

The tensile strength of the silk fibroin film molded by the method provided in Preparation Example 2 was on the order of 70-80 kPa. Increasing the content of cellulose greatly increased the strength.

The tensile strength of the silk fibroin film molded by the method provided in Preparation Example 3 is on the order of 50 KPa. However, if another divalent cation salt is used instead of calcium chloride and the immersion time is changed, the tensile strength can vary widely between 20 and 200 KPa.

Test Example 2 Addition-Added Fibroin Film

1) Squalene addition test

0.2, 0.4, 0.6, 0.8, 1.0% of squalene was added to the initial fibroin solution to make silk fibroin by the method provided in Preparation Example 1, Preparation Example 2 and Preparation Example 3 for the purpose of dissolving the fragrance and active ingredient of the cosmetic. It was.

When 0.2% of squalene was added, the tensile strength decreased by 70% compared with the case without adding the initial squalene. After that, as the amount of squalene added increased, the tensile strength slightly decreased, but there was no drastic decrease in tensile strength.

2) Polymer Addition Effect Test

Fibroin in the early stage of preparation of silk fibroin using polyethylene glycol, xanthan gum, acacia powder, hyaluronic acid, and polyvinyl alcohol, which are widely used for the purpose of improving the function of cosmetics, by the methods provided in Preparation Examples 1, 2 and 3. To the solution was added 1% of the fibroin weight, respectively. When polyethylene glycol was added, the strength decreased by 70%, and the xanthan added film showed little change. Acacia powder, hyaluronic acid, polyvinyl alcohol and the like added the film twice the strength of the first.

3) Appearance improvement test

Experiments were carried out to increase the whiteness and improve the appearance by adding magnesium oxide to the silk fibroin film. Experiments were performed by adding 2%, 4%, and 6% magnesium oxide compared to silk fibroin. On the other hand, the higher the magnesium oxide addition ratio, the higher the whiteness. However, when 2% of magnesium oxide was added, the tensile strength decreased by about one third and remained similar thereafter.

As can be seen from the above description, the hydrogel fibroin film prepared according to the manufacturing method of the present invention is excellent in moisture content and moisturizing power, suitable for cosmetic mask pack of skin adhesion and strength, and also has its own characteristics of fibroin. It is effective as a mask pack because skin can be regenerated through affinity and collagen production.

Claims (5)

A method for producing a hydrogel fibroin film for cosmetic mask pack, characterized in that the agar or alginic acid mixed with a silk fibroin aqueous solution and molded to have a plate-like form. The method of manufacturing a hydrogel fibroin film for cosmetic mask pack according to claim 1, further comprising adding a fat-soluble substance including squalene to the aqueous solution of silk fibroin. The method of manufacturing a hydrogel fibroin film for cosmetic mask pack according to claim 1, further comprising polyethylene glycol, xanthan gum, acacia powder, hyaluronic acid or magnesium oxide in the aqueous solution of silk fibroin. Fibroin film produced by the method according to any one of claims 1 to 3. The fibroin film according to claim 4, wherein the fibroin film has a plate shape using a mold.

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