KR102068173B1 - Mutifunctional bionanocomposite hydrogel, method of manufacturing thereof and use thereof - Google Patents

Abstract

The present invention is a multifunctional biodegradable nanocomposite hydrogel comprising biopolymer and metal nanoparticles, a method for preparing the same, a dressing for treating skin diseases, wounds or burns, including the biodegradable nanocomposite hydrogel, and the biodegradable It relates to a cosmetic composition for skin care comprising a nanocomposite hydrogel.

Description

Multifunctional bionanocomposite hydrogel, its manufacturing method and use thereof {Mutifunctional bionanocomposite hydrogel, method of manufacturing applications and use}

The present invention relates to a multifunctional bio nanocomposite hydrogel, a method for preparing the same, and a use thereof, and more particularly, a biodegradable nanocomposite hydrogel including biopolymer and metal nanoparticles, a method for preparing the same, and the multifunctional bionano. The present invention relates to a dressing for treating skin diseases, wounds or burns including a complex hydrogel, and a cosmetic composition for skin care comprising the multifunctional bionanocomposite hydrogel.

In recent years, the occurrence of skin diseases is rapidly increasing due to the rapid change of the environment and climate change in the world, and the demand for new functional skin protection agent is increasing due to the improvement of quality of life and the interest in skin diseases due to socioeconomic development. have. Among the products that are currently widely used as skin care products, products that impart various functionalities to it using hydrogels are widely used.

Hydrogel is a hydrophilic polymer that has a three-dimensional network structure of polymer chains and can contain a large amount of water by hydrophilic functional groups and capillary and osmotic pressures. Wound dressings, contact lenses, drug delivery systems, medical implants, It is widely used as a carrier for wastewater treatment, and recently, it is also widely used as a material of a mask pack used to replenish moisture and nutrients to the face.

Hydrogels for use in the above-mentioned fields are basically required for the ability to absorb large amounts of water, and to maintain proper physical strength and form stability.

In particular, hydrogels used for wound treatment or human body (skin) contact should be non-toxic to the human body. When used, they can prevent contamination from the outside and are easy to adhere to wounds or skin, as well as sterilized and have excellent storage properties. It should be easy to handle.

Currently, polyacrylic acid and polyvinyl alcohol are widely used as hydrogel materials. They have high water absorption and excellent morphological stability, but if they are attached to the skin for a long time, the problem of biocompatibility is somewhat lowered, such as irritating the skin and causing itching.

Recently, the use of biopolymers, which are environmentally friendly, biocompatible, and non-toxic renewable materials, has attracted attention. However, most of the raw polymer materials have a physical strength lower than that of conventional plastics, and are poor in resistance to moisture and inferior in processability.

As part of the method for improving the problem of the raw polymer material, a small amount of nanoparticles such as nanoclay, nanometal or nanofiber are mixed with the raw polymer to increase the bonding strength between the nanoparticle and the polymer, thereby enhancing the physical properties. Techniques for preparing composites are used.

The production of nanocomposite films not only increases the physical strength of raw polymer hydrogels, but also develops products with various functionalities such as antimicrobial, anti-acid and UV protection properties of nanoparticles used in the production of nanocomposites. It is expected.

For example, by adding gold nanoparticles to raw polymers, they have been developed as sustained-release drug carriers in addition to enhancement of appearance (Expert Opin. Drug Deliv. 7 (6): 753-763 (2010)), silver nanoparticles and iron nanoparticles. The addition of particles (Fe ₃ O ₄ ) has been reported to increase the water absorption of carrageenan hydrogels and to develop a sustained release drug delivery system (Biomaterials 29 (4): 487-496 (2008); Nanotechnology 27 ( 6): 085103 (2016); J. Mater. Chem. B, 1: 2874-2884 (2013), increased the pigment adsorption rate with the addition of nanoclay sodium montmorillonite (Na-MMT) and nanoclay and silver nanoparticles. It has been reported to have UV blocking and antibacterial properties when added to carrageenan (Applied Clay Science 97-98: 174-181 (2014); Materials Scienece and Engineering C 32 (8): 2349-2355 (2012); J. Mater. Chem. B, 3: 7237-7245 (2015)).

In addition, when the films were prepared by adding ZnO and CuO nanoparticles to various kinds of natural polymers, it was known that these films showed UV protection and strong antibacterial properties (Food Hydrocolloids 45: 264-271 (2015); Materials Letters 132: 307-311 (2014)).

On the other hand, an electroconductive hydrogel comprising a surface-enhanced Raman scattering patch (Korea Patent Publication No. 10-2011-0027366), a hydrogen bondable hydrogel and an electrically conductive material having a biocompatibility using a hydrogel mixed with metal nanoparticles Composite materials (Korean Patent Publication No. 10-2015-0024490) and the like are disclosed, but biodegradable nanocomposite hydrogels containing the biopolymers and metal nanoparticles of the present invention and their use are not disclosed.

Therefore, in the present invention, nanoparticles of metal or metal oxide type are produced on a large scale and dispersed and mixed in raw polymers that have been verified for safety and biocompatibility. Developed as a dressing for wound treatment applied to gauze or cotton wool, and developed as a cream-type product in which these nanoparticles were dispersed in a cream, and developed a new concept of skin disease treatment or skin protection agent that is easy to use and maintains its functionality. Was intended.

Therefore, the inventors of the present invention are regenerating and environmentally friendly every year, harmless to the human body, and the natural absorption of high moisture absorption rate of natural nanopolymers and metal nanoparticles and other environmentally friendly additives are not irritating to the skin, and has the appropriate strength and standard stability To provide a multifunctional bio nanocomposite skin care products.

An object of the present invention is to provide an environmentally friendly biodegradable multifunctional nanocomposite hydrogel utilizing biopolymers and metal nanoparticles.

Another object of the present invention is to provide a method for producing the biodegradable multifunctional nanocomposite hydrogel.

Still another object of the present invention is to provide a biodegradable nanocomposite hydrogel, which is a dressing for treating skin diseases, wounds or burns, and a cosmetic composition for skin care.

In order to solve the above problems, the present invention provides a biodegradable multifunctional nanocomposite hydrogel comprising biopolymers and metal nanoparticles.

In addition, the present invention comprises the steps of (a) adding the green polymer, metal nanoparticles and plasticizer to the solvent to dissolve, (b) homogenizing the dissolved solvent, to obtain a hydrogel solution in which the nanoparticles are homogeneously dispersed, And (c) provides a method for producing a biodegradable nanocomposite hydrogel comprising the step of gelling the hydrogel solution obtained above.

The present invention also provides a dressing to which the biodegradable nanocomposite hydrogel is applied to cotton wool or gauze.

The present invention also provides a cosmetic composition for skin care, comprising a biodegradable nanocomposite hydrogel and a cream base.

The present invention can provide a skin care article of a biocompatible multifunctional bio nanocomposite having excellent moisturizing power, large physical strength and excellent standard stability.

In addition, when manufacturing the multi-functional bio-nano complex skin care products of the present invention, wrinkles, whitening agent, lotion, acne treatment, skin disease treatment, antioxidant, analgesic anti-inflammatory, wound treatment, psoriasis treatment any one selected from the functional agent It is possible to provide a multi-functional bio nanocomposite skin care products.

In addition, the multifunctional nanomaterial and the functional material of the present invention may be mixed in a cream base to provide a functional cream preparation or a nano cosmetic product for skin application.

Figure 1 shows a manufacturing process of the zinc oxide nanoparticles (ZnO NPs).
Figure 2 shows a manufacturing process of the copper oxide nanoparticles (CuONPs).
FIG. 3A shows FE-SEM micrographs of zinc oxide (ZnO) nanoparticles, and FIG. 3B shows FE-SEM micrographs of copper oxide (CuO) nanoparticles.
FIG. 4A shows the XRD spectra of the zinc oxide nanoparticles, and FIG. 4B shows the XRD spectra of the copper oxide nanoparticles.
FIG. 5A shows FTIR spectra of zinc oxide nanoparticles, and FIG. 5B shows FTIR spectra of copper oxide nanoparticles.
Figure 6 shows the swelling degree change of the carrageenan hydrogel film according to pH.
Figure 7 shows the effect of the concentration of CaCO ₃ on the swelling degree of the carrageenan film.
8 shows the swelling degree of the nanohydrogel film.
Figure 9a is a state of the various hydrogels, Figure 9b is a carrageenan / Cu nanoparticle namocomposite film, Figure 9c is a nanocomposite hydrogel, Figure 9d shows a variety of hydrogel films.
Figure 10 shows the SEM image of the surface and cross section of the film, (1) carrageenan, (2) carrageenan-KCl, (3) carrageenan-KCl / ZnONPs 1%, (4) carrageenan-KCl / CuNPs 1%, (5) shows the appearance of ZnONPs 0.5% / CuNPs 0.5%.
Figure 11 as showing the optical properties and appearance of the carrageenan hydrogel film. (a) is light transmittance, (b) is absorbance, (c) is appearance of film, (1) carrageenan, (2) carrageenan-KCl, (3) carrageenan-KCl / ZnONPs 1%, (4) shows carrageenan-KCl / CuNPs 1%, and (5) shows ZnONPs 0.5% / CuNPs 0.5%.
Figure 12a is the antimicrobial properties of L. monocytogenes of nanocomposite carrageenan film, Figure 12b is E. coli of nanocomposite carrageenan film It shows the antimicrobial properties for.
FIG. 13 shows UV-visible spectra of various nanoparticles.
14 shows transmission electron micrographs of various nanoparticles.
15 shows the FTIR spectra of various nanoparticles.
16 shows XRD patterns of various nanoparticles.
Figure 17 shows the electron micrographs (FE-SEM micrographs) of the gelatin film and gelatin nanocomposite film.
FIG. 18 shows UV-Vis spectra of gelatin films and gelatin nanocomposite films. FIG.
19a shows the antimicrobial activity of E. coli ATCC 25922 of gelatin film and gelatin nanocomposite film, and FIG. 19b shows the antimicrobial activity of S. epidermis KCT1917 of gelatin film and gelatin nanocomposite film.
20 shows a functional nanoparticle containing cream formulation.
21 shows E of the cream formulation . It shows the antimicrobial activity against coli .

The present invention relates to a biodegradable nanocomposite hydrogel comprising biopolymers and metal nanoparticles.

In the biodegradable nanocomposite hydrogel of the present invention, the biopolymer may be one or more selected from carrageenan, gelatin, wheat protein, soy protein, whey protein, pectin, alginic acid, chitosan and agar, but is not limited thereto.

In the biodegradable multifunctional nanocomposite hydrogel of the present invention, the metal nanoparticles may be one or more selected from zinc oxide nanoparticles, copper oxide nanoparticles, nanosilver particles, nanogold particles and nanotitanium oxide particles, but is not limited thereto. It doesn't happen.

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In addition, the present invention comprises the steps of (a) adding the green polymer, metal nanoparticles and plasticizer to the solvent to dissolve, (b) homogenizing the dissolved solvent, to obtain a hydrogel solution in which the nanoparticles are homogeneously dispersed, And (c) relates to a method for producing a biodegradable nanocomposite hydrogel comprising the step of gelling the hydrogel solution obtained above.

In the method for preparing the biodegradable nanocomposite hydrogel of the present invention, the biopolymer may be at least one selected from carrageenan, gelatin, wheat protein, soy protein, whey protein, pectin, alginic acid, chitosan and agar, but is not limited thereto. It is not.

In the method for preparing the biodegradable nanocomposite hydrogel of the present invention, the metal nanoparticles may be one or more selected from zinc oxide nanoparticles, copper oxide nanoparticles, nanosilver particles, nanogold particles and nanotitanium oxide particles. It is not limited.

In the method for preparing the biodegradable nanocomposite hydrogel of the present invention, the plasticizer may be one or more selected from glycerin, polyethylene glycol, triethyl citrate, and sorbitol, but is not limited thereto.

In the method for preparing the biodegradable nanocomposite hydrogel of the present invention, the step (a) is based on 100 parts by weight of the biopolymer, 1 to 10 parts by weight of the metal nanoparticles and 25 to 40 parts by weight of the plasticizer 5,000 weight of distilled water It can be added to the part to dissolve.

In the method for preparing the biodegradable nanocomposite hydrogel of the present invention, the dissolution may be dissolved at 85 to 95 ° C, preferably at 95 ° C for 15 to 30 minutes, preferably for 20 minutes.

In the method for producing the biodegradable nanocomposite hydrogel of the present invention, the homogenization of step (a) is 500 to 2,000 rpm, preferably 1000 to 1500 rpm, for 12 to 24 hours, preferably for 15 to 20 hours. After stirring, ultrasonic waves of 20 to 100 kHz, preferably 40 to 80 kHz, may be treated by 10-20 minutes.

The present invention also relates to a dressing on which the biodegradable nanocomposite hydrogel is applied to cotton wool or gauze.

The dressing of the present invention may be for treating skin diseases, for treating wounds or for treating burns.

The dressing of the present invention, the dressing may further comprise a plant extract.

In the dressing of the present invention, the plant is Artemisia scoparia ), may be one or more selected from seaweed, persimmon seed extract, green tea extract, but is not limited thereto.

The present invention also relates to a cosmetic composition for skin care comprising the biodegradable nanocomposite hydrogel and cream base.

In the cosmetic composition for skin care of the present invention, the cream base is sufficient to use a known cream base used in the manufacture of cosmetics, a detailed description thereof will be omitted.

The cosmetic composition for skin care of the present invention may further include a grapefruit extract.

In the skin care cosmetic composition of the present invention, the grapefruit extract may be extracted from grapefruit seeds, grapefruit peel, or grapefruit seeds and shells.

In the cosmetic composition for skin care of the present invention, the grapefruit extract extraction method is sufficient to extract using a known method, such as alcohol, water and glycerol mixed extraction solvent, a detailed description thereof will be omitted.

Hereinafter, the contents of the present invention will be described in detail through experimental examples. However, these are intended to explain the present invention in more detail, and the scope of the present invention is not limited thereto.

Example 1 Preparation of Nanoparticles

One. Of ZnO nanoparticles  Produce

Nano zinc oxide (ZnO NPs) was prepared according to the process shown in Fig. 1 using NaOH as a hydrolysis agent in zinc nitride which is a raw material of zinc.

To this end, 29.75 g of 0.1 M zinc nitride was dissolved in 1000 mL of distilled water and heated to boiling point, and 40 mL of 5 M NaOH solution was added thereto and heated at 90 ° C. for 2 hours.

This produced a white precipitate, which was zinc oxide nanoparticles (ZnO NPs). The precipitate was collected by centrifugation, washed three times with distilled water and twice with anhydrous alcohol, and then dried at 70 ° C. for 6 hours to obtain a powder.

The ZnO NPs powder obtained above was sealed and stored at room temperature, and used for the preparation of the nanocomposite film.

2. Nano copper oxide ( CuO nanoparticles Manufacturing

Nano copper oxides (CuO NPs) were prepared according to the process of FIG. 2 using copper chloride as a raw material and NaOH as a hydrolysis agent.

34.96 g of 0.2 M copper chloride was dissolved in 1000 mL of distilled water and heated to boiling point, then 60 mL of 5 M NaOH solution was added, followed by heating at 90 ° C. for 2 hours, to obtain CuO NPs as a brown precipitate.

The precipitate was collected by centrifugation, washed three times with distilled water and twice with anhydrous alcohol, and then dried at 70 ° C. for 6 hours to obtain a powder.

CuO NPs powder obtained above was sealed and stored at room temperature, and used for the manufacture of nanocomposite film.

3. Characterization of Nanoparticles nanoparticles )

The microstructures of zinc oxide and copper oxide nanoparticles were investigated using field emission scanning electron microscopy (FE-SEM, S-4800, Hitachi Co., Ltd., Matsuda, Japan). To this end, zinc oxide and copper oxide nanoparticle powders were fixed with carbon tape, and then observed at an acceleration voltage of 10.0 kV.

The crystal structure of the nanoparticles was examined by measuring the X-ray diffractogram.

Cu Kα radiation (wavelength: 0.1541 nm)

Nickel Monochromator Filtration Wavelength

Acceleration to 40 kV and 30 mA

At this time, the diffraction pattern (diffraction pattern) was measured at a scanning speed of 0.4 ° / min at a diffraction angle of 2θ = 30-80.

Resolution of 4 cm ^-1 using oattenuated total reflectance-Fourier transform infrared (AT-FTIR) spectrophotometer (TENSOR 37 spectrophotometer, Billerica, MA, USA) ) Was measured in the wavelength range of 4000 ~ 500 cm ^-1 .

4. Results and Discussion

The ZnO NPs prepared in this study were white powder with a yield of 25 wt% (7.5 g of ZnO NPs / 29.75 g of zinc nitride). CuO nanoparticles, on the other hand, were dark brown powders with an output of about 30 wt% (12 g of CuO NPs / 34.96 g of copper chloride).

3A and 3B show SEM micrographs of ZnO and CuO nanoparticles. ZnO nanoparticles were spherical in size 50-50 nm and formed uniform clusters of 100-200 nm in size. CuO nanoparticles, on the other hand, had a rod shape and had a size of 100-200 nm.

4A and 4B show XRD patterns of nanoparticles. All nanoparticles show a crystalline structure and show the characteristic diffraction pits of each nanoparticle.

5A and 5B show FTIR spectra results of ZnO and CuO nanoparticles.

It is due to the formation of tetrahedral coordination of zinc at 865 cm ⁻¹ of ZnO nanoparticles, and the peak at 650-700 cm ⁻¹ is due to the stretching vibrations of ZnO nanoparticles.

On the other hand, the peaks appearing at 670 and 686 cm ⁻¹ are due to the stretch vibration of Cu—O.

Example 2 Preparation of Functional Hydrogel Using Carrageenan

1. Material

κ-carrageenan was purchased from Carogen Korea (Hwasun, Jeonnam), and copper chloride (CuCl ₂ · 2H ₂ O) and zinc nitride (N ₂ O ₆ Zn · 6 H ₂ O) were purchased from Duksan Pure Chemical Co. , Ltd., Ansan, Gyonggido, Korea) NaOH is Samchun Pure Chemical Co., Ltd., Pyeongtaek, Gyonggido, Korea), potassium chloride (KCl) and citric acid, lactic acid and glycerin are Daejung Chemicals & Metals Co., Ltd., Siheung, Gyonggido, Korea).

2. How to

To prepare a control carrageenan film, 3 g of carrageenan (2% w / v) and 0.9 g of glycerin (30% of carrageenan) were added to 150 mL of distilled water and dissolved by heating at 90C for 20 minutes, followed by Teflon film (Cole- Parmer Instrument Co., Chicago, IL, USA) was cast on a glass plate (24 cm x 30 cm) coated and dried at room temperature.

Carrageenan hydrogel film was prepared by the above method by adding 2.5 mL of 1 M KCl solution to the carrageenan film solution to promote the gel formation of carrageenan.

The carrageenan nanocomposite hydrogel film was prepared with 1 wt% zinc oxide nanoparticles (ZnO NPs), 1 wt% copper nanoparticles (CuO NPs) and 0.5 wt% ZnO NPs and 0.5 wt. Three nanocomposite films were prepared by mixing and adding% Cu NPs.

The swelling ratio, which indicates the water adsorption capacity of the film, is obtained by dipping a constant size sample (2.5 cm × 5 cm) in distilled water, removing the sample at regular intervals, removing the surface water, and then measuring the weight by the following equation 1 According to the water adsorption capacity is expressed as a percentage.

<Equation 1>

Here, W ₁ is the initial weight of the film, W ₂ is the weight after immersion.

3. Results and Discussion

3.1. Swelling ratio

In order to investigate the effect of pH on the degree of swelling of the hydrogel film, the results of different pH of the film solution were investigated.

As shown in Figure 6, in general, the degree of swelling was rapidly increased until the immersion time 15 minutes, after which the rate of increase slowed down or showed a constant value. The lower the pH, the higher the swelling degree, but the stability of the gel was slightly degraded.

In order to investigate the effect of adding CaCO ₃ on the degree of swelling, the swelling degree of the carrageenan hydrogel film was examined as shown in FIG. 7 while varying the concentration of CaCO ₃ (5, 10, 15, 20%).

As shown in Figure 7, addition of CaCO ₃ not only increased the swelling degree of the carrageenan film but also increased the stability of the gel, the highest effect was added when 5%.

Therefore, in order to enhance the stability of the film, it was determined that the pH of the carrageenan film solution was adjusted to 10, and 5% of CaCO ₃ was added.

3.2. Effect of Adding Nano Metal Particles (Zinc Oxide, Copper)

In order to impart functionality to the carrageenan hydrogel film, nanometal particles were mixed to prepare a nanohydrogel film, and the swelling degree thereof was examined as shown in FIG. 8.

As shown in FIG. 8, the swelling degree of the nanohydrogel film added with 1% ZnO NPs was increased, but the addition of Cu NPs decreased the swelling degree, and the addition of 1% Cu NPs inhibited gel formation. .

3.3. Hydrogel  Physical strength

The physical strength of the hydrogel was measured under the following conditions in a compression mode using a rheometer (Sun Rheo Meter Compac-100II, Tokyo Maruichishoji Co., Ltd., Tokyo, Japan).

Head speed: 100 mm / min

Maximum load cell: 10 kg

Tension distance: 70%.

3.4. Carrageenan Hydrogel  Effect of Nanometal Addition on Strength

Table 1 shows the results of examining the effect of the addition of nanometals on the strength of carrageenan hydrogel.

Figure 9a is a state of the various hydrogels, Figure 9b is a carrageenan / Cu nanoparticle namocomposite film, Figure 9c is a nanocomposite hydrogel, Figure 9d shows a variety of hydrogel films.

3.5. Carrageenan  Tensile Properties of Nanocomposite Films

The results of examining the tensile properties of the carrageenan nanocomposite film are shown in Table 2 below.

3.6. Carrageenan Hydrogel  Microstructure of Film

The microstructure of the surface and the cross-section of the carrageenan hydrogel film using an electron scanning microscope (SEM) is shown in FIG. 10.

In Figure 10, (1) carrageenan, (2) carrageenan-KCl, (3) carrageenan-KCl / ZnONPs 1%, (4) carrageenan-KCl / CuNPs 1%, (5) ZnONPs 0.5% / SEM images of CuNPs 0.5% are shown.

3.7. Carrageenan Hydrogel  Optical properties and appearance of the film

Transmittance, absorbance and appearance of the carrageenan hydrogel film are as shown in FIG. 11. The film added with ZnO nanoparticles was white in color, and the film added with CuO nanoparticles was brown in color.

Both ZnO nanoparticles and CuO nanoparticles-added films are light-transmissive at wavelengths in the UV-B range, confirming that these films are strongly UV-protective, which can be used for the development of skin care products. Judging.

3.8. Antimicrobial Properties of Nanocomposite Films

L. monocytogens and E. coli of nanocomposite carrageenan films The results of investigating the antimicrobial properties were as in FIG. 12A and FIG. 12B.

As shown in FIG. 12A and FIG. 12B, the carrageenan film to which the nanometals were added was gram-positive bacteria L. monocytogens and gram-negative bacteria E. coli It showed distinct antimicrobial properties against.

It is thought that the antimicrobial properties of these nanofilms can enhance their value as skin care products.

<Example 3> Development of functional bio nanocomposite film for skin disease (acne) treatment

Introduction

Acne is a skin disease that affects 85% of adolescents worldwide. Propionibacterium acne , Staphylococcus epidermidis , Escherichia coli , with Staphylococcus aureus Infection with the same bacteria not only causes disease on the skin by the fat-soluble enzymes produced by these bacteria, but also causes problems such as hormonal imbalance and immune hypersensitivity.

In addition, free radicals, caused by UV or other toxins, can damage skin cells and destroy healthy skin cells, leaving a scar on the skin. Skin ointments or oral preparations have been used to treat such skin diseases, but the use of such preparations over a long period of time may lead to the development of antibiotic resistant bacteria.

The purpose of this study was to develop bio-composite bio nanocomposite hydrogel-type skin care products to protect the skin and to treat skin diseases such as acne by preparing natural polymer hydrogel containing functional nanoparticles.

2. Materials and Methods

2.1. material

All chemicals unless otherwise specified Used from Sigma-Aldrich (St. Louis, Mo., USA). Tryptic soy broth (TSB), brain heart infusion broth (BHI) and agar powder were purchased from Duksan Pure Chemicals Co., Ltd., Ansan, Gyeonggi-do, Korea.

Microorganisms for antimicrobial test ( Escherichia coli ATCC 25922, Staphylococcus epidermis KCTC 1917, was purchased from Korean Collection for Type Cultures (KCTC, Seoul, Korea).

2.2. Scruffy  ( Artemisia scoparia ) Extract

Flameproof plant Artemisia scoparia was taken from the coast of Jeonnam in May 2014. The ground portion of the plant was steam heated and dried at 45 ° C. for 2 days, then ground using mortar and used at -70 ° C.

4 kg of dry powder was mixed with 40 L of distilled water and extracted at 121 ° C. for 30 minutes. Cucumber extract was obtained using 2 filter papers (Whatman, Maidstone, UK).

2.3. Preparation of Metal Nanoparticles

Gold, silver, copper nanoparticles, and bimetallic nanoparticles in combination of these were prepared by the method described above.

10 mL of the mugwort extract was mixed with 490 mL of distilled water while injecting nitrogen gas and heated to 70 ° C. 0.5 mL of 1 M Gold (III) chloride trihydrate (HAuCl ₄ ), silver nitride (AgNO ₃ ), and copper to prepare gold (Au NPs), silver (Ag NPs), and copper (Cu NPs) nanoparticles, respectively (II) Nitride trihydrate (CuN ₂ O ₃ ) was slowly added to the AS extract and heated until the color of the solution changed.

To prepare bimetallic nanoparticles, 0.25 mL (1 M) of each metal salt solution (HAuCl ₄ and AgNO ₃ ; HAuCl ₄ and CuN ₂ O ₃ ; AgNO ₃ and CuN ₂ O ₃ ) was added to the AS extract in the same manner as above and then heated.

The nanoparticles thus prepared were represented as AuNPs (gold), AgNPs (silver), CuNPs (copper), AuAgNPs (gold-silver), AuCuNPs (gold-copper) and AgCuNPs (silver-copper) nanoparticles, respectively.

2.4. Characterization of Nanoparticles

The formation of the nanoparticles was confirmed that the color of the solution is changed, the absorbance was measured in the 200 ~ 700 nm range using a UV-visible spectrophotometer (Mecasys Optizen POP Series UV / Vis, Seoul, Korea).

The shape and size of the nanoparticles were measured under conditions of an acceleration voltage of 120 kV using transmission electron microscopy (TEM, JEOL-1010 instrument, Tokyo, Japan).

Fourier transform infrared (FT-IR) and XRD characteristics of nanoparticles were investigated according to the method described above.

Surface charge of nanoparticles was measured by zeta potential and particle size was measured by dynamic light scattering (DLS) method using Zeta PALS-zeta potential analyzer (Brookenhaven Instruments Corporation, NY, USA). It was.

2.5. Antimicrobial Activity of Nanoparticles

Antimicrobial activity of nanoparticles against Gram-positive ( S. epidermis ) and Gram-negative ( E. coli ) pathogenic bacteria at minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using the broth microdilution method Measured.

2.6. Preparation of Gelatin / Nanoparticle Composite Film

Gelatin films and gelatin / nanoparticle composite films were prepared using a solution casting method.

For the preparation of gelatin / NPs nanocomposite film, 4 g of gelatin was added to and dissolved in the 150 mL nanoparticle solution prepared above, followed by the addition of 1.2 g of glycerol and heating at 80 ° C. for 20 minutes, followed by a horizontal glass plate. Cast to and then dried to prepare a film.

All films were used as samples after conditioning for at least 48 hours in a humidity chamber (model FX 1077, Jeio Tech Co. Ltd., Ansan, Korea) controlled at 25 ° C. and 50% RH.

The gelatin / NPs composite film thus prepared was referred to as gelatin / AuNPs, gelatin / AgNPs, gelatin / CuNPs, gelatin / AuAgNPs, gelatin / AuCuNPs, gelatin / AgCuNPs, respectively.

2.7. Characteristics of gelatin / nanoparticle composite film

2.7.1. Microstructure and Engineering Properties

The microstructure of the film was examined at an acceleration voltage of 5.0 kV using field emission scanning electron microscopy (FE-SEM, S-4800, Hitachi Co., Ltd., Matsuda, Japan).

The surface color of the film was measured using a chromameter (Konica Minolta, CR-400, Tokyo, Japan) against a white standard plate ( L = 97.75, a = -0.49, and b = 1.96) and the total color difference (total color) difference, E ) was calculated according to Equation 2 below.

<Equation 2>

The optical properties of the film were investigated using a UV-vis spectrophotometer (Mecasys Optizen POP Series UV / Vis, Seoul, Korea), and the UV protection and transparency of the film were measured at 280 nm (T ₂₈₀ ) and 660 nm (T ₆₆₀ ). Permeability was measured and investigated.

2.7.2. Tensile Properties of Film

Tensile strength (TS), elongation at break (E) and elastic modulus of the film were measured using an Instron Universal Testing Machine (Model 5565, Instron Engineering Corporation, Canton, MA) in accordance with ASTM D 882-88 standard method. , USA).

2.7.3. Film Moisture Content (MC) and Moisture Contact angle ( WCA )

The water contact angle, which is an indicator of hydrophilicity or hydrophobicity of the film, was measured using a WCA analyzer (model Phoenix 150, Surface Electro-Optics Co., Ltd., Kunpo, Korea).

The moisture content of the film was determined by weight difference after the film was dried at 105 ° C. for 24 hours.

2.7.4. Antibacterial Properties of Film

The antimicrobial activity of S. epidermis and E. coli of gelatin and gelatin / nanometal composite films was measured using the viable colony count method.

First S. Epidermis and E. coli were aseptically inoculated into 20 mL of BHI and TSB broth, respectively, and incubated at 37 ° C for 16 hours.

Cell pellets obtained by centrifuging each culture solution at 5000 rpm for 10 minutes were dissolved in 100 mL of sterile TSB and BHI broth, and then diluted 10-fold with sterile distilled water.

50 mL (10 ^{6-10 7} CFU / mL) of this solution was placed in a 100 mL conical flask containing a film sample (5 cm × 5 cm) and incubated at 37 ° C. for 12 hours. Broth diluted in the same ratio was used as a control.

 Samples were taken at 3 hour intervals and the viable cell count was measured by colony counting method. Antimicrobial experiments were performed three times using a separately prepared film.

2.7.5. Antioxidant properties of film

To investigate the antioxidant properties of the film, 200 mg of film sample was dissolved in 10 mL of distilled water to obtain a 20 mg / mL film solution, which was then used for DPPH radical scavenging assay, ABTS ⁺ radical scavenging assay. And Ferric reducing (FRAP) ability assay Antioxidants were investigated using the method.

2.8. Statistical analysis

The characteristics of the film were obtained by performing three repeated experiments, and the data were expressed as mean ± SD (standard deviation).

For statistical analysis, one-way analysis of variance (ANOVA) was performed using SPSS software (SPSS Inc., Chicago, IL, USA), and Duncan's multiple range test was performed at 5% significance level. It was.

3. Results and Discussion

3.1. Characteristics of Nanoparticles

It was confirmed that the nanoparticles were formed by changing the color of the single or two kinds of metal nanoparticle solutions prepared using the mugwort extract. That is, the solution was initially colorless, but AuNPs, AgNPs, CuNPs, AuAgNPs, AuCuNPs, AgCuNPs were formed, and the color of the solution changed to ruby red, brownish yellow, pale brown, pale ruby, brownish red, and pale yellow (Fig. 13).

This color change is due to the surface plasmon vibrations of the individual nanoparticles.

In addition, the formation of nanoparticles was investigated using a spectrophotometer. The maximum absorption wavelengths of gold, silver and copper nanoparticles were observed at 558, 424 and 344 nm, respectively. (Fig. 13).

On the other hand, the microstructure of the nanoparticles was investigated using TEM results as shown in Figure 14, all the nanoparticles were spherical, the size was 20 ~ 120nm size as follows depending on the type. AuNPs (20-30 nm), AgNPs (20-50 nm), CuNPs (<15 nm), AuAgNPs (10-20 nm), AuCuNPs (20-30 nm), AgCuNPs (80-120 nm).

However, the particle size measured by dynamic light scattering (DLS) method was 100 nm or more, as shown in Table 3.

Vibration peaks at 3300, 2931, 1749, 1635, 1550, 1456, 1240, 1033, 921, 846 cm ⁻¹ are NH or carboxylic acid of amine or OH stretch of water, CH stretch of alkane, C = of carboxylic acid O stretch, NH bend of primary amine, CC stretch of aromatic ring, CH bend of alkane, CN stretch of aromatic amine or CO s stretch of alcohol or carboxylic acid, CN stretch of aliphatic amine, OH bend of carboxylic acid, It is due to the NH of phenolic compounds.

These peaks indicate that the wormwood extract is used as a capping material for the nanoparticles.

In addition, the XRD pattern of the nano metal particles are as shown in FIG.

As shown in FIG. 16, all the nanoparticles exhibited polycrystalline characteristics and exhibited characteristic diffraction peaks of gold, silver and copper nanoparticles, respectively. All nanoparticles showed strong (111) crystalline peaks.

In order to investigate the stability of the nanoparticles in the colloidal system, the results of measuring the zeta-potential of the nanoparticles are shown in Table 3 above.

As shown in Table 3, the zeta potentials of AuNPs, AgNPs, CuNPs, AuAgNPs, AuCuNPs, AgCuNPs nanoparticles are -22.7 ± 1.4, -42.3 ± 3.4, -25.2 ± 1.8, -23.4 ± 2.6, -22.1 ± 2.7, respectively. , -16.5 ± 1.9 mV.

All nanoparticles show high zeta potential values, which means that these nanoparticles are highly stable in solution.

3.2. Nanoparticles  Antimicrobial properties

The antimicrobial properties of the nanoparticles were investigated using a micro-dilution method, and their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) results are shown in Table 4 below.

As shown in Table 4, E. coli The MIC / MBC results for AuNPs, AgNPs, CuNPs, AuAgNPs, AuCUNPs, AgCuNPs for ATCC 25922 were> 500 /> 500, 15.6 / 31.2, 250/500, 7.8 / 31.2,> 500 /> 500, 31.2 / 125 μM, respectively. S. epidermis was> 500/500, 31.2 / 125,> 500 /> 500, 7.8 / 62.5,> 500 /> 500 and 31.2 / 62.5 μM, respectively.

In general, silver nanoparticles showed stronger antimicrobial activity than copper nanoparticles, and showed stronger antimicrobial activity against gram negative bacteria than gram positive bacteria.

3.3. Characteristics of Nanocomposite Film

3.3.1. Microstructures, Surface Colors, and Optical Properties

17 shows the microstructure of the surface of these films by using a FE-SEM by preparing a gelatin film and a gelatin / nanometallic composite film.

As shown in FIG. 17, the gelatin film has a smooth and dense surface while the surface of the composite film shows a less smooth appearance. In the composite film containing AuNPs, AgNPs, and CuNPs nanoparticles, the nanoparticles were uniformly distributed, but the films in which AuAgNPs, AuCuNPs, and AgCuNPs nanoparticles were added showed a slight aggregation of nanoparticles.

The results of measuring the surface color and the light transmittance of the film are shown in Table 5 below.

Although the gelatin film shown in Table 5 was colorless and transparent, the film to which the nanometal was added showed a somewhat low transparency and a unique color according to the type of nanoparticles added.

The light absorbency and light transmittance of the gelatin film and the gelatin / nanometallic composite film were measured in the wavelength range of 200-700 nm, and are shown in FIG. 18.

The gelatin film showed a maximum absorption wavelength at 280 nm, which is due to the amino acids tyrosine and tryptophan contained in gelatin. On the other hand, gelatin / NPs nanocomposite films show a broad absorption peak in the range of 280-350 nm, which is due to phenolic compounds contained in gelatin and cucurbita extracts, and these composite films also exhibit unique peaks depending on the nanoparticles used. It was.

In general, phenolic compounds are known to have strong light absorption in the ultraviolet region. In order to investigate the UV blocking properties and transparency of these films, light transmittances were measured at 280 and 660 nm, and the results are shown in Table 5 above.

The gelatin film had a high transparency with a light transmittance (T ₆₆₀ ) of 89.3 ± 0.9% at 660 nm, but the transparency decreased with the addition of nanoparticles.

The light transmittance against ultraviolet rays (T ₂₈₀ ) was 29.4 ± 2.6% of the gelatin film, but after forming the composite film with the nanoparticles, the value was significantly lowered to 0-0.2%, indicating that the UV blocking property was greatly increased. .

3.3.2. Physical properties

The results of examining the physical properties of the gelatin film and the gelatin / nanometallic composite film are shown in Table 6 below.

As shown in Table 6, the thickness of the film was 54.0 ± 2.7 μm, and addition of nanoparticles except CuNPs did not significantly affect the thickness of the film. Tensile strength and elongation of gelatin film were 97.0 ± 6.9 MPa and 8.3 ± 1.4%, respectively.

Tensile properties were also affected by the addition of nanoparticles. There was no significant difference when AuNPs, AgNPs and CuNPs were added, but when AuAgNPs, AuCuNPs and AgCuNPs were added, there was a significant difference.

On the other hand, the elongation of the nanocomposite film did not change significantly. However, the elastic modulus (EM) of the film decreased slightly.

3.3.3. Water content and Moisture contact angle

The results of measuring the moisture content and the moisture point sensation of the gelatin film and the gelatin / nanometallic composite film are shown in Table 6 above.

The water content of the nanocomposite film was slightly lower than that of the gelatin film. The water contact angle of the gelatin film was 56.9 ± 0.7 °, and the value decreased slightly after forming the composite film with the nanoparticles.

3.3.4. Antimicrobial activity

About the S. epidermis and E. coli bacteria The antimicrobial test results of the nanocomposite film irradiated using the colony count method are shown in FIGS. 19A and 19B.

As shown in FIGS. 19A and 19B, the gelatin film and the gelatin / wormwood extract film did not exhibit antimicrobial activity, and the nanoparticle-added films showed strong antimicrobial properties except for AuNPs.

In particular, the films containing AgNPs and their alloys showed higher antimicrobial properties. Depending on the strain used, E. coli was more sensitive to AgNPs, AuAgNPs, AgCuNPs and CuNPs than S. epidermis .

This means that Gram-negative bacteria are more inhibited by these nanoparticles than Gram-positive bacteria.

3.3.5. Antioxidant

The antimicrobial properties of the gelatin / NPs nanocomposite films were investigated using DPPH, ABTS and FRAP activity assay. As shown in Table 7, the antimicrobial properties of the gelatin / NPs nanocomposite films differed depending on the type of nanoparticles used.

As shown in Table 7, among the nanoparticles used, AgNPs- was the highest in antioxidant activity, the next appeared AuNPs, and the addition of CuNPs showed the lowest antioxidant activity.

The DPPH activities of the gelatin / AgNPs, gelatin / AuNPs and gelatin / CuNPs films were 89.7%, 66.0 and 22.9%, respectively. The activity of the gelatin / bimetallic NPs films was different depending on the type of nanoparticles used.

That is, the gelatin / CuAgNPs composite film showed higher activity (50.6%) than the gelatin / CuNPs (22.4%) film. Similar results with the DPPH method were obtained for the ABTS and FRAP activity measurements.

Gelatin / nanoparticle composite film having such strong antioxidant properties and UV protection is expected to be useful in treating skin diseases by inhibiting oxidative damage of the skin and generation of free radicals.

4. Conclusion

6 kinds of mono or bimetallic nanoparticles were prepared by using the extract as a reducing agent and a capping agent, and gelatin and composite films were prepared using the same.

All gelatin / nanoparticle composite films showed UV protection.

In addition, all composite films except gelatin / AuNPs films showed strong antimicrobial properties against Gram-positive and Gram-negative bacteria causing skin diseases.

In addition, other composite films except gelatin / CuNPs films showed strong antioxidant properties.

The antimicrobial properties and sunscreen properties of such nanocomposite films can inhibit the growth and growth of microorganisms that cause skin diseases and protect the skin from ultraviolet rays. In addition, these composite films show strong antioxidant properties and are expected to be used as a treatment or protection for skin diseases such as acne.

< Application example 1> functionality  Manufacture of Cream Products containing Nanoparticles

As part of the research for the formulation diversification of skin care products, a cream product was prepared.

The cream base was provided by Mediplan Co., Ltd., GSE (Grapefruit Seed Extract) and ZnO nanoparticles were added to the cream base to prepare the following four cream-type preparations.

(1) ZnONPs 0.1%;

(2) ZnONPs 0.1% + GSE 1%

(3) ZnONPs 0.25%

(4) ZnONPs 0.25% + GSE 1%

These ingredients were first dissolved in 10 mL of sterile distilled water, then sonicated for 5 minutes, then added with 90 g of cream base and 7000 rpm using a high speed super mixer (EURO-ST PB, IKA® WERKE, Germany). Mix for 10 minutes at. Thus prepared cream was packaged in a plastic container, the appearance is as shown in FIG.

The antimicrobial activity of E. coli was investigated using the agar diffusion method in FIG. 21. The cream formulations containing GSE and ZnO nanoparticles showed distinct antimicrobial activity against E. coli .

When manufacturing a functional cream type product, a functional substance which is known to have a whitening effect and an anti-wrinkle effect can be added to enhance the range of its functionality. 2 to 5% by weight of niacin amide may be added for the whitening effect, and 0.04% to adenosine may be added for the wrinkle improvement effect.

< Application example 2> wound  Manufacture of dressings for treatment or burn treatment

The multifunctional (antibacterial, antioxidant) biodegradable nanohydrogel developed in the present invention was applied to gauze or cotton wool to prepare a dressing for wound treatment or burn treatment.

As described above, although described with reference to a preferred embodiment of the present invention, those skilled in the art various modifications and variations of the present invention without departing from the spirit and scope of the present invention described in the claims below. It will be understood that it can be changed.

The present invention can provide a functional bio-nanocomplex hydrogel, which is friendly to the human body, having excellent moisturizing ability, large physical strength, and excellent standard stability.

In addition, in the preparation of the bio-nanocomplex hydrogel of the present invention, a functional agent is selected so as to include any functional agent selected from a wrinkle remover, a whitening agent, a lotion, an acne treatment, a skin disease treatment, an antioxidant, an analgesic anti-inflammatory agent, a wound treatment, and a psoriasis treatment. Since it is possible to provide a bio-nanocomplex hydrogel comprising a, it is possible to contribute to the development and product development of the industry requiring the functional agent there is industrial applicability.

In addition, the biodegradable nanocomposite material having antimicrobial and biocompatibility may be applied to cotton wool or gauze to be used as a dressing for wound treatment or to treat skin diseases such as inflammation and acne.

In addition, by adding a functional nano material and a known functional material such as a whitening effect and an anti-wrinkle effect to the cream base, the functional nano material developed in the present invention may be used as a functional cream or nano cosmetic product for skin application.

Claims (19)

delete delete delete delete delete delete delete delete delete delete delete delete A dressing comprising a biodegradable nanocomposite hydrogel comprising biopolymer and metal nanoparticles on cotton wool or gauze, and further comprising one or more plant extracts selected from Artemisia scoparia and seaweed,
And the metal nanoparticles are one or two or more selected from zinc oxide nanoparticles, copper oxide nanoparticles, nanosilver, nanogold, and nanotitanium oxide. The dressing according to claim 13, wherein the dressing is for treating skin diseases, for treating wounds or for treating burns. delete delete delete delete delete

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