KR20170089427A - Nanofiber scaffold having moisturising and skin regeneration effect and method for preparing thereof - Google Patents

Abstract

The present invention relates to a nanofiber supporter having excellent moisturizing and skin regenerating performance and to a manufacturing method thereof. The nanofiber supporter according to the present invention has a remarkable moisturizing effect and adhesion ability, and especially, when the nanofiber supporter is wetted with water, alginate and spirulina are secreted from the nanofiber and has moisturizing performance, which is, a skin condition improving effect. Furthermore, the nanofiber supporter of the present invention has excellent biocompatibility, can be utilized as various cosmetic products, and specifically, can be variously used as cosmetics, skin-beauty sheets, and the like.

Description

(Nanofiber scaffold having moisturizing and skin regeneration effect and method for preparing the same)

More particularly, the present invention relates to a nanofiber support having a multi-layered structure including alginate and spirulina and having a moisturizing ability, an adhesive ability and a skin condition improving effect, And a manufacturing method thereof.

Nanofibers have a large surface area and high porosity. The smaller the diameter of the nanofiber, the larger the ratio of the area to the volume. The large surface area not only sufficiently constitutes the wet state but also has a three-dimensional structure similar to the structure of the extracellular matrix, which facilitates drug release at the wound site and is effective in wound regeneration.

Nanofibers can be produced using polymers having biocompatibility and biodegradability. Examples of the biocompatible and biodegradable polymer include natural polymers such as collagen, gelatin, alginate, hyaluronic acid and chitosan, polyethylene glycol, polylactic acid, polyglycolic acid, Synthetic polymers such as polyesters of poly (D, L-lactic acid-co-glycolic acid), poly (caprolactone) and poly (hydroxybutyrate) can be used.

Meanwhile, a method of manufacturing nanofibers includes self-assembly, phase separation, electrospinning, and the like.

Nanofibers made by double electrospinning are formed from electrospinning by uniaxial extension of viscoelastic jets of polymer solution or melt. The procedure uses an electric field to create jets of one or more electrically charged polymer solutions from the surface of the solution to the surface of the collector. When a high voltage is applied to the polymer solution (or melt), the filled extrudate of the solution is drawn in the direction of the fixed collector. The extrudate is stretched and bent in the inward direction of the coil, which coagulates as the solvent evaporates in order for the precipitate on the grounded collector to take the form of a nanofiber with a diameter in the nanometer range. The nanofibers thus prepared can be used in drug delivery systems or tissue engineering fields, and are applicable to a variety of fields such as clothing, electrochemical, and environmental fields.

On the other hand, alginate is a material extracted from natural materials such as impression materials and brown algae, and has a copolymer form of D-mannuronic acid and L-glutaric acid. It has a characteristic of being easily deformed into various formulations with sticky viscous components have. These alginates are generally characterized by relatively easy hydrogel formation through the use of divalent cations, that is, ionic cross-linking using a cross-linking agent. The cross-linking agent is bound to D-mannuronic acid and L- The hydrogel can be formed to stabilize the alginate. However, when the alginate and the cation solution are not sufficiently mixed with each other during ionic crosslinking, they do not exhibit uniform physical properties. Further, when the crosslinking agent is used, toxicity is exhibited in the living body. Therefore, biocompatibility There is a limit to fall.

In addition, microalgae are single cell organisms that produce photosyntheses themselves as producers of marine ecosystems. Spirulina is a kind of green algae among microalgae. It is rich in proteins, essential fatty acids, vitamins and minerals, and has been used as a food resource for mankind and has been widely used in animal feed and aquaculture. Recently, however, microalgae have been found to have various physiologically active substances, and they are attracting attention in biotechnology fields such as food nutrition, skin aesthetics, pharmacy, and medicine. There are hundreds of thousands of microalgae, and they produce various physiologically active substances depending on the growing environment. Especially microalgae growing in extreme environment such as deep sea lava and lava zone, new functional materials not found in the land are found.

Therefore, microalgae can be considered as a report of various physiologically active substances. And microalgae are easy to mass-produce. Indeed, in the industrial sector, microalgae have been cultured in large quantities for food or aquaculture. In recent years, in order to utilize microalgae as biomass in the field of bio-energy, studies for increasing the production of microalgae are actively carried out, and the industrial value of microalgae will be further increased in the future. Therefore, research that utilizes microalgae as biomaterials will lead the source technology for securing biologically active materials derived from marine life, leading the super high value-added industry through the combination of two high value-added industries, namely tissue engineering industry and marine bio-industry It is significant in that it can be. Moreover, microalgae will become more important as depletion of land resources as marine resources and enforcement of environmental regulations.

Korean Patent No. 10-1182736

Accordingly, the inventors of the present invention prepared nanofiber scaffolds having excellent moisturizing ability, adhesion ability and skin condition improving effect through the combination of porous polymer, alginate and spirulina while studying nanofiber scaffolds having excellent moisture-absorbing ability and adhesion ability, Completed.

Accordingly, an object of the present invention is to provide a method for producing a porous membrane, comprising: (S1) dissolving a porous polymer in a solvent to prepare a first spinning solution; (S2) electrospinning the first spinning solution to form a first nanofiber layer; And (S3) dissolving alginate and spirulina in a solvent to prepare a second spinning solution; And (S4) forming a second nanofiber layer by electrospinning a second spinning solution on the first nanofiber layer by a second electrospinning method. .

Another object of the present invention is to provide a nanofiber support having a multi-layer structure manufactured by the above method and a cosmetic article comprising the same.

In order to achieve the above object, the present invention provides a method for producing a porous membrane, comprising the steps of: (S1) dissolving a porous polymer in a solvent to prepare a first spinning solution; (S2) electrospinning the first spinning solution to form a first nanofiber layer; And (S3) dissolving alginate and spirulina in a solvent to prepare a second spinning solution; And (S4) forming a second nanofiber layer by electrospinning a second spinning solution on the first nanofiber layer. The present invention also provides a method of manufacturing a nanofiber support having a multilayer structure.

The present invention also provides a nanofiber support having a multi-layer structure prepared by the above method.

The present invention also provides a cosmetic composition for improving skin condition comprising the nanofiber support.

The present invention also provides a skin care sheet comprising the nanofiber support.

The nanofiber support according to the present invention has a remarkable moisturizing effect and adhesion ability. Especially, when the nanofiber support is wetted with water, alginate and spirulina are secreted from the nanofiber and have a moisturizing ability, that is, a skin condition improving effect . Furthermore, the nanofiber support of the present invention is excellent in biocompatibility and can be utilized as various cosmetic products. Specifically, the nanofiber support can be variously applied to cosmetics, skin care sheets, and the like.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view showing a method for producing a nanofiber support having a multi-layer structure; FIG.
FIG. 2 is a view showing the appearance of the nanofiber support according to the present invention using SEM. FIG.
FIG. 3 is a graph showing diameters derived from the appearance of a nanofiber support according to the present invention. FIG.
FIG. 4 is a graph showing contact angles derived from external observation of a nanofiber support according to the present invention. FIG.
5 is a view showing the appearance of the nanofiber support according to the present invention wetted with water using SEM.
FIG. 6 is a graph showing the result of confirming the moisture resistance of the nanofiber support according to the present invention through water content measurement. FIG.
FIG. 7 is a graph showing the result of confirming the efficiency of secretion of the nanofiber support according to the present invention by measuring the amount of spirulina secretion. FIG.
8 is a graph showing the result of confirming the moisture and adhesion of the nanofiber support according to the present invention through the measurement of adhesion.
FIG. 9 is a graph showing the results of confirming the cell viability of the nanofiber support according to the present invention through MTT analysis. FIG.

The present invention provides a method for producing a nanofiber support having a multi-layer structure.

Hereinafter, the present invention will be described in detail.

In the present invention, the nanofiber support having a multilayer structure relates to a nanofiber support having a single layer or more layer structure, and preferably has a two-layer structure.

In the present invention, the nanofiber support having a multi-layered structure comprises (S1) preparing a first spinning solution by dissolving a porous polymer in a solvent as shown in FIG. 1; (S2) electrospinning the first spinning solution to form a first nanofiber layer; (S3) dissolving alginate and spirulina in a solvent to prepare a second spinning solution; And (S4) forming a second nanofiber layer by electrospinning a second spinning solution on the first nanofiber layer.

In the present invention, the nanofiber scaffold generally has a high radial voltage during electrospinning, and when the spinning distance is long, the diameter of the nanofiber is narrowed. Depending on the radial voltage, scattering distance, And physical properties are precisely controlled.

In the present invention, the porous polymer may be any polymer capable of easily and rapidly absorbing water, regardless of its form. More specifically, the polymer may be collagen, gelatin, hyaluronic acid, chitosan, laminin, keratin, Alginate, fibronetin, polycaprolactone (PCL), polyglycolic acid (PGA), polylactic acid (PLA), polylactic acid-glycolic acid copolymer (PLGA), poly-epsilon -caprolactone ), Polyamino acid, polyanhydride, polyorthoester, polyurethane and the like can be used, and preferably collagen, hyaluronic acid, chitosan, alginate, fibronectin, polycaprolactone And it is more preferable that it is polycaprolactone, but the present invention is not limited thereto.

In the step (S1), the porous polymer is preferably contained in 10 to 20 parts by weight, and more preferably 13 to 18 parts by weight, based on the solvent in the preparation of the first spinning solution.

The solvent for the preparation of the first spinning solution in the step (S1) may be an organic solvent. The organic solvent may be tetrahydrofuran (THF), dimethylformamide (DMF), chloroform, Acetone, methylene chloride, ethyl acetate and the like. A mixed solvent of tetrahydrofuran and dimethylformamide may be used, but the present invention is not limited thereto.

In the step (S2), the primary electrospinning of the nanofiber scaffold of the present invention is carried out at a spinning distance of 12 to 25 kV and a spinning speed of 0.1 to 5 ml / h while maintaining a spinning distance of 10 to 25 cm It is characterized in that it is carried out in the electrospinning condition for 20 to 40 minutes, more preferably in the electrospinning condition of radiation voltage 13 to 18 kV and spinning speed 0.5 to 3 ml / h while maintaining the radiation distance of 13 to 22 cm It can be carried out for 20 to 40 minutes.

The second spinning solution may further contain polyethylene oxide (PEO) in step (S3). The PEO may be included in an amount of 1 to 10 parts by weight, preferably 3 to 5 parts by weight, based on the weight of the solvent. But is not limited thereto.

In step (S3), alginate is mainly composed of impurities (water-soluble sodium alginate and triethanol amine alginate) and brown algae. The alginate is contained in an amount of 1 to 10 parts by weight based on 100 parts by weight of the solvent during the production of the nanofiber support By weight, more preferably 2 to 8 parts by weight. If the amount is less than 1 part by weight, the effect of alginate may not be exhibited. If the amount is more than 10 parts by weight, the viscosity of the solution becomes excessively strong, which is difficult to form into nanofibers upon electrospinning.

In the present invention, spirulina is a marine bacterium derived from the sea and is characterized in that it contains a pigment protein picocyanin. The spirulina may be naturally cultivated, artificially processed and marketed without restriction. The above-mentioned pyrosisin promotes the secretion of serine protease when applied to wound or surgical site and promotes the fibrinolytic activity, thereby regenerating the skin of the dermis by promoting the movement of fibroblasts The nanofiber support according to the present invention, which is manufactured by including the above composition, has remarkable improvement in skin condition, especially moisturizing and skin regeneration ability, by spirulina secreted by contact with water.

In addition, in the present invention, the form of the spirulina is not limited, and it is preferable to formulate the extract in powder form, but the present invention is not limited thereto. In addition, the spirulina extract used in the present invention may be extracted with water, a water-soluble alcohol, or a mixed solvent thereof, but is not limited thereto. The water-soluble alcohol may be ethanol or methanol. Examples of the extraction method include filtration, hot water extraction, immersion extraction, reflux cooling extraction, ultrasonic extraction, and the like, and extraction by 1 to 5 times of hot water extraction may be used. The extraction can be performed at a temperature of 20 to 40 캜 for 12 to 48 hours, but is not limited thereto. After the extraction, the mixture is filtered according to a conventional method, and then concentrated under reduced pressure and dried to obtain spirulina powder. At this time, the vacuum concentration may be performed using a vacuum decompression concentrator or a vacuum rotary evaporator. The drying may be performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying.

In step (S3), the amount of spirulina is preferably 0.01 to 10 parts by weight, more preferably 0.01 to 5 parts by weight based on 100 parts by weight of the solvent used. When the content is less than 0.01 part by weight, the moisture resistance may be deteriorated. When the content is more than 5 parts by weight, the viscosity and the hydrophilicity of the solution become excessively strong, and the composition is difficult to be formulated into a nanofiber form upon electrospinning.

The solvent for preparing the second spinning solution in step (S3) may be water, tetrahydrofuran (THF), dimethylformamide (DMF), chloroform, acetone, methylene chloride, ethyl acetate and the like. A mixed solvent of water, tetrahydrofuran and dimethylformamide is preferably used, and it is more preferable to prepare it in an aqueous solution. In the present invention, a small amount of organic solvent may be used in order to dissolve the porous polymer in the preparation of the first spinning solution, but it is preferable to use an aqueous solution when preparing the spinning solution as a whole. When an aqueous solution is used, It is possible to exhibit a remarkable effect when used as a beauty patch or the like.

In the step (S4), the secondary electrospinning in the preparation of the multi-layered structure of the nanofiber support according to the present invention is performed under the conditions of 12 to 20 kV of voltage, 0.01 to 5 ml / h of solution spinning rate and 10 to 25 cm of spinning distance More preferably 20 to 40 minutes, more preferably 12 to 18 kV, irradiation speed of the solution is 0.1 to 4 ml / h and irradiation distance is 13 to 18 cm.

In step (S4), a crosslinking agent such as CaCl ₂ is used for stabilizing the alginate contained in the fiber. However, in the present invention, the crosslinking agent is not used after step (S4). However, the present invention is not limited thereto. When the crosslinking agent is not used, the alginate can be uniformly dispersed on the surface of the nanofiber support according to the present invention wetted with water, so that the adhesion and moisture resistance of the nanofiber support of the present invention are remarkable.

On the other hand, the use of the crosslinking agent indicates that the nanofiber support prepared according to the manufacturing method of the present invention is more stable than the general nanofiber support, and at the same time, the application of the nanofiber does not show any toxicity due to the chemical substance It is characterized by being biocompatible.

In addition, the present invention provides a nanofiber support having a multi-layer structure manufactured by the method of manufacturing the nanofiber support.

In the present invention, the nanofiber support of the present invention preferably has a multilayer structure, and more preferably has a two-layer structure. Accordingly, the nanofiber support of the present invention preferably has a diameter of 30 to 200 nm, But is not limited to. If the diameter is less than 30 nm, it is likely to be easily broken by external force. If the diameter is more than 200 nm, spirulina emission may be inhibited. In the case of a multilayer structure of two or more layers, the thickness is preferably 30 to 200 nm, more preferably 30 to 100 nm.

In the present invention, the nanofiber scaffold is characterized in that it has excellent moisturizing ability, that is, skin improving effect and adhesion ability.

In the present invention, the nanofiber support may further comprise at least one known active ingredient for enhancing the effect of improving the moisture-retaining ability and the adhesion performance, if necessary.

The present invention also provides a cosmetic composition for skin condition improvement comprising the nanofiber support.

In the present invention, the improvement of the skin condition is intended to make the condition of the skin health and beauty including the skin tone, the wrinkles of the skin, the elasticity of the skin and the skin texture better. Specifically, But is not limited thereto.

In the present invention, the cosmetic composition may further contain conventional auxiliary agents and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments, perfumes and the like which are commonly used in cosmetic compositions in addition to the nanofibers. For example, the cosmetic composition may further contain an auxiliary component such as glycerin, butylene glycol, polyoxyethylene hardened castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate, allantoin and the like .

In the present invention, since the cosmetic composition is basically applied to the skin, it can be made into any formulation conventionally manufactured by referring to the cosmetic composition of the related art. For example, they may be formulated as solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays, But is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritive cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a mask pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, the carrier component may include an animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, have.

When the formulation of the present invention is a powder or a spray, the carrier component may include lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder and the like. Particularly in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane, dimethyl ether, and the like.

When the formulation of the present invention is a solution or an emulsion, the carrier component may include a solvent, a solubilizing agent, and an emulsifying agent. Specific examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid esters of sorbitan, and the like.

When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol, propylene glycol or the like as a carrier component; Suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, trachant, and the like.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, and the like.

The present invention also provides a skin care sheet comprising the nanofiber support.

The skin-care sheet may be a mask pack, a patch, or the like.

In the present invention, the skin-beauty sheet can be produced by a conventional method. For example, the nanofiber and the galactomisses fermented filtrate, purified water, Carbopol # 940 (2%), 1,3 Pentaerythritol, Pentaerythritol, Butylene glycol, NATRASOL 250HHR, Propylene glycol, Allantoin, DL-Panthenol, Glycerin, HC-60, PHENONIP, MP, PF (W80402) , 1%), HYAL-JOINT (1%), SKIN SILK OH4, aloe extract, GPC, ceramide, BSASM and silk protein.

For example, the mask pack can be manufactured as follows. First, purified water, carbomer, and butylene glycol were mixed and dissolved homogeneously while stirring at a speed of 1,500 rpm for 60 minutes. The aquatic raw materials are then dissolved and the fragrance and preservative raw materials are dissolved while warming to 70-75 占 폚. The mixture is emulsified and mixed at a rate of 2,000 to 3,000 rpm for 30 minutes while being added to the perfume and preservative mixture raw material mixture, and then mixed at a rate of 1,500 to 2,000 rpm for 20 minutes to prepare a mask stock solution. The nanofibers of the present invention are added to the mask stock solution in an amount of 1 to 50% by weight based on the total weight of the undiluted solution, mixed thoroughly for 15 minutes, and filtered through 100 mesh to prepare a mask stock solution. Next, the mask sheet used in a conventional mask pack is filled with the mask stock solution, and then mask-bonded using a mask sealer to produce a mask pack.

The redundant contents are taken into consideration in the complexity of the present specification, and terms not otherwise defined herein have the meanings commonly used in the art to which the present invention belongs.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. It is to be understood, however, that the invention is not limited to the disclosed embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

Example 1: Preparation of spirulina extract

Spirulina powder (Nature's Care Inc., Belrose, Australia) was added to 1 L of distilled water and stirred for one day. The Spirulina solution was then centrifuged at 100 xg for 5 min and the supernatant was collected in another bottle. The supernatant was separated by filtration under reduced pressure with a Whatman filter paper, and the supernatant was lyophilized. The spirulina extract solution prepared through the series of steps was stored at 20.degree. C. in powder form.

Example 2. Fabrication of a nanofiber support having a multi-layer structure

Polycaprolactone was used as a kind of porous polymer. First, PCL nanofibers were fabricated using electrospinning technology. To this end, 1.5 g of polycaprolactone (PCL) was dissolved in 10 mL of a solvent mixture of tetrahydrofuran (THF) and dimethylformamide (DMF) in a volume ratio of 7: 3 to prepare a 15% (w / v) 1 spinning solution. 15 ml of the first spinning solution was injected through an 18 G needle at a rate of 1 mL / hr. During the injection of the first spinning solution, the needle and collector ) At a distance of 15 cm and a voltage of 15 kV was applied to form a nanofiber first nanofiber layer.

Next, 0.4 g of alginate and 0.5 g of polyethylene oxide (PEO) were added to 10 mL of distilled water as a solvent, and 0, 1.41, 2.35 and 3.76 parts by weight (w / v) of Spirulina extract powder were added to the solvent A second spinning solution was prepared. Thereafter, the second spinning solution was sprayed at a rate of 0.3 mL / hr on the first nano fiber layer using a 22 G needle, and the spinning solution was sprayed for 30 minutes A second nanofiber layer was formed in nanofiber form by applying a voltage of 15 kV between the needle and the collector at a distance of 15 cm. The formed nanofibers of the two-layer structure were allowed to stand overnight in a vacuum state, and then the organic solvent was removed to prepare a two-layered nanofiber support according to the present invention. Separately, the nanofiber support was treated with CaCl ₂ at a concentration of 2% (w / v) to prepare a cross-linked nanofiber support. A schematic diagram showing the series of steps is shown in Fig.

Example 3. Identification of Physical Properties of Nanofiber Support by SEM Observation

The diameter of the nanofiber support, which is the physical property of the multi - layered nanofiber support, was determined by SEM (scanning electron microscope) observation. Then, the contact angle of the water drop on the nanofiber support was measured using a contact angle analyzer. First, the nanofiber support prepared in Example 2 was dried in a vacuum chamber and then plated with platinum. Thereafter, observation was carried out using a SEM at a distance of 15 cm at 15 kV, and the analysis was performed by Image J. The results are shown in FIG. 2, and the result of converting the diameter of the fiber is shown in FIG. 3 占 퐇 of water drops were dropped on each nanofiber sample and contact angle was measured. The result is shown in Fig. Specifically, FIG. 2 (a) shows the results for PCL, 2b for alginate, 2c for alginate and spirulina 1.41 parts, 2d for alginate and spirulina 2.35 parts, and 2e for alginate and 3.76 parts for spirulina.

As shown in Fig. 2, the surface of the patch and the diameter of the fiber were confirmed through SEM observation. As shown in Fig. 3, diameters of the nanofibers were 1843 ± 547, 173 ± 25, 77 ± 18, 73 ± 17 and 63 ± 14 nm, and the diameters of the nanofiber scaffolds prepared by adding 1.41 parts by weight of spirulina Was confirmed to be the longest, and 3.76 parts by weight of spirulina was added to confirm that the diameter of the nanofiber support prepared was the shortest.

As shown in FIG. 4, the contact angles of the nanofibers were 121.06 ± 5.56, 70.01 ± 6.13, 55.39 ± 1.49, 36.15 ± 8.66, and 25.41 ± 7.85 °, respectively. And it was confirmed that the hydrophilicity was increased as the addition amount of spirulina was increased.

Example 4. Determination of moisture resistance of nanofiber support by measuring water content

A moisture content measurement was performed to confirm whether the moisture content confirmed in Example 3 was maintained over time. First, the nanofiber support prepared in Example 2 was weighed to a size of 2 cm x 2 cm before treating the distilled water, and the weight was measured. The nanofibers were immersed in a Petri dish of 60 nm in distilled water for 30 minutes. After that, the wet nanofibers were taken out, and the weight of the nanofibers was measured after removing the water formed on the surface of the fibers. The weight of the nanofibers before the distilled water treatment was compared with the weight of the nanofibers after the treatment. The results are shown in FIG. 5, and the hydroscopic degree was measured by the following equation.

As shown in FIG. 5, the water content value representing the ratio of the weight of water to the total weight was compared. As a result, it was confirmed that the nanofiber support containing spirulina had a higher water content than the non-crosslinked nanofiber support Respectively. Particularly, when 2.35 parts by weight of spirulina was added, it was confirmed that it had the most remarkable water content, that is, moisture.

In addition, in order to confirm the moisture resistance of the nanofiber support according to the present invention, the appearance of the nanofiber support was observed by SEM after 30 minutes passed by immersing the nanofiber support in water. The results are shown in Fig.

As shown in Fig. 6, the PCL nanofiber support, a nanofiber made of a simple porous polymer, did not show a large change on the surface immediately after being wetted with water. However, as described in Example 2, it was confirmed that the nanofiber support prepared by adding alginate and spirulina to the multi-layered support filled in voids between fibers when immersed in water. This indicates that the alginate and spirulina of the two layers of the multi-layered nanofiber support melt to expose the PCL nanofiber, which is a porous polymer of the first layer, and simultaneously, the molten alginate and spirulina fill the spaces between the fibers. As a result, it can be confirmed that the alginate and spirulina, which are sandwiched between the fibers, exhibit remarkable moisturizing power.

Example 5: Determination of the secretion efficiency of nanofiber scaffolds by measuring changes in secretion amount of spirulina

The secretion efficiency of the nanofiber scaffold of the present invention was measured by measuring the amount of spirulina secretion, which is one of the nanofiber support materials. First, the nanofiber support prepared in Example 1 was cut into a size of 1 cm x 1 cm, and then 16 pieces of the nanofiber support were added to a 24 well plate in each well. Thereafter, 1 ml of distilled water was added to the wells, followed by allowing to stand for 30 minutes. Then, the solution of each well was transferred into a 96-well plate in 200 μl by a micropipette, and the procedure was repeated three times. The solution in the wells was then quantitatively determined using the absorbance at 620 nm. The results are shown in Fig.

As shown in FIG. 7, it was confirmed that the nanofiber support having cross-linking with CaCl ₂ showed no optical density. In addition, it was confirmed that, in the case of the nanofiber support having no cross-linking with CaCl ₂ , the spirulina release increased with increasing the weight ratio of spirulina.

Example 6. Determination of Moisture and Adhesion of Nanofiber Support by Adhesion Measurement

The nanofiber support of the multi-layered structure according to the present invention was attached to the actual skin, and the moisture and adhesion strength thereof was confirmed. First, the nanofiber support prepared in Example 1 was cut into a size of 2 cm x 2 cm, and then wetted with distilled water, which was attached to the inside of the arm. Immediately after attachment of the nanofiber support to the arm and after 30 minutes of attachment were visually observed and compared, the results are shown in FIG.

As shown in FIG. 8, it was confirmed that the adhesion was decreased in all samples over time. CaCl ₂ And the adhesion of the nanofiber support prepared without addition of spirulina was remarkably decreased to such an extent that the nanofiber support did not adhere completely to the arm. On the other hand, it was confirmed that as the addition amount of spirulina increased, the adhesion also increased.

Example 7. Determination of cytotoxicity on nanofiber scaffolds by MTT assay

At the cellular level, MTT analysis was performed to confirm the cytotoxicity of the nanofiber scaffold according to the present invention. MTT assays were performed using a keratinocyte cell line (HaCaT). The HaCaT was cultured in a cell culture flask in DMEM-H medium containing 10% FBS (fetal bovine serum) and 1% antibiotic-antifungal agent in an incubator (5% The cultured HaCaT was inoculated into a 24-well plate at a concentration of 310 ⁴ cells / well. These cells were replaced with serum-free medium (DMEM-H containing 1% antibiotic-antifungal agent) and cultured in an incubator. One day later, MTT analysis was performed in a manner commonly practiced in the art. The optical density (OD) of the solution was measured using microplate liters at 540 nm. The results are shown in Fig.

As shown in FIG. 9, it was confirmed that the cell survival rate was increased by addition of spirulina, compared with the nanofiber support prepared by PCL, which is a simple porous polymer, and the nanofiber support prepared by adding only alginate thereto. That is, the nanofiber support of the present invention has high biocompatibility.

In general, the nanofiber support according to the present invention has a high moisture content, a high spirulina secretion efficiency, and a high adhesion and adhesion ability. Thus, the biocompatible nanofiber support of the present invention has a high water content, Cosmetic products, skin care sheets, and the like.

All experiments were repeated three times and the mean and standard deviation values of the data were measured using a Sigma plot. In addition, the difference of each test group was confirmed using the Student t-test, and the p-value shows significance of the difference value (* p <0.05, ** p <0.01, *** p <0.001).

Claims (13)

(S1) dissolving the porous polymer in a solvent to prepare a first spinning solution;
(S2) electrospinning the first spinning solution to form a first nanofiber layer; And
(S3) dissolving alginate and spirulina in a solvent to prepare a second spinning solution; And
(S4) forming a second nanofiber layer by electrospinning a second spinning solution on the first nanofiber layer;
Wherein the nanofibrous support comprises a plurality of nanofibers.
The method according to claim 1,
Wherein the multi-layer structure is a two-layer structure.
The method according to claim 1,
In step (S1), the porous polymer may be selected from the group consisting of collagen, gelatin, hyaluronic acid, chitosan, laminin, keratin, alginate, fibronetin, polycaprolactone (PCL), polyglycolic acid , Polylactic acid (PLA), polylactic acid-glycolic acid copolymer (PLGA), poly-epsilon -caprolactone (PCL), polyamino acid, polyanhydride, polyorthoester, and polyurethane ). &Lt; / RTI &gt; The method of claim &lt; RTI ID = 0.0 &gt; 1, &lt; / RTI &gt;
The method according to claim 1,
Wherein the step (S3) further comprises adding polyethylene oxide (PEO) to produce a second spinning solution.
The method according to claim 1,
Wherein the alginate is contained in an amount of 1 to 10 parts by weight based on 100 parts by weight of the solvent in the step (S3).
The method according to claim 1,
Wherein the spirulina is contained in an amount of 0.01 to 10 parts by weight based on 100 parts by weight of the solvent in the step (S3).
The method according to claim 1,
Wherein the solvent in step (S3) is water.
The method according to claim 1,
In the step (S4), the electrospinning is performed by applying a voltage of 12 to 20 kV at a speed of 0.3 to 5 ml / h while maintaining a distance of 10 to 25 cm. Gt;
The method according to claim 1,
Wherein the crosslinking agent is not used after the step (S4).
A nanofiber scaffold of multilayer structure prepared by the process according to any one of claims 1 to 9.
A cosmetic composition for improving skin condition comprising the nanofiber support of claim 10.
12. The method of claim 11,
Wherein the skin condition improving agent is a moisturizing agent.
A skin-beauty sheet comprising the nanofiber support of claim 10.

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